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Objective: To compare and evaluate the clinical esthetic effect of angle screw channel abutment and personalized zirconia adhesive abutment for single crown restoration in esthetic area. Methods: A total of 44 patients (21 males and 23 females), aged (37.4±13.5) years (18-67 years) who completed single crown restoration in the esthetic area of the Department of Oral Implantology, Dalian Stomatological Hospital from January 2018 to June 2022 were retrospectively selected. A total of 44 implants were inserted. According to the abutment selected for final restoration, the patients were divided into angle screw channel abutment group and personalized zirconia bonding abutment group, with 22 patients and 22 implants in each group. The implant survival rate, complication rate, pink and white esthetic score and marginal bone resorption were compared between the two groups. Results: Follow-up to 12 months after final restoration, implant survival rates were 100% (22/22) in both groups, and there was no statistically significant difference in the incidence of mechanical complications between the two groups [9% (2/22) in the angle screw channel abutment group and 0(0/22) in the personalized zirconia bonding abutment group, χ 2 =2.10, P= 0.148]. In the follow-up appointment 12 months after final restoration, the pink esthetic score of the angle screw channel abutment group (12.95±1.05) was significantly better than that of the personalized zirconia bonding abutment group (11.45±2.02) ( t= 3.10, P= 0.003). There was no significant difference in white esthetic scores between the two groups ( t= 1.27, P= 0.212). There was no significant difference in the marginal bone resorption between the two groups ( t= 0.32, P= 0.749). Conclusions: When a single implant supported restoration is delivered in the esthetic area of the anterior maxilla, high implant survival rate and stability of the marginal bone can be obtained by using the angle screw channel abutment or the personalized zirconia bonding abutment. The clinical efficacy of the angle screw channel system is reliable, and it will provide clinicians with a new treatment option.

Here, we reported the diagnosis and treatment of a case of HIV infected person complicated by an extremely rare infection with Mycobacterium celatum . Due to the similarity of homologous sequence regions between Mycobacterium celatum and Mycobacterium tuberculosis complex, the identification of conventional Mycobacterium species was incorrect, which was corrected after first-generation 16S rRNA sequencing. This report aimed to improve the clinical understanding of Mycobacterium celatum infection and the level of differential diagnosis between non-tuberculous mycobacterial disease and tuberculosis.

Objective: To assess the value of using flat-sided culture tubes for preparing chromosomes through chorionic villi (CV) and amniotic fluid (AF) cell cultures during prenatal diagnosis., Methods: From February to March 2020, 157 CV samples and 147 AF samples subjected to prenatal diagnosis at the Maternal and Child Health Care Hospital of Guangxi Zhuang Autonomous Region were selected as the study subjects. For each sample, one flat-sided tube and one flask culture were set up by following the standard protocols. The methods were evaluated by comparing the cell growth, experimental process, quality of chromosome preparation and costs., Results: The success rates for the culturing of CV and AF samples by the flat-sided culture tube method were 97.45% (153/157) and 97.96% (144/147), respectively. By contrast, the success rates for the conventional flask method were 98.72% (155/157) for CV and 98.64% (145/147) for AF samples. No significant difference was found between the two methods (P > 0.05). The average harvest time required by the flat-sided culture tube method was 8.45 days for CV and 9.43 days for AF cultures, whilst the average harvest time for conventional flask method was 9.05 days and 9.54 days, respectively. The flat-sided culture tube method for CV had required significantly shorter average harvest time than the conventional method (P < 0.001). No statistical significant difference was found in the average harvest time for AF by the two methods (P > 0.05). The conventional culturing method had required three containers with two sample transfers. By contrast, the flat-sided culture tube method was carried out in one tube without any sample transfer. The average total amount of medium used was 3.91 mL for each flat-sided culture tube and 6.26 mL for each conventional flask., Conclusion: The flat-sided culture tube method can provide a simple, cost-effective and error-reducing procedure for the CV and AF samples culture during prenatal diagnosis.

Objective: To investigate the awareness status of human papilloma virus (HPV) among college students and relevant influencing factors in order to provide references and suggestions for the prevention of HPV related diseases., Methods: Based on the random sampling method, 5 412 college students in Shandong Province were surveyed by questionnaire, including the general demographic characteristics of the subjects, knowledge about HPV and related diseases caused by HPV infection., Results: In the final analysis, 1 777 male students and 3 653 female students were included in the final analysis. Among them, 40.5 percent of male students were unaware of HPV, compared with 35.8 percent of female students, the difference was statistically significant (P<0.001). The average score of HPV-related knowledge cognition was 13.87 points, of which the male and female cognitive scores were 13.4 and 14.1 respectively, with a statistically significant difference (P<0.01). The cognitive scores of students in different grades and major backgrounds have significant differences. The cognitive scores of medical students were significantly higher than those of non-medical students (15.66 vs 12.58,P<0.01). Multi-factor logistics analysis showed that female students, senior students, college students with medical professional background, and college students who obtained knowledge about HPV from doctors or hospital or school related lectures had higher cognition of HPV., Conclusion: College students, especially male students, lower grade students and students with non-medical background have a low cognition degree of HPV, suggesting that in the prevention and treatment of HPV-related diseases, it is necessary to strengthen the health education of this population.

Objective: This study optimizes three-dimensional(3D)culture conditions of L02 cells using response surface methodology(RSM) based on the VitroGel system to construct the hepatocytes model in vitro., Methods: L02 cells were 3D cultured by the VitroGel system. The appropriate level of three key factors(concentration of inoculated cells, culture time and dilution degree of the hydrogel) was determined by single-factor experiment, and the optimal conditions of 3D culture of L02 cells based on the VitroGel system were determined by RSM. During the detection process, the optical density(OD) value of cell viability was used as the detection index, and the cell viability was detected using the cell counting kit-8(CCK-8) assay. The proliferative performance and viability of L02 cells was measured by fluorescent staining assay., Results: The selected optimal culture conditions by RSM were as follows: concentration of inoculated cells was 1.1 × 10~5/mL, culture time was 9.5 days, and dilution degree of hydrogel was 1∶3.7. The result shows that under optimal conditions, the predicted OD value of cell viability was 2.17 and measured 2.13 with a relative error of 1.84%, indicating that the condition was suitable and reliable. The fluorescent staining and dead and live cells detection results showed the 3D hepatocytes model was successfully constructed., Conclusion: The optimal conditions for 3D culture of L02 cell based on the VitroGel system were determined by RSM, and a hepatocytes model with high cellular activity was successfully constructed.

Objective: To investigate the anatomic zone localization based on biparametric magnetic resonance imaging (bpMRI) for the prediction of the risk degree in patients with prostate cancer. Methods: A total of 92 patients with prostate cancer confirmed by radical surgery in First Affiliated Hospital, Air Force Medical University, from January 2017 to December 2021 were collected. All patients underwent bpMRI (non-enhanced scan and DWI). According to ISUP grade, those patients were divided into low-risk group [≤grade 2, n =26, aged 71 (64.0, 5.2) years] and high-risk group[≥grade 3, n =66, aged 70.5 (63.0, 74.0) years]. The interobserver consistency test for ADC values was evaluated using the intraclass correlation coefficients (ICC). The differences in total prostate specific antigen (tPSA) between the two groups were compared and the χ 2 test was used to compare the differences in the risk of prostate cancer in the transitional and peripheral zone. Independent correlation factors for prostate cancer risk were analyzed by logistic regression using high and low risk of prostate cancer as dependent variables, including factors such as anatomical zone, tPSA, apparent diffusion coefficient mean (ADC mean ), apparent diffusion coefficient minimum (ADC min ) and age. Receiver operating characteristic (ROC) curves were plotted to assess the efficacy of the combined models of anatomical zone, tPSA, and anatomical partitioning+tPSA for diagnosing prostate cancer risk. Results: The ICC values of the ADC mean and ADC min between the observers were 0.906 and 0.885, respectively, with good agreement. The tPSA in the low-risk group was lower than that in the high-risk group [19.64 (10.29, 35.18) ng/ml vs 72.42 (24.79, 187.98) ng/ml; P <0.001]; the risk of prostate cancer in the peripheral zone was higher than that in the transitional zone, and the difference was statistically significant ( P <0.01). Multifactorial regression showed that anatomical zones ( OR =0.120, 95% CI :0.029-0.501, P =0.004) and tPSA ( OR =1.059, 95% CI :1.022-1.099, P =0.002) were risk factors for prostate cancer risk. The diagnostic efficacy of the combined model (AUC=0.895, 95% CI : 0.831-0.958) was better than the predictive efficacy of the single model for both anatomical partitioning (AUC=0.717, 95% CI :0.597-0.837) and tPSA (AUC=0.801, 95% CI : 0.714-0.887) ( Z =3.91, 2.47; all P <0.05). Conclusions: The malignant degree of prostate cancer in peripheral zone was higher than that in transitional zone. Combination of anatomic zone located by bpMRI and tPSA can be used to predict the risk of prostate cancer before surgery, expected to provide support for patients to develop personalized treatment strategies.

Objective: To establish a inductively coupled plasma mass spectrometry method for the determination of trace cobalt and tungsten in human urine. Methods: The authors used 1% nitric acid solution as diluent in October-December 2021, the sample dilution factor and internal standard element were optimized by single factor rotation experiment, and the difference between the working curve and the standard curve was compared. Results: The method uses working curve to determine cobalt and tungsten in urine, the linear range of this method was 0.0~10.0 μg/L, the correlation coefficient was 0.999 9, the detection limits respectively were 0.005 μg/L (cobalt) and 0.09 μg/L (tungsten), the recoveries of samples respectively were 87.0%~100.2% (cobalt) and 89.4%~104.8% (tungsten), the relative standard deviations respectively were 0.4%~4.4% (cobalt) and 0.6%~3.8% (tungsten) . Conclusion: A simple and rapid method for determination of cobalt and tungsten in urine has been established. This method has the advantages of simple operation, high sensitivity, low detection limit and good stability. It is suitable for determination of cobalt and tungsten in urine of all kinds of people.

