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Vitiligo is a skin depigmentation disorder. GATA3 expression is downregulated in vitiligo patients, and its role and regulatory mechanism in vitiligo are unclear. GATA3 and HMGB1 levels were detected by qRT-PCR in peripheral blood cells of vitiligo patients and healthy controls, as well as H 2 O 2 -treated PIG1 cells. Their expression correlation was assessed by Pearson analysis. qRT-PCR, MTT assay, Ki67 immunostaining, flow cytometry, ELISA and Western blot were applied to determine GATA3 expression, cell survival, cell proliferation, cell apoptosis, melanin contents, and melanin-related protein expressions. The cellular distributions of HMGB1 and its deacetylation levels were detected by Western blot. The binding of GATA3 to SIRT3 promoter and effects on SIRT3 expression and HMGB1 deacetylation was determined by dual-luciferase assay, ChIP assay, and Western blot. GATA3 was decreased, and HMGB1 was increased in vitiligo. Pearson correlation assay showed that they were negatively correlated. H 2 O 2 significantly inhibited cell survival, proliferation, melanin secretion, and melanin-related protein expressions but remarkably increased cell apoptosis. GATA3 overexpression could distinctly reverse the effects of H 2 O 2 through decreasing HMGB1 expression and retained HMGB1 in nuclear due to the decreased HMGB1 acetylation. GATA3 bound to the SIRT3 and subsequently decreased H 2 O 2 -induced HMGB1 acetylation. Overexpressing HMGB1 or knockdown of SIRT3 could reverse the effects of GATA3 overexpression. GATA3 inhibited H 2 O 2 -induced injury in PIG1 cells and enhanced melanin secretion by SIRT3-regulated HMGB1 deacetylation, which might provide new evidence to treat vitiligo., (© 2022 Japanese Dermatological Association.)

Asthma is a chronic inflammatory disease of the respiratory system. Maresin-2 (MaR2) is biosynthesized from docosahexaenoic acid (DHA) by macrophages, display strong anti-inflammatory and pro-resolving activity. To investigate the therapeutic effect and mechanism of MaR2 on asthmatic mice induced by ovalbumin (OVA) in conjunction with the adjuvant aluminum hydroxide. Twenty four female BALB/c mice were randomly divided into control, OVA, OVA + MaR2, and OVA + dexamethasone (Dexa) groups. MaR2 or Dexa were given as a treatment for OVA-induced asthma. Serum, bronchoalveolar alveolar lavage fluid (BALF) and lung tissue were collected for further analysis. The Pathological changes of lung tissue, proportion of inflammatory cells in BALF, levels of inflammatory cytokines in BALF or serum, oxidative stress indices, and the protein concentration of ASC, MPO, Ly-6G, ICAM-1, NLRP3 and Caspase-1 in lung tissues were evaluated. Compared with the OVA group, both OVA + MaR2 and OVA + Dexa group had reduced inflammation and mucus secretion in lung tissue, number of inflammatory cells in BALF, levels of related inflammatory cytokines in serum or BALF, and expressions of ASC, MPO, Ly-6G, ICAM-1, NLRP3 and Caspase-1 proteins in lung tissue. In addition, the oxidative stress was alleviated as indicated by decreased MDA, and elevated SOD and GSH. MaR2 has an obvious protective effect on OVA-induced bronchial asthma in mice, in a similar manner as Dexa. The mechanism may be related to the inhibition of the Th2 type immune response, the NLRP3 inflammasome activation and oxidative stress., (Copyright © 2022. Published by Elsevier Ltd.)

Abstract: There is a scarcity of research into the impact of medication beliefs on adherence in patients with non-dialysis chronic kidney disease (CKD). This study is to determine the psychometric properties of the Chinese version of the Beliefs about Medicines Questionnaire (BMQ)-Specific among patients with non-dialysis CKD stages 3-5, and to assess the beliefs of CKD patients and their association with medication adherence.A cross-sectional study was conducted in CKD patients who recruited at the nephrology clinics of Xi'an Central Hospital, Xi'an, Shaanxi, China. The original BMQ-Specific was translated into Chinese. The internal consistency and test-retest reliability of the Chinese version of the BMQ-Specific scale were assessed, while exploratory and confirmatory factor analyses were also applied to determine its reliability and validity. The Kruskal-Wallis test and multiple ordered logistic regression were performed to identify the relationship between beliefs about and adherence to medication among CKD patients.This study recruited 248 patients. Cronbach's α values of the BMQ-Specific necessity and concern subscales were 0.826 and 0.820, respectively, with intraclass correlation coefficients of 0.784 and 0.732. Factor analysis showed that BMQ-Specific provided a good fit to the two-factor model. The adherence of patients was positively correlated with perceived necessity (r = 0.264, P < .001) and negatively correlated with concern (r = -0.294, P < .001). Medication adherence was significantly higher for the accepting group (high necessity and low concern scores) than for the ambivalent group (high necessity and concern scores; β = -0.880, 95% confidence interval [CI] = -1.475 to -0.285), skeptical group (low necessity and high concern scores; β = -2.620, 95% CI = -4.209 to -1.031) and indifferent group (low necessity and concern scores; β = -0.918, 95% CI = -1.724 to -0.112).The Chinese version of BMQ-Specific exhibited satisfactory reliability and validity for use in patients with non-dialysis CKD stages 3-5 and has been demonstrated to be a reliable screening tool for clinicians to use to predict and identify the non-adherence behaviors of patients., Competing Interests: The authors have no conflicts of interests to disclose., (Copyright © 2022 the Author(s). Published by Wolters Kluwer Health, Inc.)

