Objective The aim of this study was to detect the expression of miR-383-5p in bladder cancer tissues and bladder cancer 5637 cells,BIU-87 cells,TCCSUP cells and HT-1376 cells,and to explore the effects of miR-383-5p on the prolif-eration,invasion,migration and apoptosis of bladder cancer cells by targeting CIP2A.Methods The expression of miR-383-5p was detected by qRT-PCR in human bladder cancer tissues and their corresponding adjacent tissues,5637 cells,BIU-87 cells,TCCSUP cells,HT-1376 cells,human bladder transitional epithelial cells.BIU-87 cells with low miR-383-5p expression were selected for subsequent experiments.BIU-87 cells were divided into the blank group(normal culture),miR-383-5p NC group(negative control,transfected with miR-383-5p negative control),miR-383-5p mimic group(transfected with miR-383-5p mimic),and miR-383-5p mimic+pc-CIP2A group(co-transfected with miR-383-5p mimic and CIP2A overexpression plasmid pc-CIP2A).CCK-8 kit was used to detect the viability of BIU-87 cells in each group;Flow cytometry was used to detect apoptosis of BIU-87 cells;Transwell assay was used to measure cell invasion ability of BIU-87 cells;Scratch assay was used to measure cell migration ability of BIU-87 cells;Western blot was used to determine the expression of proteins related to apoptosis,invasion(MMP-2,MMP-9),and CIP2A/PP2A in BIU-87 cells;The dual luciferase assay was used to verify the targeting relationship between miR-383-5p and CIP2A in BIU-87 cells.Results The expression of miR-383-5p was low in bladder cancer tissues and bladder cancer cells.Compared with the blank group,BIU-87 cells in the miR-383-5p mimic group showed a significant increase the level of miR-383-5p(0.91±0.10 vs.1.67±0.24,P<0.01)and a significant decrease in the expression of CIP2A protein(1.32±0.17 vs.0.45±0.03,P<0.001),the cell viability,invasion,migration abilities,the expression of proteins related to invasion(MMP-2,MMP-9),and the expression of Bcl-2 protein[(100.00±4.36)% vs.(32.15±2.65)% ,(150.20±12.95)vs.(82.35±7.01),(77.91±3.63)% vs.(46.12±2.54)% ,1.02±0.11 vs.0.22±0.04,1.03±0.18 vs.0.21±0.04,1.01±0.14 vs.0.27±0.05,P<0.001];The apoptosis rate,the expression of caspase-3 and Bax proteins related to apoptosis,and PP2A expression were significantly increased[(14.02±2.29)% vs.(38.21±3.20)% ],0.81±0.11 vs.1.78±0.24,0.83±0.12 vs.1.72±0.24,0.27±0.02 vs.0.95±0.16,P<0.001].Compared with the miR-383-5p mimic group,BIU-87 cells in the miR-383-5p mimic+pc-CIP2A group significantly increased the cell viability,invasion,migration abilities,the expression of proteins related to invasion,and the expression of Bcl-2 protein[(32.15±2.65)% vs.(50.18±3.77)% ,(82.35±7.01)% vs.(116.30±13.70),(46.12±2.54)% vs.(58.43±3.15)% ,0.22±0.04 vs.0.60±0.08,0.21±0.04 vs.0.5 8±0.06,0.27±0.05 vs.0.64±0.08,P<0.05];The apoptosis rate,the expression of caspase-3,Bax,and PP2A was signifi-cantly reduced in the miR-383-5p mimic+pc-CIP2A group[(38.21±3.20)% (23.15±2.74)% ,1.78±0.24 vs.1.25±0.21,1.72±0.24 vs.1.23±0.18,0.95±0.16 vs.0.60±0.13,P<0.05].The results of dual luciferase experiments showed a corresponding tar-geting relationship between miR-383-5p and CIP2A.Conclusion Increasing the expression of miR-383-5p can inhibit the prolif-eration,invasion and migration of bladder cancer BIU-87 cells,and enhance the ability of apoptosis,which may be achieved by targe-ted regulation of CIP2A.