Early weaned piglets suffer from oxidative stress and enteral infection, which usually results in gut microbial dysbiosis, serve diarrhea, and even death. Rice bran oil (RBO), a polyphenol-enriched by-product of rice processing, has been shown to have antioxidant and anti-inflammatory properties both in vivo and in vitro. Here, we ascertained the proper RBO supplementation level, and subsequently determined its effects on lipopolysaccharide (LPS)-induced intestinal dysfunction in weaned piglets. A total of 168 piglets were randomly allocated into four groups of seven replicates (42 piglets each group, (21±1) d of age, body weight (7.60±0.04) kg, and half males and half females) and were given basal diet (Ctrl) or basal diet supplemented with 0.01% (mass fraction) RBO (RBO1), 0.02% RBO (RBO2), or 0.03% RBO (RBO3) for 21 d. Then, seven piglets from the Ctrl and the RBO were treated with LPS (100 μg/kg body weight (BW)) as LPS group and RBO+LPS group, respectively. Meanwhile, seven piglets from the Ctrl were treated with the saline vehicle (Ctrl group). Four hours later, all treated piglets were sacrificed for taking samples of plasma, jejunum tissues, and feces. The results showed that 0.02% was the optimal dose of dietary RBO supplementation based on diarrhea, average daily gain, and average daily feed intake indices in early weaning piglets. Furthermore, RBO protected piglets against LPS-induced jejunal epithelium damage, which was indicated by the increases in villus height, villus height/crypt depth ratio, and Claudin-1 levels, as well as a decreased level of jejunal epithelium apoptosis. RBO also improved the antioxidant ability of LPS-challenged piglets, which was indicated by the elevated concentrations of catalase and superoxide dismutase, and increased total antioxidant capacity, as well as the decreased concentrations of diamine oxidase and malondialdehyde in plasma. Meanwhile, RBO improved the immune function of LPS-challenged weaned piglets, which was indicated by elevated immunoglobulin A (IgA), IgM, β‍‍-defensin-1, and lysozyme levels in the plasma. In addition, RBO supplementation improved the LPS challenge-induced dysbiosis of gut microbiota. Particularly, the indices of antioxidant capacity, intestinal damage, and immunity were significantly associated with the RBO-regulated gut microbiota. These findings suggested that 0.02% RBO is a suitable dose to protect against LPS-induced intestinal damage, oxidative stress, and jejunal microbiota dysbiosis in early weaned piglets.

Objective: To analyze the clinical characteristics and prognosis of primary and secondary pancreatic diffuse large B-cell lymphoma (DLBCL) . Methods: Clinical data of patients with pancreatic DLBCL admitted at Shanghai Rui Jin Hospital affiliated with Shanghai Jiao Tong University School of Medicine from April 2003 to June 2020 were analyzed. Gene mutation profiles were evaluated by targeted sequencing (55 lymphoma-related genes). Univariate and multivariate Cox regression models were used to evaluate the prognostic factors of overall survival (OS) and progression-free survival (PFS) . Results: Overall, 80 patients were included; 12 patients had primary pancreatic DLBCL (PPDLBCL), and 68 patients had secondary pancreatic DLBCL (SPDLBCL). Compared with those with PPDLBCL, patients with SPDLBCL had a higher number of affected extranodal sites ( P <0.001) and had higher IPI scores ( P =0.013). There was no significant difference in the OS ( P =0.120) and PFS ( P =0.067) between the two groups. Multivariate analysis indicated that IPI intermediate-high/high risk ( P =0.025) and double expressor (DE) ( P =0.017) were independent adverse prognostic factors of OS in patients with pancreatic DLBCL. IPI intermediate-high/high risk ( P =0.021) was an independent adverse prognostic factor of PFS in patients with pancreatic DLBCL. Targeted sequencing of 29 patients showed that the mutation frequency of PIM1, SGK1, BTG2, FAS, MYC, and MYD88 in patients with pancreatic DLBCL were all >20%. PIM1 ( P =0.006 for OS, P =0.032 for PFS) and MYD88 ( P =0.001 for OS, P =0.017 for PFS) mutations were associated with poor OS and PFS in patients with SPDLBCL. Conclusion: There was no significant difference in the OS and PFS between patients with PPDLBCL and those with SPDLBCL. IPI intermediate-high/high risk and DE were adverse prognostic factors of pancreatic DLBCL. PIM1, SGK1, BTG2, FAS, MYC, and MYD88 were common mutations in pancreatic DLBCL. PIM1 and MYD88 mutations indicated worse prognosis.

Liucheng county, as a suburb of Liuzhou City in Guangxi province, has a prominent ozone (O 3 ) pollution problem; however, there have been no relevant analyses of the cause of local O 3 pollution reported. In order to investigate the causes of O 3 pollution, an online observation of 116 VOCs with a time resolution of 1 h was carried out in Liucheng county from October 1 st to 15 th , and the sensitivity of ozone to the relative changes in the NO x and VOCs was analyzed. The results showed that the average value of φ ［total volatile organic compounds (TVOCs)］ during the observation period was 27.52×10 -9 , and the average value of φ (TVOCs) during the polluting process (October 1 st to 6 th ) was 32.15×10 -9 , which was 32.79% higher than that of the non-pollution process (October 8 th to 15 th ). In terms of species concentration, oxygenated volatile organic compounds (OVOCs) contributed the highest, accounting for 43.70%, followed by alkanes (23.00%), aromatics (11.75%), and halocarbons (7.35%). In terms of ozone formation potential (OFP), OVOCs contributed the highest (41.96%) to OFP, followed by aromatics (32.60%) and alkenes (17.92%). During the observation period, VOCs mainly came from motor vehicle emissions (32.44%), biomass combustion sources (29.31%), solvent use sources (16.43%), plant sources (11.34%), and chemical industry emissions (10.49%). The contribution ratios of solvent use sources and plant sources in the pollution process increased by 28.58% and 28.53%, respectively. The EKMA curve shows that, during the observation period, Liucheng county was in a synergistic control area for VOCs and nitric oxide (NO x ). Therefore, in the high ozone-occurrence autumn of Liucheng county, the key will be to reduce both VOCs and NO x emissions.

Prostate cancer is the most common malignant tumor in male urinary system, and the morbidity and mortality rate are increasing year by year. Traditional imaging examinations have some limitations in the diagnosis of prostate cancer, and the advent of molecular imaging probes and imaging technology have provided new ideas for the integration of diagnosis and treatment of prostate cancer. In recent years, prostate-specific membrane antigen (PSMA) has attracted much attention as a target for imaging and treatment of prostate cancer. PSMA ligand positron emission tomography (PET) has important reference value in the diagnosis, initial staging, detection of biochemical recurrence and metastasis, clinical decision-making guidance and efficacy evaluation of prostate cancer. This article briefly reviews the clinical research and application progress on PSMA ligand PET imaging in prostate cancer in recent years, so as to raise the efficiency of clinical applications.

Objective: To evaluate the quality of life and influencing factors of patients with herpes zoster (HZ) seen in hospitals. Methods: Based on Zoster Brief Pain Inventory (ZBPI) and Five-level EuroQol Five-dimensional Questionnaire (EQ-5D-5L), a cross-sectional survey was conducted to evaluate the pain severity and quality of life of 332 HZ cases seen in 22 hospitals of Lu'an City (Anhui Province), Zibo City (Shandong Province) and Tongchuan City (Shaanxi Province) from October to December 2021. The censored least absolute deviations (CLAD) model was used to analyze the related factors affecting the changes of patients' health utility values. Results: The 45.5% of 332 HZ cases were male. The median ( Q 1 , Q 3 ) age was 59 (50, 68) years. 59.64% of them assessed by ZBPI had moderate to severe pain in the past 24 hours (worst pain score≥5), and that of PHN cases was 84.8%(39/46). 77.7% (258/332), 77.4% (257/332) and 74.1% (246/332) of all patients reported that pain interfered with sleep, mood and general activities, respectively. Aging [β 40-49y (95% CI )=-0.11 (-0.15, -0.08); β 50-59y (95% CI )=-0.03 (-0.05, 0.00); β 60-69y (95% CI )=-0.09 (-0.12, -0.06); β 70-90y (95% CI )=-0.16 (-0.19, -0.12)], working status (unemployed) [β farmer (95% CI )=0.15 (0.13, 0.18); β retirees (95% CI )=0.21 (0.18, 0.24); β employee (95% CI )=0.13 (0.10, 0.16) ], complications[β PHN (95% CI )=-0.08 (-0.13, -0.04); β other complications (95% CI )=-0.12 (-0.15, -0.08)], within 30 days after onset [β(95% CI )=-0.01 (-0.03, 0.01)] and treatment [β other complications (95% CI )=-0.09 (-0.11, -0.06)] were related factors for the decline of health utility value (all P values <0.05). Conclusions: More than half of the patients with HZ had moderate to severe pain in the past 24 hours, which had a serious negative impact on the physical and mental health of the patients. Elderly patients, acute patients and patients with complications had lower health utility values and worse health status. We suggest that eligible people be vaccinated with HZ vaccine as soon as possible.

Purpose: To construct graphene oxide/methacrylated gelatin(GO/GelMA) hydrogel and to investigate its mechanical property and biocompatibility., Methods: 50, 100, 200 μg/mL GO was added to GelMA and mixed thoroughly. The micromorphology of hydrogels was observed under scanning electron microscope(SEM). Compression text was used to assess lastic modulus, CCK-8 and live/dead cell staining was conducted to assess the viability of rat bone marrow-derived mesenchymal stem cells (rBMSCs). The data were analyzed using SPSS 23.0 software package., Results: SEM showed a highly porous structure of the hydrogel. After adding GO, the elastic modulous increased (100, 200 μg/mL), swelling ratio increased(100 μg/mL) and expansion ratio decreased(200 μg/mL). The viability increased in 50 μg/mL GO and decreased in 100 and 200 μg/mL GO., Conclusions: Proper concentration of GO can increase the mechanical property and biocompatibility. 50 μg/mL GO/GelMA was suggested in bone tissue engineering given priority to biocompatibility.

Objective: In daily life, the movement of the neck will cause certain deformation of the blood vessel and the stent. This study explores the quantitative influence of the torsion deformation of the blood vessel on the mechanical properties of the stent., Methods: In the finite element simulation software Abaqus, the numerical simulation of the crimping and releasing process of the stent, the numerical simulation of the torsion process of the blood vessel with the stent, and the numerical simulation of the pressure loading process of the outer wall of the blood vessel were carried out., Results: After the stent was implanted, when a load was applied to the outer surface of the blood vessel wall, when the applied load did not change, as the torsion angle increased, the smallest cross-sectional area in the blood vessel decreased., Conclusions: After the stent is placed, when the external load is fixed, the radial support capacity of the stent will decrease as the torsion angle increases.