Accumulating evidence demonstrates that the nucleus tractus solitarii (NTS) neurons serve as central respiratory chemoreceptors, but the underlying molecular mechanisms remain undefined. The present study investigated the expression of acid-sensitive ether-à-go-go-gene-like (Elk, Kv12) channels in the NTS of mice. Immunofluorescence staining was used to observe the distribution and cellular localization of the Kv12 channels in NTS neurons. Western blot and quantitative real-time PCR (qPCR) were used to evaluate protein and mRNA expression levels of Kv12 channels. The results showed that all of the three members (Kv12.1, Kv12.2, Kv12.3) of the Kv12 channel family were expressed in NTS neurons, and their expressions were co-localized with paired-like homeobox 2b gene (Phox2b) expression. The expression of Kv12.1 mRNA was the largest, whereas the expression of Kv12.3 was the least in the NTS. The results suggest Kv12 channels are expressed in Phox2b-expressing neurons in the NTS of mice, which provides molecular evidence for pH sensitivity in Phox2b-expressing NTS neurons.

Polyanion-type phosphate materials are highly promising cathode candidates for next-generation batteries due to their excellent structural stability during cycling; however, their poor conductivity has impeded their development. Isostructural and multivalent anion substitution combined with carbon coating is proposed to greatly improve the electrochemical properties of phosphate cathode in sodium-ion batteries (SIBs). Specifically, multivalent tetrahedral SiO 4 4- substitute for PO 4 3- in Na 3 V 2 (PO 4 ) 3 (NVP) lattice, preparing the optimal Na 3.1 V 2 (PO 4 ) 2.9 (SiO 4 ) 0.1 with high-rate capability (delivering a high capacity of 82.5 mAh g -1 even at 20 C) and outstanding cyclic stability (≈98% capacity retention after 500 cycles at 1 C). Theoretical calculation and experimental analyses reveal that the anion-substituted Na 3.1 V 2 (PO 4 ) 2.9 (SiO 4 ) 0.1 reduces the bandgap of NVP lattice and enhanced its structural stability, Na + -diffusion kinetics and electronic conductivity. This strategy of multivalent and isostructural anion substitution chemistry provides a new insight to develop advanced phosphate cathodes., (© 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Understanding the impact of tillage erosion on soil organic carbon (SOC) and nitrogen (N) fractions is essential for targeted soil conservation in mountainous and hilly areas. However, little is known about this issue. In this study, we selected a tillage erosion-dominated hillslope from the Sichuan Basin, China, to determine the effect of tillage erosion on particulate OC (POC), dissolved OC (DOC), light fraction OC (LFOC), ammonium N (NH 4 + -N), nitrate N (NO 3 - -N) and alkali-hydrolysable N (AN). Additionally, we investigated the microbial activities in relation to soil C and N dynamics, including soil microbial biomass, β-glucosidase and urease activities. Tillage erosion induced serious soil loss in upper slope positions and soil deposition in lower slope positions. The observations of the various labile OC fraction distributions across the hillslope suggest that tillage erosion exerts less impact on DOC and LFOC dynamics but a notable effect on POC. The distribution pattern in total organic carbon under tillage erosion mainly depends on POC redistribution. The POC redistribution is a major factor affecting microbial activities. The AN is more prone to the tillage erosion impact than NH 4 + -N and NO 3 - -N. Effective soil conservation measures should be taken to weaken the adverse impacts of tillage erosion on POC and AN redistribution in sloping farmlands.

Objective: High-risk (HR) human papillomavirus (HPV) is one of the major causes for most anal and penile cancers and oropharyngeal cancers in men, and vaccination against HPV is recommended for the prevention of these cancers. Data on HPV infection in Chinese men is still limited, which requires further investigation to guide vaccine development and assess the effectiveness of HPV vaccines. We thus studied the HR-HPV genotype distribution in HPV-infected men in Northern China., Patients and Methods: Genital specimens were obtained from male patients (≥18 years old) at the clinic for sexually transmitted infections of the Shandong Provincial Hospital between January 2016 and December 2018. Samples were analyzed for 15 HR-HPV genotypes, and 2 low-risk HPV (LR-HPV) genotypes using a multiplex real-time quantitive polymerase chain reaction (qPCR) assay., Results: Of 1,163 participants enrolled, 426 patients were diagnosed as verruca acuminata (CA) and 737 were asymptomatic men. The overall prevalence of HPV infection was 42% (489/1,163), and 27.4% (319/1,163) were positive for HR-HPV. HPV 16 (5.2%, 61/1,163) was the most common HR genotype overall, followed by HPV 52 (4.6%, 54/1,163), 51 (4.3%, 50/1,163), 18 (4.1%, 48/1,163), and 39 (4.0%, 47/1,163). Genotypes 16, 52, 39, 51, and 18 were most prevalent in CA patients, and 16, 51, 18, 59, and 39 in asymptomatic men. Prevalence of genotypes 31, 33, and 45 covered by the 9-valent HPV prophylactic vaccine was low in the assessed region., Conclusions: HPV 16, 52, 51, 18, 39, and 59 were the most common HR genotypes detected in men in Northern China. Importantly, HPV 39, 51, and 59 are not currently covered by either the 4-valent or 9-valent HPV vaccines.

