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Objective: To explore effectiveness of minimally invasive internal fixation with locking plates for mid-shaft clavicle fractures., Methods: Between October 2022 and August 2023, 28 patients with mid-shaft clavicle fractures were treated by minimally invasive internal fixation with locking plates. There were 10 males and 18 females with a mean age of 46.2 years (range, 18-74 years). The fractures were caused by traffic accident in 16 patients, sports-related injury in 7 patients, and other injuries in 5 patients. According to Robinson classification, the fractures were classified as type 2A1 in 1 case, type 2A2 in 6 cases, type 2B1 in 15 cases, and type 2B2 in 6 cases. The interval between fracture and operation ranged from 5 hours to 21 days (median, 1.0 days). The pain visual analogue scale (VAS) score was 8.1±1.6. The VAS score at 3 days after operation and the occurrence of complications after operation were recorded. During follow-up, X-ray films were re-examined to observe the healing of the fracture; the shoulder joint function was evaluated according to the Constant-Murley score at 6 months, and the length of the incision scar (total length of the distal and proximal incisions) was measured., Results: All operations were successfully completed without any subclavian vascular or nerve damage. All incisions healed by first intention. The VAS score was 1.2±0.7 at 3 days after operation, and there was a significant difference in VAS score between pre- and post-operation ( t =8.704， P< 0.001). At 1 week after operation, the patient's shoulder was basically painless, and they resumed normal life. All patients were followed up 12-20 months (mean, 13.3 months). X-ray films showed that the bone callus began to form at 2-4 months after operation (mean, 2.7 months). There was no delayed healing or non healing of the fracture, and no loosening or fracture of the internal fixators during follow-up. At 6 months after operation, the mean total incision length was 1.5 cm (range, 1.1-1.8 cm); no patient complained of numbness or paresthesia on subclavicular region or anterior chest wall. The Constant-Murley score of shoulder joint function was 93-100 (mean, 97.6)., Conclusion: Minimally invasive internal fixation with locking plates is a good surgical method for treating mid-shaft clavicle fractures, with simple operation, minimal trauma, good postoperative results, and high satisfaction.

A miniaturized, low-cost high-intensity focused ultrasound device is developed for the problems of cross-contamination and uneven sample fragmentation in conventional ultrasound devices. This device generates ultrasonic waves through a concave spherical self-focusing piezoelectric ceramic piece, and creates a cavitation effect in the focusing area to achieve sample fragmentation. The feasibility of the device is demonstrated by physical simulation, then a driving circuit with adjustable power is designed and manufactured to generate 0 ~ 22.4 W acoustic power, and finally paraffin-embedded sample dewaxing experiments are performed to verify the validation of the device. The experimental results show that the dewaxing efficiency and safety of the high-intensity focused ultrasound device is significantly better than those of traditional chemical methods, and this device is comparable with commercial ultrasonic instruments. In summary, the high-intensity focused ultrasound device is expected to be applied in automated nucleic acid extraction and purification equipment and has a broad application prospect in the field of sample pre-processing.

Objective: To analyze the difference in the gene expression, amino acid and carnitine levels in the cervical secretions between the endometria of pre-receptive and receptive stages, with an aim to provide clues for identifying new molecular markers for endometrial receptivity., Methods: Fifty nine infertile women treated at the Department of Reproductive Medicine of Linyi People's Hospital from January 6, 2020 to January 31, 2022 were selected as as the study subjects, which were matched with 3 pairs (6 cases) of infertile women preparing for embryo transfer based on factors such as age, body mass index, and length of infertility. Endometrial tissue samples were collected for gene transcription and expression analysis. Twenty five women who had become pregnant through assisted reproductive technology were selected as the control group, and 28 non-pregnant women receiving ovulation monitoring at the Outpatient Department were enrolled as the case group. Status of endometrial receptivity was determined by ultrasonography. In the former group, endometrial tissues were sampled for sequencing, and GO and KEGG database enrichment analysis of differentially expressed genes was carried out. In the latter group, cervical secretions were collected, and amino acid and carnitine levels were measured by mass spectrometry. Statistical analysis was carried out using rank sum test, t test and chi-square test with SPSS v25.0 software., Results: No difference was found in the clinical data of the patients with regard to age, body mass index, infertility years, AMH, FSH, LH, E2, and type of infertility. Compared with the receptive endometrial tissues, there were 100 significantly up-regulated genes and 191 significantly down-regulated genes in the pre-receptive endometrial tissue, with the most significantly altered ones being HLA-DRB5 and MMP10. The biological processes, molecular functions and pathways enriched by more differentially expressed genes in GO and KEGG were mainly immune regulation, cell adhesion and tryptophan metabolism. Analysis of secretion metabolism also revealed a significant difference in the levels of amino acids and carnitine metabolites between the two groups (P < 0.05), in particular those of Alanine, Valine, 3-hydroxybutyrylcarnitine (C4OH) + malonylcarnitine (C3DC)/captoylcarnitine (C10)., Conclusion: A significant difference has been discovered in the levels of gene transcription and protein expression in the endometrial tissues from the pre-receptive and receptive stages. The levels of amino acids and carnitine, such as Alanine, Valine, 3-hydroxybutyryl carnitine (C4OH)+malonyl carnitine (C3DC)/caproyl carnitine (C10), may be associated with the receptive status of the endometrium, though this need to be verified with larger samples.

NaHCO 3 -activated buckwheat biochar was studied, and an optimal biochar of 0.25N-BC [ m (NaHCO 3 ): m (buckwheat bark)=0.25:1]was selected. SEM, BET, XRD, Raman, FTIR, and XPS methods were applied to analyze the effects of NaHCO 3 on the physicochemical properties of buckwheat biochar. The adsorption properties and mechanism of NaHCO 3 -activated buckwheat biochar for iopamidol(IPM), a nonionic iodol X-ray contrast agent, were also investigated. The results showed that compared with buckwheat skin biochar(BC), NaHCO 3 -activated biochar had higher structural defects(surface area and pore volume increased, respectively, from 480.40 m 2 ·g -1 and 0.29 cm 3 ·g -1 to 572.83 m 2 ·g -1 and 0.40 cm 3 ·g -1 , with I D / I G being 1.22 times that of BC), the carbon and oxygen functional groups on the BC surface changed significantly, and the polarity increased [(N+O)/C from 0.15 to 0.24]. The maximum adsorption capacity of 0.25N-BC for IPM was 74.94 mg·g -1 , which was 9.51 times that of BC(7.88 mg·g -1 ). The pseudo-second-order adsorption kinetics and Langmuir and Freundlich isotherm models could well fit the adsorption of 0.25N-BC for IPM. The adsorption processes were mainly chemical, monolayer, and heterogeneous multilayer adsorption. Pore filling, hydrogen bonding, π-π, and n-π interactions were the main mechanisms of 0.25N-BC adsorption for IPM. Comparing the activated buckwheat biochar by different bases [KOH, Na 2 CO 3 , NaHCO 3 , KHCO 3 , and Ca(HCO 3 ) 2 ], 0.25N-BC exhibited high adsorption capability and short equilibrium time and could effectively remove the IPM residue in the actual water(secondary sedimentation tank effluent and lake). The removal rate of IPM remained at 74.91% after three adsorption-desorption cycles. The results showed that NaHCO 3 -activated buckwheat biochar was a green, effective, and sustainable adsorbent for the removal of iodine-containing organic matter.

Bone homeostasis is based on the dynamic balance of bone formation and bone resorption. An imbalance in bone homeostasis is a major contributor to many skeletal diseases, including osteoporosis. Changes in the composition and diversity of the gut microbiota (GM) are supposed to have a significant impact on bone homeostasis and are closely correlated with changes in bone mass and bone microarchitecture. The "gut-immune" axis, which is formed by the interaction between the host intestinal immune system and GM, is essential for maintaining bone homeostasis, as well as regulating the body's immunological response and maintaining immune homeostasis throughout the intestine and body. The article reviews recent advances in the study of GM, the immune system, and their synergistic impact on bone homeostasis.

