Objective To evaluate the factors associated with recurrence of chronic hepatitis B (CHB) after nucleoside analogs (NAs) withdrawal.Methods A literature search from PubMed,Wanfang data,CQVIP,CNKI,Duxiu and SinoMed was conducted to identify studies on the recurrence of CHB after NAs withdrawal.Meta-analysis was performed using RevMan 5.3.Random-effects or fixed-effects model was performed based on the heterogeneity.Weighted mean difference (WMD) was used to assess the continuous data,and odds ratio (OR) was used to assess the dichotomous data.Publication bias was evaluated with Egger' s regression test using Stata SE 11.0.Results A total of 20 case-control studies were included in this analysis.The recurrence rates were 21.0％,30.4％,33.2％ in HBeAg-positive CHB patients,and 26.5％,34.1％,50.1％ in HBeAg-negative patients after NAs withdrawal for 3 months,6 months and 1 year,respectively.For patients treated with lamivudine,the recurrence rates of CHB were 21.0％,28.0％,34.3％ at 3-,6-and 12-month after NAs withdrawal.Meta analysis demonstrated that among HBeAg-positive CHB patients,the average age (WMD =7.36,95％ CI:5.72-9.00,Z =8.81,P ＜0.01) and baseline HBV DNA level (WMD =0.26,95％ CI:0.05-0.46,Z =2.44,P =0.01) were higher in recurrence group,while antiviral treatment duration was shorter in recurrence group (WMD =-3.12,95％ CI:-4.56--1.68,Z =4.26,P ＜ 0.01),and the rate of recurrence was higher in patients with liver cirrhosis (OR =2.59,95％ CI:1.33-5.04,Z =2.79,P ＜ 0.01).Among HBeAg negative patients,the average age of patients in recurrence group was higher than that in non-recurrence group (WMD =5.90,95％ CI:1.57-10.23,Z =2.67,P ＜ 0.01),and no difference was observed in other factors between recurrence and non-recurrence patients.Among patients treated with lamivudine,the average age (WMD =7.68,95％ CI:5.02-10.34,Z =5.66,P ＜0.01) and baseline HBV DNA level (WMD =0.26,95％ CI:0.05-0.46,Z =2.44,P =0.01) were higher,while antiviral treatment duration was shorter in recurrence group (WMD=-2.11,95％CI:-3.85--0.38,Z=2.39,P＜0.01),and the rate of recurrence was higher in patients with liver cirrhosis (OR =2.59,95％ CI:1.33-5.04,Z =2.79,P ＜ 0.05).Conclusion Among HBeAg-positive and lamivudine-treated patients,age,baseline HBV DNA level,antiviral treatment duration and liver cirrhosis are associated with the recurrence of CHB after NAs withdrawal; while for HBeAg-negative patients,age is the only risk factor.

Objective To screen promoter binding protein of cyclin B2 by using human liver cDNA library, and investigate the expression and regulation mechanism of cyclin B2 gene. Methods By using cyclin B2 biotinylated promoter DNA as the selective molecule, the T7select human liver cDNA library was biopanned and positive clones were selected. After screening, positive plaques were performed to amplify for inserted DNA fragment, and the DNA fragment was cloned into pGEM-Teasy vector. Results Sequence analysis was performed in 20 positive plaques, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 6 coding sequences were obtained, all of which were known ones. Conclusion Cyclin B2 promoter binding proteins were screened. The results will be useful for further study the expression and regulation mechanism of cyclin B2.

Objective To clone and identify human genes transactivated by homo sapiens HCV FTP2 by constructing a cDNA subtractive library with suppression subtractive hybridization tech- nique.Methods Suppression subtractive hybridization(SSH)and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV FTP2.The mRNA was iso- lated from HepG2 cells transfected pcDNA3.1(-)-HCV FTP2 and pcDNA3.1(-)empty vector re- spectively,and SSH method was employed to analyze the differentially expressed DNA sequence be- tween the two groups.After digestion with restriction enzyme Rsa I,small-size cDNAs were ob- tained.Then tester cDNA was divided into two groups and ligated to the specific adaptor I and adap- tor 2 respectively.After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR and then was subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain DH5?.Futhermore,the cDNA was se- quenced and analyzed in GenBank with Blast search after PCR.Results The subtractive library of genes transactivated by HCV FTP2 was constructed successfully.The amplified library contains 71 positive clones.Colony PCR shows that 56 clones contain 200～1000 hp inserts.Sequence analysis was performed in 24 clones randomly,and the full length sequences were obtained with bioinformatics method.Altogether 20 coding sequences in total were obtained,consisting of 19 known and 1 un- known.Conclusion The obtained sequences may be target genes transactivated by HCV FTP2,and some genes coding proteins involved in cell cycle regulation,metabolism and cell apoptosis.

