Objective:To investigate the changes of podocyte circadian rhythm in the high-glucose environment and diabetic nephropathy (DN) mouse model and the protective effect of melatonin on podocyte injury in DN.Methods:Primary podocytes were cultured in vitro and divided into 3 groups: control group, high glucose (30 mmol/L) group and high glucose (30 mmol/L) treated with melatonin (0.1 mmol/L or 0.5 mmol/L) group. The podocytes were collected every 4 hours for 24 hours, synchronized with dexamethasone (100 nmol/L) for 2 hours, which was recorded as zeitgeber time 0 point. Male C57BL/6J mice aged 6-8 weeks and weighing about 20 g were randomly (randomized block) assigned to three groups: control group, DN (high-fat diet+streptozotocin 120 mg/kg intravenously injected) group, and DN (high-fat diet+streptozotocin 120 mg/kg intravenously) treated with melatonin (20 mg/kg intragastric treatment) group (melatonin group). Real-time quantitative PCR was used to detect the mRNA levels of rhythm genes in podocytes. Western blotting was used to detect the protein expression levels of circadian clock genes (Clock and Bmal1), podocyte marker proteins (Nephrin, Synaptopodin, WT1, and Desmin) and autophagy-related proteins (Beclin1, LC3Ⅱ/Ⅰ and P62). Immunofluorescence staining was used to detect the protein expression level of WT1, and immunohistochemistry was used to analyze the protein expression levels of P62 and cleaved-caspase-3 in renal tissues of mice. The pathological changes of renal glomerulus were observed under electron microscope. Results:(1) Dexamethasone reseted the expression and rhythmic oscillation of circadian clock genes in podocytes. The circadian rhythmic oscillations of Clock and Ck1e mRNA in the high glucose group were flattened compared to the control group, and the circadian rhythmic oscillations in Clock and Ck1e mRNA expression were partial recovery in high glucose treated with melatonin group (all P<0.05). (2) Compared with the control group, the protein expression levels of Nephrin, Synaptopodin and WT1 were lower while Desmin was higher in the high glucose group (all P<0.05). The protein expression levels of Nephrin, Synaptopodin and WT1 were higher and the protein expression level of Desmin was lower in the high glucose treated with melatonin (0.5 mmol/L) group compared with the high glucose group (all P<0.05). (3) The invivo experimental results showed that compared with the DN group, melatonin group had higher protein expression levels of glomerular Nephrin and WT1, and lower urinary albumin/creatinine ratio, width of foot process and thickness of glomerular basement membrane (all P<0.05). The protein expression levels of Beclin1 and LC3Ⅱ/Ⅰ in the DN group were lower than those in the control group, and the protein expression level of P62 was higher than that in the control group (all P<0.05). Compared with the DN group, the protein expression levels of Beclin1 and LC3Ⅱ/Ⅰ in the melatonin group were significantly higher, and the protein expression level of P62 was lower (all P<0.05). Conclusions:Melatonin can partially restore the circadian rhythm of clock genes in high-glucose environment, improve autophagy and alleviate injury in podocytes.

AIM: To observe the effects of Tangshen formula (TSF) treatment on lipid efflux and uptake in sodium palmitate (PA) induced RAW264.7 macrophages. METHODS: After 200 μmol/L PA induced RAW264.7 macrophages, TSF and PGC-1α-siRNA were given to intervene respectively. The lipid content in the cells was detected by ELISA kit; intracellular lipid droplet deposition was detected by BODIPY 493/503 and Filipin staining. Western blot and Real-time PCR were used to detect the expression of PGC-1α, LXR, ABCA1 and CD36. RESULTS: TSF diminished the levels of TC, TG and intracellular lipid droplet deposition in PA-induced RAW264.7 macrophages. Western blot and Real-time PCR analysis showed that TSF could up-regulate the expression of PGC-1α, LXR, ABCA1 and down-regulate the expression of CD36. Furthermore, silencing PCG-1α by SiRNA significantly suppressed the effects of upregulating the expression of PGC-1α, LXR and ABCA1, and downregulating the CD36 expression with TSF treatment. CONCLUSION: TSF may extenuate intracellular lipid droplet deposition in macrophages by upregulating cholesterol efflux through activating the PGC-1α/LXR/ABCA1 pathway and inhibiting lipid uptake through down-regulateing the expression of CD36.