Objective The aim of this study was to detect the expression of miR-383-5p in bladder cancer tissues and bladder cancer 5637 cells,BIU-87 cells,TCCSUP cells and HT-1376 cells,and to explore the effects of miR-383-5p on the prolif-eration,invasion,migration and apoptosis of bladder cancer cells by targeting CIP2A.Methods The expression of miR-383-5p was detected by qRT-PCR in human bladder cancer tissues and their corresponding adjacent tissues,5637 cells,BIU-87 cells,TCCSUP cells,HT-1376 cells,human bladder transitional epithelial cells.BIU-87 cells with low miR-383-5p expression were selected for subsequent experiments.BIU-87 cells were divided into the blank group(normal culture),miR-383-5p NC group(negative control,transfected with miR-383-5p negative control),miR-383-5p mimic group(transfected with miR-383-5p mimic),and miR-383-5p mimic+pc-CIP2A group(co-transfected with miR-383-5p mimic and CIP2A overexpression plasmid pc-CIP2A).CCK-8 kit was used to detect the viability of BIU-87 cells in each group;Flow cytometry was used to detect apoptosis of BIU-87 cells;Transwell assay was used to measure cell invasion ability of BIU-87 cells;Scratch assay was used to measure cell migration ability of BIU-87 cells;Western blot was used to determine the expression of proteins related to apoptosis,invasion(MMP-2,MMP-9),and CIP2A/PP2A in BIU-87 cells;The dual luciferase assay was used to verify the targeting relationship between miR-383-5p and CIP2A in BIU-87 cells.Results The expression of miR-383-5p was low in bladder cancer tissues and bladder cancer cells.Compared with the blank group,BIU-87 cells in the miR-383-5p mimic group showed a significant increase the level of miR-383-5p(0.91±0.10 vs.1.67±0.24,P<0.01)and a significant decrease in the expression of CIP2A protein(1.32±0.17 vs.0.45±0.03,P<0.001),the cell viability,invasion,migration abilities,the expression of proteins related to invasion(MMP-2,MMP-9),and the expression of Bcl-2 protein[(100.00±4.36)% vs.(32.15±2.65)% ,(150.20±12.95)vs.(82.35±7.01),(77.91±3.63)% vs.(46.12±2.54)% ,1.02±0.11 vs.0.22±0.04,1.03±0.18 vs.0.21±0.04,1.01±0.14 vs.0.27±0.05,P<0.001];The apoptosis rate,the expression of caspase-3 and Bax proteins related to apoptosis,and PP2A expression were significantly increased[(14.02±2.29)% vs.(38.21±3.20)% ],0.81±0.11 vs.1.78±0.24,0.83±0.12 vs.1.72±0.24,0.27±0.02 vs.0.95±0.16,P<0.001].Compared with the miR-383-5p mimic group,BIU-87 cells in the miR-383-5p mimic+pc-CIP2A group significantly increased the cell viability,invasion,migration abilities,the expression of proteins related to invasion,and the expression of Bcl-2 protein[(32.15±2.65)% vs.(50.18±3.77)% ,(82.35±7.01)% vs.(116.30±13.70),(46.12±2.54)% vs.(58.43±3.15)% ,0.22±0.04 vs.0.60±0.08,0.21±0.04 vs.0.5 8±0.06,0.27±0.05 vs.0.64±0.08,P<0.05];The apoptosis rate,the expression of caspase-3,Bax,and PP2A was signifi-cantly reduced in the miR-383-5p mimic+pc-CIP2A group[(38.21±3.20)% (23.15±2.74)% ,1.78±0.24 vs.1.25±0.21,1.72±0.24 vs.1.23±0.18,0.95±0.16 vs.0.60±0.13,P<0.05].The results of dual luciferase experiments showed a corresponding tar-geting relationship between miR-383-5p and CIP2A.Conclusion Increasing the expression of miR-383-5p can inhibit the prolif-eration,invasion and migration of bladder cancer BIU-87 cells,and enhance the ability of apoptosis,which may be achieved by targe-ted regulation of CIP2A.