Objective To investigate the clinical characteristics and risk factors of bacterial liver abscess (BLA) complicated with septicemia.Methods Fifty two BLA patients complicated with septicemia admitted in our hospital from January 2011 to December 2015 were retrospectively reviewed;and 52 cases of BLA without septicemia admitted at the same period were randomly selected as control group.The clinical manifestations, laboratory and radiographic findings, clinical outcome of these patients were analyzed.Logistic regression analysis was used to study the clinical features and risk factors of BLA complicated with septicemia.Results Compared to the control group, the BLA with septicemia group had higher prevalence rates in diabetes mellitus, malignant tumors, jaundice, albumin <35 g/L, BUN≥8.2 mmol/L, hyperglycemia, multiple abscesses and abscesses size ≥10 cm(P<0.05 or <0.01).The blood culture showed that K.pneumoniae(63.3%) was the most commonly isolated pathogen, followed by E.coli(16.7%).Univariate analysis revealed that diabetes mellitus(OR=2.200,95%CI 1.042-4.646), malignant tumors (OR=3.667,95%CI 1.023-13.143), albumin <35 g/ L(OR=2.800,95%CI 1.009-7.774), BUN≥8.2 mmol/L(OR=3.167,95%CI 1.265-7.929), hyperglycemia(OR=3.400,95%CI 1.254-9.216), multiple abscesses(OR=2.667,95%CI 1.043-6.815), abscesses size≥10 cm (OR=5.000,95%CI 1.096-22.820) were positively associated with bacterial liver abscess complicated with septicemia.Multivariate Logistic regression showed that abscesses size≥10 cm (OR=14.016,95%CI 1.354-145.070) was an independent risk factor for complication of with septicemia.Conclusion septicemia is a common complication for bacterial liver abscess, clinically effective measures shauld be taken to prevent and control risk factors associated with septicemia.

Objective To investigate the inhibiton effect of 4‐styrenesulfonic acid‐co‐maleic acid(PSM ) in HIV‐1 .Methods The inhibition effect of different doses of PSM on HIV‐1 in susceptible cells GHOST (3) X4/Hi5 was observed by Luciferase ,and so did the inhibitory effect of PSM on JR‐FL、HXB2、CNE6 ,CNE30 ,CNE50 ,CNE55 .The cellular toxicity of PSM on the VK2/E6E7 was also evaluated by CCK8 kit .The transcript level of tight junction proteins (ZO‐1 ,E‐cadherin and Occludin) of HEC‐1‐A were analyzed by qRT‐PCR .And then observed the effect of PSM on expression of genitourinary epithelial cells HEC‐1‐A ,so we could e‐valuated the effect of integrity of local mucosal indirectly .Results The results showed that PSM exhibited potent antiviral activity against a broad spectrum of HIV‐1 major isolates with different genotypes and biotypes (EC50 value of JR‐FL ,HXB2 ,CNE6 , CNE30 ,CNE50 ,CNE55 were 5 .78 ,0 .77 ,1 .85 ,3 .15 ,1 .70 ,2 .27 μg/mL respectively) .Meanwhile ,it had less cytotoxicity on VK2/E6E7 .qRT‐PCR showed that no obvious restrain effect on expression of ZO‐1 was observed and PSM increased the level of tran‐scription of E‐cadherin and Occludin .Conclusion PSM may be a potential agent for the prevention of HIV‐1 infection .

