Objective To investigate the role and implication of Engrailed-2 (EN2) in human hepatocellular carcinoma (HCC) and the effect of silencing EN2 genes on the proliferation and invasion of HepG2 cells by RNA interference.Methods Immunohistochemistry was used to detect the expression of EN2 in the HCC tissues and corresponding non-cancerous adjacent liver tissues in 126 patients with HCC.The expressions of EN2 and the relationship between EN2 expression and clinicopathological features and prognosis were analyzed using the Chi-square test.The survival curve was drawn using the Kaplan-Meier method and the survival was analyzed using the log-rank test.EN2-small interfering RNA (siRNA) was transfected into HepG2 cell lines mediated by LipofectamineTM 2000,and the expression of EN2 were detected by Western blotting assay.The cell proliferation and invasion were measured by methyl thiazol tetrazolium (MTT) and Transwell assays.Results EN2 positive expression rate was high (62.7％) in HCC tissue but low in the corresponding peritumoral tissue (23.5％,P ＜ 0.01).The high expression of EN2 was strongly correlated with tumor size,metastasis and AJCC TNM stage.The cumulative 5-year survival rate was 33.9％ in the low EN2 expression group,whereas it was 5.1％ in the high EN2 expression group (P ＜ 0.01).Expression of EN2 in EN2 siRNA group was significantly lower than that in control siRNA group and control group (P ＜ 0.05),respectively.The cell proliferation and invasion ability were significantly reduced in the EN2 siRNA group.Conclusion The expression of EN2 is highly up-regulated in HCC tissues and down-regulation of EN2 could inhibit proliferation and invasion of HepG2 cells,which indicating that EN2 is involved in the process of HCC carcinogenesis and progression and may serve as a biomarker for predicting prognosis of HCC patients.

Encephalomyocarditis virus (EMCV) ,named JZ1202 ,was isolated from domesticated boar in Henan ,China . We performed the full-length genome sequencing and molecular characteristic analysis of the isolated strain .Results showed that the full-genome sequence of EMCV JZ1202 generated a sequence of 7 735 bp in length ,and had 81 .2%-99 .9% nucleotide identity with other reference strains from different animals ,but 99 .4% with Chinese reference from pig .The phylogenetic tree was constructed based on the full-length genome;ORF and VP1 gene sequences identified EMCV was divided into G1 ,G2 and G3 groups ;the strain JZ1202 belongs to G1 with other Chinese reference strains .Results identified that the EMCV infection could cause severe clinical symptoms in domesticated boar .Big regional differences exist in EMCV and the transmission is limit-ed in a range of area .However ,cross infection and prevalence of EMCV disease between domesticated boar and mice might ex-ist .Mutation of some amino acid may occurred in EMCV infected domesticated boar .

Objective To study plasmid-mediated transfer,plasmid replicon typing,and genetic environment of blaNDM-1 gene in Enterobacteraerogenes(E.aerogenes).Methods E.aerogenes HN-NDM0711 was used as the subject of this research,the transferable properties of plasmid were analyzed by conjugation testing,conjugant was performed stability testing,plasmid type was determined by PCR-based replicon typing (PBRT),downstream and upstream of blaNDM-1 were sequenced using chromosome walking method,genetic context was analyzed by BLASTN and BALSTP,as well as annotated using Vector NTI 11.5.1 software,sequence pipeline graph was made,the sequence was submitted to Genbank through software Banklt.Results The conjugation testing of E.aerogenes pHN-NDM0711 was positive,after positive conjugant was conducted 4-day passage,minimal inhibitory concentrations (MICs) of imipenem and meropenem to all the cloned strains didn't change,blaNDM-1 were all positive.The replicon type was IncA/C;blaNDM-1 gene was localized between ISAba14 and IS91,at upstream of the blaNDM-1,class 1 integron and Tn3 transposon were identified,class 1 integron contained a new mosaic structure of a drug-resistant resistance gene cassette.Conclusion E.aerogenes pHN-NDM071 1,bearing blaNDM-1 gene in IncA/C plasmid,derived from gene recombination under different antimicrobial selection pressure.Antimicrobial use in clinical,industrial and agricultural area should be strictly controlled,so as to reduce the emergence of such bacteria.

Obej ctive Knockout mice are widely used in the studies of joint diseases .This article investigated the effects of joint processing methods , collagenase types ,and collagenase digestion time on the number of primary fibroblast -like synoviocytes (FLSs) obtained from mice. Methods The hind legs of 6 of the 12 male mice were cut open from the hip joints , but not those of the other 6.FLSs were isolated using the type-Ⅳcollagenase digestion method and purified by differential digestion .Cell morphology was observed under the inverted microscope .The type, viability, and purity of the cells were determined by flow cytometry . Rse ults Significantly fewer FLSs were obtained from the mice with the hind legs cut open ( 19 133 ±115 ) than from those without (24 933 ± 503) (P<0.05).The numbers of FLSs collected from the cell suspension at 1, 2, 3, 4, 5,6 , and 7 hours after digestion were 700 ±300 , 600 ±100 , 15 200 ±900 , 5100 ±800 ,2700 ±300 , 900 ±200, and 300 ±100, respectively, the highest at 3 hours. There were statistically significant differences in the total number of FLSs obtained by type-Ⅳ and type-Ⅱ collagenase digestions (24900 ±500v s 18 100 ±400, P<0.05). Conclusion For in virt o culture of primary mouse FLSs, it is recommended that the hip joints be not cut open, and type-Ⅳcollagenase be used with cell sus-pension at 2-6hours after digestion .