Objective To investigate and predict the molecular targets and mechanism of Huanglian Jiedu Decoction (黄连解毒汤, HLJDD) in the treatment of Corona Virus Disease 2019 (COVID-19) through network pharmacology and molecular docking analysis. Methods The chemical constituents and action targets of HLJDD were retrieved on Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), SymMap v2, Encyclopedia of Traditional Chinese Medicine (ETCM), a High-throughput Experiment- and Reference-guided Database of Traditional Chinese Medicine (HERB), and Traditional Chinese Medicine Integrated Database (TCMID). UniProt and GeneCards were used to query the target genes that corresponding to the active compounds, and then a compound-target network was constructed using Cytoscape 3.7.2. Gene Ontology (GO) database was used to annotate GO functions. Kyoto Encyclopedia of Genes and Genomes (KEGG) was used to predict the possible mechanisms of active compounds. The Database for Annotation, Visualization and Integrated Discovery (DAVID) was used to analysis the tissue enrichment. The main active compounds in HLJDD are molecularly docked with their corresponding related targets. Results Seventy-six compounds were screened and 458 corresponding targets in the network were obtained. Gene annotation showed that the targets were involved mainly in 1 953 biological processes. 884 signaling pathways was enriched, involving signaling by interleukins, cytokine signaling in immune system, generic transcription pathway, and RNA polymerase II transcription. The targets mainly distributed in the lung, liver, and placenta, involving a variety of immune cells, such as T cells and B cells. The molecular docking results showed that core compounds such as wogonin, berberine, and baicalein had high affinity with tumor necrosis factor (TNF), insulin (INS), and tumor protein 53 (TP53). Conclusion The active compounds in HLJDD may have a therapeutic effect on COVID-19 through regulating multiple signal pathways by targeting genes such as vascular endothelial growth factor A (VEGFA), INS, interleukin-6 (IL-6), TNF, caspase-3 , TP53, and mitogen-activated protein kinase 3 (MAPK3).

Objective:To compare the difference between the combined diagnostic effect of plasma Septin9(SEPT9) and polyligand Syndecan-2(SDC2) methylation with four serum tumor markers in the diagnosis of colorectal cancer.Methods:In this study, 128 patients who were treated in the affiliated Hospital of Xuzhou Medical University from March to December in 2019 were selected for a case-control study.All the subjects were examined by gastroenteroscopy.According to the pathological results, they were divided into three groups: colorectal cancer group( n=74) and colorectal adenoma group( n=7). The patients with no abnormal or inflammatory polyps or proliferative polyps examined by gastroenteroscopy were taken as the control group( n=47). The methylation levels of SEPT9 gene and SDC2 gene were detected by Roche Lightcycler 480 II real-time fluorescence quantitative polymerase chain reaction, and the concentrations of alpha-fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 were detected by Roche Cobas 8000 electrochemiluminescence instrument.Chi-square test was used to compare the positive rate of each marker in the three groups.Medcalc was used to draw the receiver operating characteristic curve (ROC) curve of the subjects′ working characteristic curve, and the value of each index in the diagnosis of colorectal cancer was analyzed. Results:The positive rates of SEPT9 gene and SDC2 gene methylation were 81.1%(60/74) and 67.6%(50/74) respectively in colorectal cancer group, and increased to 85.1%(63/74) after combined detection.The positive detection rates of alpha-fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 in colorectal cancer group were 1.4%(1/74), 33.8%(25/74), 6.8%(5/74) and 13.5%(10/74), respectively.When the four tumor markers were detected together, the positive detection rates were only increased to 43.2%(32/74), except for AFP and carbohydrate antigen 125(χ 2=3.847, 2.430, all P>0.05). The differences were statistically significant (χ 2=48.230, 30.487, 43.285, 3.847, 8.788, 6.988, 8.722, all P<0.05). The area under the curve (AUC) of SEPT9 methylation, SDC2 methylation, alpha fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199 were 0.854 (0.781, 0.910), 0.795 (0.715, 0.861), 0.575 (0.485, 0.662), 0.685 (0.597, 0.764), 0.603 (0.513, 0.689) and 0.631 (0.541, 0.715), respectively.The AUC of combined detection of two DNA methylation markers was better than that of alpha fetoprotein, carcinoembryonic antigen, carbohydrate antigen 125 and carbohydrate antigen 199, and the differences were statistically significant (alpha fetoprotein: Z=4.990, P<0.001; carcinoembryonic antigen: Z=3.743, P<0.001; carbohydrate antigen 125: Z=4.951, P<0.001; carbohydrate antigen 199: Z=3.983, P<0.001). The combined detection of two kinds of gene methylation was better than the combined detection of four kinds of serum markers in the diagnosis of colorectal cancer, and the difference was statistically significant ( Z=3.334, P<0.001). Conclusion:The combined detection of SEPT9 gene and SDC2 gene methylation in plasma is more suitable for non-invasive diagnosis of colorectal cancer than the combined detection of 4 serum tumor markers.