BACKGROUND : The musk glands of adult male Chinese forest musk deer ( Moschus berezovskii Flerov, 1929) (FMD), which are considered as special skin glands, secrete a mixture of sebum, lipids, and proteins into the musk pod. Together, these components form musk, which plays an important role in attracting females during the breeding season. However, the relationship between the musk glands and skin of Chinese FMD remains undiscovered. Here, the musk gland and skin of Chinese FMD were examined using histological analysis and RNA sequencing (RNA-seq), and the expression of key regulatory genes was evaluated to determine whether the musk gland is derived from the skin. METHODS : A comparative analysis of musk gland anatomy between juvenile and adult Chinese FMD was conducted. Then, based on the anatomical structure of the musk gland, skin tissues from the abdomen and back as well as musk gland tissues were obtained from three juvenile FMD. These tissues were used for RNA-seq, hematoxylin-eosin (HE) staining, immunohistochemistry (IHC), western blot (WB), and quantitative real-time polymerase chain reaction (qRT-PCR) experiments. RESULTS : Anatomical analysis showed that only adult male FMD had a complete glandular organ and musk pod, while juvenile FMD did not have any well-developed musk pods. Transcriptomic data revealed that 88.24% of genes were co-expressed in the skin and musk gland tissues. Kyoto Encyclopedia of Genes and Genomes (KEGG) signaling pathway analysis found that the genes co-expressed in the abdomen skin, back skin, and musk gland were enriched in biological development, endocrine system, lipid metabolism, and other pathways. Gene Ontology (GO) enrichment analysis indicated that the genes expressed in these tissues were enriched in biological processes such as multicellular development and cell division. Moreover, the Metascape predictive analysis tool demonstrated that genes expressed in musk glands were skin tissue-specific. qRT-PCR and WB revealed that sex-determining region Y-box protein 9 ( Sox9 ),Caveolin-1 ( Cav-1 ), andandrogen receptor ( AR ) were expressed in all three tissues, although the expression levels differed among the tissues. According to the IHC results, Sox9 and AR were expressed in the nuclei of sebaceous gland, hair follicle, and musk gland cells, whereas Cav-1 was expressed in the cell membrane. CONCLUSIONS : The musk gland of Chinese FMD may be a derivative of skin tissue, and Sox9, Cav-1, and AR may play significant roles in musk gland development.

Objectives: The intestinal microbial characteristics of patients with simple cerebral infarction (CI) and CI complicated with Type 2 diabetes mellitus (CI-T2DM) are still not clear. This study aims to analyze the differences in the variable characteristics of intestinal flora between patients simply with CI and CI-T2DM., Methods: This study retrospectively collected the patients who were admitted to the Affiliated Hospital of Putian University from September 2021 to September 2022. The patients were divided into a CI group ( n =12) and a CI-T2DM group ( n =12). Simultaneously, 12 healthy people were selected as a control group. Total DNA was extracted from feces specimens. Illumina Novaseq sequencing platform was used for metagenomic sequencing. The Knead Data software, Kraken2 software, and Bracken software were applied for sequencing analysis., Results: At phylum level, the average ratio of Firmicutes, Bacteroidetes, and Proteobacteria in the CI-T2DM group were 33.07%, 54.80%, and 7.00%, respectively. In the CI group, the ratios of each were 14.03%, 69.62%, and 11.13%, respectively, while in the control group, the ratios were 50.99%, 37.67%, and 5.24%, respectively. There was significant differences in the distribution of Firmicutes ( F =6.130, P =0.011) among the 3 groups. At the family level, compared with the CI group, the relative abundance of Eubacteriaceae ( t =8.062, P <0.001) in the CI-T2DM group was significantly increased, while Corynebacteriaceae ( t =4.471, P <0.001), Methanobacteriaceae ( t =3.406, P =0.003), and Pseudomonadaceae ( t =2.352, P =0.028) were decreased significantly. At the genus level, compared with the CI group, there was a relative abundance of Cutibacterium ( t =6.242, P <0.001), Eubacterium ( t =8.448, P <0.001), and Blautia ( t =3.442, P =0.002) in the CI-T2DM group which was significantly increased. In terms of Methanobrevibacter ( t =3.466, P =0.002), Pyramidobacter ( t =2.846, P =0.009) and Pseudomonas ( t =2.352, P =0.028), their distributions were decreased significantly in the CI-T2DM group. At the species level, compared with the CI group, the relative abundance of Cutibacterium acnes ( t =6.242, P <0.001) in the CI-T2DM group was significantly increased, while Pseudomonas aeruginosa ( t =2.352, P =0.028) was decreased significantly. Still at the genus level, linear discriminant analysis effect size (LEfSe) analysis showed that the distributions of Pseudomonas and Blautia were determined to be the most significantly different between the CI-T2DM and the CI group. At the species level, the total number of operational taxonomic units (OTUs) in the 3 groups was 1 491. There were 169, 221, and 192 kinds of OTUs unique to the CI-T2DM, CI, and control group, respectively., Conclusions: From phylum level to species level, the composition of intestinal flora in the patients with CI-T2DM is different from those in the patients simply with CI. The change in the proportion of Firmicutes , Bacteroidetes and Proteus compared with the healthy population is an important feature of intestinal flora imbalance in the patients with CI and with CI-T2DM. Attention should be paid to the differential distribution of Bacteroides monocytogenes and butyrate producing bacteria.

Polyurethane (PUR) plastics is widely used because of its unique physical and chemical properties. However, unreasonable disposal of the vast amount of used PUR plastics has caused serious environmental pollution. The efficient degradation and utilization of used PUR plastics by means of microorganisms has become one of the current research hotspots, and efficient PUR degrading microbes are the key to the biological treatment of PUR plastics. In this study, an Impranil DLN-degrading bacteria G-11 was isolated from used PUR plastic samples collected from landfill, and its PUR-degrading characteristics were studied. Strain G-11 was identified as Amycolatopsis sp. through 16S rRNA gene sequence alignment. PUR degradation experiment showed that the weight loss rate of the commercial PUR plastics upon treatment of strain G-11 was 4.67%. Scanning electron microscope (SEM) showed that the surface structure of G-11-treated PUR plastics was destroyed with an eroded morphology. Contact angle and thermogravimetry analysis (TGA) showed that the hydrophilicity of PUR plastics increased along with decreased thermal stability upon treatment by strain G-11, which were consistent with the weight loss and morphological observation. These results indicated that strain G-11 isolated from landfill has potential application in biodegradation of waste PUR plastics.

With the escalation of plastic bans and restrictions, bio-based plastics, represented by polylactic acid (PLA), have become a major alternative to traditional plastics in the current market and are unanimously regarded as having potential for development. However, there are still several misconceptions about bio-based plastics, whose complete degradation requires specific composting conditions. Bio-based plastics might be slow to degrade when it is released into the natural environment. They might also be harmful to humans, biodiversity and ecosystem function as traditional petroleum-based plastics do. In recent years, with the increasing production capacity and market size of PLA plastics in China, there is an urgent need to investigate and further strengthen the management of the life cycle of PLA and other bio-based plastics. In particular, the in-situ biodegradability and recycling of hard-to-recycle bio-based plastics in the ecological environment should be focused. This review introduces the characteristics, synthesis and commercialization of PLA plastics, summarizes the current research progress of microbial and enzymatic degradation of PLA plastics, and discusses their biodegradation mechanisms. Moreover, two bio-disposal methods against PLA plastic waste, including microbial in-situ treatment and enzymatic closed-loop recycling, are proposed. At last, the prospects and trends for the development of PLA plastics are presented.

Objective: To explore the effect of microRNA-424-5p (miR-424-5p) on the drug resistance of diffuse large B-cell lymphoma (DLBCL) cells by regulating the programmed death receptor-1 (PD-1)/programmed death ligand-1 (PD-L1) signaling pathway., Methods: Human DLBCL cell line CRL2631 cells were induced to construct CRL2631-CHOP resistant cell line. RT-qPCR and Western blot were used to detect the expression levels of MiR-424-5p , PD-L1 mRNA and protein, and multidrug resistance gene-1 (MDR-1) protein in CRL2631 cells and CRL2631-CHOP cells, respectively. The target genes of MiR-424-5p was verified by dual luciferase reporter assay. The miRNA simulation/interference technology and thiazole blue (MTT) method were used to detect the resistance of CRL2631 cells and CRL2631-CHOP cells to CHOP., Results: Compared with CRL2631 cells, the drug resistance of CRL2631-CHOP cells to CHOP and the levels of MDR-1 protein ( P <0.05), PD-L1 mRNA and protein in the cells were significantly increased (both P <0.001), while the relative level of MiR-424-5p was significantly reduced ( P <0.001). The result of the dual luciferase reporter assay showed that PD-L1 was the direct downstream target gene of MiR-424-5p ( P <0.001). After transfection of MiR-424-5p inhibitor, the resistance of CRL2631 cells to CHOP drugs increased, and the expression level of MDR-1 protein ( P <0.01), PD-L1 mRNA and protein also increased significantly (both P <0.01). After transfection of MiR-424-5p mimics, the resistance of CRL2631-CHOP cells to CHOP drugs decreased, and the expression level of MDR-1 protein ( P <0.001), PD-L1 mRNA and protein also decreased significantly (both P <0.001). Overexpression of PD-L1 could reverse the inhibitory effect of upregulating MiR-424-5p on PD-L1 ( P <0.001)., Conclusion: Down-regulation of MiR-424-5p enhances the drug resistance of DLBCL cells by regulating the PD-1/PD-L1 signaling pathway.