Objective: To study the composition of gut microbiota of Tibetan, Mongolian and Zhuang children., Methods: A total of 100 school-age children aged 10-12 were randomly selected from three ethnic groups in 2016, the Tibetan in Bayi County, Tibet, the Mongolian in Zha Qi, Inner Mongolia, and the Zhuang in Shanglin County, Nanning, Guangxi. Fecal samples were collected for high-throughput sequencing and bioinformatics analysis to compare the structural differences of gut microbiota of children from the three ethnic groups in the above areas., Results: The result of α diversity analysis showed that the diversity and richness of gut microbiota of children from three ethnic groups in the surveyed area were different in pairwise comparison. The dominant phyla and genera of Tibetan children&apos;s gut bacteria are Firmicutes(54.01%), Bacteroidetes(31.13%), Prevotella(15.72%) and Faecalibacterium(15.46%). The dominant phyla and genera of Mongolian children&apos;s gut bacteria are Firmicutes(54.89%), Bacteroidetes(17.85%), Proteobacteria(15.99%), Faecalibacterium(20.84%) and Escherichia(17.52%). While in Zhuang children, the dominant phyla and genera are Firmicutes(50.86%), Bacteroidetes(34.74%), Prevotella(18.40%), Bacteroides(17.78%) and Faecalibacterium(14.68%)., Conclusion: The composition and structure of gut microbiota of Tibetan and Zhuang children in the three regions are similar, while the structure of gut microbiota of Mongolian children is obviously different from those of the first two regions.

The extent of D3 lymphadenectomy for right colon cancer, especially the medial border of central lymph node dissection remains controversial. D3 lymphadenectomy and complete mesocolon excision (CME) are two standard procedures for locally advanced right colon carcinoma. D3 lymphadenectomy determines the medial border according to the distribution of the lymph nodes. The mainstream medial border should be the left side of superior mesenteric vein (SMV) according to the definition of D3, but there are also some reports that regards the left side of superior mesenteric artery (SMA) as the medial border. In contrast, the CME procedure emphasizes the beginning of the colonic mesentery and the left side of SMA should be considered as the medial border. Combined with the anatomical basis, oncological efficacy and technical feasibility of D3 lymph node dissection, we think that it is safe and feasible to take the left side of SMA as the medial boundary of D3 lymph node dissection. This procedure not only takes into account the integrity of mesangial and regional lymph node dissection, but also dissects more distant lymph nodes at risk of metastasis. It has its anatomical basis and potential oncological advantages. However, at present, this technical concept is still in the exploratory stage in practice, and the related clinical evidence is not sufficient.

This study aims to explore the targets of ginsenosides in brain based on drug affinity responsive target stability(DARTS) technology. Specifically, DARTS technology was combined with label-free liquid chromatography tandem mass spectrometry(LC-MS) to screen out the proteins in the brain that might interact with ginsenosides. Based on the screening results, adenylate kinase 1(AK1) was selected for further confirmation. First, the His-AK1 fusion protein was yielded successively through the construction of recombinant prokaryotic expression vector, expression of target protein, and purification of the fusion protein. Biolayer interferometry(BLI) was employed to detect the direct interaction of Rg&#95;1, Re, Rb&#95;1, Rd, Rh&#95;2, F1, Rh&#95;1, compound K(CK), 25-OH-PPD, protopanaxa-diol(PPD), and protopanaxatriol(PPT) with AK1, thereby screening the ginsenoside monomer or sapogenin that had strong direct interaction with the suspected target protein AK1. Then, the BLI was used to further determine the kinetic parameters for the binding of PPD(strongest interaction with AK1) to His-AK1 fusion protein. Finally, molecular docking technology was applied to analyze the binding properties between the two. With DARTS and LC-MS, multiple differential proteins were screened out, and AK1 was selected based on previous research for target verification. Fusion protein His-AK1 was obtained by prokaryotic expression, and the response(nm) of Re, Rg&#95;1, Rd, Rb&#95;1, Rh&#95;1, Rh&#95;2, F1, PPT, PPD, 25-OH-PPD, and CK with His-AK1 was respectively 0.003 1, 0.001 9, 0.042 8, 0.022 2, 0.013 4, 0.037 3, 0.013 9, 0.030 7, 0.140 2, 0.016 0, and 0.040 8. The K&#95;(on), K&#95;(off), and K&#95;D values of PPD and His-AK1 were determined by the BLI as 1.22×10~2 mol~(-1)·L·s~(-1), 1.04×10~(-2) s~(-1), 8.52×10~(-5) mol·L~(-1). According to the molecular docking result, PPD bound to AK1 with the absolute value of the docking score of 3.438, and hydrogen bonds mainly formed between the two. Thus, AK1 is one of the protein action sites of ginsenosides in the brain. The direct interaction between ginsenoside metabolite PPD and AK1 is the strongest.

Objective: To investigate the role and mechanism of long non-coding RNA (lncRNA) C9ORF139 targeting micro RNA(miR)-24-3P/TAOK1 in regulating the proliferation of acute myeloid leukemia (AML) cells. Methods: AML cells HL-60 and THP-1 were purchased from the Chinese Academy of Sciences and divided into 4 groups:group A was negative control group (siNC group), group B was interference C9ORF139 group (siC9ORF139 group), group C was siC9ORF139+miR-24-3p inhibitor group, and group D was miR-24-3P+TAOK1 overexpression group (oe-TAOK1 group). Real-time fluorescence quantitative reverse transcription PCR was used to detect the expression levels of AML cell lines of HL-60 and THP-1 in four groups. Cell Counting Kit-8 assay was performed to measure cell proliferation. Flow cytometry was applied to analyze cell apoptosis. Transwell test was applied to detect cell migration and invasion ability. Western blot was used to detect p-serine/threonine kinase (p-raf) and p-mitogen activation proteinkinase (p-MEK), p-extracellular regulatory protein kinase (p-ERK) expression. The luciferase reporter gene plasmid was constructed to verify the binding ability of C9ORF139,miR-24-3P and TAOK1.Nude mice were inoculated with subcutaneous tumor cells of HL-60 (group A) and HL-60 (group B). Results: After the C9ORF139 gene was knocked down and cultured for 120 h, The cell proliferation ability (0.62±0.02, 0.82±0.02), migration ability (0.22±0.03, 0.05±0.01), invasion ability (0.20±0.02, 0.13±0.03) of group B were all lower than that of group A (1.30±0.02, 1.83±0.07; 0.99±0.02, 0.99±0.02; 1.00±0.01, 1.00±0.01) (all P <0.05). When co-transfected with miR-24-3 inhibitor, cell proliferation ability, migration ability and invasion ability were all higher in group B (all P <0.05). When co-transfected with miR-24-3P and oe-TAOK1 plasmid, cell proliferation ability, migration ability and invasion ability were all higher than group B (all P <0.05).When the C9ORF139 gene in the cells was knocked down, the apoptosis level of group B (28.56±8.07, 17.74±1.91) were higher than those of group A (0.31±0.27, 2.49±0.33)(all P <0.05); when co-transfected with miR-24-3P inhibitor, the apoptosis level (2.34±0.09, 3.06±0.06) were lower than those in group B (all P <0.05); when co-transfected with miR-24-3P and oe-TAOK1 in the plasmid group, the apoptosis level (2.16±1.29, 4.80±0.37) were also lower than those of group B (all P <0.05). In HL-60 and THP-1 cells, when C9ORF139 was not mutated, the luciferase activity of miR-24-3P group was lower than that of the miR-NC group ( P <0.05). When the binding site with miR-24-3p in C9ORF139 sequence was mutated, the luciferase activity in miR-24-3p group was equivalent to that in miR-NC group ( P> 0.05).When TAOK1 was not mutated; the luciferase activity of miR-24-3P group was lower than that of group A ( P <0.05). When the binding site with miR-24-3p in TAOK1 sequence was mutated, the luciferase activity in miR-24-3p group was equivalent to that in miR-NC group ( P> 0.05).When the C9ORF139 gene in HL-60 cells was knocked down and cultured for 72 h, the phosphorylation expression levels of Raf, MEK and ERK molecules in group B were significantly lower than those in group A (all P <0.05). By day 14, the tumor volume in the group A was greater than the tumor cell volume in the group B [(284.49±57.61) vs (125.70±18.64) mm 3 , P =0.017]. The tumor weight of HL-60 in group A was heavier than that of group B [(847.80±159.36) vs (408.40±113.16) mg, P =0.001]. Conclusions: LncRNA C9ORF139 regulates TAOK1 by sponging miR-24-3P to promote the proliferation, invasion and migration of acute myeloid leukemiacell.In vivo experiments have confirmed that the expression of C9ORF139 can promote the growth of subcutaneous tumors in AML nude mice.

Objective: To investigate the difference expression of circular RNA (circRNA) in acute myeloid leukemia (AML) by using bioinformatics method., Methods: The microarray chip data of AML was searched and downloaded from the Gene Expression Omnibus (GEO) of the National Center for Bioinformatics (NCBI). The differences between AML samples and control samples were analyzed by R software. The interaction between deregulated circRNA, miRNA and mRNA were predicted by miranda software and miRTarBase software. The circRNA-miRNA-mRNA regulatory network was constructed by using the cytoHubba plugin based on the Cytoscape software., Results: A total of 203 differential expression of circRNAs were finally collected, including down-regulated 161 circRNAs and up-regulated 42 circRNAs. CircRNA/miRNA/mRNA interaction network was constructed through software prediction. hsa&#95;circ&#95;0001080, hsa&#95;circ&#95;0004511, hsa&#95;circ&#95;0054211, hsa&#95;circ&#95;0001944 may be positively regulated the gene expression in AML., Conclusion: Abnormal expression of circRNA in AML may become a new target for AML treatment.