Objective To investigate the effect of functional training on knee pain,functional movement screen (FMS) score and balance in Chinese elite fencing athletes with patellar tendinopathy.Methods Twenty-four fencing athletes with a diagnosed patellar tendinopathy were randomized into a treatment group (TG) and a control group (CG),each of 12.Both groups were given routine physical therapy,while TG received motor function training in addition for eight weeks.Both groups completed the numerical rating scale (NRS),FMS and balance test before and after the intervention.Results After the intervention,the average PRS and FMS of TG were 2.08± 1.24 and 16.25±0.97 respectively,which significantly outperformed those of TG before the intervention and those of CG after the intervention (P＜0.05).Moreover,TG indicated superior results in parameters of static postural balance including center of pressure,total length of swinging pathway,maximal length of swinging pathway,and area of swinging pathway when compared to TG before the intervention and CG after the intervention(P＜0.05).Conclusion The motor functiontraining is effective in improving functional movement and balance in elite fencing athletes with patellar tendinopathy.

Objective The aim of this study is to dynamically investigate peripheral blood lymphocyte subsets in fever with thrombocytopenia syndrome (SFTS) patients at different stages,to evaluate the influence of these changes in the infection process.Methods Case-control study was used in the research.Twelveconfirmedthrombocytopeniasyndromevirus ( SFTSV ) infectedpatientswere enrolled.According to SFTS prevention guide issued by Chinese Ministry of Health,these patients were divided into two groups,recovery group and death group.For each group,dynamic profiles of the CD3 + T cells,CD4 + helper T cells,CD8 + cytotoxic T cell and CD3 - CD16 + CD56 + natural killer cells were tested by flow cytometry.Meanwhile, the relationshipsbetween these dynamicchanges and liver function,leukocytes,and platelets were analyzed respectively.Two independent-samples t test was used to compare the difference of the peripheral blood lymphocyte subsets count between the SFTS patients and healthy control.Small sample was analyzed by Mann-Whitney U test.Results In the early stage of infection,Th cells in peripheral blood of recovery group were significantly reduced and Th/Tc ratio was reversed.On day 5,7,9 of post infection,Th cell counts in peripheral blood were (740.9 ± 6.4),(836.2 ± 272.3 ) and ( 1083.6 ± 319.7 ) cells/μl respectively,which were significantly lower than health control ( 1351.4 ± 295.1 ) cells/μl ( t value was -2.883,-4.235,-2.145 respectively,all P ＜0.05).Tc cell counts were significantly more than healthy controls (690.1 ± 194.8) cells/μl through the course,which were ( 1006.3 ±356.5),(1166.4±242.4),(1102.4±245.9),(991.3±205.1) and (886.5±154.5) cells/μl on day 7,9,11,13,15 of the course (t value was 3.312,5.661,4.574,3.874,2.382,all P＜0.05).NK cells were decreased significantly from the ninth day of the course.Associated with abnormal changes of cell subsets,WBC and PLT decreased significantly,and serum ALT,AST,LDH and CK etc.were higher than normal level.With the disease recovery,the abnormality above was gradually improved.In contrast,death cases showed significant decrease in T and Th cells compared with health control (P ＜ 0.05).On day 7,8,9 of the course,the counts of total T cell were (735.9 ± 359.9),(724.9 ± 125.9),(845.3 ± 389.3) cells/μl and the counts of Th cell were ( 533.2 ± 246.9 ),( 532.1 ± 105.7 ),( 551.7 ± 86.9 ) cells/μl,significantly lower than healthy control ( 1727.9 ± 230.2 ) cells/μl and ( 1351.4 ± 295.1 ) cells/μl,with statistically differences (z value was - 2.828, - 2.342,- 2.342 and - 2.828, - 2.342, - 2.342,all P ＜ 0.05 ).On day 7,8,9 of the course,the numbers of NK cell in death group were ( 1141.8 ± 415.5),( 1047.2 ±68.4),( 1276.3 ±545.3) cells/μl,which were significantly more than health group (470.7 ± 242.2) cells/μl,with statistically differences (z value was - 2.180,- 2.335,- 2.258,all P ＜0.05).Conclusions SFTSV infection can induce cell immunity damage.The changes of lymphocyte subsets are associated with clinical classification and prognosis.Significant reduction of T cell and CD4 +cell in peripheral blood are accompanied with significant increase of NK cell,which may be a pivotal indicator of poor prognosis and play an important role in making appropriate strategy in clinical treatment.( Chin J Lab Med,2012,35:826-831 )