On January 22, 2020, China National Center for Bioinformation (CNCB) released the 2019 Novel Coronavirus Resource (2019nCoVR), an open-access information resource for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). 2019nCoVR features a comprehensive integra-tion of sequence and clinical information for all publicly available SARS-CoV-2 isolates, which are manually curated with value-added annotations and quality evaluated by an automated in-house pipeline. Of particular note, 2019nCoVR offers systematic analyses to generate a dynamic landscape of SARS-CoV-2 genomic variations at a global scale. It provides all identified variants and their detailed statistics for each virus isolate, and congregates the quality score, functional annotation,and population frequency for each variant. Spatiotemporal change for each variant can be visualized and historical viral haplotype network maps for the course of the outbreak are also generated based on all complete and high-quality genomes available. Moreover, 2019nCoVR provides a full collection of SARS-CoV-2 relevant literature on the coronavirus disease 2019 (COVID-19), including published papers from PubMed as well as preprints from services such as bioRxiv and medRxiv through Europe PMC. Furthermore, by linking with relevant databases in CNCB, 2019nCoVR offers data submission services for raw sequence reads and assembled genomes, and data sharing with NCBI. Collectively, SARS-CoV-2 is updated daily to collect the latest information on genome sequences, variants, hap-lotypes, and literature for a timely reflection, making 2019nCoVR a valuable resource for the global research community. 2019nCoVR is accessible at https://bigd.big.ac.cn/ncov/.

Objective To study the CT and MRI findings and corresponding pathological basis of non-epithelial ovarian cancer (NEOC).Methods The CT,MRI and clinical data of 74 patients with NEOC proved by surgical pathology were reviewed retrospectively.CT plain scan was performed in 57 cases,and CT enhancement scan was performed in 52 cases;MRI plain scan was performed in 17 cases,and MRI enhancement scan was performed in 11 cases;CT and MRI scans were performed together in 3 cases.Results Among the 74 patients,there were 23 mature teratomas (25 lesions),2 immature teratomas,8 strumas,10 yolk sac tumors,6 dysgerminomas,7 granulosa cell tumors,11 thecomas,5 fibromas and 2 sertoli-leydig cell tumors.97.3% (72/74)of the lesions were unilateral.There were 88.0% (22/25)and 92.0% (23/25)detection rate of calcification and fat in teratomas,respectively.On contrast enhancement,the immature teratoma showed significant enhancement,whereas the mature teratoma showed mild enhancement.On plain CT,strumas presented as multilocular cystic-solid masses,with calcification in 5 cases.After contrast agent administration,the solid portions showed marked enhancement.Tortuous vessels were seen in 7 yolk sac tumors.The linear hypointensity in dysgerminoma on T2WI showed marked enhancement.The granulosa cell tumors were cystic-solid masses,and 4 cases with increased estradiol.Solid mass with mild enhancement were present in thecomas and fibromas,and there were 8 cases with ascites.CT findings of 2 sertoli-leydig cell tumors were cystic-solid masses with significant enhancement of the cystic wall on contrast CT.Conclusion NEOC have some certain CT and MRI characteristics.Teratomas often contain calcification and fat.Strumas:multilocular cystic-solid masses.Yolk sac tumor:tortuous vessels.Dysgerminoma:marked linear enhancement.