O -linked β - N -acetylglucosamine ( O -GlcNAc), an important post-translational modification of proteins, plays an essential role in maintaining normal biological activities of organisms. Studies have shown that the disruption of O -GlcNAc homeostasis is closely associated with various human diseases. Therefore, large-scale enrichment and identification of O -GlcNAc proteins is important for exploring diagnostic biomarkers of clinical diseases. O -GlcNAcylation is substoichiometric, and its glycosidic bond is unstable; hence, the enrichment and identification of O -GlcNAc proteins remains a challenge. Recently, metabolic labeling technology with per- O -acetylated unnatural monosaccharides has been introduced to enrich O -GlcNAc proteins or peptides in cells. However, azidosugars can also react with the thiol group of cysteine to form cysteine thiol-azidosugar artificial modification as a byproduct in cell metabolism, which interferes with the identification of O -GlcNAc proteins or peptides. Therefore, the development of a methodology for the specific and complete removal of cysteine thiol-azidosugar artificial modification is necessary. O -Mesitylenesulfonylhydroxylamine (MSH) is an oxidizing and aminating reagent with great potential to oxidatively eliminate the cysteine thiol modification to dehydroalanine. Hence, it is worthwhile to probe whether MSH can eliminate cysteine thiol-azidosugar artificial modification. Two cysteine thiol-azidosugar artificial modifications were synthesized successfully by incubating thiol standard peptides with tetraacetylated N -azidoacetylgalactosamine (Ac 4 GalNAz) in sodium carbonate buffer (200 mmol/L, pH 10) at 37 ℃ for 90 min. Then, the reaction conditions for MSH oxidative elimination were optimized to completely remove cysteine thiol-azidosugar artificial modification. Sodium phosphate buffer (50 mmol/L, pH 8.0) was selected to protect the O -GlcNAc modification due to its mild nature. After extensive investigation, the optimized reaction conditions were established as 95 ℃ (reaction temperature) and 30 min (reaction time). Both cysteine thiol-azidosugar artificial modifications could be removed completely under these conditions. Furthermore, two azide-labeled O -GlcNAc (N 3 - O -GlcNAc) peptides were used to assess whether MSH destroyed the O -GlcNAc modification at the same time. The results showed that the two N 3 - O -GlcNAc peptides were stable after treatment with MSH at 95 ℃ for 30 min. In short, with excess MSH, the N 3 - O -GlcNAc peptides were stable, but the cysteine thiol-azidosugar artificial modification was exhausted in sodium phosphate buffer (50 mmol/L, pH 8.0) at 95 ℃ for 30 min. Moreover, both O -GlcNAc modification and cysteine thiol-azidosugar artificial modification exist in cell metabolism. This method can not only remove cysteine thiol-azidosugar artificial modification but also ensure the stability of O -GlcNAc modification. There is growing evidence that O -GlcNAcylation mainly occurs in proteins in the cytoplasm and nucleus, and that most O -GlcNAc proteins are involved in important biological signaling pathways. The nuclear and cytoplasmic proteins incubated with Ac 4 GalNAz in HeLa cells were selected as a model system. After the nuclear and cytoplasmic proteins were digested into peptides, MSH was applied to remove cysteine thiol-azidosugar modification. In addition, 51 peptides for the elimination of the cysteine thiol-azidosugar artificial modification were identified, indicating that MSH can remove the interference of cysteine thiol-azidosugar artificial modification in cell metabolism. Biotin probe and streptavidin dynabeads were subsequently used to label and enrich N 3 - O -GlcNAz peptides in cell metabolism. Finally, 157 O -GlcNAc peptides attributed to 130 proteins were identified. To better understand the functional roles of O -GlcNAc proteins, gene ontology analysis was performed. Cell component analysis showed that the identified O -GlcNAc proteins were mainly distributed inpostsynaptic density, cytoplasm, and condensed nuclear chromosome. The proteins were mostly involved in biological processes, including cell division, excitatory postsynaptic potential, and microtubule-based movement. The proteins responsible for transferase activity, transferring acyl groups, histone acetyltransferase activity, and microtubule binding were highly enriched, indicating that O -GlcNAc proteins play important roles in cells. In summary, this work developed an approach to enrich O -GlcNAc peptides precisely in metabolic labeling by removing cysteine thiol-azidosugar artificial modification with MSH. This methodology provides a new strategy for the application of metabolic labeling technology with unnatural monosaccharides in glycoproteomics analysis.

Objective: To select and optimize the conditions of urine cobalt determination by graphite furnace atomic absorption spectroscopy and establish a method for urine cobalt determination. Methods: In April 2020, the matrix modifier and spectrum wavelength were selected by mathematical statistics method, the heating procedure of graphite furnace was optimized, the variance of orthogonal test results was analyzed, and the working curve was quantified by external standard method. Results: The results showed that the diammonium hydrogen phosphate was a matrix modifier with a wavelength of 240.7 nm, and the optimized graphite furnace heating procedure showed a good linear relationship at 5-80 μg/L. The correlation coefficient was 0.9991. The detection limit was 0.9 μg/L, the lowest detection concentration was 1.8 μg/L. The recovery was 94.0%-101.4%, and the precision was 2.6%-5.8%. Conclusion: The orthogonal design method can efficiently and scientifically optimize the optimal combination of the various factor levels of graphite furnace atomic absorption spectrometry. The determination method established based on this has high precision and good accuracy, and can be applied to the determination of cobalt in urine.

N -Glycosylation of proteins, an important post-translational modification in eukaryotic cells, plays an essential role in the regulation of cell adhesion, migration, signal transduction, and apoptosis. Abnormal changes in protein glycosylation are closely related to the occurrence of many critical diseases, including diabetes, tumors, and neurological, kidney, and inflammatory diseases. A non-invasive type of liquid biopsy, urine sampling has the advantage of reducing the complexity of proteomic analysis. This facilitates the design of large-scale and continuous or multi-time point sampling strategies. However, the dynamic range of urinary protein abundance is relatively large, owing to individual differences and physiological conditions. Currently, there is a lack of specialized research on individual differences, physiological fluctuations, and physiological abundance ranges of urinary N -glycoproteins in large healthy populations. Therefore, it is difficult to accurately distinguish individual differences and normal physiological fluctuations from changes caused by disease; this poses a great challenge in disease marker research. Liquid chromatography-mass spectrometry (LC-MS) is an analytical technique widely used for the large-scale profiling of proteomes in biological systems, and the enrichment of N -glycopeptides is a prerequisite for their detection by MS.In this study, we established an approach based on hydrophilic interaction chromatography (HILIC) by optimizing the activation, cleaning, and elution processes of the enrichment method, for instance through the optimization of particle size and solvent composition, and investigated the identification number, selectivity, and stability of N -glycoprotein/ N -glycopeptide enrichment under different experimental conditions. We found that N -glycoproteins and N -glycopeptides were highly enriched in a trifluoroacetic acid system with 5-μm filling particles in the HILIC column. On this basis, we analyzed the levels of N -glycoproteins/ N -glycopeptides in urine samples. The consistency of N -glycoprotein/ N -glycopeptide levels in urine samples taken from the same healthy person for five consecutive days was investigated by correlation analysis. This analysis revealed that the urinary N -glycoproteome of the same healthy person was relatively stable over a short period of time. Next, urinary samples from 20 healthy male volunteers and 20 healthy female volunteers were enriched for N -glycoproteins/ N -glycopeptides, which were profiled by MS through qualitative and quantitative analyses. Screening and functional analysis of differential proteins were then carried out. A total of 1016 N -glycoproteins and 2192 N -glycopeptides were identified in the mid-morning urine samples of the 40 healthy volunteers. A label-free quantitation strategy was used to investigate the fluctuation range of the physiologically abundant urinary N -glycopeptides. The abundance of urinary N -glycopeptides spanned across approximately five orders of magnitude. Subsequently, gender differences in the N -glycosylation levels of urinary proteins were also explored in healthy people. Functional analysis of the N -glycoproteins that exhibited gender differences in abundance was performed. Based on multivariate statistical analysis, 206 differentially expressed proteins ( p <0.05, fold change (FC)> 4) were identified. In females, we found 175 significantly down-regulated N -glycoproteins and 31 significantly up-regulated N -glycoproteins with respect to males. The expression levels of N -glycopeptides between the two groups suggested a clear gender difference. To investigate the biological processes and functions of these proteins, gene ontology (GO) analysis was performed on the N -glycoproteins/ N -glycopeptides differentially expressed between males and females. Metabolic pathway analysis was also carried out based on the kyoto encyclopedia of genes and genomes (KEGG). Differentially expressed N -glycoproteins were mostly associated with platelet degranulation, extracellular region, and ossification. The top three relevant pathways were glycan biosynthesis and metabolism, metabolism of cofactors and vitamins, and lipid metabolism. Overall, sex may be an important factor for urinary N -glycoproteome differences among normal individuals and should be considered in clinical applications. This study provides relevant information regarding the function and mechanisms of the urinary glycoproteome and the screening of clinical biomarkers.

Objective: To investigate the expression of microRNA-29 a(miR-29 a) in the human steatotic hepatocyte model and the mechanism of targeting silent mating type information regulation 2 homolog-1(Sirt1)to regulate fat deposition of steatotic hepatocyte., Methods: The nonalcoholic fatty liver cell model was induced by a mixture of oleic acid and palmitic acid. After successful validation model, the expression of miR-29 a and Sirt1 was measured by PCR. The target genes of miR-29 a was predicted in biological system. MiR-29 a mimic and miR-29 a inhibitor were transfected into hepatocytes, and then established the human steatotic hepatocyte model, the result of oil red O staining and triglyceride(TG)lipid content were observed, the expression of Sirt1 mRNA and protein were detected by qRT-PCR and Western blotting, respectively., Results: The steatosis hepatocyte model was successfully established. Compared with control group, the relative expression of miR-29 a and triglyceride increased significantly(P&lt;0. 01), while the relative expression of Sirt1 decreased significantly(P&lt;0. 01) in the model group. Sirt1 was a target gene of miR-29 a. After transfection, the lipid droplet and the deposition of fat increased obviously in miR-29 a mimic group than those in the control group. TG content in miR-29 a mimic group increased significantly(P&lt;0. 05), the expression of miR-29 a increased significantly(P&lt;0. 01), while the expression of Sirt1 mRNA decreased significantly(P&lt;0. 05), and the expression of Sirt1 protein showed a downtrend. On the contrary, after the inhibition of miR-29 a expression, the lipid droplets in miR-29 a inhibitor group were relatively reduced, the fat deposition was alleviated. The TG content was significantly decreased(P&lt;0. 05), the expression of miR-29 a in the cells was effectively inhibited(P&lt;0. 01), while the expression of Sirt1 mRNA was significantly increased(P&lt;0. 05), and Sirt1 protein was on an upward compared with the control group., Conclusion: The expression of miR-29 a is significantly increased in the nonalcoholic fatty liver cell model. Upregulation of miR-29 a negatively regulates the expression of Sirt1, thus promoting fat deposition of steatotic hepatocyte.