Objective:To study the mechanistic role of myeloid-specific Notch1 knockout inhibiting STING signaling to regulate hepatocyte lipophagy.Methods:A mouse model of nonalcoholic steatohepatitis (NASH) was established using a high-fat diet (HFD) and mouse bone marrow-derived macrophages (BMMs). Primary hepatocytes were isolated to construct a co-culture system. Twelve Notch1 FL/FL mice were randomly divided into two groups: the Notch1 FL/FL + normal diet (NCD) and the Notch1 FL/FL + HFD group. Further, 12 Notch1 M-KO mice were randomly divided into two groups: Notch1 M-KO + NCD, and Notch1 M-KO + HFD group.Serum alanine aminotransferase (sALT), total cholesterol (TC) and triglyceride (TG) were collected from mice serum samples. Liver tissue samples were collected for H&E staining, immunofluorescence (IF), Western blot and qRT-PCR. Tumor necrosis factor (TNF)-α was detected in the supernatant by enzyme-linked immunosorbent assay (ELISA). The comparison of inter group data was conducted using a t-test. Results:The mouse NASH model, mouse BMMs co-culture system, and primary hepatocytes were successfully constructed. Compared with the Notch1 FL/FL + HFD group, the Notch1 M-KO + HFD group showed a significant increase in serum ALT [(250.02 ± 58.21) U/L vs (370.70 ± 54.57) U/L, t = 3.705, P = 0.004], TG [(29.90 ± 3.54) mg/g vs (43.83 ± 8.56) mg/g, t = 3.685, P = 0.004], and TC [(33.70 ± 8.43) mg/g vs (90.53 ± 12.53) mg/g, t = 9.917, P < 0.001]. HE staining of liver tissue showed remarkable balloon-like alterations in liver cells, while IF staining demonstrated increased macrophage infiltration ( t = 7.346, P < 0.001). Compared with the hepatocyte group co-cultured with Notch1 FL/FL BMMs, the BODIPY probe showed a significant increase in lipid droplet (LDs) deposition in liver cells in the Notch1 M-KO group ( t = 3.835, P < 0.001). The co-localization of lysosomal associated membrane protein 1 (LAMP1), LDs ( t = 7.103, P < 0.001), microtubule-associated protein light chain 3 (LC3) -II/LC3-I ( t = 5.0, P = 0.007), and autophagy associated gene 12 (Atg12) ( t = 28.36, P < 0.001) had decreased expression, while P-62 had increased expression ( t = 3.253, P = 0.03), indicating a decrease in autophagic flow. Additionally, LC3 and LDs colocalization decreased ( t = 5.24, P = 0.0003), indicating reduced lipophagy. Compared with the Notch1 FL/FL group, the Notch1 M-KO BMMS mouse group showed an increase in the expression of p-STING ( t = 5.318, P = 0.006), p-TANK1 binding kinase 1 (TKB1) ( t = 6.467, P = 0.002), p-interferon regulatory factor 3 (IRF3) ( t = 14.61, P < 0.001), and p-P65 ( t = 12.7, P = 0.002) protein, accompanied by mRNA expression of the inflammatory mediators interferon (IFN)-β ( t = 7.978, P < 0.001), TNFα ( t = 8.496, P = 0.001), interleukin-1 β (IL-1 β) ( t = 4.7, P < 0.001), and CXCL-10 ( t = 4.428, P = 0.001). The STING gene was knocked out in the BMMs Notch1 M-KO mice using CRISPR/Cas9. Compared with the CRISPR-Control group, the expression of P-TKB1 ( t = 2.909, P = 0.044), p-IRF3 ( t = 10.96, P < 0.001), p-IRF3 ( t = 10.96, P < 0.001), and p-P65 ( t = 7.091, P = 0.002) proteins was lower in the STING-KO BMMs group. The release of TNF-α in the supernatant was decreased (732.3 ± 129.35 pg/ml vs. 398.17 ± 47.15 pg/ml, t = 4.204, P = 0.014). However, in hepatocytes co-cultured with STING-KO BMMs, LC3-II/LC3-I ( t = 7.546, P = 0.001) increased, p-62 ( t = 10.96, P < 0.001) expression decreased, autophagic flow increased, and the colocalization of LC3 and LDs increased, lipophagy increased, and LDs deposition decreased. Conclusion:Myeloid-specific Notch1 knockout can activate macrophages STING signaling, increase the expression of inflammatory mediator genes, inhibit the occurrence of autophagy flow and lipophagy in hepatocyte cells, and aggravate LDs deposition and NASH progression.