Objective:To explore the resilience of nurses against COVID-19 in Xixi Hospital of Hangzhou and to analyze its influencing factors.Methods:From February 4th to 5th 2020, this study investigated 193 nurses against COVID-19 at Xixi Hospital of Hangzhou selected by convenience sampling with the general questionnaire, Connor-Davidson Resilience Scale (CD-RISC) and the General Self-Efficacy Scale (GSES) . Pearson correlation and multiple linear regression were used to analyze the influencing factors of resilience.Results:Among 193 nurses, the total score of resilience was (62.60±13.70) ; the scores of three dimensions, optimism, self-reliance and tenacity were (9.59±2.37) , (21.62±4.68) and (31.40±7.74) respectively. The total score of GSES was (28.33±3.55) . Simple correlation showed that there were statistical differences in the total score of CD-RISC and dimension score of tenacity among nurses with different genders, experience of participating in emergent public health events, self-conscious readiness degree and confidence in fulfilling the task ( P<0.05) . Pearson correlation showed that resilience had a positive correlation with the general self-efficacy with a statistical difference ( r=0.474, P<0.01) . Multiple regression analysis demonstrated the influencing factors of nurses against COVID-19 were the self-conscious readiness degree, confidence in fulfilling the task and the general self-efficacy which totally explained 27.6% of the variance. Conclusions:Nurses against COVID-19 have the low level of the resilience which needs to be improved. We should pay more attention to the high-risk population, and provide a positively and effectively psychological intervention to improve the resilience of nurses against COVID-19.

Objective ToexploretherelationshipbetweenSchizasgradeofthenerverootwithintheduralsacandtheduralsac cross-sectionalarea(DSCA)ofthelumbarspineaswellastheclinicalsignificance.Methods 3.0T MRIexaminationofthelumbar spineof89patientswithlunbarspinestenosis(LSS)from May2016toSeptember2017intheaffiliatedhospitalofNantongUniversitywere collected.Twoexperienceddoctorsindependently measuredthekyphosisdegreeofthethoracolumbarspine,theDSCAofthe2-5 lumbarlevels,vDSCA,dDSCA,andevaluatedSchizasgradeofthenerverootforfourdegradsofA,B(gradeB1:DCSA≥100 mm2, gradeB2 :DCSA<100 mm2 ),CandDaccordingtozygopophysisconnectingline,andfinallyconductedthetestof Kappa consistency.DSCA wasdividedintothreegroupsof≤75 mm2,76-99 mm2and≥100 mm2,andχ2 wasadoptedtoexaminetherateineachSchizas grade.Schizasgradewithd/vvalue(dDSCA/vDSCA)andthekyphosisdegreeofthethoracolumbarspinewerecomparatedbyttest. Forthecorrelationcoefficient,S pear m an analysis wasadopted.Results In89cases with173lumbarlevels,schizasgradeofthenerve rootwere52,51,32and38levelsforgradeA-DrespectivelyI.nDSCA≤75mm2group,SchizasCandDwere18.5%and21.9%respectively, whichweresignificantlyhigherthanthoseforgradeAandB(0% and3.5%,P<0.01);InDSCA=76-99mm2group,Schizasgrade AandBwere8.7% and17.9%,whichweresignificantlyhigherthanthoseofgradeCandD (0% and0%,P<0.05and0.01);In DSCA≥100mm2group,therewere0% and0%forSchizasgradeCandD,whichweresignificantlylowerthanthoseforgradeAand B(21.4% and8.1%,P<0.0SchizasgradesofA-Dgroups,d/vaveragevalueswere0.64±0.29,0.48±0.22,0.42±0.20and0.34±0.11 respectively,in whichgradeCand D weresignificantlylower thanthoseofgradeAandB(P<0.01).Thecorrelationcoefficientof SchizasgradewiththeDSCAandd/vvalueswere0.83and0.87 respectively(P<0.01).Thekyphosisdegreeofthethoracolumbar spinewas(158.7±15.9)°inSchizasgradeB1,and (167.8±11.2)°inothergrades(t=4.37,P<0.05).Conclusion Theclassification ofnerverootSchizasgradeishighlyrelatedtoDCSA,andbothofthemaretheindicatorsforjudgingwhetherthelumbarspinalis stenosisornormal.TheSchizasgradeismoreconvenientandquicker;InordertoavoidconflictwithDCSA,SchizasBshouldbedividedintoB1 andB2 Whenitisusedtodeterminewhetherhavestenosis.