In "shotgun" proteomics strategy, the proteome is explained by analyzing tryptic digested peptides using liquid chromatography-mass spectrometry. In this strategy, the retention time of peptides in liquid chromatography separation can be predicted based on the peptide sequence. This is a useful feature for peptide identification. Therefore, the prediction of the retention time has attracted much research attention. Traditional methods calculate the physical and chemical properties of the peptides based on their amino acid sequence to obtain the retention time under certain chromatography conditions; however, these methods cannot be directly adopted for other chromatography conditions, nor can they be used across laboratories or instrument platforms. To solve this problem, in recent years, deep learning was introduced to proteomics research for retention time prediction. Deep learning is an advanced machine-learning method that has extraordinary capability to learn complex relationships from large-scale data. By stacking multiple hidden neural networks, deep learning can ingest raw data without manually designed features. Transfer learning is an important method in deep learning. It improves the learning process a new task through the transfer of knowledge from an already-learned related task. Transfer learning allows models trained using large datasets to be utilized across conditions by fine-tuning on smaller datasets, instead of retraining the whole model. Many retention time prediction methods have been developed. In the process of training the model, the sequences of peptides are encoded to represent peptide information. Deep learning considers the relationship between the characteristics of the peptides and their corresponding retention times without the need for manual input of the physical and chemical properties of the peptides. Compared with traditional methods, deep learning methods have higher accuracy and can be easily used under different chromatography conditions by transfer learning. If there are not enough datasets to train a new model, a trained model from other datasets can be used as a replacement after calibration with small datasets obtained from these chromatography conditions. While the retention times of modified peptides can also be predicted, the predictions are inadequate for complex modifications such as glycosylation, and this is one of the main problems to be solved. The predicted retention times were used to control the quality of peptide identification. With high accuracy, the predicted retention times can be considered as actual retention times. Therefore, the difference between predicted and observed retention times can serve as an effective and unbiased quantitative metric for evaluating the quality of peptide-spectrum matches (PSMs) reported using different peptide identification methods. Combined with fragment ion intensity prediction, retention time prediction is used to generate spectral libraries for data-independent acquisition (DIA)-based mass spectrometry analysis. Generally, DIA methods identify peptides using specific spectrum libraries obtained from data-dependent acquisition (DDA) experiments. As a result, only peptides detected in the DDA experiments can be present in the libraries and detected in DIA. Furthermore, it takes a lot of time and effort to build libraries from DDA experiments, and typically, they cannot be adopted across different laboratories or instrument platforms. In contrast, the pseudo spectral libraries generated by retention times and fragment ion intensity prediction can overcome these shortcomings. The pseudo spectral libraries generate theoretical spectra of all possible peptides without the need for DDA experiments. This paper reviews the research progress of deep learning methods in the prediction of retention time and in related applications in order to provide references for retention time prediction and protein identification. At the same time, the development direction and application trend of retention time prediction methods based on deep learning are discussed.

Ribonucleic acid (RNA) rarely exists alone in the cell. RNAs interact with a variety of proteins and form RNA-protein complexes (RP-complexes) in every step of their life cycle, from transcription to degradation. These RP-complexes play key roles in regulating a variety of physiological processes. Defects in the composition and function of RP-complexes have been associated with many diseases, including metabolic disorders, muscular atrophy, autoimmune diseases, and cancer. It is hence evident that deciphering the highly complex interaction network of RNA-binding proteins (RBPs) and their RNA targets will provide a better understanding of disease development and lead to the discovery of new targets for cancer therapy. Large-scale identification of RP-complexes at the omics level is a prerequisite for obtaining insights into the complex RNA-protein interaction network. As the first step in omics-wide decoding of RP-complexes, enrichment and purification of RP-complexes is a highly challenging task. Recently, intensive efforts have been undertaken to better enrich and identify RP-complexes. Generally, the enrichment strategies can be classified into two major categories: in vitro and in vivo. Although it has been successfully applied in many studies, the in vitro transcribed bait RNA lacks modifications or structural similarity compared with its natural counterpart. Further, since the proteins relocate and remodel after cell lysis, the use of cell lysates as a protein source may result in capturing false interacting proteins that bind non-physiologically with the bait RNA. Finally, weak interactions between the non-covalently bound proteins and RNA require mild washing to remove non-specific binding, which needs careful optimization. However, substantial sample loss is inevitable. To overcome the disadvantages of in vitro approaches, in vivo cross-linking strategies that "freeze" natural RNA-protein complexes in intact cells via covalent cross-linking have become increasingly popular. The in vivo methods allow RNA to interact with proteins in the intracellular environment. Therefore, the RP-complexes formed under physiological conditions are more biologically relevant than those obtained by in vitro methods. We herein summarize recent in vivo methodological advances in the large-scale enrichment and identification of RP-complexes, including cross-linking and immunoprecipitation (CLIP) and related methods, click chemistry-assisted methods, and organic phase separations. CLIP involves irradiating living cells with 254-nm ultraviolet (UV) light to establish covalent bonds between RNA and proteins. This enables CLIP to purify RNAs bound to a specific RBP under conditions that are stringent enough to prevent co-purification of nonspecifically bound proteins or free RNAs. Since the original study, multiple variant protocols have been derived to increase both efficiency and convenience. Photoactivatable ribonucleoside-enhanced-CLIP (PAR-CLIP) introduces a variation in the crosslinking strategy. Cells were preincubated with photoactivatable ribonucleosides 4-thiouridine (4SU) or 6-thioguanosine (6SG), which enables protein-RNA crosslinking with 365-nm UV-A irradiation. It increases the efficiency of cross-linking between RNA and RBPs and is particularly valuable for studying the interactions between RBPs and nascent RNA. Using a click chemistry-assisted strategy, an alkyne modified uridine analog, 5-ethynyluridine (EU), was incorporated into nascent RNAs via metabolic incorporation in living cells. Combined with UV irradiation-based cross-linking, the alkyne-functionalized RNA and the bound proteins were purified in a poly A-independent fashion by the highly selective bioorthogonal copper (I)-catalyzed azide-alkyne cycloaddition using azide-modified beads. Thus, full lists of both coding and non-coding RNAs with their interacting proteins can be purified, which is a major methodological advance. Organic phase separation methods exploiting the physicochemical difference between cross-linked RP-complexes and free RNA and proteins do not require metabolic-based alkyne labeling or polyA-based RNA capture. Each method has unique strengths and drawbacks, which makes it important to select optimal approaches for the biological question being addressed. We hope that this review points out the current limitations and provides future directions to facilitate further development of methods for large-scale investigation of RP-complexes.

Objective: To investigate the mechanisms of anti-apoptosis and immune evasion in drug-resistant leukemia cells mediated by STAT3, further to explore the possible mechanism of leukemia relapse caused by minimal residual., Methods: Drug-resistance leukemia cell line was established by transfecting pcDNA3.1-STAT3 into K562 cells (K562/STAT3). The expression of STAT3, BAX and NKG2D ligands (MICA and ULBP1) in K562/-cells, K562/STAT3 were detected by Western blot and/or RQ-PCR. Cells apoptosis and the killing effect of NK cells on leukemia cells were detected by flow cytometry., Results: The expression of the total STAT3, STAT3 phosphorylation in K562/STAT3 was significantly increased, and P-gp mRNA expression was increased also significantly (P<0.005). In K562/STAT3 cells, the expression of pro-apoptotic BAX (P=0.005) was significantly lower, and the number of apoptotic cells (P=0.002) induced by adriamycin was significantly decreased as compared with those in K562/- cells. After K562/STAT3 cells were treated by STAT3 inhibitor (SH-4-54), the expression of BAX mRNA (P=0.017) was significantly higher and the number of apoptotic cells (P=0.005) was significantly increased. The MICA and ULBP1 mRNA expression in K562/STAT3 cells was significantly lower than that in K562/- cells, and also for MICA and ULBP1 protein (MICA and ULPB1 mRNA: P<0.0001, MICA protein: P=0.001, ULPB1 protein: P=0.022). After K562/STAT3 cells were treated with STAT3 inhibitor (SH-4-54), the expression of MICA mRNA and protein was increased (mRNA: P=0.001, protein: P=0.002), but ULBP1 mRNA and protein showed no significantly change (mRNA: P=0.137, protein: P=0.1905). The cytotoxicity of NK cells to K562/STAT3 cells was susceptible as compared with K562/- (P=0.002), but the cytotoxicity of K562/STAT3 cells to NK cell could be recovered by STAT3 inhibitor (P=0.006)., Conclusion: STAT3 phosphorylation can inhibits cell apoptosis and promotes cell immune escape. STAT3 inhibitors can promote the apoptosis of leukemia cells and increase their sensitivity to NK cells.

Stress distribution of denture is an important criterion to evaluate the reasonableness of technological parameters, and the bite force derived from the antagonist is the critical load condition for the calculation of stress distribution. In order to improve the accuracy of stress distribution as much as possible, all-ceramic crown of the mandibular first molar with centric occlusion was taken as the research object, and a bite force loading method reflecting the actual occlusal situation was adopted. Firstly, raster scanning and three dimensional reconstruction of the occlusal surface of molars in the standard dental model were carried out. Meanwhile, the surface modeling of the bonding surface was carried out according to the preparation process. Secondly, the parametric occlusal analysis program was developed with the help of OFA function library, and the genetic algorithm was used to optimize the mandibular centric position. Finally, both the optimized case of the mesh model based on the results of occlusal optimization and the referenced case according to the cusp-fossa contact characteristics were designed. The stress distribution was analyzed and compared by using Abaqus software. The results showed that the genetic algorithm was suitable for solving the occlusal optimization problem. Compared with the reference case, the optimized case had smaller maximum stress and more uniform stress distribution characteristics. The proposed method further improves the stress accuracy of the prosthesis in the finite element model. Also, it provides a new idea for stress analysis of other joints in human body.

To improve and perfect the quality standards and propose recommendations for the revision of quality standards for Andrographis Herba and its processed slices in Chinese Pharmacopoeia(ChP)(2020 edition) based on the problems and limitations in ChP(2015 edition). TLC identification method with andrographolide and control herbs as references was established using silica gel G thin layer plate, with chloroform-methylbenzene-methanol(8∶1∶1) as developing solvent, and 10% sulfuric acid ethanol solution as colour-developing agent. This method has good reproducibility, strong specificity and high sensitivity. As compared with the original method in ChP 2015, this method has better development effect and clearer spots. Based on the previous research, a quantitative analysis of multi-components by single-marker(QAMS) method with andrographolide as the internal reference substance was developed to simultaneously determine the contents of 4 diterpene lactones: andrographolide(S), neoandrographolide(A), 14-deoxyandrographolide(B), and dehydroandrographolide(C). The relative correction factors of f&#95;(A/S), f&#95;(B/S), and f&#95;(C/S) were determined as 1.12, 0.79, and 0.63, respectively. The relative retention time of t&#95;(A/S), t&#95;(B/S), and t&#95;(C/S) was 1.95, 2.18, and 2.25, respectively. According to the content determination results in 46 batches of crude drugs and 38 batches of processed slices, it was stipulated that the total contents of 4 diterpene lactones should not be less than 1.5% and 1.2% in crude drugs and processed slices, respectively. As compared with the original method in ChP 2015, the present QAMS method could not only reduce the detection cost and improve the efficiency, but also can be used to evaluate the quality of Andrographis Herba and its processed slices more comprehensively and objectively. Diterpene lactones are generally recognized as the effective components in Andrographis Herba, and their contents in leaves were much higher than those in stems. However, almost all of the current commercial processed slices are processed from stems, so their quality is gene-rally poor and the efficacy is hard to be guaranteed. Therefore, the weight percentage of leaves should be added into the inspection items of the processed slices and it should not be less than 25%.