Objective To investigate the distribution and antimicrobial resistance profiles of the common pathogens isolated from urine from 2015 to 2021 in the CHINET Antimicrobial Resistance Surveillance Program.Methods The bacterial strains were isolated from urine and identified routinely in 51 hospitals across China in the CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021.Antimicrobial susceptibility was determined by Kirby-Bauer method,automatic microbiological analysis system and E-test according to the unified protocol.Results A total of 261 893 nonduplicate strains were isolated from urine specimen from 2015 to 2021,of which gram-positive bacteria accounted for 23.8％(62 219/261 893),and gram-negative bacteria 76.2％(199 674/261 893).The most common species were E.coli(46.7％),E.faecium(10.4％),K.pneumoniae(9.8％),E.faecalis(8.7％),P.mirabilis(3.5％),P.aeruginosa(3.4％),SS.agalactiae(2.6％),and E.cloacae(2.1％).The strains were more frequently isolated from inpatients versus outpatients and emergency patients,from females versus males,and from adults versus children.The prevalence of ESBLs-producing strains in E.coli,K.pneumoniae and P.mirabilis was 53.2％,52.8％and 37.0％,respectively.The prevalence of carbapenem-resistant strains in E.coli,K.pneumoniae,P.aeruginosa and A.baumannii was 1.7％,18.5％,16.4％,and 40.3％,respectively.Lower than 10％of the E.faecalis isolates were resistant to ampicillin,nitrofurantoin,linezolid,vancomycin,teicoplanin and fosfomycin.More than 90％of the E.faecium isolates were ressitant to ampicillin,levofloxacin and erythromycin.The percentage of strains resistant to vancomycin,linezolid or teicoplanin was＜2％.The E.coli,K.pneumoniae,P.aeruginosa and A.baumannii strains isolated from ICU inpatients showed significantly higher resistance rates than the corresponding strains isolated from outpatients and non-ICU inpatients.Conclusions E.coli,Enterococcus and K.pneumoniae are the most common pathogens in urinary tract infection.The bacterial species and antimicrobial resistance of urinary isolates vary with different populations.More attention should be paid to antimicrobial resistance surveillance and reduce the irrational use of antimicrobial agents.

Objective To understand the distribution and changing resistance profiles of clinical isolates of Enterococcus in hospitals across China from 2015 to 2021.Methods Antimicrobial susceptibility testing was conducted for the clinical isolates of Enterococcus according to the unified protocol of CHINET program by automated systems,Kirby-Bauer method,or E-test strip.The results were interpreted according to the Clinical & Laboratory Standards Institute(CLSI)breakpoints in 2021.WHONET 5.6 software was used for statistical analysis.Results A total of 124 565 strains of Enterococcus were isolated during the 7-year period,mainly including Enterococcus faecalis(50.7％)and Enterococcus faecalis(41.5％).The strains were mainly isolated from urinary tract specimens(46.9％±2.6％),and primarily from the patients in the department of internal medicine,surgery and ICU.E.faecium and E.faecalis strains showed low level resistance rate to vancomycin,teicoplanin and linezolid(≤3.6％).The prevalence of vancomycin-resistant E.faecalis and E.faecium was 0.1％and 1.3％,respectively.The prevalence of linezolid-resistant E.faecalis increased from 0.7％in 2015 to 3.4％in 2021,while the prevalence of linezolid-resistant E.faecium was 0.3％.Conclusions The clinical isolates of Enterococcus were still highly susceptible to vancomycin,teicoplanin,and linezolid,evidenced by a low resistance rate.However,the prevalence of linezolid-resistant E.faecalis was increasing during the 7-year period.It is necessary to strengthen antimicrobial resistance surveillance to effectively identify the emergence of antibiotic-resistant bacteria and curb the spread of resistant pathogens.