Objective To investigate the effects of NANOG/deleted in breast cancer 1 (DBC1)pathway on biological behavior of gastric cancer cells.Methods From May 2014 to May 2015,25 patients who underwent gastric cancer resection were selected.The expression of NANOG and DBC1 was detected by real time reverse transcription polymerase chain reaction (RT-PCR) and Western blotting in N-tera,SGC-7901,HGC-27,MKN-45,MGC803,NCI-N87,BGC823 cell lines,normal gastric epithelial cell line GES-1 and gastric cancer tissues.The proliferation,apoptosis and colony formation ability of MKN-45 cells in short hairpin (sh)NANOG-1,shNANOG-2,sh-control and shDBC1 groups were determined by MTT assay,flow cytometry and colony forming assay.The effects on the expression of the two genes in MKN-45 cells were verified by polymerase chain reaction (PCR) in shNANOG,sh-control,shDBC1 and shDBC1+NANOG groups and the effects of down regulation of DBC1 on cell biological behavior were further investigated.The differences in gene expression profile after interference which were screened by gene chips and bioinformatics were analyzed.The mechanism of NANOG regulating DBC1 was explored by Dual-luciferase assay.T test was used for two groups comparison while one-way analysis of variance was for multiple groups.Results NANOG and NANOG mRNA were highly expressed in N-tera cells,which were 1.02±0.08 and 0.95 ±0.03,respectively,and the expressions in SGC-7901,HGC-27,MKN-45 and NCI-N87 cell lines were 0.67±0.03 and 0.64±0.04,0.58±0.02 and 0.28±0.02,0.83±0.03 and 1.04 ± 0.05,and 0.61 ± 0.02 and 0.64 ± 0.08,respectively;no expression was detected in normal gastric epithelial cell line GES-1,and the expressions in MKN-45 cells were the highest in gastric cancer cells (F=21.51 and 85.53,both P＜0.01).The expression of DBC1 in HGC-27,MGC803,NCIN87,SGC-7901,BGC823 and MKN-45 cells were 0.37±0.02,0.33±0.02,0.42±0.01,0.58±0.04,0.33±0.05,and 0.87±0.02,respectively;while there was no expression of NANOG,NANOG mRNA and DBC1 in GES-1 cells.The expression of NANOG mRNA and DBC1 was detected in gastric cancer tissues of 24.0％ (6/25) patients.Compared with that of the sh control group,the apoptosis rates of MKN-45 cells in the shNANOG-1,shNANOG-2 groups were increased ((2.24±0.17)％ vs (6.03±0.24) ％ and (6.95 ± 0.38) ％),and the difference was statistically significant (F =81.18,P ＜ 0.01).Compared with that of the sh-eontrol group,the colony forming abilities of MKN-45 cells in the shNANOG-1 and shNANOG-2 groups were significantly decreased (172.03±6.35 vs 74.32±5.32 and 53.08±3.82),and the difference was statistically significant(F=171.61,P＜0.01).The results of PCR showed that compared with that of sh-eontrol group,the expression levels of NANOG mRNA and DBC1 mRNA in shNANOG group were lower (1.04±0.05 vs 0.54±0.03,1.08±0.08 vs 0.42±0.03),the level of DBC1 mRNA in shDBC1 group was lower (1.08±0.08 vs 0.50±0.04),and the differences were statistically significant (t=9.15,7.37 and 6.06,all P＜0.01).The expression level of NANOG mRNA in shDBC1 + NANOG group was higher (1.04 ± 0.05 vs 3.01 ± 0.08),while the expression level of DBC1 mRNA was lower (1.08 ± 0.08 vs 0.71 ± 0.06),and the difference was statistically significant (t=-20.22 and 3.74,both P＜0.05).The expression level of DBC1 mRNA in shDBC1±NANOG group was higher than that in shDBC1 group (0.71±0.06 vs 0.50±0.04),and the difference was statistically significant (t=4.00,P＜0.05).Bioinformatic analysis showed that DBC1 gene promoter region had the potential NANOG protein binding sites.Dual-luciferase assay indicated NANOG played the role in transcription activation in DBC1 promoter regions.Conclusion NANOG and DBC1 are highly expressed in various gastric cancer cell lines.NANOG may affect the proliferation,apoptosis and colony formation of MKN-45 cells by regulating the expression of DBC1.NANOG/DBC1 pathway may be a promising new target of gastric cancer treatment.