Objective: To investigate the effect of teriparatide on the differentiation of MC3T3-E1 cells in high-glucose microenvironment and explore the possible mechanism., Methods: MC3T3-E1 cells cultured in normal glucose or high-glucose (25 mmol/L) medium were treated with 10 nmol/L teriparatide with or without co-treatment with H-89 (a PKA inhibitor). CCK-8 assay was used to detect the changes in cell proliferation, and cAMP content in the cells was determined with ELISA. Alkaline phosphatase (ALP) activity and mineralized nodules in the cells were detected using ALP kit and Alizarin red staining, respectively. The changes in cell morphology were detected by cytoskeleton staining. Real-time PCR was used to detect the mRNA expressions of PKA, CREB, RUNX2 and Osx in the treated cells., Results: The treatments did not result in significant changes in proliferation of MC3T3-E1 cells ( P > 0.05). Compared with the cells in routine culture, the cells treated with teriparatide showed significantly increased cAMP levels ( P < 0.05) with enhanced ALP activity and increased area of mineralized nodules ( P < 0.05). Teriparatide treatment also resulted in more distinct visualization of the cytoskeleton in the cells and obviously up-regulated the mRNA expressions of PKA, CREB, RUNX2 and Osx ( P < 0.05). The opposite changes were observed in cells cultured in high glucose. In cells exposed to high glucose, treatment with teriparatide significantly increased cAMP levels ( P < 0.05), ALP activity and the area of mineralized nodules ( P < 0.05) and enhanced the clarity of the cytoskeleton and mRNA expressions of PKA, CREB, RUNX2 and Osx; the effects of teriparatide was strongly antagonized by co-treatment with H-89 ( P < 0.05)., Conclusion: Teriparatide can promote osteoblast differentiation of MC3T3-E1 cells in high-glucose microenvironment possibly by activating the cAMP/PKA/CREB signaling pathway.

Objective: To investigate the dietary fiber intake status and analyze the relationship between dietary fiber and glucose metabolic disorder of the elderly in China., Methods: Data were collected from the participants of Chinese adult chronic diseases and nutrition surveillance in 2015. General information were collected by standardized questionnaires, anthropometric index and blood pressure of respondents were measured according to standard method. Food intake was collected by three consecutive day 24 h dietary recalls, dietary fiber was calculated through China food composition. Fasting venous blood were collected to measure glucose, HbA1C, and other related biochemical index. Subjects were divided into three groups: normal glucose(NG, normal glucose), pre-diabetes(Pre-DM, pre-diabetes mellitus) and diabetes(T2DM, type 2 diabetes mellitus). Multiple logistic regression model was used to analyze the relationship between dietary fiber intake and Pre-DM as well as T2DM., Results: A total of 20 996 elderly people aged 60 years and above were included. There 10 773 cases were males(51.3%) and 10 223 cases were females(48.7%), the age of both gender were(68.21±6.26) years and(67.67±6.26) years. A total of 6526 cases of pre-diabetes were detected in 20 996 elderly participants with detection rate of 31.1%. There 3274 cases were male and 3252 cases were female, the detection rates of both genders were 30.4% and 31.8%, respectively. While 1572 participants were detected as T2DM(784 of males and 788 of females), the detection rate of T2DM was 7.5%, 7.3% for males and 7.7% for females. There were significant differences in mean age, BMI, waist circumference, systolic blood pressure, diastolic blood pressure, TC, TG, HDL-C, LDL-C, HbA1c level among different dietary intake groups(P&lt;0.005). With the increase of dietary fiber intake, the proportion of overweight and obesity, central obesity and dyslipidemia showed an increasing trend(P&lt;0.05), and the proportion of hypertension showed a decreasing trend(P&lt;0.000 1). After adjusting for potential confounding factors, compared to participants with lowest fiber intake, participants in subgroups of lower, minor lower, and higher fiber intake were associated with decreased risk of pre-diabetes, the OR and 95%CI were(OR=0.911, 95%CI 0.835-0.993), (OR= 0.861, 95%CI 0.790-0.938) and(OR= 0.913, 95%CI 0.838-0.994), respectively. However, there was only a statistically significant negative association between the higher intake of dietary fiber and T2DM(OR = 0.848, 95%CI 0.726-0.991)., Conclusion: Dietary fiber intake was negatively related with diabetes and pre-diabetes mellitus. The risk of glucose metabolic disorder was decreased with the increase of dietary fiber intake.

Objective: To explore the relationship between preoperative blood indexes, clinicopathological characteristics and prognosis of patients with laryngeal squamous cell carcinoma（LSCC）, and to establish a nomogram for predicting survival and recurrence time of LSCC patients. Methods: The clinical data of 530 patients with LSCC who received primary surgical treatment were analyzed retrospectively. The patients were randomly divided into training group and testing group according to the ratio of 7∶3. The nomograms of overall survival（OS） and recurrence-free survival（RFS） were constructed based on Cox regression model, and the nomograms were verified and compared with TNM stage. Results: The results of multivariate analysis showed that age, operation mode, tumor diameter, TNM stage, cervical lymph node metastasis, fibrinogen level and systemic immune inflammation index were significantly correlated with OS, while operation mode, tumor diameter, TNM stage, cervical lymph node metastasis, international normalized ratio, neutrophil to lymphocyte ratio and platelet to lymphocyte ratio were significantly correlated with RFS. The nomograms were constructed based on the above factors. The C-indexes of the nomograms of OS and RFS in the training group were 0.755 and 0.687, respectively, which were higher than those of TNM stage（0.657 and 0.582）. In the testing group, the C-indexes of OS and RFS nomograms were 0.717 and 0.646, respectively, which were higher than those of TNM stage（0.599 and 0.528）. The area under the receiver operating characteristic curves of these nomograms were also higher than TNM stage. The calibration curves showed that the models had good consistency. Decision curve analysis（DCA） showed that these nomograms had higher clinical benefit than TNM stage. Conclusion: Based on the independent prognostic factors, the predictive nomograms of OS and RFS at 1, 3 and 5 years after LSCC were constructed, which have certain clinical significance in guiding the individualized diagnosis and treatment of LSCC., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

A patient complained of progressive visual acuity decline in the left eye for 3 years was admitted. The appearance of the patient was bilateral microcornea. After ocular ultrasonography, ultrasound biomicroscopy, etc, the patient was diagnosed as sclerocornea of the left eye with open-angle glaucoma. Trabeculectomy of the left eye was performed after drug treatment failed, and there were no surgical complications. The intraocular pressure of the left eye was normal and the anterior chamber was well formed 6 months after surgery.

Due to the large scale of mining and smelting activities, considerable amounts of heavy metals are discharged into the environment and accumulate in the sediment of rivers and lakes. The combined pollution of heavy metals and the intrinsic phosphorus in sediment calls for novel remediation technologies. In this study, lanthanum-modified zeolite (LMZ) was employed as an inactivation agent for the immobilization of phosphorus, zinc, and lead in sediments. The adsorption capacities as well as the inactivation performance of LMZ for P, Zn, and Pb were investigated, and the adsorption mechanisms were explored via desorption experiments, X-ray photoelectron spectroscopy (XPS), and X-ray diffraction (XRD). The results indicated that the adsorption maximums of LMZ for P, Zn, and Pb were 53.76, 27.70, and 123.45 mg·g -1 , respectively. Pre-adsorption of Zn and Pb had a negligible effect on the P adsorption by LMZ, whereas the adsorption of Zn and Pb were inhibited significantly by the pre-adsorption. P, Zn, and Pb in the sediment were transformed to more stable or less bioavailable forms by dosing 0.83% and 1.66% weight percentages of LMZ. It was found that P, Zn, and Pb were adsorbed through the formation of inner-sphere complexes. Further, desorption experiments and XRD patterns suggested that electrostatic attraction and surface precipitation also contributed to the adsorption of Zn and Pb, respectively.

Objective: To study the molecular mechanism of PI3K-Ⅲ like functional domain inducing programmed cell death of leukemia cell line K562., Methods: The purified PI3K-Ⅲ like functional domain protein was obtained by Pichia pastoris expression system. MTT assay and colony-forming assay were used to detect the effects of PI3K-Ⅲ like functional domain protein on K562 cell proliferation. The effects of PI3K-Ⅲ like functional domain protein on apoptosis and cell cycle of on K562 cells were detected by flow cytometry. The ultrastructural changes were detected by transmission electron microscopy. The expression of caspase-3 was detected by ELISA. The protein expressions of ATG4B, Beclin-1, Bcl-2 and LC3-II were evaluated by Western blot., Results: PI3K-Ⅲ like functional domain protein could inhibit the proliferation and clony formation of K562 cells, which was significantly higher than the control group (P<0.05). In the experimental group, apoptosis and autophagosome were shown in K562 cells. The proportion of cells in G 0 /G 1 phase increased significantly, while in S phase decreased significantly. Cell growth mostly stagnated in G 0 /G 1 phase, which was significantly different from the control group (P<0.05). With the increase of concentration, the expression of caspase-3 protein increased significantly compared with the control group (r=0.966, P<0.05). The expression of ATG4B and beclin-1 appeared from increase to decrease, LC3-II increased while Bcl-2 decreased at different time points., Conclusion: PI3K-Ⅲ like functional polypeptide could induce programmed cell death of leukemia cell K562. Beclin-1/Bcl-2 and caspase pathway may be involved in this way, which suggesting meant autophagy and apoptosis may work together at the same time.