Objective: To investigate the effect of electroacupuncture (EA) at "Dazhui" (GV14) and "Ciliao" (BL32) on rats with bladder detrusor hyperreflexia (DH) after supersacral spinal cord transection, as well as the mechanism of EA in improving the urinary function by regulating the expression of Wnt-1, β-catenin and Neurogenin 1(Ngn1)., Methods: A total of 48 female Sprague-Dawley rats were randomly divided into sham-operation group, model control group, EA group, and EA control group, with 12 rats in each group. T10 spinal cord transection (SCT) was performed by surgery. The Basso, Beattie and Bresnahan (BBB) score was used to evaluate the motor function of SCT rat, and the Crede technique was used to assist urination. After the urine volume became stable, the urodynamic test was used to determine whether a rat model of DH was successfully established. The rats in the EA group were given EA at GV14 and BL32, and those in the EA control group were given EA (10 Hz/50 Hz, 20 min) at the acupuncture points at 1 cm next to GV14 and BL32 at both sides alternatively. EA was performed once a day for one week. Urodynamic parameters were used to evaluate urinary function. Western blot and immunohistochemistry were used to measure the expression of Wnt-1 and β-catenin in the spinal cord, and immunofluorescence assay was used to measure the expression of Ngn1 in the spinal cord., Results: The BBB score of the model control group significantly decreased compared with that of the sham-operation group( P <0.01), and the EA group was significantly higher than the model control group and the EA control group. Compared with the sham-operation group, the model control group had significant increases in bladder base pressure, maximum pressure, and leak point pressure ( P <0.01) and significant reductions in maximum bladder capacity and compliance ( P <0.01). Compared with the model control group, the EA group had significant reductions in bladder base pressure, maximum pressure, and leak point pressure ( P <0.01) and significant increases in maximum bladder capacity and compliance ( P <0.01, P <0.05). Compared with the EA group, the EA control group had significant increases in bladder base pressure, maximum pressure, and leak point pressure ( P <0.01) and significant reductions in maximum bladder capacity and compliance ( P <0.01, P <0.05). Compared with the sham-operation group, the model control group had significant increases in the protein expression of Wnt-1 and β-catenin ( P <0.05, P <0.01) and a signi-ficant reduction in the protein expression of Ngn1 in the spinal cord ( P <0.01). Compared with the model control group, the EA group had significant increases in the protein expression of Wnt-1, β-catenin and Ngn1 in the spinal cord ( P <0.01). Compared with the EA group, the EA control group had significant reductions in the protein expression of Wnt-1, β-catenin, and Ngn1 in the spinal cord ( P <0.01)., Conclusion: EA at GV14 and BL32 can significantly improve urinary function in rats with bladder DH due to SCT, partially by activating the Wnt/β-catenin signaling pathway and promoting the protein expression of Wnt-1, β-catenin and Ngn1.

Objective: To examine genetic variants of familial hematuria (FH) associated genes in 3 families with hematuria with probands initially diagnosed with IgA nephropathy (IgAN). Methods: A retrospective analysis was performed on the clinical data, laboratory tests and genetic test results of three children with hematuria and the probands in three families with hematuria. The families were ascertained at the Department of Pediatrics, Fuzhou General Hospital of Nanjing Military Command from August 2014 to May 2018. Results: The proband of Family One, an 8-year-old boy, manifested gross hematuria. His renal biopsy pathology revealed IgAN. His father also manifested hematuria. Genetic testing showed that the proband and his father carried a heterozygous variant of the CFHR5 gene,533A>G (Asn178Ser). The child of Family Two, a 4-year-old girl, manifested hematuria. Her father, the proband of the family, was 36 years old, and manifested hematuria, proteinuria, high-frequency sensorineural deafness and renal insufficiency. He was diagnosed as IgAN according to clinical manifestations, renal pathology and routine immunohistochemistry without renal biopsy electron microscopy, renal tissue type Ⅳ collagen α3, α4, α5 chains immunofluorescence and skin type Ⅳ collagen α5 chain immunofluorescence. Genetic testing showed that the girl carried a heterozygous variant of the COL4A5 gene,566G>T (Gly189Val), and her father carried the hemizygous variant. The child of Family Three, a 7-year-old girl, manifested hematuria and proteinuria. Her mother, the proband of the family, was 34 years old, and manifested hematuria and proteinuria as well. The proband was diagnosed as IgAN by the same method used for Family Two. The girl's grandfather died of uremia at the age of 44. Genetic testing showed that the girl and her mother carried a heterozygous variant 539G>A (Gly180Glu)in COL4A5 gene. Conclusions: The variant of the CFHR5 gene identified in Family One is of uncertain signifance, and the two variants of the COL4A5 gene identified in Families Two and Three are pathogenic. The probands of Families Two and Three are diagnosed as Alport syndrome. The study suggests that clinicians should examine genetic variants of FH associated genes in families with hematuria when the probands were diagnosed as IgAN by their clinical manifestations, renal pathology and routine immunohistochemistry.

Metallic lithium protection plays a crucial role on improving the electrochemical properties of Li-anode-based batteries. Herein, for an advanced Li//graphite dual-ion battery, constructing a robust and conductive film of carbon nanofibers on a Li anode effectively achieves dendrite-free Li growth and hence significantly enhances the long-life cyclic stability.

Aim: The aim of the study was to explore the growth inhibitory effect of myricanol 5-fluorobenzyloxy ether (5FEM) and the underlying mechanism in human leukemic cells HL-60., Materials & Methods: 5FEM was obtained by chemical modification of myricanol with fluorobenzyloxy ether at the OH(5) position. The cytotoxicity, cell apoptosis, cell cycle and the expression of key apoptosis-related genes in HL-60 were evaluated., Results & Conclusion: 5FEM can significantly inhibited growth of HL-60 cells, increased the G2/M population and upregulated the expression of Bax, Fas, FasL, caspase-9 and p21 and downregulated that of Bcl-2 and survivin. The results enhance our understanding of 5FEM and aid the discovery of novel myricanol derivatives as potential antitumor agents.