Objective To detect the expression of interlenkin-22 (IL-22) and relative CD4+ T cell subsets in patients with ulcerative colitis (UC),and to explore their roles in the pathogenesis of UC.Methods Thirty-five adult UC patients were enrolled in this study and 35 healthy subjects were taken as control.Plasma IL-22 level was quantified by ELISA.The percentages of Th1,Th17 and Th22 cells in peripheral blood were determined by flow cytometry.The relationships of these results and disease activity were analyzed.Results Plasma IL-22 levels in UC patients [ (354.12±104.22 ) pg/ml ]were significantly higher than that of healthy controls ( P＜0.05 ),and the levels increased significantly in severely active patients.The percentages of Th17 cells in UC patients [ (2.36±0.94) ％ ]were elevated compared to healthy controls ( P＜0.05 ),and the percentages increased significantly in moderately active and severely active patients.The percentages of Th22 cells in UC patients [ (2.27±0.87 ) ％ ]were elevated compared to healthy controls (P±0.05),and the percentages increased significantly in severely active patients.The percentage of Th1 cells was not significantly different between UC patients and normal controls.ConclusionOur resuits demonstrate elevated IL-22 correlated to Th17 and Th22 cells may play an important role in the immunopathogenesis of UC.

Objective To assess the prognosis and effect on renal function of pediatric urolithiasis caused by melamine-contaminated milk powder (PUMMP) in a long-term follow-up.Methods One hundred and two of 8335 children (≤ 6-year-old) with history of consuming melamine-contaminated milk powder screened in our hospital were followed up for eighteen months after diagnosis. Urinary system ultrasonography, urinalysis, urinary microprotein profiles [microalbumin (ALBU), immunoglobulin G (IgG), and N-acetyl-β-D-glucosidase (NAG)], urinary melamine and cyanuric acid were examined in the first visit and at the end of follow-up. Results Follow-up was completed in 91 children and the stone was excreted in 82 children (90.1%).Stones less than 5 mm in diameter were most vulnerable to discharge, and stones larger than 10 mm could not be expelled without interventions. At the end of follow-up, no melamine or cyanuric acid was found in the urine samples of 74 patients. Urinalysis showed that incidences of proteinuria, microscopic hematuria and leukocyturia were 0%, 5.1% and 2.0%, which were significant different from those in the first visit (Pproteinutria=0.123, Phemnatuna=0.038 and Pleukocyhuris=0.005).Urinary microprotein profiles revealed that some children whose urinalysis was normal still presented glomerular and renal tubular injury and the abnormal rates were 8.8% and 12.1%respectively. The glomerular injury was mainly related to persistent stone, male and younger.Conclusions 90.1% of children with PUMMP passes urinary stones at the end of follow-up.Stone size is the major risk factor of discharge. No melamine or cyanuric acid is found in the urine of children. After eighteen months, glomerular and renal tubular injury is still found in some patients. Further follow-up is necessary.