Microplastics (MPs) have been detected in many ecosystems, such as the ocean, land and the atmosphere. A large number of MPs in urban sewage are trapped in the activated sludge by sewage treatment plants, but tens of thousands of MPs 'escape' the treatment and are discharged into the nature. Meanwhile, most of the MPs are transferred into the activated sludge during sewage treatment, and the sludge will be further used in agriculture, leading to secondary pollution of the MPs. Through literature research, we summarized the sources, distribution and hazards of MPs in the environment, the treatment of MPs with activated sludge, and the treatment methods of residual MPs in activated sludge, and summed up the potentials of biotechnology and synthetic biology in the genetic modification of key bacteria in activated sludge to endow them with MPs-degrading ability. The conclusion is expected to serve as a reference for optimizing the biodegradation of MPs in wastewater treatment plants.

Objective: To investigate the distribution of HIV-1 genetic subtypes and pretreatment drug resistance (PDR) among men who have sex with men (MSM) from 19 cities of 6 provinces in China. Methods: From April to November 2019, 574 plasma samples of ART-naive HIV-1 infected MSM were collected from 19 cities in Hebei, Shandong, Jiangsu, Zhejiang, Fujian, and Guangdong provinces, total ribonucleic acid (RNA) was extracted and amplified the HIV-1 pol gene region by nested polymerase chain reaction (PCR) after reverse transcription. Then sequences were used to construct a phylogenetic tree to determine genetic subtypes and submitted to the Stanford drug resistance database for drug resistance analysis. Results: A total of 479 samples were successfully amplified by PCR. The HIV-1 genetic subtypes included CRF01&#95;AE, CRF07&#95;BC, B, CRF55&#95;01B, CRF59&#95;01B, CRF65&#95;cpx, CRF103&#95;01B, CRF67&#95;01B, CRF68&#95;01B and unrecognized subtype, which accounted for 43.4%, 36.3%, 6.3%, 5.9%, 0.8%, 0.8%, 0.4%, 0.4%, 0.2% and 5.5%, respectively. The distribution of genetic subtypes among provinces is statistically different ( χ 2 =44.141, P <0.001). The overall PDR rate was 4.6% (22/479), the drug resistance rate of non-nucleoside reverse transcriptase inhibitors, nucleoside reverse transcriptase inhibitors, and protease inhibitors were 3.5% (17/479), 0.8% (4/479) and 0.2% (1/479), respectively. The PDR rate of recent infections was significantly higher than that of long-term infections ( χ 2 =4.634, P =0.031). Conclusions: The HIV-1 genetic subtypes among MSM infected with HIV-1 from 19 cities of 6 provinces in China are diverse, and the distribution of subtypes is different among provinces. The overall PDR rate is low, while the PDR rate of recent infections was significantly higher than that of long-term infections, suggesting the surveillance of PDR in recent infections should be strengthened.

Objective: To explore the genetic basis of three children with unexplained developmental delay/intellectual disability (DD/ID)., Methods: Peripheral blood samples were collected from the patients and subjected to chromosomal microarray analysis (CMA)., Results: Patient 1 was found to harbor a 190 kb deletion at 9q34.3, which encompassed most of EHMT1 (OMIM 607001), the key gene for Kleefstra syndrome (OMIM 610253). Patients 2 and 3 were siblings. CMA showed that they have shared four chromosomal copy number variations (CNVs) including a deletion at 9q34.3 which spanned 154 kb and 149 kb, respectively, and encompassed the EHMT1 and CACNA1B (OMIM 601012) genes. The remaining 3 CNVs were predicted to be with no clinical significance., Conclusion: Microdeletions at 9q33.4 probably underlay the pathogenesis of DD/ID in the three children, for which EHMT1 may be the key gene.

Objective: To study the difference in dielectric properties (relative permittivity and electrical conductivity) between normal human esophageal tissues and esophageal cancer tissues resected in esophageal cancer surgery for rapid identification of esophageal cancer during surgery., Methods: An open- end coaxial probe in the frequency range of 50 MHz to 4 GHz was used for dielectric measurement of the normal tissues and tumor tissues immediately after resection in 51 esophageal cancer cases. The difference in dielectric characteristics of the tissues was analyzed in the full frequency range, and the measurement data at 6 specific frequencies (64, 128, 298, 433, 915, and 2450 MHz) were analyzed statistically. The Cole-Cole model was used for data fitting in the entire frequency band and the measured values were compared with reported values., Results: In the frequency range of 50 MHz- 4 GHz, the dielectric measurement values of esophageal cancer tissues were significantly higher than those of normal esophageal tissues. At the 6 specific frequencies, the dielectric properties also differed significantly between normal esophageal tissues and esophageal cancer tissues ( P < 0.01). The Cole-Cole model achieved a good fitting result of the measured data. The measured values in this study were lower than the reported values., Conclusion: There are significant differences in the dielectric properties between normal esophageal tissue and tumor tissue, which may provide a new theoretical basis for rapid identification of esophageal cancer during surgery.

With the transformation and revolution of the global plastics recycling system, recycling and upcycling of mixed plastics waste not only reduces the carbon emissions of plastics during its life cycle, but also addresses its potential ecological and environmental hazards. This article summarizes an international cooperation project, "MIXed plastics biodegradation and UPcycling using microbial communities" (MIX-UP) which was funded by the National Natural Science Foundation of China and the European Union (NSFC-EU) in 2019. The consortium of MIX-UP consists of 14 partners from European Union and China. Focusing on the global issue of "plastics pollution", this Sino-European MIX-UP project took the mixed waste of petroleum-based plastics (PP, PE, PUR, PET and PS) and bio-based plastics (PLA and PHA) as starting materials for biotechnological conversion into value-added, sustainable biomaterials. MIX-UP has three subprojects: 1) identification of plastics biodegradation pathway and design & engineering of key degrading elements, 2) construction and functional regulation of microbial consortia/enzyme cocktails with high-efficiency for degradation of plastics mixtures, 3) strategy of design and utilization of plastics degradation products for production of high value materials. Through NSFC-EU complementary and cross-disciplinary cooperation, MIX-UP proposes the engineering of a new-to-nature biological route for upcycling, a low carbon and sustainable bio-treatment that is different from the traditional physico-chemical treatment, which will empower the recycling industry to a new dimension. The implementation of the project will not only help to promote innovation and development in the field of biotechnology in China, but also contribute to the achievement of China's carbon neutral goal.

Biodegradation of polyurethane (PUR) pollutants by microorganisms has received widespread attention currently. Identification of microorganisms capable of efficiently degrading PUR plastics is a key point. In this study, a strain P10 capable of degrading PUR was isolated from the plastic wastes, and identified as a bacterium belonging to the genus of Brevibacillus based on colony morphology and 16S rDNA phylogenetic analysis. Brevibacillus sp. P10 was capable of degrading 71.4% of waterborne polyurethane (Impranil DLN) after 6 days growth in MSM medium with DLN as a sole carbon source. In addition, strain P10 can use commercial PUR foam as the sole carbon source for growth. Brevibacillus sp. P10 can degrade 50 mg PUR foam after 6 days growth in MSM medium supplemented with 5% (V/V) LB after optimization of degradation conditions. This indicates that Brevibacillus sp. P10 has potential to be used in biodegradation of PUR waste.

Polycyclic aromatic hydrocarbons (PAHs) are a class of common environmental pollutants that pose threats to human health. In this study, a mesophilic bacterial strain CFP312 (grown at 15-37 °C, optimal at 30 °C) was isolated from PAHs-contaminated soil samples. It was identified as Moraxella sp. by morphological observation, physiological and biochemical test, and 16S rRNA gene phylogeny analysis. This is the first reported PAHs degrading strains in Moraxella. Degradation analysis showed that 84% and 90% of the loaded phenanthrene (400 mg/L) were degraded within 48 h and 60 h, and the degradation rates reached 1.21 and 1.29 mg/(L·h), respectively. During the degradation of phenanthrene, phenanthrene-3,4-dihydrodiol was detected as an intermediate. Based on this, it was proposed that double oxygenation at the positions 3 and 4 of phenanthrene was the first step of biodegradation. Adaptability of strain CFP312 to different enhanced phenanthrene-degradation systems was tested in aqueous-organic system, micellar aqueous system, and cloud point system. Strain CFP312 showed good adaptability to different systems. In addition, the bacterium can rapidly degrade the phenanthrene in contaminated soil in slurry-aqueous system, indicating great potential in environmental remediation.

Objective: To investigate the role of G-protein coupled receptor Smoothened (Smo) in regulating proliferation and migration of adult neural stem cells (ANSCs) and explore the underlying mechanism., Methods: Cultured ANSCs were treated with purmorphamine (PM, an agonist of Smo) or cyclopamine (CPM, an inhibitor of Smo), and the changes in cell proliferation migration abilities were assessed using cell counting kit-8 (CCK8) assay and wound healing assay, respectively. The mRNA expressions of membrane receptor Patched 1 (Ptch1), Smo, glioma-associated oncogene homolog 1 (Gli1), axon guidance cue slit1 (Slit1) and brain-derived neurotrophic factor (BDNF) in the treated cells were detected using real-time quantitative PCR (RT-PCR)., Results: PM significantly promoted the proliferation ( P < 0.01) and migration of ANSCs ( P < 0.01), and up-regulated the mRNA expressions of Ptch1, Smo, Gli1, Slit1 and BDNF. Treatment with CPM significantly inhibited the proliferation and migration of ANSCs., Conclusion: Modulating Smo activity can positively regulate the proliferation and migration of ANSCs possibly by regulating the expressions of BDNF and Slit1.