As a representative part of the oral system and masticatory robot system, the modeling method of the dental model is an important factor influencing the accuracy of the multi-body dynamic model. Taking the right first molars of the masticatory robot as the research object, an equivalent model, point-contact higher kinematic pair composed of v-shaped surface and sphere surface, was proposed. Firstly, the finite element method was used to analyze the occlusal dynamics of the original model in three static contact cases (intrusive contact, centric occlusion, and extrusive contact) and one dynamic chewing case, and the expected bite force was obtained. Secondly, the Hertz contact model was adopted to establish the analytical expression of the bite force of the equivalent model in three static contact cases. The normal vectors and contact stiffness in the expression were designed according to the expected bite force. Finally, the bite force performance of the equivalent model in three static contact cases and one dynamic chewing case was evaluated. The results showed that the equivalent model could achieve the equivalent bite force of 8 expected items in the static contact cases. Meanwhile, the bite force in the early and late stages of the dynamic chewing case coincides well with the original model. In the middle stage, a certain degree of impact is introduced, but it can be weakened by subsequent trajectory planning. The equivalent modeling scheme of the dental model proposed in this paper further improves the accuracy of the dynamic model of the multi-body system. It provides a new idea for the dynamic modeling of other complex human contacts.

Objective: To analyze the clinical features and death-related risk factors of COVID-19. Methods: We enrolled 891 COVID-19 patients admitted to the Affiliated Hospital of Jianghan University from December 2019 to February 2020, including 427 men and 464 women. Of the 891 cases, 582 were severe or critical, including 423(73%)severe and 159 (27%) critical cases. We compared the demographics, laboratory findings, clinical characteristics, treatments and prognosis data of the 582 severe patients. Univariate and multivariate logistic regression analysis was conducted to explore the risk factors associated with death in COVID-19 patients. Results: The 582 severe patients included 293 males and 289 females, with a median age of 64(range 24 to 106). Sixty-three patients died, including 45 males and 18 females, with a median age of 71(range 37 to 90). The average onset time of the 582 patients was 8 days, of whom 461 (79%) had fever, 358 (62%) dry cough, 274 (47%) fatigue. There were 206 cases with shortness of breath (35%), 155 cases with expectoration (27%), 83 cases with muscle pain or joint pain (14%), 71 cases with diarrhea (12%), and 29 cases with headache (4%). Underlying diseases were present in 267 (46%) patients, most commonly hypertension (194, 33%), followed by diabetes (69, 12%), coronary atherosclerotic heart disease (37, 6%), tumor (18, 3%), and chronic obstructive pulmonary disease (5, 1%). Chest CT showed bilateral lung involvement in 505 patients (87%). Upon admission, the median lymphocyte count of the 582 patients was 0.8(IQR, 0.6-1.1)×10(9)/L, the median D-dimer was 0.5 (IQR, 0.4- 0.8) mg/L, the median N-terminal brain natriuretic peptide precursor (NT-proBNP) was 433 (IQR, 141- 806) pg/L, and the median creatinine was 70.3 (IQR, 56.9-87.9) μmol/L. The death group had a median lymphocyte count of 0.5 (0.4-0.8)×10(9)/L, D-dimer 1.1 (0.7-10.0)mg/L, N-terminal brain natriuretic peptide precursor 1479(893-5 087) pg/ml, and creatinine 89.9(67.1-125.3) μmol/L. Multivariate logistic analysis showed that increased D-dimer ( OR : 1.095, 95% CI: 1.045-1.148, P< 0.001), increased NT-proBNP ( OR : 4.759, 95% CI: 2.437-9.291, P< 0.001), and decreased lymphocyte count ( OR : 0.180, 95% CI: 0.059-0.550, P= 0.003) were the risk factors of death in COVID-19 patients. Conclusions: The average onset time of severe COVID-19 was 8 days, and the most common symptoms were fever, dry cough and fatigue. Comorbidities such as hypertension were common and mostly accompanied by impaired organ functions on admission. Higher D-dimer, higher NT-proBNP, and lower lymphocyte count were the independent risk factors of death in COVID-19 patients.

Objective: To investigate the effects of serum immunoglobulin A/complement factor 3 (IgA/C3) ratio and glomerular C3 staining on clinical prognosis in patients with IgA nephropathy. Methods: From January 1st, 2007 to December 30th, 2016, a total of 519 patients with biopsy-proven IgA nephropathy (IgAN) in West China Hospital were retrospectively reviewed and divided into four groups based on serum IgA/C3 ratio and glomerular C3 staining: group A with IgA/C3 ratio ≥3.046 (median) and glomerular C3 staining ≥2 ( n= 151), group B with IgA/C3 ratio ≥3.046 and glomerular C3 staining<2 ( n= 109), group C with IgA/C3 ratio<3.046 and glomerular C3 staining ≥2 ( n= 119), and group D with IgA/C3 ratio<3.046 and glomerular C3 staining<2 ( n= 140). Clinical data, pathological characteristics and the primary endpoint [≥ 50% decline in estimated glomerular filtration rate (eGFR) and/or end-stage renal disease (ESRD)]were collected. Clinical prognosis and relevant risk factors were analyzed among the four groups. Results: Totally, 519 patients (298 males, 57.4%) with an average age of (33.6±10.9) years were recruited and followed up for (43.4±21.6) months. The rate of complete remission plus partial remission was 74.2% (112/151), 74.3% (81/109), 72.3% (86/119), 81.4% (114/140) in group A, B, C, D, respectively. Meanwhile, The rate of ESRD was highest in group A (14.6% vs 9.2%, 13.4%, 8.6%). Renal outcome (patients reached the endpoint) was worse in group A and C compared with group B and D (15.2%, 16.0 vs 8.3%, 7.9%). Moreover, 80-month renal survival rate was significantly worse in group A (84.8%) than that in group B and D (91.7% and 92.1%), but no statistical significant difference was found between group A and B ( P (AB)=0.085; P (AD)=0.028). There was no significant difference of renal survival rate between group A and C (84.8% vs 84.0%, P= 0.896). Multivariate Cox model showed that hypertension ( HR =2.753, 95 %CI : 1.452-5.217, P= 0.002), serum creatinine ( HR =1.011, 95 %CI : 1.008-1.014, P< 0.001), and tubular atrophy/interstitial fibrosis (T1/T2) ( HR =6.595, 95 %CI : 3.107-13.999, P< 0.001) were independent predictors of poor renal survival. Conclusion: Serum IgA/C3 ratio and glomerular C3 staining are predictors of renal clinical prognosis in patients with IgA nephropathy.

Protein N -glycosylation plays a crucial role in the folding, transportation, and localization of proteins, and participates in many important biological processes such as receptor activation and signal transduction. An increasing number of studies have shown that abnormal protein glycosylation is closely related to various diseases. Therefore, N -glycosylated proteins are potential candidates for become new biomarkers or drug targets. The current research strategy for N -glycosylated proteins is to first digest the protease into peptides and then identify them by mass spectrometry. In sample preparation processing, enrichment and separation of N -glycopeptides have become the vital step for glycoproteomic analysis. However, because of their low abundance and poor ionization, mass spectrometric identification of N -glycopeptides in complex samples remains a challenging task. Therefore, the development of a selective enrichment strategy for N -glycopeptides from complex biological samples is necessary. In this work, new functional nanomaterials were prepared through a simple, convenient, and efficient two-step "Au-S" reaction. In short, the "Au-S" bond was first used to load nano-gold wires on the surface of molybdenum disulfide (MoS 2 ), and then to connect these wires to 4-mercaptophenylboronic acid (4-MPB). The resulting nanomaterial named MoS 2 /Au/4-MPB was successfully prepared by serial functionalization of ultra-thin two-dimensional MoS 2 , nano-gold wires and 4-MPB for the efficient enrichment of N -glycopeptides. The layered structure of molybdenum disulfide nanomaterials can provide numerous modifiable sites for the reaction, allowing for convenient modification of the nano-gold wires. The functional group 4-MPB has high affinity for N -glycopeptides, and it can selectively enrich them in biological samples. In order to evaluate the N -glycopeptide enrichment performance of MoS 2 /Au/4-MPB, standard proteins human immunoglobulin G (IgG) and bovine serum albumin (BSA) trypsin digests were used. Thus, facilitated interactions with N -glycopeptides for boric acid-based retention could be expected. Low femtomolar detection sensitivity, 1:1000 enrichment selectivity, and 100 μg/mg loading capacity were achieved for N -glycopeptide enrichment. In the application of this material to biological samples, exosomes were chosen as the research objects. On the one hand, exosomes serve as a new class of carriers for intercellular communication and drug delivery. Almost all types of cells can secrete exosomes. On the other hand, urine is another potential source for liquid biopsy samples in addition to blood. Obtaining urine samples is a less invasive process, and urine samples are easier to handle and save as compared to blood samples. Exosomes can also be detected in urine. Cell-specific components such as proteins, lipids, and nucleic acids can provide diverse information, which is of great significance in disease diagnosis and treatment. However, there are only a few studies on urinary exosomes N -glycoprotein group. In this research, N -glycoproteins were successfully identified in urine exosomes. In three replicate experiments, 536, 515 and 487 N -glycosylated peptides were identified from 279, 270 and 279 N -glycoproteins, respectively. After de-duplication of the three sets of data, a total of 768 N -glycopeptides corresponding to 377 N -glycoproteins were identified. This result indicates that the novel nanocomposite has good enrichment selectivity and sensitivity for N -glycopeptides enrichment in complex biological samples, and this method provides a new method for glycoproteomics research.