Objective To observe the effect of glycosides on serum indexes in elderly patients with primary nephrotic syndrome.Methods A total of 280 elderly patients with primary nephrotic syndrome were randomly divided into control group and observation group,140 cases in each group.Patients in the control group were treated with conventional western medicine,and those in the observation group were treated with additional glycosides.The efficacy and 24 h urinary protein,serum adiponectin,serum albumin,serum creatinine and immunological parameters,carbohydrate antigen levels before and after treatment were compared between two groups.Results The clinical control rate and total effective rate in the observation group were significantly higher than those in the control group (P ＜ 0.05).4,8 weeks after treatment,24 h urinary protein,serum adiponectin,serum creatinine of observation group were significantly lower than those of the control group,while the serum albumin was significantly higher than the control group (P ＜ 0.05).The serum levels of IgA,IgG and IgG/IgM in the observation group were significantly higher than those in the control group,while the IgM level was significantly lower than that in the control group (P ＜ 0.05).The serum levels of CA125,CA153 and CA199 in the observation group were significantly lower than those in the control group (P ＜ 0.05).Conclusion Glycosides can effectively increase the clinical remission rate of primary nephrotic syndrome in elderly patients,reduce the loss of albumin and protect renal function,and delay the progression of NS.

Objective To observe the effect of glycosides on serum indexes in elderly patients with primary nephrotic syndrome.Methods A total of 280 elderly patients with primary nephrotic syndrome were randomly divided into control group and observation group,140 cases in each group.Patients in the control group were treated with conventional western medicine,and those in the observation group were treated with additional glycosides.The efficacy and 24 h urinary protein,serum adiponectin,serum albumin,serum creatinine and immunological parameters,carbohydrate antigen levels before and after treatment were compared between two groups.Results The clinical control rate and total effective rate in the observation group were significantly higher than those in the control group (P ＜ 0.05).4,8 weeks after treatment,24 h urinary protein,serum adiponectin,serum creatinine of observation group were significantly lower than those of the control group,while the serum albumin was significantly higher than the control group (P ＜ 0.05).The serum levels of IgA,IgG and IgG/IgM in the observation group were significantly higher than those in the control group,while the IgM level was significantly lower than that in the control group (P ＜ 0.05).The serum levels of CA125,CA153 and CA199 in the observation group were significantly lower than those in the control group (P ＜ 0.05).Conclusion Glycosides can effectively increase the clinical remission rate of primary nephrotic syndrome in elderly patients,reduce the loss of albumin and protect renal function,and delay the progression of NS.

Objective To provide the reference basis for reducing the occurrence of nutritional risk by analyzing possible risk factors for nutritional risk status and investigating the relationship between preoperative nutritional risk status and prognosis.Methods We retrospectively analyzed the clinical data of 894 patients(nutritional risk group of 491 cases,non-nutritional risk group of 403 cases) with esophageal cancer resection.The univariate analysis was used to analyze the relationships between nutritional risk status with postoperative complications and length of hospitalization.And the multiple Logistic regression model analysis was applied to analyze the risk factors of nutritional risk status.Results The nutritional risk group had a higher rate of postoperative complications (33.60 ％ vs.19.60 ％,U =-3.429,P =0.001),higher incidence of serious complications (23.01％ vs.8.68％,U =-3.611,P =0.000) and longer hospital stays [(37.20 ± 13.89) days vs.(31.69 ± 13.49) days,t =4.124,P =0.000] than that of non-nutritional risk group.The results of the multiple logistic regression analysis showed that the preoperative nutritional risk factors were associated with the patients' age (OR =1.58,95％ CI:1.101-2.268),number of symptoms entries(OR =7.97,95 ％ CI:6.071-10.463),symptom severity (OR =0.26,95％ CI:0.186 -0.385),and dietary intake (OR =0.62,95 ％ CI:0.482-0.813),P ＜ 0.05 for all.Conclusion The older patients with more severe symptoms and poor diet are more likely to suffer from nutritional risk.Prolonged hospital stay and postoperative complications easily happen in patients with nutritional risk.So patients with preoperative nutritional risk should be given timely and effective nutrition intervention measures,in order to reduce postoperative complications and length of hospitalization.