Objective: To evaluate the clinical and molecular pathologic features of uterine inflammatory myofibroblastic tumor (UIMT). Methods: Six UIMT cases collected at Department of Pathology, Shanghai First Maternity and Infant Hospital, School of Medicine, Tongji University from 2019 to 2020. They were analyzed for their general characteristics and clinicopathologic features. ALK rearrangements were detected by fluorescence in situ hybridization. Results: The age of the six patients ranged from 14 to 65 years, the tumors ranged in size from 2.5 to 6.0 cm. The masses were intramural or submucosal in location. Most of them (4/6) were white with yellow foci, and two (2/6) were white with tan foci. Other features noted included a soft or firm appearance. The fasciitis-like pattern of UIMT had myxoid stroma around the spindle cells and inflammatory cells. The ganglion-like pattern showed either fascicular or storiform architecture with diffuse growth. Nuclear atypia was mild or moderate. Mitoses ranged from 2 to 4 per 10 high-power fields. Five tumors were ALK-positive with granular cytoplasmic staining by immunohistochemistry. ALK rearrangements were detected in five cases but was absent in one case. Conclusions: UIMT is an intermediate grade soft tissue tumor, a minority may present with extrauterine spread and/or recurrence. The tumors are composed of spindled cells. The main differentials include smooth muscle tumors and endometrial stromal tumors and their morphology may overlap with that of UIMT. Immunohistochemical positivity for ALK or FISH testing for ALK rearrangements can help in the diagnosis.

Objective: To summarize and analyze the feasibility, safety and efficacy of parapharyngeal space surgery assisted by coblation and endoscopic system with transoral approach. Methods: The data of 20 patients with parapharyngeal space tumors were retrospectively analyzed. All the patients underwent CT and/or MRI examination before surgery, and all underwent transoral approach assisted by coblation and endoscopic systems. The patients were followed up strictly after the operation, with a follow-up time of 8-56 months and the median follow-up time of 28 months. Results: Among the 20 patients, 18 （90%） were pathologically benign tumors and 2 （10%） were malignant tumors. The maximum tumor diameter was （4.4±1.6） cm, the operative time was （79.00±30.03） min, the intraoperative blood loss was （23.63±22.20） mL, and the postoperative pain VAS score was 2.8±1.4. There were 17 cases complete resection, and 3 cases of relapse, including 1 patient who died after distant metastasis of synovial sarcoma postoperative complications occurred in 2 cases, hoarseness in 1 case of neurofibroma and tongue extension deflection in 1 case of schwannoma. Conclusion: Coblation assisted endoscopic system for the treatment of parapharyngeal space tumors with transoral approach has no cervical scar, which is a satisfaction for the patients, less intraoperative bleeding, short operative time, mild postoperative reaction and quick recovery. However, external approach is still recommended for primary malignant lesions, extensive or highly vascularized lesions, tumors located on the lateral side of the internal carotid artery, less than 2 cm from the skull base, or lateral invasion of the deep lobe of the parotid gland, or a pleomorphic adenoma is considered or is found to be too large to be completely resected preoperatively or intraoperatively., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Editorial Department of Journal of Clinical Otorhinolaryngology Head and Neck Surgery.)

Consolidated bioprocessing (CBP) is a multi-step process in a bioreactor, which completes hydrolase production, enzymatic hydrolysis, and microbial fermentation. It is considered to be the most promising process for the production of second-generation biofuels because of its simple steps and low cost. Due to the complexity of lignocellulose degradation and the butanol synthesis pathway, few wild microorganisms can directly utilize lignocellulose to synthesize butanol. With the development of synthetic biology, single-bacterium directly synthesizes butanol using lignocellulose by introducing a butanol synthesis pathway in the cellulolytic Clostridium. However, there are still some problems such as heavy metabolic load of single bacterium and low butanol yield. Co-culture can relieve the metabolic burden of single bacterium through the division of labor in different strains and can further improve the efficiency of butanol synthesis. This review analyzes the recent research progress in the synthesis of biobutanol using lignocellulose by consolidated bioprocessing from both the single-bacterium strategy and co-culture strategy, to provide a reference for the research of butanol and other biofuels.

Objective: To investigate the effect of zoledronate (ZOL) on osteoclast differentiation and bone resorption under high glucose, and the regulation mechanism of p38 mitogen activated kinase (p38 MAPK) signaling pathway in this process., Methods: RAW264.7 cells were divided into four groups: low group, high group, low+ZOL group and high+ZOL group after induced into osteoclasts. Cell proliferation activity was determined by MTT assay. The migration of RAW264.7 cells were examined Optical microscopy. Immunofluorescence microscopy was used to observe the cytoskeleton and sealing zones of osteoclasts. After adding group 5: high + ZOL + SB203580 group, trap staining was used to identify the number of positive osteoclasts in each group. The number and area of resorption lacunae were observed by SEM. The mRNA and protein expression of osteoclast related factors were detected by real-time PCR and Western blotting., Results: The cells in the 5 groups showed similar proliferative activity. High glucose promoted the migration of RAW264.7 cells ( P < 0.05), inhibited the clarity of cytoskeleton and the formation of sealing zones in the osteoclasts. Exposure to high glucose significantly lowered the expressions of p38 MAPK, p-p38 MAPK, NFATc1, CTSK and TRAP, and inhibited osteoclast differentiation and bone absorption ( P < 0.05). Treatment with ZOL obviously suppressed the migration ability of RAW264.7 cells, further reduced the clarity of the cytoskeleton, inhibited the formation of sealing zones of the osteoclasts, lowered the expressions of p38 MAPK, p-p38 MAPK, NFATc1, CTSK, and TRAP ( P < 0.05), and inhibited osteoclast differentiation and bone absorption. Treatment with SB203580 obviously inhibited osteoclast differentiation and bone resorption and the expressions of P38 MAPK, p-p38 MAPK, NFATc1, CTSK and TRAP ( P < 0.05)., Conclusions: High glucose inhibits osteoclast differentiation and bone resorption. ZOL inhibits osteoclast differentiation and bone resorption in high-glucose conditions by regulating p38 MAPK pathway, which can be a new pathway for ZOL to regulate diabetic osteoporosis.

Objective To investigate SIRT1-PGC-1α signaling pathway-mediated effect of hyperoxia on mitochondrial function in A549 human alveolar epithelial cells and its possible mechanism. Methods Human alveolar epithelial cells in logarithmic growth phase were randomly divided into control group and hyperoxia group. The control group was cultured in a 37DegreesCelsius, 50 mL/L CO 2 saturated humidity incubator, and the hyperoxia group was treated with 950 mL/L O 2 . Following 24-hour culture, Mito SOX TM staining was used to detect the level of mitochondrial reactive oxygen species (Mito-ROS) and JC-1 staining to detect the mitochondrial membrane potential. Real-time quantitative PCR was performed to detect the mitochondrial DNA content and the mRNA levels of SIRT1, PGC-1α, nuclear respiratory factor 1 (NRF1) and mitochondrial transcription factor A (TFAM), and Western blotting to detect the protein levels of SIRT1, PGC-1α, NRF1 and TFAM. Results Compared with the control group, the Mito-ROS of the hyperoxia group increased significantly, while the membrane potential decreased obviously; the mitochondrial DNA content of the hyperoxia group went down, and the mRNA and protein expression of SIRT1, PGC-1α, NRF1 and TFAM dropped. Conclusion Hyperoxia induces mitochondrial dysfunction in human alveolar epithelial cells by inhibiting the expression of SIRT1 and PGC-1α.

We used CRISPR/Cas9 to delete plin1 of 3T3-L1 preadipocyte, to observe its effect on lipolysis in adipocytes and to explore regulatory pathways. We cultured 3T3-L1 preadipocytes, and the plin1 knockout vectors were transfected by electroporation. Puromycin culture was used to screen successfully transfected adipocytes, and survival rates were observed after transfection. The optimized "cocktail" method was used to differentiate 3T3-L1 preadipocytes. The glycerol and triglyceride contents were determined by enzymatic methods. The changes in lipid droplet form and size were observed by Oil red O staining. The protein expression of PLIN1, PPARγ, Fsp27, and lipases was measured by Western blotting. RT-PCR was used to measure the expression of PLIN1 and lipases mRNA. After the adipocytes in the control group were induced to differentiate, the quantity of tiny lipid droplets was decreased, and the quantity of unilocular lipid droplets was increased and arranged in a circle around the nucleus. Compared with the control group, the volume of unilocular lipid droplets decreased, and the quantity of tiny lipid droplets increased after induction of adipocytes in the knockout group. The expression of PLIN1 mRNA and protein in the adipocytes was significantly inhibited (P<0.05); glycerol levels increased significantly (0.098 4±0.007 6), TG levels decreased significantly (0.031 0±0.005 3); mRNA and protein expression of HSL and ATGL increased (P<0.05); PPARγ and Fsp27 expression unchanged in adipocytes. The above results indicate that the knockout of plin1 enhances the lipolysis of 3T3-L1 adipocytes by exposing lipids in lipid droplets and up-regulating lipases effects.