Objective: To investigate the prevalence and characteristics of pathogenic variants in complement genes in Han Chinese children with atypical hemolytic uremic syndrome (aHUS). Method: Eleven Han Chinese children with aHUS, including 9 boys and 2 girls aged between 1 year and 4 months and 13 years, were investigated in Department of Pediatrics, Fuzhou General Hospital, from November 1998 to February 2014. Analysis of variants of all the exons of 10 complement genes (CFH, MCP, CFI, C3, CFB, CFHR1, CFHR2, CFHR3, CFHR4 and CFHR5), including 25 bases from 3' end and 25 bases from 5' end, was performed in the 11 cases by targeted sequence capture and next generation sequencing. Significant variants detected by next generation sequencing were confirmed by Sanger sequencing. To understand pathogenicity of variants found in the captured genes, we investigated genetic conservation by multiple protein sequence alignment among different species, and analyzed whether the variants were located in protein domains or not, and investigated functional significance by functional computational prediction methods. Result: Twenty-seven percent of Han Chinese children with aHUS carried pathogenic variants in the 10 complement genes. Pathogenic variant CFB 221G>A (R74H) was detected in Patient 3 and Patient 9, which was not found in parents of Patient 3' , and was found in healthy father of patient 9. Pathogenic variant CFHR5 242C>T (P81L) was found in Patient 2, and was found in healthy father of patient 2. However, no pathogenic variants in genes CFH, MCP, CFI, C3, CFHR1, CFHR2, CFHR3 and CFHR4 were identified. Conclusion: Pathogenic variants in the 10 complement genes were identified in 3/11 of Han Chinese children with aHUS in our study and CFB was the most frequently mutated gene.

This work describes the preparation of a PEGylated niosomes-mediated drug delivery systems for Paeonol, thereby improving the bioavailability and chemical stability of Paeonol, prolonging its cellular uptake and enhancing its synergistic anti-cancer effects with 5-Fu. PEGylated niosomes, which are prepared from biocompatible nonionic surfactant of Spans 60 and cholesterol, and modified with PEG-SA. Pae-PEG-NISVs were evaluated in vitro and in vivo. The cytotoxicity of Pae-PEG-NISVs was investigated against HepG2 cells. Fluorescence microscope was used to detect the apoptotic morphological changes. Growth inhibition assays were carried out to investigate whether Pae-PEG-NISVs could enhance the antiproliferative effects of Pae co-treated with 5-FU on HepG2 cells. The optimized Pae-PEG-NISVs had mean diameters of approximately 166 nm and entrapment efficiency (EE) of 61.8%. Furthermore, the in vitro release study of Paeonol from PEGylated niosomes exhibited a relatively prolonged release profile for 12 h. Pharmacokinetic studies in rats after i.v. injection showed that Pae-PEG-NISVs had increased elimination half-lives (t 1/2 , 87.5 versus 17.0 min) and increased area under the concentration-time curve (AUC 0-t , 38.0 versus 19.48 μg/ml*min) compared to Paeonol solution. Formulated Paeonol had superior cytotoxicity versus the free drug with IC 50 values of 22.47 and 85.16 μg/mL at 24 h on HepG2 cells, respectively, and we found that low concentration of Pae-PEG-NISVs and 5-Fu in conjunction had obviously synergistic effect. Our results indicate that the PEG-NISVs system has the potential to serve as an efficient carrier for Paeonol by effectively solubilizing, stabilizing and delivering the drug to the cancer cells.

BACKGROUND Cutaneous squamous cell carcinoma (cSCC) is the second most common type of non-melanoma skin cancer (NMSC) globally. The aims of this study were to further systematically clarify the potential association of rs833061 (-460 C>T) and rs1570360 (-1154 G>A), two SNPs of VEGF, with the risk of cSCC and the prognostic impacts on cSCC patients. MATERIAL AND METHODS This hospital-based case-control study analyzed peripheral venous blood collected from 100 cSCC patients and 124 healthy controls, and gathered personal information on patients. Genotypes of the VEGF gene -460C>T and -1154G>A polymorphism were detected using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method. Different distributions of allele frequencies and genotype in the case and control group were measured, comparing different genotype differences in the survival of patients with cSCC. RESULTS Distributions of allele frequencies and genotype of -460 C>T in the case and control group were statistically different; the TT + CT genotype was significantly correlated with a decrease risk of cSCC (OR=0.36, 95% CI=0.21-0.63, P<0.001). There was no difference in the distribution of allele frequencies and genotype of -1154 G>A between control and case groups. For -1154460C>T, the CC genotype was an adverse factor, associated with a significant decrease in the survival status of cSCC patients (P<0.001). For VEGF-1154 G>A, the AA genotype was significantly correlated with the reduced overall survival in cSCC patients, with the mean survival time of 23.88 months (P=0.009). CONCLUSIONS The VEGF gene -460 C>T polymorphism and -1154 G>A polymorphism may serve as potential genetic markers for the risk and prognosis of cSCC., Competing Interests: of conflict of interest None of the authors have conflicts of interest to report with regard to this manuscript.

Plasma exchange (PE) for the treatment of ricin toxicity has not been previously reported. Here we describe the use of PE to treat children who experienced ricin toxicity after ingesting castor beans. Seven children (median age: 8.1 years) who consumed castor beans (median: 5 beans) were treated with PE. All had bradycardia and sinus arrhythmia, and most had experienced episodes of vomiting and/or diarrhea. PE settings were blood flow, 50-80 mL/min; PE rate, 600-800 mL/h; volume of exchange, 1440-1950 mL. Median time from ingestion to PE was 73 h. All clinical symptoms disappeared and vital signs rapidly returned to normal after PE; no severe organ dysfunction occurred. All children were discharged and recovered uneventfully. Concentrations of all serum biochemical parameters significantly decreased immediately after PE. Some, but not all, of these parameters were also significantly decreased at 48 and 72 h after PE compared with before PE. Our findings suggest that PE can be an effective early intervention in the treatment of ricin toxicity due to castor bean ingestion., (© 2014 Wiley Periodicals, Inc.)