Masticatory robots have a broad application prospect in the field of denture material tests and mandible rehabilitation. Mechanism type of temporomandibular joint structure is an important factor influencing the performance of the masticatory robot. In view of the wide application of elastic components in the field of the biomimetic robot, an elastic component was adopted to simulate the buffering characteristics of the temporomandibular joint disc and formed the elastic temporomandibular joint structure on the basis of point-contact high pair. Secondly, the influences of the elastic temporomandibular joint structure (on mechanism degree, kinematics, dynamics, etc.) were discussed. The position and velocity of the temporomandibular joint were analyzed based on geometric constraints of the joint surface, and the dynamic analysis based on the Lagrange equation was carried out. Finally, the influence of the preload and stiffness of the elastic component was analyzed by the response surface method. The results showed that the elastic temporomandibular joint structure could effectively guarantee the flexible movement and stable force of the joint. The elastic joint structure proposed in this paper further improves the biomimetic behavior of masticatory robots. It provides new ideas for the biomimetic design of viscoelastic joint discs.

Objective: To find the risk assessment model of [di (2-ethylhexyl) phthalate, DEHP] fitting well for producing companies. Methods: The Cluster Sampling method was used in selecting a DEHP producing company in the occupational health and workplace environment monitoring study conducted between July and August in 2017. Data was collected by site evaluating and workplace environment monitoring. According to GBZ/T 298-2017《Technical Guidelines for Occupational Health Risk Assessment of Chemical Hazards in the Workplace》, a qualitative assessment method and a semi-quantitative comprehensive index method were chosen as the models and were compared. Results: The occupational health risk of the two methods rated 4 (high risk) and 2 (negligible risk) . Conclusion: The semi-quantitative comprehensive index method is more comprehensive and accurate when used to assess the occupational health risk caused by DEHP.

Objective: To observe the steatosis and inflammatory effects of L02 hepatocytes induced by different concentrations of sodium oleate., Methods: L02 cells were incubated with sodium oleate in different concentration for 24 h, and the cell viability was detected. L02 cells were respectively cultured with 75, 150, 300 μmol/L sodium oleate for 24 h. The lipid accumulation of the cells was observed by oil red staining. The content of triglyceride in the cells was detected, the IL-6 content in the cell supernatant was detected. The expression of SIRT1 and nuclear factor-κB(NF-κB) was detected by Western Blot. The expression of Toll-like receptor 2 and Toll-like receptor 4(TLR2 and TLR4) on the cell surface was detected by flow cytometry., Results: With the increase of sodium oleate concentration, the cell viability decreased, the cell growth inhibition rate increased. The content of triglyceride in L02 cells treated with 75, 150 and 300 μmol/L sodium oleate was significantly higher than that in the control group(P&lt;0. 01, P&lt;0. 001, P&lt;0. 001), the IL-6 in the supernate was significantly higher than that in the control group(P&lt;0. 05, P&lt;0. 01, P&lt;0. 001). The result of oil red staining showed that the lipids in the cells were obviously accumulated after sodium oleate treatment. The expression of TLR2 in L02 cells treated with 150 and 300 μmol/L sodium oleate was significantly higher than that in the control group(P&lt;0. 05, P&lt;0. 001). There was no significant increase in TLR4 expression after sodium oleate intervention in L02 cells. The expression of SIRT1 protein in the sodium oleate group was lower than that in the control group, and the expression of NF-κB p65 was higher than that in the control group through Western Blot result., Conclusion: L02 hepatocyte steatosis caused by sodium oleate may be associated with TLR2/NF-κB mediated inflammatory pathway.

Objective: To establish a method for simultaneous determination of 20 flavonoid compounds from honey samples using solid phase extraction coupled with ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS), which was applied to study the difference of flavonoid compounds in honey samples from different botanical origins., Methods: The honey samples were extracted with ultra-water solution by ultrasound, and purified with an Oasis MAX column, and then separated on a Waters ACQUITY UPLC HSS T3(2. 1 mm×100 mm, 1. 8 μm) and eluted with the mobile phase consisted of 0. 1% formic acid in water and acetonitrile in a gradient program, and analyzed by mass spectrometry equipped with an electrospray ionization source, operating in the positive or negative ionization modes, and performed in the multiple reaction monitoring using external standards., Results: Twenty flavonoid compounds showed good linearity with coefficients higher than 0. 9969; The recoveries of spiked honey samples in the range of 76. 2%-108. 6% at the concentrations of 100, 250 and 500 μg/L; The relative standard deviations were 0. 50%-4. 34% and 2. 51%-6. 71%(n=6)for inter-day and intra-day precisions, respectively. It was found that the variations of flavonoid compounds in 16 types of honey samples from different botanical origins differed largely. Among them, quercetrin content in fennel honey was the highest(165 ng/g). High contents of luteolin in red eucalyptus and manuka honey were 186 ng/g and 304 ng/g, respectively. High contents of kaempferol in buckwheat, fennel and manuka honeys were from 113 to 257 ng/g. High contents of chrysin in snow lotus honey, manuka, sunflower honey and lavender Honey were from 136 to 213 ng/g. The content of pinocembrin(473 ng/g) and pinobanksin(684 ng/g) in manuka honey were much higher than that in other honeys., Conclusion: The proposed method is simple for pretreatment, also sensitive, accurate and producible for quantitative of 20 flavonoid compounds in different honey samples, which provide more scientific data and application basis for quality analysis.

Objective: To investigate the effect of nutrition package on intestinal flora of infants aged 6-24 months., Methods: A total of 118 infants in Songxian County and Ruyang County of Henan Province, 60 infants in Songxian County with nutrition package coverage, and 58 infants in Ruyang County of the control county were selected for the long-term effects of early childhood nutrition package intervention project in 2018. All infant faeces were collected, and the composition and abundance of intestinal flora were analyzed and compared by high-throughput sequencing technology., Results: The Alpha diversity of intestinal flora in the nutrition package intervention group was significantly higher than that in the control group. Firmicutes were on the rise in the nutrition package intervention group of infants aged 6-24 months. The abundance of Prevotella in the intervention group was significantly lower than that in the control group. The abundance of Faecalibacterium and Bacteroides in the nutrition package treatment group for infants aged 12 to 17 months was significantly higher than that of the control group, while the abundance of Bifidobacterium and Prevotella was significantly lower than that of the control group. Lactobacillus was significantly less abundant in the 18-24 month old infant nutrition package intervention group than in the control group., Conclusion: Nutrition package can improve the diversity of intestinal flora, also play an important role in promoting the balance of intestinal flora of infants.

Objective: To detect ASXL1 gene variants among patients with myelodysplastic syndrome (MDS) and explore their correlation with variants of other genes and clinical features of patients., Methods: For 149 patients with MDS, genomic DNA was amplified by PCR and subject to direct sequencing to identify variants of ASXL1, U2AF1, SF3B1, DNMT3A, TET2, IDH1/2, NPM1, FLT3-ITD and C-KIT genes., Results: ASXL1 variants were found among 37 patients (24.8%). Other commonly mutated genes included U2AF1 (22.8%), TET2 (11.4%), DNMT3A (9.4%), NPM1 (8.1%) and SF3B1 (6.0%). The frequency of concurrent U2AF1 and TET2 variants among patients with ASXL1 variants was slightly higher than that of wild-type patients. No significant difference was found in median age, MDS subtype, karyotype, peripheral leukocytes, hemoglobin, platelet levels, and bone marrow blast counts between the ASXL1-variant and the wild-type groups (P> 0.05). Twenty-nine patients harboring ASXL1 variants were followed up, 37.9% progressed to acute myeloid leukemia (AML). The rate of transformation in ASXL1-variant group was significantly higher than the wild-type group (37.9% vs. 14.1%, P< 0.01)., Conclusion: ASXL1 showed a high frequency of variant among MDS patients, which was frequently accompanied with U2AF1 and TET2 variants. Compared with the wild type group, patients with ASXL1 variants were more likely to progress to AML.

Objective: To investigate the mutation of RUNX1 gene in patients with myelodysplastic syndrome (MDS) and its correlation with other gene mutations and some clinical parameters., Methods: The mutations of RUNX1, DNMT3A, TET2, IDH1/2, NPM1, FLT3-ITD and C-KIT in 170 patients with MDS were detected by direct and indirect sequencing of genomic DNA-PCR amplification products., Results: The RUNX1 mutation was found in 23 patients (13.5 %, 23/170). Among the 170 patients, other most frequent mutation was TET2 (11.2%, 19/170), followed by mutations in DNMT3A (9.4%, 16/170), NPM1 (8.2%, 14/170), IDH2 (4.1%, 7/170)、FLT3-ITD (2.9%, 5/170), IDH1 (1.7%, 3/170) and c-KIT (0.58%, 1/170). The most common coexisting mutations were TET2 (5/23). The RUNX1-mutated group showed significantly higher leukocyte levels, higher percentages of blast cells, higher incidences of leukemia transformation and lower platelet counts in comparison with RUNX1 non-mutation group (P＜0.05). whereas there were no statistically significant difference in age, MDS subtype, karyotype and hemoglobin level between 2 groups (P＞0.05). Seventeen patients harboring RUNX1 mutations were followed up and almost 47.05% (8/17) of the patients progressed into acute myeloid leukemia (AML). The rates of transformation into AML in ASXL1-mutation group was significantly higher than that in ASXLL- non-mutation group (47.05% vs 11.7%) (P=0.001)., Conclusion: The incidence of RUNX1 mutation is high in MDS patients. The RUNX1-mutated patients have higher leukocyte level, higher percentages of blast cells, higher incidences of leukemia transformation and lower platelet count.

Objective: Improvement the method of nitrogen oxide sampling, analysis and detection in workplace air. Methods: Absorption and collection with sodium carbonate-sodium bicarbonate buffer solution, analysised by ion chromatography, anion column AS23, column temperature 30 ℃. Results: The linearity of the nitrite ion in the corresponding mass concentration range 0~90.0 μg/ml was good (Nitrogen dioxide in 0~120.0 mg/m(3)) , correlation coefficient>0.999 5. The detection limit was 0.16 μg/ml, The minimum detection concentration was 0.11 mg/m(3) with the sampling volume 7.5 L. Conclusion: The method has good accuracy, high sensitivity, with directing injection and no pre-treatment, and can popularize in general laboratories, and can satisfy the determination of nitrogen oxides in the air of workplace.