Objective To prepare the high-affinity and high-specificity mouse anti-human IgE monoclonal antibody (mAb) as a candidate immunosorbent. Methods BALB/c mice were immunized using recombinant antigen IgECepsilon2-4. Coated ELISA plate with IgECepsilon2-4 was used to screen positive cell lines by indirect ELISA, then coated ELISA plate with natural IgE, IgG, IgM, IgA, IgD to detect the affinity and specificity of serum-free culture supernatant of positive hybridoma cells with natural IgE. The cell line stably secreting specific anti-IgE monoclonal antibodies was expanded and inoculated into the abdominal cavity of BALB/c mice to prepare mAbs. The secreted mAbs were identified by ELISA kit followed by the identification of mAb subtypes. Results The 29 positive hybridoma cells were obtained after five cell fusions, of which 11 strains had strong affinity with natural IgE and 2 strains did not cross-react with other immunoglobulins 3E9 and 7B4. Conclusion The study successfully prepared mAbs against human IgE with high titer, affinity and specificity.

Statistics show that 76.74% (4 688) of 6 109 patients with chronic wounds are over 50 years of age; the proportion of patients with underlying diseases in all age groups above 50 years ranges from 78.25% to 100.00%. Among the underlying diseases of chronic wound patients, the top four diseases are diabetes mellitus, cardiovascular and cerebrovascular diseases, hypertension, and respiratory diseases. The above underlying diseases and ages of patients are the susceptibility factors of coronavirus disease 2019 released by National Health Commission of China. It is an unavoidable fact that the patients with chronic wounds have to go to the hospital for treatment prescribed by the physician. At the same time, we found that quite a few patients preferred going far afield in choosing hospital for treatment due to various reasons. During the prevention and control of coronavirus epidemic, this " go far afield" style of seeking medical treatment may increase the exposure risk during travelling. Accordingly, we convened 36 wound care clinics in different regions in Shanghai to implement the " Five Measures" encouraging the patients with chronic wounds to seek medical treatment nearby. The principle of this operation is that when seeking medical treatment, patients with chronic wounds should try their best to reduce the travel distance as much as possible to minimize the exposure risk during the epidemic period, which will in turn support the campaign of epidemic prevention and control.

Objective: To provide data support for the study of pathogenic mechanism of SARS-CoV-2 at the molecular level, and provide suitable candidate targets for vaccine, antibody and drug research and development through comparative analysis for structural characteristics and epitopes of S protein of SARS-CoV-2 and SARS-CoV., Methods: Based on the reference sequences of S protein, physical and chemical properties, hydrophobicity, signal peptide, transmembrane region, domain, secondary structure, tertiary structure analysis and antigenic epitopes prediction were carried out. Meanwhile, the tissue expression, related pathways and reactome pathways of angiotensis Ⅰ converting enzyme 2 (ACE2) and C-type lectin domain family 4 member M (CLEC4M) receptors were analyzed., Results: The amino acid sequence of S protein of SARS-CoV-2 and SARS-CoV has a 75.80% consistency. The structural characteristics of the two coronaviruses are highly consistent, but the secondary structure and tertiary structure of SARS-CoV-2 is not as obvious as SARS-CoV. ACE2 and CLEC4M are expressed in alimentary system, heart, kidney, lung and placenta. The main related the pathways of renin-angiotensin system, protein digestion and absorption pathway, and the reactome pathways of metabolism of angiotensinogen to angiotensins, GPCR ligand binding, are related to typical symptoms of coronavirus disease 2019 induced by SARS-CoV-2. Three pairs of highly or completely homologous epitopes of S protein were obtained. The 600-605, 695-703 and 888-896 amino acid residues in SARS-CoV-2 were highly homologous with 586-591, 677-685 and 870-878 amino acid residues in SARS-CoV, respectively., Conclusions: The similarity of S protein of SARS-CoV-2 and SARS-CoV determines that they have similar infection patterns and clinical manifestations. The candidate epitopes with high reliability can provide reference for virus diagnosis and vaccine development.

The " exploration of treatment technology of chronic wound with sinus tract supported by endoscope and related auxiliary methods" study had been conducted by Wound Healing Center of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine. The feasibility and effectiveness of this technique have been confirmed, and good clinic results have been achieved. In order to further promote the new technology and its related research, the theoretical knowledge and technical experience accumulated in the early stage are summarized as diagnosis and treatment standard for treatment with endoscopy technique in chronic wounds with sinus tract, including four parts: the applicable subjects, the diagnosis and treatment procedures and classification criteria, the healing criteria, and the risk assessment and prevention measures. The purpose of this standard is to facilitate the application of standardized endoscopy technique, to make the most of its technological advantages, prevent risks, and provide a reference for the official version of the diagnosis and treatment standard.

Objective: To investigate the prognostic value of N-terminal B-type natriuretic peptide (NT-proBNP) on all-cause mortality in heart failure patients with preserved ejection fraction (HFpEF) at real world scenarios. Methods: Patients who met the diagnostic criteria of HFpEF in the China National Heart Failure Registration Study (CN-HF) were divided into death and survival groups. The demographic data, physical examination, results of the first echocardiography, laboratory results at admission, complications, drug use and clinical outcomes were obtained from CN-HF. The univariate Cox proportional hazard model was used to screen the variates that might predict prognosis, and then the covariates with statistical significance were included in the multivariate Cox regression model to analyze the predictive value of baseline NT-proBNP on all-cause death. Spearman correlation analysis was used to evaluate the relationship between NT-proBNP and estimated glomerular filtration rate (eGFR), so as to further explore the predictive value of the interaction between renal dysfunction and NT-proBNP on death. Since NT-proBNP did not obey the binary normal distribution, it was expressed by the natural logarithm of NT-proBNP (LnNT-proBNP). Results: A total of 1 846 HFpEF patients were enrolled in this study, with an average age of 71.5 years, 1 017 males(55.1%), median NT-proBNP 860 ng/L, and median eGFR 73.9 ml·min -1 ·1.73m -2 . After a median follow-up of 34 months, 213 (11.5%) patients died. Patients in the death group were older, with higher NYHA classification Ⅲ-Ⅳ ratio, longer hospital stay, higher serum potassium and NT-proBNP level, prevalence of complications of diabetes mellitus, arrhythmia and atrial fibrillation, use of angiotensin receptor antagonist(ARB), mineralocorticoid receptor antagonists (MRA), diuretic and digoxin was significantly higher in death group than in survival group. Body mass index (BMI), diastolic blood pressure, left ventricular ejection fraction (LVEF), hemoglobin, serum cholesterol(TC), serum triglycerides (TG) and eGFR, and use of angiotensin converting enzyme inhibitors (ACEI), statins and aspirin were lower in death group than in survival group. Univariate Cox regression analysis showed that NT-proBNP was a predictor of all-cause death in HFpEF patients ( HR =2.522, 95 %CI 2.040-3.119, P< 0.001). Multivariate Cox regression analysis showed that the elevated NT-proBNP remains as the independent predictor of all-cause death in patients with HFpEF ( HR =1.230, 95 %CI 1.049-1.442, P= 0.011) after adjusting for age, BMI, diastolic blood pressure, LVEF, hemoglobin, serum potassium, serum sodium, TC, serum high-density lipoprotein cholesterol (HDL-C), TG, eGFR, atrial fibrillation, as well as the treatment of ACEI/ARB, MRA, diuretics and digoxin. Spearman correlation analysis showed that LnNT-proBNP was negatively correlated with eGFR ( r= -0.361, P< 0.001), but there was no interaction between NT-proBNP and renal dysfunction in predicting death in HFpEF patients ( P> 0.05). Conclusion: The elevated level of NT-proBNP at admission is an independent predictor of all-cause mortality in HFpEF patients.