The aim of this study was to investigate the selection of plasma exchange (PE) parameters and the safety of children with severe ricinism. The PE parameters and heparin dosage in 7 children with severe ricinism were recorded, and changes in the patients' vital signs and coagulation function were monitored before and after PE. All patients successfully completed PE. The speed of blood flow was 50-80 mL/min, speed of exchange flow was 600-800 mL/h, and isolating rate of blood plasma was 12.5-19.05%. Transmembrane pressure was stable at approximately 100 mmHg, and venous pressure was stable at approximately 95 mmHg. The first dose of heparin was 0.39 ± 0.04 mg/kg, and the maintaining heparin dose was 0.40 ± 0.05 to 0.22 ± 0.03 mg·kg(-1)·h(-1). During the PE process, mean arterial pressure, heart rate, respiratory rate, and pulse oxygen saturation were steady. After PE, the activated partial thromboplastin time and thrombin time prolonged to 2-3 times greater than that before PE. However, no bleeding tendency was seen. For children with severe ricinism, the choice of PE to eliminate the toxin from blood, tissues, and organs was safe and effective.

Concentrated turtle aquaculture effluent poses an environmental threat to water bodies, and therefore needs to be treated prior to disposal. This study was conducted to assess the effect of multi-soil-layer (MSL) systems treating turtle aquaculture effluent with adding different amounts of sludge. Four MSL systems were constructed with dry weight ratios of sludge with 0%, 5%, 10%, and 20% (MSL 1, MSL 2, MSL 3, and MSL 4, respectively). The turtle aquaculture effluent had an average chemical oxygen demand (COD), ammonia nitrogen (NH4(+)-N) and total nitrogen (TN) concentration of 288.4, 213.4, and 252.0 mg/L, respectively. The COD/TN (C/N) ratio was 1.2. The results showed that the four MSL systems could effectively treat the COD, NH4(+)-N, and TN, and MSL 4 showed significantly improved NH4(+)-N removal efficiency, suggesting the potential of sludge addition to improve the turtle aquaculture effluent treatment. The average COD, TN, and NH4(+)-N removal efficiencies of MSL 4 were 70.3%, 66.5%, and 72.7%, respectively. To further interpret the contribution of microorganisms to the removal, the microbial community compositions and diversities of the four MSL systems were measured. Comparisons of the denaturing gradient gel electrophoresis (DGGE) profiles revealed that the amount of nitrifying bacteria and diversity in MSL 4 were higher than those in the other three systems. We concluded that adding 20% of sludge improved the NH4(+)-N removal and stability of the system for nitrification, due to the enrichment of the nitrifying bacteria in MSL 4.

Objective: Steroid-resistant nephrotic syndrome (SRNS) with MYO1E mutations has been identified as autosomal recessive focal segmental glomerulosclerosis (FSGS). To date, only two homozygous mutations in the MYO1E gene were reported in three families with FSGS. This study aimed to examine mutations in the MYO1E gene in children with familial SRNS in the Han Chinese ethnic group., Methods: Between 2005 and 2010, peripheral blood samples were collected from the probands, their siblings and parents of four families with autosomal recessive SRNS in the Han Chinese ethnic group. Four probands were studied from nine patients. The mutational analysis of MYO1E was performed by polymerase chain reaction and direct DNA sequencing. Fifty-nine healthy volunteers with normal urine analysis were included as controls., Results: Twenty-five MYO1E variants in the prohands from 4 families with SRNS were identified in this study. Among them, 24 variants were found in NCBI dbSNP. One heterozygous mutation IVS21-85G>A was found in the prohand from Family D, whereas it was absent in 59 normal Chinese controls. No splice site change caused by IVS21-85G>A was reported by analysis with NetGene2., Conclusions: MYO1E mutations are not a major cause of Chinese familial SRNS in this study.

Mutations in the Wilms' tumor suppressor gene (WT1) can lead to syndromic forms of steroid-resistant nephrotic syndrome (SRNS) such as Denys-Drash or Frasier syndrome and can cause isolated SRNS. A mutation within WT1 is a frequent cause of sporadic isolated SRNS in girls. In a worldwide cohort of girls, the rate of occurrence was 10.8%. Previous reports have indicated that in Chinese girls, the detection rate of WT1 mutations is 16.7% for early onset isolated nephrotic syndrome. The detection rate of WT1 mutations in Chinese girls with sporadic isolated SRNS is unknown. We examined WT1 mutations in 14 Chinese girls with sporadic isolated SRNS using polymerase chain reaction and direct sequencing and studied a control group of 38 boys with sporadic isolated SRNS. We identified a WT1 mutation in 1 of 14 (7.1% detection rate) Chinese girls with sporadic isolated SRNS. No mutations occurred in WT1 in the remaining 13 girls or the control group. Our investigation supports the necessity of genetic examination for mutations in WT1 in girls with sporadic isolated SRNS.

Objective: To study the effects of oxymatrine (OM) on the expressions of pro-collagen I (PC I), pro-collagen II (PC III), fibronectin (FN), matrix metalloproteinase-1 (MMP-1) mRNA of fibroblasts from keloid (KFb), hyperplastic scar (HFb), and normal skin (NFb), and to compare with hydrocortisone (HC)., Methods: The primary KFb, HFb and NFb were derived from patients and cultured in vitro using tissue block culture method. The fibroblasts were treated with 500 microg/mL OM, 2 microg/mL HC, or without any medicine (as the control). The mRNA expressions of PC I, PC III, FN, MMP-1 of the fibroblasts were detected using RT-PCR., Results: Under the normal condition, when compared with NFb, the mRNA expressions of PC I of KFb and HFb increased by 31.7% and 34.2% (both P < 0.05). Besides, the mRNA expression of PC III of KFb increased by 44.9% (P < 0.01). OM down-regulated the mRNA expressions of FN and PC I of HFb by 18.8% and 23.6% respectively (both P < 0.05). HC decreased the mRNA expressions of FN and PC I of HFb by 26.8% and 43.6% respectively (P < 0.05, P < 0.01). Meantime, OM up-regulated the mRNA expression of MMP-1 of KFb by 21.8% (P < 0.05)., Conclusions: OM suppressed the synthesis of extracellular matrix (ECM) possibly through down-regulating the mRNA expressions of PC I and FN. Compared with HC, OM could promote the degradation of ECM through inducing the MMP-1 mRNA expressions of KFb. Therefore, OM could be potentially used in treatment of hypertrophic scar and keloid.