With the increasing depth of proteomic identification, quantitative accuracy and increasing analytical speed, new challenges are being encountered in the development of proteomics methods. Traditional proteomics methods are time-consuming and have low sensitivity and poor accuracy; hence, they do not satisfy the new demands in proteomics research. Preparation of novel materials with specific functions via chemical and biochemical routes or by methods based on electricity, magnetism, heat, and photoirradiation is the key to overcome the limitations of traditional analytical techniques and promote active research in the field of proteomics. This paper reviews the recent advances in the application of functional materials in proteomics researches.

Objective: To explore the coexisting mutations in NPM1 mutated elderly patients with acute myeloid leukemia(AML). Methods: The clinical data of 152 elderly adults(aged≥60 years) and 49 young adults(aged 18-45 years) with AML between June 2013 and December 2018 in outpatient and hospitalized patients of Changzhou Second People's Hospital and Wuxi Second People's Hospital were retrospectively analyzed. A total of 51 gene mutations were detected using targeted next-generation sequencing (NGS) and sanger sequencing. The general clinical characteristics, the occurrence of coexistence gene mutations, the correlation between coexistence gene mutations and some clinical parameters, and the initial induction remission rate between elderly and young adult AML patients with NPM1 mutations were analyzed and compared. Results: NPM1 mutations were detected in 46 of 152 elderly AML patients. Thirty eight patients (82.6%) with NPM1 mutations carried other gene mutations at the same time, among whom 8 patients (17.4%) carried NPM1 mutations alone, while 14(30.4%) carried 2, 16 (34.8%) carried 3, and 8 (17.4%) carried ≥ 4 mutations. NPM1 mutations frequently co-occurred with FLT3-ITD15 cases (32.6%) , DNMT3A10 (21.7%) , TET26 (13.4%) and FLT3-TKD5 (10.9%) . Compared with young adults with NPM1 mutations, elderly patients had higher TP53, FLT3-TKD rates, lower incidence of DNMT3A, RAS mutation (all P< 0.05) and lower coexistence rate of 4 gene mutations ( P= 0.002).The presence of ≥ 4 mutations was found to be significantly associated with higher white blood level than those in patients with single, double and 3 mutations coexisted in elderly adults AML patients(all P< 0.05). With the increase of the amount of mutations, the complete remission(CR) rate decreased gradually after the initial induction. Patients who carried 3 or more mutations showed a lower CR rate than those with single gene mutations (all P< 0.05) . Patients who carried>4 genes also showed a significantly lower CR rate than those with double gene mutations ( P= 0.031). Patients with FLT3-ITD mutations exhibited higher white blood level and lower CR rate than that in nonmutant type group (all P< 0.05). The CR rate of patients with DNMT3A mutation was also significantly lower than that with nonmutant type ( P= 0.033). However, patients with FLT3-TKD mutations showed a higher platelet level than that with nonmutant type ( P= 0.019). There was no significant difference in CR rate and peripheral blood cell level between TET2 mutated and nonmutant type. Conclusion: NPM1 mutated elderly patients with AML commonly show additional mutations, and the amount and type of coexisting mutations have an influence on the clinical features and CR rate of elderly patients with AML.

Exosomes are vesicles secreted by many types of cells through exocytosis, and their sizes range from 30 to 200 nm. Exosomes consist of a lipid bilayer membrane, containing a number of bioactive molecules, e. g., proteins, ribose nucleic acid (RNA), and deoxyribo nucleic acid (DNA) derived from the cell of origin. As intercellular communication carriers, exosomes participate in many physiological and pathological processes. Because of the complexity of body fluids, as well as the small size and low density of exosomes, the isolation of exosomes is an essential and challenging step before subsequent analysis and functional studies. This review summarizes the advances in the analytical approaches, characterization methods, biological functions and clinical applications of exosomes, with particular emphasis on exosomes isolation techniques.

Objective: To explore the coexisting mutations in IDH-mutated acute myeloid leukemia(AML) and its relation with partial clinical parametrs., Methods: The exon 4 mutation of IDH1/2 gene was screened by using genome DNA-PCR combined with sanger sequencing, 51 targeted gene mutations in the patients with IDH1/2 mutation were detected by using high throughput DNA sequencing combined with sanger sequencing., Results: Among 358 patients, the IDH1/2 mutation was found in 46 cases including IDH1 mutation in 35 cases and IDH2 mutation in 11 cases, 97.87%(45/46) patients with IDH1/2 mutation simultaneously carried other gene mutations including 8(17.8%) cases with mutation of double gene, 17(37.8%) cases with mutation of 3 genes and 20(44.4%) cases with mutation of ≥ 4 genes. The mutation frequency of each patient averaged 3.52 times. In mutation of accompanied genes, the common genes were NPM1(n=29, 63.0%), next DNMT3A(n=25, 54.3%), FLT3-ITD(n=7, 15.2%), TET2(n=5, 10.9%) and NRAS(n=5, 10.9%). The average WBC level of patients with NPM1 mutation in IDH1 mutation group was higher than that of patients in wild type group(P<0.05). The complete remission (CR) rate of patients with DNMT3A mutation was significant lower than that of patients with wild type (30% vs 80%, P<0.01). The presence of ≥ 4 mutations was found to be significantly associated with higher white blood level than that in the patients with double mutations(P<0.05)., Conclusion: More than 95% AML patients with IDH1/2 mutation commonly show additional mutations. The number and the type of IDH coexisting mutations have certain effect on the clinical features and CR rate.

Objective: To investigate the effect of icarin/attapulgite/collagen type Ⅰ/polycaprolactone (ICA/ATP/Col Ⅰ/PCL) composite scaffold in repair of rabbit tibia defect., Methods: The ICA/20%ATP/Col Ⅰ/PCL (scaffold 1), ICA/30%ATP/Col Ⅰ/PCL (scaffold 2), 20%ATP/Col Ⅰ/PCL (scaffold 3), and 30%ATP/Col Ⅰ/PCL (scaffold 4) composite scaffolds were constructed by solution casting-particle filtration method. The structure characteristics of the scaffold 2 before and after cross-linking were observed by scanning electron microscopy, and the surface contact angles of the scaffold 2 and the scaffold 4 were used to evaluate the water absorption performance of the material. The in vitro degradation test was used to evaluate the sustained-release effect of the scaffold 2. Thirty male Japanese white rabbits, weighing (2.0±0.1) kg, were randomly divided into groups A, B, C, D, and E, 6 in each group. After making a 1 cm- diameter bilateral tibial defects model, group A was the defect control group without any material implanted. Groups B, C, D, and E were implanted with scaffolds 3, 4, 1, and 2 at the defect sites, respectively. At 4, 8, and 12 weeks after operation, the repairing effects of 4 scaffolds were observed by gross observation, histological observation of HE and Masson staining, and immunohistochemical staining of osteogenic specific transcription factor (runt-related transcription factor 2, RUNX2), osteogenic related transcription factor [Osterix (OSX), Col Ⅰ, osteopontin (OPN)]., Results: Scanning electron microscopy observation showed that the scaffolds were all porous. The structure of the material was loose before and after cross-linking. The surface contact angle showed that the scaffold was hydrophobic, and the scaffold 2 was more hydrophobic than scaffold 4. The sustained-release effect in vitro showed that the drug could be released in a micro and long-term manner. In the animal implantation experiment, the gross observation showed that the defects were significantly smaller in groups D and E than in groups A, B, and C at 4 and 12 weeks after operation. HE and Masson staining showed that the defect of group A was full of connective tissue at 4 weeks after operation, a large number of fibers were seen in groups B and C, and the new bone formation was observed in groups D and E. The increase of new bone was observed in each group at 8 weeks after operation. The defect of group A was still dominated by connective tissue at 12 weeks after operation, and a small amount of new bone tissue was observed in groups B and C, and a large number of new bone tissue was observed in groups D and E, especially in group E, and most of the materials degraded. Immunohistochemical staining showed that the expressions of RUNX2 and OSX in the new tissues of groups D and E were significantly higher than those of the other groups at 4 weeks after operation. The expression of RUNX2 decreased at 8 and 12 weeks after operation. After 8 weeks and 12 weeks, the expressions of Col Ⅰand OPN increased than in 4 weeks. And the expressions of Col Ⅰ and OPN in the new tissues of groups D and E were significantly more than those of the other groups., Conclusion: ICA/ATP/Col I/PCL composite scaffolds have good porosity and biocompatibility, can promote bone formation, and have good bone regeneration and repair effect.

Objective: To carry out mutation analysis for patients with myelodysplastic syndromes (MDS) and a normal karyotype., Methods: Targeted capture and next-generation sequencing (NGS) was carried out using a customized 49-gene panel. FLT3 internal tandem duplication (FLT3-ITD), CALR, NPM1 and CEBPA mutations were detected by PCR and Sanger sequencing., Results: Sixty-two patients (80.5%) were found to harbor at least one mutation. Each patient has carried 2.21 mutations in average. Coexistence of ≥ 3 mutations was common (43.7%). The most commonly mutated genes were RUNX1 (23.4%, 18/77), ASXL1 (18.2%, 14/77), NPM1 (15.6%, 12/77), U2AF1 (15.6%, 12/77), DNMT3A (11.7%, 9/77). Patients with SF3B1 mutations were significantly older than those with ASXL1 mutations (P=0.023). Mutations of the DNMT3A gene were significantly associated with the blood platelet level compared with BCOR mutations (P=0.02). No significant difference was found in the number and rate of mutations between those under or above 60-year-old. Among 67 patients with clinical follow-up, 20 (29.8%) has transformed to acute myeloid leukemia, and the time of transformation has ranged from 1 to 44 months, with a average of 5.3 months. RUNX1, U2AF1 and FLT3 mutations are associated with leukemic transformation., Conclusion: Coexistence of ≥ 3 mutations are frequent among patients with normal-karyotype MDS. Certain mutations are associated with age and leukemic transformation.

Two types of biodegradable polymers, polycaprolactone (PCL) and poly-3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV), were used as a denitrification slow-release carbon source and a microbial carrier. By comprehensively comparing their performances in denitrification, carbon release, surface morphology, and material composition as well as their microbial community characteristics, the PHBV was determined as the better performer. It had a shorter denitrification start time, a higher denitrification rate, a lower residual organic matter concentration, and a more stable and sustained denitrification performance than PCL. This is because its surface was rough and contained large amounts of hydrophilic groups such as C-O and C=O, which is easily attached and degraded by microorganisms. As a result, the microorganisms on its surface were diverse. The dominant ones were identified with heterotrophic denitrification potentials, such as Thiothrix , Pseudomonas , Zoogloea , Flavobacterium , and Dechloromonas . Therefore, PHBV is suitable as a carbon source medium for tertiary nitrogen removal.