Objective: To investigate the association of venous blood neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) and levels of interleukin-17 (IL-17) and IL-6 at 2 weeks after admission with the occurrence of post-stroke depression (PSD) in patients with first-ever acute ischemic stroke., Methods: The hospitalized patients with first-ever acute ischemic stroke were recruited consecutively from the Department of Neurology of Yijishan Hospital from March, 2015 to September, 2017. The demographic and baseline clinical data on admission and the imaging data were collected. The diagnosis of PSD was established in line with DSM-IV (SCID-I-R) at 3 months during the follow-up. The severity of PSD was assessed using the Hamilton Depression Scale (HAMD-17). Multivariate logistic regression analysis was used to investigate the correlation of NLR, PLR, IL-17, and IL-6 with PSD in these patients., Results: A total of 376 patients with acute ischemic stroke were enrolled, including 224 male patients (59.57%) and 152 female patients (40.43%), whose mean age was 61.37±10.34 years. Of these patients 104 (27.66%) were found to have PSD. Univariate analysis showed that gender, years of education, BMI, widowhood, NIHSS score, MMSE score, mRS score, and laboratory indexes (NLR, PLR, IL-17, and IL-6) were all significantly correlated with PSD (all P &lt; 0.05). Multivariate logistic regression analysis showed that, after adjusting for the compounding factors, the third quartile of NLR ( P &lt; 0.001), the third quartile of PLR ( P =0.002), IL-17 ( P =0.025) and IL-6 ( P =0.016) were independent factors that predicted the occurrence of PSD., Conclusions: Elevated NLR and PLR at admission and levels of IL-17 and IL-6 at 2 weeks after admission are all independent predictors of the occurrence of PSD at 3 months after stroke.

The aim of this study is to investigate the effect of the chimeric intron in different directions on the expression of the nerve growth factor (NGF) in recombinant Chinese hamster ovary (CHO) cells. The chimeric intron that contained the splice sequence of the first intron of the human β-globin and the human immunoglobulin heavy chain variable region intron was used. NGF gene was cloned into the expression vectors containing the chimeric intron in the forward or reverse direction, followed by transfecting into CHO cells, and screened under G418 to produce the stable transfected CHO cells. Fluorescence quantitative PCR, ELISA, and Western blotting were performed to detect the recombinant NGF gene expression in CHO cells. The results showed that the chimeric introns could significantly enhance the expression of NGF in recombinant CHO cells. Moreover, the enhancing effect on NGF expression level by the intron in the forward direction showed stronger than that of the reverse direction both at mRNA and protein level. In conclusion, the chimeric intron could increase NGF expression in stably transfected CHO cells and the effect is associated with the direction of the intron insertion.

Objective: To evaluate the effect of malaria vector control measures so as to provide the evidence for consolidating the control effectiveness of malaria and carrying out the vector surveillance in Suizhou City., Methods: The distributions of Anopheles sinensis and An. anthropophagus were investigated by the combination of retrospective review and field survey. The changes of density and population of mosquito vectors were investigated and compared among various years., Results: From 1985 to 1996, both An. sinensis and An. anthropophagus were found in 18 towns, and An . anthropophagus mosquitoes accounted for 52.3% of the total Anopheles mosquitoes. Only An. sinensis mosquitoes were found in other 26 towns. In 2003 and 2004, An. anthropophagus mosquitoes were found in the original 18 towns, and they accounted for 47.0% and 38.1% respectively, but in 2005, An. anthropophagus mosquitoes were not found in this city. However, the density of An. sinensis presented an upward trend., Conclusions: An. anthropophagus population has gradually disappeared in Suizhou City, and currently, An. sinensis is the main Anopheles population, which might be the malaria vector in suitable conditions. Therefore, the surveillance and control of Anophe les vector should be strengthened in order to consolidate the achievements of malaria elimination.

Objective: By comparing the mechanics of human auricular cartilage, polyurethane elastic material, and high density polyethylene material (Medpor), to produce theoretical proof on choosing optimal artificial auricular scaffold materials., Methods: The experimental materials were divided into 3 groups with 6 samples in each: the auricular cartilage group (group A), the polyurethane elastic material group (group B), and the Medpor group (group C). With an Instron5967 mechanical testing machine, compression and tensile testing were performed to respectively measure values of compression parameters (including yield stress, yield load, elastic modulus, yield compressibility, compressibility within 2 MPa, and compression stress within 10% strain) and values of tensile parameters (including yield stress, yield load, elastic modulus, yield elongation, elongation within 2 MPa, tensile stress within 1% strain) for comparison., Results: Compression testing: no obvious yield points were observed in the whole process in samples of group B, while obvious yield points were observed in samples of groups A and C. There was no significant difference between groups A and C with respect to yield stress and yield load ( P >0.05); while the yield compressibility in group C was significantly lower than that in group A ( P <0.05) and the elastic modulus in group C was significantly higher than that in group A ( P <0.05). There was a significant difference with respect to compressibility within 2 MPa of materials among the 3 groups ( P <0.05), the high, medium, and low values go to groups B, A, and C respectively. The compression stress within 10% strain in group C was significantly higher than that in groups A and B ( P <0.05), and there was no significant difference between that in groups A and B ( P >0.05). Tensile testing: the materials in group B had extremely high tensile strength. The yield stress in groups A and B was significantly higher than that in group C ( P <0.05), and the elastic modulus and tensile stress within 1% strain were significantly lower than those in group C ( P <0.05); but no significant difference was found between those in groups A and B ( P >0.05). There was no significant difference with respect to yield load among the 3 groups ( P >0.05); but there was significant difference with respect to yield elongation among the 3 groups ( P <0.05), and the high, medium, and low values go to groups B, A, and C respectively. The elongation within 2 MPa in group B was significantly higher than that in groups A and C ( P <0.05), and there was no significant difference between that in groups A and C ( P >0.05)., Conclusion: Compared with the Medpor, the polyurethane elastic material is a more ideal artificial auricular scaffold material.

Objective: To establish a rapid and accurate "on/off" switch technique consisted of 3'-phosphorothioate-modified allele-specific primers and exo+ polymerase to screen the G719S and T790M mutations of epidermal growth factor receptor (EGFR) gene. The switch was used to identify cervical cancer patients who are sensitive to tyrosine kinase inhibitor (TKI)., Methods: Allele-specific primers targeting recombinant wild-type and mutation-type templates were designed with 3' terminal phosphorothioate modification. Two-directional primer extension was carried out using Pfu polymerase. The G719S and T790M mutations were detected by the technique among cervical cancer tissues. The results were verified by Sanger sequencing., Results: No mutation was detected among the 80 cervical cancer cases, and the results were consistent with that of Sanger sequencing. No significant difference was found between the frequencies of the G719S and T790M mutations between the patient and the control groups (P> 0.05)., Conclusion: A sensitive "on/off" switch technique for detecting the two EGFR mutations was established. The G719S and T790M mutations are not associated with cervical cancer.

Objective: To investigate the effects of oridonin (ORI) on the proliferation and apoptosis of human multiple myeloma cell line H929 and its possible mechanism., Methods: H929 cells were exposed to ORI 0、4、8、12、16、20、24、28、32 μmol/L for 12, 24 and 36 hours respectively. The prolifcration inhibitory effect of ORI on H929 cells was determined by MTT assay and then the working concentrations of ORI were determined. The morphological changes and apoptosis of H929 cells were observed by TUNEL (TdT-mediated dUTP Nick-End Labeling) and fluorescence microscopy. The apoptosis rate of H929 cells was detected by flow cytometry with Annexin V-FITC/PI staining. The protein expressions of pro-caspase-3, BCL-2，p-PI3K, p-Akt, BAX, Cleaved PARP and p-JNK, p-ERK and p-p38 in H929 cells were detected by Western blot., Results: Compared with the control group, the proliferation of H929 cells treated with the ORI of 8-16 μmol/L was significantly inhibited and the apoptosis of H929 cells was obviously increased in dose- and time-dependent manners. As for morphological changes, the characteristics of apoptotic cells were presented in H929 cells treated with ORI for 24 hours. The protein levels of pro-caspase-3, BCL-2，p-PI3K, p-Akt were down-regulated with increasing of ORI concentration（r=0.9861, r=0.9725, r=0.9413, r=0.9373）, while the BAX, Cleaved PARP and p-JNK, p-ERK and p-p38 were up-regulated（r=0.9178, r=0.8877, r=0.882, r=0.9645, r=0.8623）., Conclusion: The ORI possesses anti-myeloma effects, can inhibit the proliferation and induce the apoptosis of H929 cell line in vitro. Its potential mechanism may be related with up-regulating the MAPK and down-regulating the PI3K/Akt signal pathways.

Flower medicinal materials usually refer to Chinese medicinal materials with a complete flower,inflorescence,or part of a flower as the different medicinal parts,they have an important share in the Chinese herbal medicine market and appeared frequently in Chinese medicine prescriptions. Firstly,the species and regional distribution of the flower medicinal materials resources in China were briefly summarized. Secondly,the characteristics,yield,producing area and origin distribution of the main flower medicinal materials in Henan province were discussed. Finally,the present situation and the main problems of the flower medicinal materials industry in Henan province were comprehensively analyzed,and the corresponding industrial development countermeasures were put forward.This research was intended to provide decision-making demonstration and scientific basis for the rational exploitation and utilization of resources,breeding of new varieties,planting division,production layout and the healthy and sustainable development of the flower medicinal materials industry in Henan province.