Background: Enterovirus 71 (EV71) is a highly infectious agent that plays an etiological role in hand, foot, and mouth disease. It is associated with severe neurological complications and has caused significant mortalities in recent large-scale outbreaks. Currently, no effective vaccine or specific clinical therapy is available against EV71., Methods: Unmodified 21 nucleotide small interfering RNAs (siRNAs) and classic 2'-modified (2'-O-methylation or 2'-fluoro modification) siRNAs were designed to target highly conserved 5' untranslated region (UTR) of the EV71 genome and employed as anti-EV71 agents. Real-time TaqMan RT-PCR, western blot analysis and plaque assays were carried out to evaluate specific viral inhibition by the siRNAs., Results: Transfection of rhabdomyosarcoma (RD) cells with siRNAs targeting the EV71 genomic 5' UTR significantly delayed and alleviated the cytopathic effects of EV71 infection, increased cell viability in EV71-infected RD cells. The inhibitory effect on EV71 replication was sequence-specific and dosage-dependent, with significant corresponding decreases in viral RNA, VP1 protein and viral titer. Appropriate 2'-modified siRNAs exhibited similar RNA interference (RNAi) activity with dramatically increased serum stability in comparison with unmodified counterparts., Conclusion: Sequences were identified within the highly conserved 5' UTR that can be targeted to effectively inhibit EV71 replication through RNAi strategies. Appropriate 2'-modified siRNAs provide a promising approach to optimizing siRNAs to overcome barriers on RNAi-based antiviral therapies for broader administration.

Since the identification of the NPHS1 gene, which encodes nephrin, various investigators have demonstrated that the NPHS1 mutation is a frequent cause of congenital nephrotic syndrome (CNS); it is found in 98% of Finnish children with this syndrome and in 39-80% of non-Finnish cases. In China, compound heterozygous mutations in the NPHS1 gene have been identified in two Chinese families with CNS. To our knowledge, however, whether or not NPHS1 is the causative gene in sporadic Chinese CNS cases has not been established. We identified a homozygous mutation of NPHS1, 3250insG (V1084fsX1095), in a Chinese child with sporadic CNS. This finding leads us to suggest that NPHS1 mutations are also present in sporadic Chinese CNS cases. This gives additional support for the necessity for genetic examination of mutations in the NPHS1 gene in Chinese children with sporadic CNS.

Objective: To develop a fluorescence imaging-based novel system for quick screening of antitumor compounds in vitro., Methods: The antitumor activity of 26 components from Lindera aggregate were determined by relative number of viable cell labelled with fluorescein diacetate (FDA) in multiwell plates after exposure to these 26 different components. Then, the linearity and precision of this method were validated. The structures of active compounds in components with strong antitumor activity were deduced by LC/MS., Results: The linearity of this method for cells stained with FDA was validated (r² = 0.9858) in the range of 0-10⁴ cells per well, and the in-plate precision was 9.41 %. Two of 26 components from Lindera aggregate showed significant inhibition effect on proliferation of HepG2 cells (inhibition rate >90%)., Conclusion: This proposed rapid and reliable approach can be used for screening compounds with antitumor activity from Traditional Chinese Medicine in vitro. The major active compound of Lindera aggregate was putatively identified as norboldine by LC/MS analysis.

In the present study, A fluorescent imaging-based high-throughput screening method was developed for identifying anti-migratory compounds with 96-well Transwell plates. The correlation, precision and stability of this method were examined and the incubation time of dye Hoechst 33342 in addition to migration time was optimized. In addition, The inhibitory activity of anti-cancer drug paclitaxel on tumor cell migration was assayed and an IC50 value of 0.717 micromol x L(-1) was obtained. Using this method, 24 components from Rhizoma Alismatis were screened and one component with anti-migration activity was found. These results show that the new proposed method with good precision, stability and linear range has the potential to assay the inhibitory activity of anticancer compounds.

Aim: To prepare, identify and apply anti-human adenoviru(HAdv)neutralization monoclonal antibody(mAb)., Methods: BALB/c mice were immunized with live human adenovirus type3(HAdv-3) strain intranarially. Sp2/0 cells were fused with the spleen cells harvested from BALB/c mice. The chromosomal amounts of the hybridoma cells were analyzed by colchicine. A commercially available mouse mAb isotyping kit was used to identify the isotype of this mAb. Clones secreting specific monoclonal antibody were screened by indirect enzyme linked immunosorbent assay (ELISA), Western blot and indirect immunofluorescent assay. Infected animal model was established, and the protective effect of mAb was studied., Results: The fusion rate was 86%, and the positive rate was 51.4%. One of the hybridoma cell was identified(1A4), the chromosomal amounts of was 98, the subtype mAb type belonged to IgG2a/kappa, and the titer of the mAb secreted by the strain in ascite reached more than 10(-5);. The specificity of the mAb was proved by ELISA , Western blot and indirect immunofluorescent assay. This mAb had protective effect on animal infected by HAdv-3., Conclusion: The anti-adenovirus mAb which have neutralization activities was successfully prepared. The mAb recognized the hexon subunit and had protective effect on animal infected by HAdv-3.