[ ABSTRACT] AIM:To evaluate the clinical application of single nucleotide polymorphism array ( SNP array) in prenatal diagnosis for screening the abnormality of women with Down’ s syndrome ( DS) .METHODS:The amniotic fluid samples ( n=312) collected by amniocentesis for the DS screening abnormality women were tested by karyotyping and SNP array analysis, respectively.The findings of karyotyping and SNP array analysis were compared.RESULTS:Two cases of trisomy 21 were identified by karyotyping and SNP array analysis, but SNP array analysis failed to identify 6 cases of chro-mosome balanced structural rearrangement.SNP detected 176 cases copy number variants ( CNVs) in 303 cases normal karyotype were detected by SNP, including 106 benign CNVs, 61 variants of unknown significance (VOUS), 9 de novo CNVs, and none of them was pathogenic.The distribution difference of CNVs in DS screening positive group and DS screening positive plus advanced maternal age group was not statistically significant ( P>0.05) .Furthermore, we reported 14 kinds of CNVs for the first time in population.CONCLUSION:SNP array can further assure chromosome microdupli-cation/microdeletion.In normal karyotype fetus of prenatal diagnosis, SNP can detect some clinical significant CNVs.

Objective To investigate the diagnostic value ofbiomarkers in cerebrospinal fluid in differentiating meningeal carcinomatosis (MC) and tubercular meningitis (TBM).Methods The clinical features and cerebrospinal fluid (CSF) test results of 34 patients with MC and 44 patients with TBM,admitted to our hospital from January 1994 to August 2014,were retrospectively analyzed and compared.Results The main clinical manifestations of MC included headache and bowel and bladder dysfunction,and those of TBM were fever and headache.The cell population of MC and TBM patients increased and the number of leucocyte of TBM patients was significantly larger than that of MC patients (P＜0.05);the CSF of all the patients presented with decreased levels of glucose and chloride,and increased protein level,and higher protein in MC patients was noted than that in TBM patients (P＜0.05).The biomarkers in CSF of MC and TBM patients increased to varying degrees,and the biomarkers,including carcinoembryonic antigen (CEA),cancer antigen (CA) 125,CA 199,CA 153 and CYFRA21-1,in MC group were significantly higher than those in TBM group (P＜0.05).The neuron-specific enolase level in the TBM patients of Ⅲ grade was significantly higher than that in the MC patients (P＜ 0.05).Conclusion The detections of tumor biomarkers in cerebrospinal fluid is a tool in differentiating MC and all stages of TBM,especially with the combined detections of CEA,CA125,CA199 and CYFRA21-1.

OBJECTIVETo study the feasibility of using methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) for the detection of DNA methylation in placenta tissue.

METHODSFor blood cells from 13 non-pregnant women and 9 euploid placenta, the ratios of DNA methylation were evaluated for 4 genes including CGI149, CGI113, HLCS and ACTB with MS-MLPA and bisulfite sequencing, respectively.

RESULTSThe methylation ratio of the ACTB gene was 0-0.1 for the blood cells when the digestion control was completely digested. The cutoff value for the methylation ratio of MS-MLPA has been determined as 0.1. For the 9 placenta samples, results of MS-MLPA and bisulfite sequencing were concordant for all of the four genes.

CONCLUSIONMS-MLPA is an effective alternative to bisulfite sequencing for the assessment of methylation ratios in placental tissues.

Actins ; genetics ; Adult ; Carbon-Nitrogen Ligases ; genetics ; CpG Islands ; genetics ; DNA Methylation ; Endosomal Sorting Complexes Required for Transport ; genetics ; Feasibility Studies ; Female ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Placenta ; metabolism ; Pregnancy ; Reproducibility of Results ; Ribosomal Proteins ; genetics ; Young Adult