A novel protein encoded by the open reading frame 4 (ORF4) was recently discovered in porcine circovirus type 2 (PCV2). However, little is known about the interaction proteins of ORF4 which hindered better understanding the biological functions of ORF4 in the life cycle of PCV2. In the present study, the ORF4 was inserted into the multiple cloning site of pCMV-N-Flag-GST, yielding recombinant plasmid pCMV-N-Flag-GST-ORF4. The recombinant plasmid was transfected into 293T cells and the intracellular interaction complex of ORF4 were enriched and separated by GST pull-down and SDS-PAGE, sequentially. The potential interacting proteins of PCV2 ORF4 were stained with silver and identified by mass spectrometry (MS). Finally, five candidate ORF4-interacting proteins, including Serine/threonine-protein phosphatase 6 catalytic subunit, alpha cardiac muscle 1, actin, SEC14-like protein 5 and myosin 9 were identified. These results would benefit a better understanding of the biological function of ORF4 in PCV2 infected cells.

Objective: To assess the associations of indoor fine particulate matter (PM(2.5)) from outdoor and indoor sources with heart rate (HR) and heart rate variability (HRV) in patients with chronic obstructive pulmonary disease (COPD) of Beijing. Methods: A total of 40 male patients in a stable stage of COPD were recruited from a hospital in a panel study in Beijing with 5 consecutive days of measurement for each subject. General information and disease history of the participants from questionnaires were obtained prior to the study. HR and HRV were repeatedly examined using dynamic electrocardiograph. HRV included standard deviation of all NN intervals (SDNN), root mean square of successive differences between adjacent NN intervals (rMSSD), total power (TP) power in the low-frequency band (LF) and the high-frequency band (HF). Iron was used as tracer element to separate indoor-originated PM(2.5) and outdoor-originated PM(2.5). Mixed-effect models were applied to assess the associations of outdoor-originated PM(2.5) or indoor-originated PM(2.5) and health effects. Results: The P (50) ( P (25), P (75)) values of daily indoor PM(2.5), indoor-originated PM(2.5) and outdoor-originated PM(2.5) were 50.9 (26.8, 122.7), 16.0 (1.9, 43.7) and 27.3 (13.5, 61.8) μg/m(3), respectively. The mean± SD of concentrations of real-time indoor PM(2.5), indoor-originated PM(2.5) and outdoor-originated PM(2).5 were (61.5±58.8), (25.3±39.1) and (36.2±42.7) μg/m(3), respectively. Compared with outdoor-originated PM(2.5), indoor-originated PM(2.5) had significant associations with HRV and HR. Each 10 μg/m(3) increase at 4 h indoor-originated PM(2.5) and outdoor-originated PM(2.5) moving average was associated with 3.4% (95 %CI : -4.7%, -2.1%) and 0.6% (95 %CI : -2.0%, -0.8%) reduction in TP ( P< 0.001). Each 10 μg/m(3) increase at 12 h indoor-originated PM(2.5) moving average was associated with 7.6% (95 %CI : -10.1%, -5.1%), 4.7% (95 %CI : -6.7%, -2.7%), 3.3% (95 %CI : -4.2%,-2.4%) and 3.0% (95 %CI : -4.5%, -1.5%) reduction in HF, LF, SDNN and rMSSD, respectively. Each 10 μg/m(3) increase at 12 h outdoor-originated PM(2.5) moving average was associated with 0.7% (95 %CI : -2.7%, -1.4%), 0.2% (95 %CI : -1.9%, 1.4%), 0.7% (95 %CI : -1.4%, -0.1%) and 0.2% (95 %CI : -1.3%, 0.9%) reduction in HF, LF, SDNN and rMSSD, respectively ( P< 0.001). Each 10 μg/m(3) increase at 8 h indoor-originated PM(2.5) and outdoor-originated PM(2.5) moving average was associated with 0.7% (95 %CI : 0.4%, 1.0%) and 0.4% (95 %CI : 0.2%, 0.6%) increase in HR. Conclusion: Exposure to indoor-originated PM(2.5) was more strongly associations with HRV indices and HR compared with outdoor-originated PM(2.5) in male COPD patients.

The correct thoughts and principles of diagnosis and treatment of chronic refractory wounds need to be formulated. Through the relevant domestic and international consensus and based on clinical experience, the Thoughts and principles of diagnosis and treatment of chronic refractory wounds in China is proposed. It is considered that in the diagnosis and treatment of chronic refractory wounds, in the case of fully understanding the patient's medical history, the following thoughts and principles should be complied in order. (1) Pay attention to the cleanliness of the wound after being cleaned. (2) Reasonably perform debridement to avoid being " excessive" or " not thorough". (3) Reasonably perform examination, diagnosis, and differential diagnosis of pathogenic factors. (4) Treat according to etiology. (5) Find comorbidities and prevent adverse outcomes. (6) Select the correct wound treatment method reasonably and timely. When the conservative wound care treatment is considered, pay attention to embodying the concept of etiological treatment, treat the wound according to the principles of safety, phase, selectivity, and effectiveness, and make a reasonable choice of continuing conservative treatment or surgical treatment in time after completing the preparation of the wound bed. When surgical treatment is considered, pay attention to the selection of reasonable surgical method and donor site, pay attention to the healing rate of surgical wound site and the outcome of donor site, and give reasonable protection to the wound site after surgery. (7) Carry out rehabilitation treatment after wound healing and related health education.

Objective: To understand the HIV and syphilis infection and related treatment status of low-fee female sex workers (FSWs) in 3 provinces of China. Methods: Four cross-sectional survey data of low-fee FSWs from six cities (counties) in Guangxi, Yunnan and Hunan Province between October 2012 and July 2015 were obtained from the national science and technology major special project intervention study for reducing sexually transmitted diseases (STDs) and acquired immunodeficiency syndrome (AIDS) in low-fee FSWs' database, which included social demographic characteristics, sexual service characteristics and related medical care seeking behaviors, etc. A total of 2 050 subjects were included in the database. Results: The age of the subjects was (35.16±9.76) years old, with a minimum age of 15 and a maximum age of 67. Those who use condoms every time in commercial sex accounted for 58.9% ( n= 1 206). Among the reasons of not using condom, the proportion of client reluctant to use was the highest (81.0% ( n= 682)). Only 38.1% ( n= 782) was tested for HIV in the last six months. HIV confirmed positive rate was 6.8% ( n= 139), previous positive accounts for 76.3% ( n= 106). Rate of antiviral therapy was 55.4% ( n= 77). By the end of 2015, the loss rate of antivirus treatment was 18.2% ( n =14). Those who self-reported symptoms of sexually transmitted diseases in the last 6 months accounted for 9.4% ( n =191). 50.3% ( n= 96) of reporters chose to go to formal hospitals, 23.0% ( n= 44) chose to go to private clinics and 20.4% ( n= 39) chose their own medication. The syphilis infection rate was 13.5% ( n= 277), among them, 91.3% ( n= 253) were asymptomatic. Conclusion: Among low-fee FSWs, the rates of HIV and syphilis infection are higher, the condom consistent use rate, HIV antibodies and syphilis test rate are lower. In this group, active seeking medical idea is poor, the rate of anti-virus treatment and the rate of seeking medical treatment in formal medical institutions is low.

Epigenetic modification, especially histone modification, plays an important role in maintaining plant genome stability, regulating gene expression and promoting regeneration in vitro. MtSERK1 is an important marker gene involved in establishing of embryogenic callus during in vitro regeneration of Medicago truncatula. In order to understand the regulation Epigenetic modification, especially histone modification, plays an important role in maintaining plant genome stability, regulating gene expression and promoting regeneration in vitro. MtSERK1 is an important marker gene involved in establishing of embryogenic callus during in vitro regeneration of Medicago truncatula. In order to understand the regulation relationship between dynamic histone modification and MtSERK1s expression during the processes of in vitro organogenesis, the expression of MtSERK1 was analyzed by qRT-PCR, and the modification status of H3K9me2, H3K4me3 and H3K9ac in the promoter region and different regions included in the gene body was analyzed by chromatin immunoprecipitation (ChIP). We found expression activation of MtSERK1 was related to the dynamic changes of histone H3K4me3 and H3K9ac in the 5' and 3' regions. This study will provide important theoretical guidance for understanding of the regulatory mechanism of MtSERK1 and also for establishing efficient genetic transformation system of Medicago truncatula.

Objective: To prepare and purify the rabbit anti-TgMIC16 polyclonal antibody, so as to apply it in subcellular localization., Methods: New Zealand white rabbits were immunized with purified recombinant TgMIC16 mixing with the same volume of Freund's adjuvant for three times, respectively. The rabbit serum was collected on the 14th day after the last immunization. The polyclonal antibody in rabbit serum was purified with Protein A affinity purification column. ELISA and Western blotting were used to detect the antibody titer and specificity of polyclonal antibody. The polyclonal antibody was used to the localization of TgMIC16 by the immunofluorescence method., Results: Indirect ELISA showed that the antibody titer was 1∶512 000. Western blotting showed that the recombinant TgMIC16 protein was recognized by the specific polyclonal antibody. IFA showed that TgMIC16 was located in the microneme of Toxoplasma gondii ., Conclusions: The rabbit anti-TgMIC16 is prepared and purified, and successfully applied to immunofluorescence localization of TgMIC16 in T. gondii .