Objective: To investigate the features and clinical implications of event related potential (ERP) and cognitive-processing in patients with depression, anxiety or both anxiety and depression, Methods: A total of 34 patients with anxiety (HAMA>14, HAMD<17), 33 patients with depression (HAMD>17, HAMA<14), 54 patients with both anxiety and depression (HAMA>14, HAMD>17), and 37 health volunteers were recruited in the study. The event related potential were recorded by Nihon Kohden instruments., Results: Compared with the healthy volunteers, the latency of P3a and P3b was longer (P<0.05) and the amplitude of N2-P3b was lower in the patients with anxiety (P<0.05); the Amplitude of N2-P3b was lower (P<0.05) in the patients with depression; the latency of P3a was longer and the amplitude of N2-P3b was lower (P<0.05) in the patients with both anxiety and depression. The latencies of P3a and P3b in the patients with anxiety and both anxiety and depression were longer than in the patients with depression(P<0.05). The latency of N2 in the patients with anxiety and depression was longer than in the patients with anxiety or depression (P<0.05)., Conclusion: The features of ERP in patients with depression are different from those with anxiety, which provides an electroneurophysiology basis for the clinical diagnosis and differential diagnosis for anxiety and depression.

Objective: To isolate the prevalent strain of enterovirus 71 (EV71) in Xi'an area in 2008, and compare the concordance of viral isolation, reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescent technique in detecting EV71, find the fast and effective method for detection, and analyze the differences between the EV71 strains isolated from Xi'an and Fuyang, Anhui., Method: Virus isolation and RT-PCR were carried out on vesicle fluid and throat swab specimens that were collected from the patients with hand-foot-and-mouth disease, RD and HEp-2 cell lines were used for viral isolation. The virus was identified by using immunofluorescence technique. Nucleotide sequencing was performed on positive product of RT-PCR, and compared with EV71 isolated from Fuyang in 2008, then submitted to Genbank., Result: Among the 56 samples of throat swab inoculated on RD and HEp-2 cells, the positive rates were 5.4% (3/56) and 1.8% (1/56), respectively. Among the 56 samples of vesicle fluid inoculated on RD and HEp-2 cells, the positive rates were 12.5% ( 7/56 ) and 5.4% (3/56), respectively. Cytopathic effect of RD and HEp-2 cells appeared on days 7 and 10, respectively. The positive rates of RT-PCR on throat swab and vesicle fluid samples were 21.4% (12/56) and 33.9% (19/56), respectively. Cytopathic effect was found in cell culture for 14 cases and immunofluorescence, showed that 9 of them were infected with EV71. The authors obtained the EV71 strain prevalent in Xi'an during 2008. The nucleotide sequence was submitted to the NCBI Genbank and gained the accession number EU812461., Conclusion: The EV71 in Xi'an prevalent during 2008 may have a weaker epithelial tropism. Comparison of the EV71 strain isolated from Xi'an with EU703812, EU703813 and EU703814 isolated from Fuyang, Anhui showed that the homology was 97%-98%. RT-PCR is an important method for rapid detection of EV71.

Objective: To evaluate the influence and therapeutic effect of self-improving based group psychotherapy which bases on pathopsychology mechanism of social anxiety disorder (SAD) in denfense mechanisms for the client with SAD., Methods: 70 psychotherapy clients and inpatients with SAD were involved in this group psychotherapy voluntarily. Every group had six to eight clients with SAD and two psychotherapists. Psychotherapy was conducted once a week with 2 to 2. 5 hours for a period of 8 weeks. The evaluation of the therapeutic effect contains the subjective perception of the clients with SAD, Liebowitz Social Anxiety Scale (LSAS), and Defense Style Questionnaire (DSQ)., Results: 65 clients with SAD completed the therapy. The clients reported self and social functions were improved after 8 weeks therapy. The total score of LSAS after the therapy was significantly lower than that of before the therapy (Z = -5.673, P=0.000). A significant decrease in immature defense machanism and in neurotic defense machanism were observed (Z = -4.866, -2.973; P=0.000, 0.003 respectively). And a trend of increasing the use of mature defense machanism was also observed (Z = -2.780, P=0.005)., Conclusion: Self-improving based group psychotherapy can encourage the clients with SAD accept themselves and cure the social anxiety symptoms. And there was a increasing use of mature defense machanism. These imply the clinic application value of the studied group psychotherapy.

Background: Determination of the mitotic index in sections of endometrium stained with haematoxylin and eosin (H&E) is difficult and time-consuming. We assessed the value of two mitotic markers as immunocytochemical reagents for measuring mitotic rates in endometrium., Methods: Mitotic protein monoclonal antibody 2 (MPM-2) and anti-phosphorylated histone H3 (Phospho H3) were applied to paraffin sections of rhesus macaque and human endometrium., Results: In estrogen-treated macaque endometrium the mean +/- SEM mitotic indices were: H&E 1.5 +/- 0.25%, Phospho H3 antibody 1.02 +/- 0.23% and MPM-2 antibody 0.69 +/- 0.17%; these were not statistically significantly different, but the Phospho H3 antibody gave a stronger and cleaner signal than the MPM-2 antibody. Comparisons were made between a computer-determined Phospho H3 index, the H&E-determined mitotic index and the Ki-67 index in samples of human endometrium across the cycle. All revealed that the highest proliferative rate occurred during the follicular phase, but the Phospho H3 and the mitotic indices were more highly correlated (R(2) = 0.89, P < 0.001) than the Ki-67 and mitotic indices (R(2) = 0.74, P < 0.05)., Conclusions: The exceptionally high contrast staining and the excellent correlation between the Phospho H3 and mitotic indices validates the specificity of the Phospho H3 antibody as a new tool for the assessment of endometrial mitotic activity.