Objective:To analyze the characteristics of plasmids in KPC-2-producing Serratia marcescens ( S. marcescens) isolates. Methods:Four carbapenem-resistant S. marcescens strains were isolated from four patients admitted to the hepatobiliary ward of Affiliated Cancer Hospital of Zhengzhou University in 2016. BD Phenix-100 was used to identify the strains and detect the minimum inhibitory concentrations (MICs). Homology analysis was performed using pulsed-field gel electrophoresis (PFGE). The modified Hodge test was used to detect the phenotypes of carbapenemase. PCR and gene sequencing were used to detect the types of carbapenem resistance genes. The transferability of plasmids was detected by conjugation test. The characteristics of plasmids were analyzed by genomic alignment method after whole genome sequencing. DNAMAN V9 software was used to compare the amino acid sequences of the replication initiation proteins. A phylogenetic tree was constructed with neighbor-joining method using MEGA7.0. Results:All of the four S. marcescens strains were resistant to carbapenem antibiotics. They were highly homologous according to PFGE. Hodge test results were all positive and the carbapenemase genotype was blaKPC-2. Conjugation test results were positive. The plasmid was a circular DNA of 42 742 bp in length. It had the similar skeleton of incX6 plasmid and the similar amino acid sequence of replication initiation protein. Moreover, it and incX6 plasmid were at the same node in the phylogenetic tree. The blaKPC-2 was located in the core of drug resistance, which was composed of insertion elements including Tn3 family transposons, recombinant enzyme genes, △ISKpn6 and ISKpn27. Conclusions:The plasmid was incX6-like. The blaKPC-2 gene was located in the transposon of △Tn6296. More attention should be paid to the bacteria carrying KPC-2 in incX plasmids.

Objective:To systematically evaluate the efficacy of different kinds of smoking cessation drugs by network Meta-analysis.Methods:Literature was retrieved from PubMed, Web of Science, Embase, Cochrane Library, CBM, CNKI, VIP, Wan fang database, from the establishment of the database to November 2022, and randomized controlled trials (RCT) about bupropion, varenicline, nicotine replacement therapy (NRT) versus placebo in the treatment of smoking patients were collected. After data extraction from included literature which met inclusion criteria, and quality evaluation with Cochrane 5.1 risk bias evaluation tool, network Meta-analysis was performed by Stata15.1 software.Results:A total of 19 RCTs, involving 6106 patients and three interventions measures (bupropion, varenicline, NRT) and one control measure (placebo) were included. The results of network Meta-analysis showed that in terms of short-term abstinence rate, varenicline [ OR=4.21, 95% CI (2.32, 7.63)], bupropion [ OR=2.81, 95% CI(1.05, 7.54)] were better than placebo ( P<0.05). The surface under the cumulative ranking area (SUCRA): varenicline (90.2%)>bupropion (64.8%)>NRT (41.7%)>placebo (3.2%). In terms of the long-term abstinence rate, varenicline [ OR=3.06, 95% CI (1.59, 5.90)], NRT [ OR=3.39, 95% CI (2.20, 5.21)] were better than placebo ( P<0.05). SUCRA: varenicline (83.8%)>NRT (73.9%)>bupropion (37.2%)>placebo (5.2%). Conclusion:The existing evidence shows that compared with bupropion, NRT, varenicline has the best effect on quitting smoking, but more high-quality randomized trial evidence is needed for verification.

Objective    To analyze whether hypernatremia within 48 hours after cardiac surgery will increase the incidence of delirium which developed 48 hours later after surgery (late-onset delirium). Methods    We conducted a retrospective analysis of 3 365 patients, including 1 918 males and 1 447 females, aged 18-94 ( 60.53±11.50) years, who were admitted to the Department of Cardiothoracic and Vascular Surgery of Nanjing First Hospital and underwent cardiac surgery from May 2016 to May 2019. Results    A total of 155 patients developed late-onset delirium, accounting for 4.61%. The incidence of late-onset delirium in patients with hypernatremia was 9.77%, the incidence of late onset delirium in patients without hypernatremia was 3.45%, and the difference was statistically different (P<0.001). The odds ratio (OR) of hypernatremia was 3.028 (95% confidence interval: 2.155-4.224, P<0.001). The OR adjusted for other risk factors including elderly patients, previous history of cerebrovascular disease, operation time, cardiopulmonary bypass time, lactate, hemoglobin≥100 g/L, prolonged mechanical ventilation, left ventricular systolic function, use of epinephrine, use of norepinephrine was 1.524 (95% confidence interval: 1.031-2.231, P=0.032). Conclusion    Hypernatremia within 48 hours after cardiac surgery may increase the risk of delirium in later stages.