Recently, scientists interested in diseases of the human brain have paid much attention to the endothelin group of peptides. Under normal conditions they are found in some types of neurons and in endothelial cells of microvessels but not in glial cells. This review focuses on the endothelin peptides and their involvement in various brain diseases. Particular attention is paid to their expression in reactive astrocytes seen in many pathological conditions of the human brain. Endothelin-1 is a very potent vasoconstrictor which may be involved in the vasospasm occurring in subarachnoid haemorrhage. Intracerebral injection or application to cerebral arteries in animals will cause a focal necrosis, apparently due to severe vasoconstriction. Reactive astrocytes occurring in cases with infarcts, lacunae, Alzheimer's disease, progressive multifocal leukoencephalopathy (PML) and subacute sclerosing panencephalitis (SSPE) express endothelin-like immunoreactivity. Astrocytes in vitro may produce, store and release endothelins. To some extent astrocytes grown in vitro mimic reactive astrocytes in vivo since in cultures astrocytes are removed from their natural environment which may trigger reactive responses. Therefore, in vivo reactive astrocytes may produce, store and release endothelins just as in vitro. If endothelins are released from reactive astrocytes they may act as mitogens and may influence microcirculation by inducing vasoconstriction of intracerebral arterioles. In such ways endothelins may contribute to the final lesions seen in cases with infarcts, lacunae, traumatic conditions, Alzheimer's disease and inflammatory diseases of the brain.

In order to investigate the expression of endothelin-like immunoreactivity in astrocytes of viral infections of the human brain the avidin-biotin peroxidase complex method and a polyclonal antiserum were used. Autopsy material was obtained from 5 cases of herpes simplex encephalitis, two of progressive multifocal leukoencephalopathy (PML) and two of subacute sclerosing panencephalitis (SSPE). All the 5 herpes simplex encephalitis cases presented groups of immunoreactive astrocytes around necrotic, inflammatory lesions. The PML cases exhibited a large number of immunoreactive astrocytes in and around lesions of the white matter. The cases of SSPE disclosed numerous, markedly stained fibrillary immunoreactive astrocytes; they were most abundant in degenerated regions of the white matter. The processes and peripheral cytoplasm of giant astrocytes in the PML cases contained immunoreactive material but the perinuclear region was devoid of such material. In the herpes simplex and the SSPE cases immunoreactivity was present throughout the cytoplasm and processes of reactive fibrillary astrocytes. Many nerve cells in the cerebral cortex, hippocampus, cerebellum and pons of control cases exhibited endothelin-like immunoreactivity but this occurred in only exceptional astrocytes of control cases. Endothelin-like immunoreactivity was not present in the oligodendrocytes and vascular endothelial cells of controls and cases of virus infection. The expression of endothelin-like immunoreactivity in astrocytes in human viral diseases reflects probably an increased intracellular content of endothelin. If this peptide is released from such astrocytes, it may act as a mitogen and by inducing constriction of arterioles it may influence the microcirculation.

The avidin-biotin peroxidase complex method and a polyclonal antiserum were used to investigate the expression of endothelin-like immunoreactivity in human autopsy cases presenting cerebral infarcts and lacunes. Cases with recent infarcts showed loss of immunoreactivity in neurons of the infarcts. Immunoreactive granules started to appear in astrocytic endfeet of microvessels of the border zones. Later on, numerous reactive astrocytes presented immunoreactivity in the entire cell body. Cases with old infarcts had a more widespread presence of astrocytes with endothelin-like immunoreactivity around the lesions. Macrophages particularly of old infarcts showed a strong immunoreactivity. Cases with old lacunes presented numerous reactive fibrillary astrocytes with endothelin-like immunoreactivity in the cell body and the processes. Astrocytes of controls showing endothelin-like immunoreactivity were very rare and such cells were usually confined to the molecular layer of the cerebral cortex. The endothelin-like immunoreactivity in astrocytes and macrophages of cases with cerebrovascular diseases probably reflects an increased intracellular content of endothelin. If this compound is released from such cells it may act as a mitogen and influence microcirculation by inducing vasoconstriction of intracerebral arterioles.-, (1994 Lippincott Williams & Wilkins.)

The avidin-biotin peroxidase complex method and a polyclonal antiserum were used to investigate the distribution of endothelin-1-like immunoreactivity of cerebral astrocytes in autopsy cases of Alzheimer's disease compared with controls. The cases of Alzheimer's disease presented numerous astrocytes with intense endothelin-1-like immunoreactivity of the cell body often extending into the finest ramifications of the cell processes. Absorption of the antiserum by the corresponding antigen eliminated this immunostaining. The immunoreactive astrocytes were most consistently present in the subcortical white matter of the cerebral hemispheres and the folia of the cerebellum. The immunoreactive cells were often located in small clusters close to blood vessels. Five of the seven cases showed immunoreactive astrocytes in the molecular layer of the cerebral cortex and three of the seven cases presented regions in which immunoreactive astrocytes appeared to be located in the periphery of plaques. The pons contained small groups of immunoreactive astrocytes in five of the cases. The cerebellum had such cells in six of the seven investigated patients. Immunoreactive astrocytes were very rare in control cases without cerebral disease. Many nerve cells in the cerebral neocortex, hippocampus, cerebellum and pons of Alzheimer cases and controls exhibited endothelin-1-like immunoreactivity. Oligodendrocytes and endothelial cells of blood vessels of controls and Alzheimer cases did not show such immunoreactivity. The expression of endothelin-1-like immunoreactivity in astrocytes of Alzheimer's disease probably reflects an increased content of endothelin-1. If endothelin-1 is released from such astrocytes it may reach smooth muscle cells of the intracerebral blood vessels and disturb micro-circulation since this compound is a most powerful vasoconstrictor peptide.

The expression of endothelin-1-like immunoreactivity in astrocytes of the human brain was investigated by the avidin-biotin-peroxidase complex method in post mortem material. A marked immunoreaction was present in reactive astrocytes around infarcts, lacunes, traumatic injuries, the lesions of progressive multifocal leuco-encephalopathy and in the cerebral cortex and white matter of Alzheimer's disease. The brains of patients who had neither history nor signs of cerebral disease exhibited only occasional immunoreactive astrocytes. A hypothesis is presented that endothelin-1 may be released from reactive astrocytes in many organic diseases of the human brain with considerable pathogenic consequences. It is known from experimental investigations that endothelin-1 may for instance cause severe vasoconstriction resulting in cell injury and that it may act as a growth factor for glial cells.