OBJECTIVE To investigate the distribution and the resistance rate of Shigella in Anhui Province to guide the choice of antibacterials. METHODS Ninety one strains of Shigella were cultured in Sep 2005.The groups were identified by biochemical and serologic tests.Susceptibility of 91 strains of Shigella in Anhui to various antibiotics was tested using standardized custom dilution MIC panels according to CLSI(2005) guidelines. RESULTS There were 57 strains of Shigella flexneri,31 strains of S.sonnei and 2 strains ofS.boydii among 91 strains of Shigella.The resistance rates of Shigella to cefoperazone/sulbactam and piperacillin/tazobactam were remarkably lower than to other third generation cephalosporins.The susceptible rates to carbopenems were 100%.The resistance rates to ciprofloxacin lactate and pazufloxacin mesilate were 27.47% and 32.97%,respectively. CONCLUSIONS There is a certain resistance rate of the Shigella to fluoroquinolones and the third generation cephalosporins.More attention should be paid to the surveillance and control of such resistance.

Objective To detect the integration of hepatitis B virus (HBV) DNA in HBVrelated human hepatocellular carcinomas (HCC). Methods Extracted DNA from the liver tissue samples and amplified by nested polymerase chain reaction (PCR) with specially designed U-base primers. According to the known genes and human Alu repeat sequences (Alu repeat) , primers were designed respectively. Integrated clones combined target HBV DNA and the adjacent cell gene sequences were established by PCR and products were sequenced by biotechnology companies.Accurate locations of HBV genes integrated in the human genomes were analyzed by national center for biotechnology information (NCBI) basic local alignment search tool (BLAST) and Map Viewer search. Results In 24 HBsAg positive HCC samples, 15 cases showed the integrations of HBV fragment. And the other 8 samples didn't show any evidence of integration. Among 14 samples with integration, forward insertions of HBV DNA into the host chromosomal DNA were found in 10 samples and reverse insertions were found in 8 samples while both forward and reverse insertions were found in 5 samples. Analysis from viral-cellular junctions suggested that the integrations were all happened with truncated viral DNA and could be in any locus of X gene. Conclusion HBV DNA integration is not distributed evenly throughout the host genome.

Objective To investigate the prevalence of plasmid-mediated quinolone resistance ( PMQR ) determinants [ qnr,aac ( 6' ) -Ib-cr and qepA ]and mutations in quinolone resistance-determining regions (QRDRs) of gyrA and parC and their association with fluoroquinolone susceptibility in clinical isolates of Serratia marcescens in Anhui.Methods The minimum inhibition concentration ( MIC ) of 104 strains of S.rnarcescens collected from various clinical specimens from 34 hospitals during 2005 to 2010 were determined by agar dilution method.The qnr,aac (6')-Ib,qepA,gyrA and parC genes were screened by polymerase chain reaction (PCR) in 31 strains resistant to ciprofloxacin,and positive results were subsequently confirmed by sequencing.The conjugation experiments were performed for qnr and aac(6')-Ib-cr positive strains.The MIC of S.marcescens isolates,recipient strains and conjugants were tested by agar dilution method for quinolones and other antimicrobial agents.Results Six strains of the 31 S.marcescens isolates harboured qnr and/or aac(6')-Ib-cr genes.Among those 6 strains,2 strains harboured a qnrB6 gene,1 harboured a qnrS2 gene,and 4 harboured aac( 6' ) -Ib-cr,whereas no qnrA-,qnrC- or qnrD-positive isolate was detected.None of the 31 isolates carried the qepA gene.Mutations in the QRDR of gyrA and parC genes were detected in 9 and 7 isolates,respectively.The conjugation experiments were successfully carried out in 5 isolates of 6 PMQR determinants-postive strains.The MIC of conjugants for quinolones were increased evidently compared to recipient strains.Conclusions Chromosome and plasmid-mediated resistance determinants play an important role in quinolone resistance in clinical isolates of S.marcescens.And more important is that the PMQR determinants can be horizontal transmitted.It is necessary to continuously survey and watch for the spread of PMQR in S.marcescens in public health control program.

Objective To investigate the profile and antibiotic resistance of bacteria in patients with ascites infection in intensive care unit (ICU) patients in order to provide a reference for rational clinical use of antibiotics. Methods A retrospective analysis was conducted. The bacteria isolated from ascetic fluid patients admitted from January 1st, 2004 to October 31st, 2015 to ICU of the First Affiliated Hospital of Anhui Medical University were identified, and their susceptibility to antibiotics was analyzed. Patients, who were admitted from January 1st, 2004 to December 31st, 2009 were assigned to group A, and patients admitted afterwards were assigned to group B. Results A total of 637 specimens of ascetic fluid were examined, with 185 positive culture (29.0%) during the 12 years, and 203 strains of bacteria were found. Among them 126 strains (62.1%) of gram-negative bacteria (G-), 54 (26.6%) of gram-positive bacteria (G+) and 23 (11.3%) strains of fungi were found. Compared the result of group B with that of group A, the proportion of G- bacteria was increased [71.2% (99/139) vs. 44.2% (27/64)], and that of G+ decreased [17.3% (24/139) vs. 46.9% (30/64)] in group B. The difference was statistically significant (χ2 = 20.34, P = 0.001). The main pathogenic bacteria were G-, and Enterobacteriaceae was the most common pathogenic bacteria in intra-abdominal infection of ICU patients. The isolation rate of Escherichia coli and Klebsiella pneumoniae(35.7%, 10.3%) ranked in the first and third in G- bacteria, respectively. The resistant rate of Escherichia coli against penicillin and third generation cephalosporin were > 95.0% and > 73.3%, and it showed a sensitive rate of 70% to β-lactam/inhibitor, amikacin and minocycline, and a higher sensitivity to carbapenems and tigecycline (11.1%, 0). Forty-eight strains of non-fermentation bacteria were found with a rate of 23.7%. The positive rates of Acinetobacter baumannii in groups A and B were 7.8% (5/64) and 23.7% (33/139), respectively, and they ranked first among non-fermentation bacteria. Twenty strains (62.5%) multidrug-resistant Acinetobacter baumannii were found. Acinetobacter baumannii showed a resistance rate of 84.6% to cefoperazone/sulbactam, 35.3% to minocycline, and 53.3% to tigecycline. Candida albicans was the most commonly isolated fungus in intra-abdominal infections (87.5%). No strains resistant to common antifungal drugs were isolated. Conclusions G- bacteria was the main pathogen in intra-abdominal infection in patients with ascites. Non-fermenters showed an increasing trend of producing infection, and the proportion of multidrug-resistant Acinetobacter baumannii infection increased year by year, and more attention should be taken by attending doctors.

ObjectiveTo analyze the clinical distribution and antimicrobial resistance profile of Serratia marcescens(S. marcescens), and to provide the scientific evidence supporting clinical diagnosis and treatment.MethodsThe antimicrobial susceptibility test was performed in 104 strains of S. marcescens by agar dilution method. The results were judged according to the criteria recommended by Clinical and Laboratory Standards Institute (CLSI) 2010.The data were analyzed by chi square test. Results The majority of S. marcescens were isolated from sputum specimens,accounting for 59.6％ (62/104). The bacteria were most frequently isolated from department of respiratory (33.7％,35/104),followed by intensive care unit (23.1％,24/104),department of gerontology (16.3％, 17/104). The results of antimicrobial susceptibility test showed that the resistance rates of S.marcescens against ampicillin,gentamicin and cephazolin were high,which were 90.4％,86.5％ and 79.8％,respectively; those against the 3rd generation of cephalosporins were 24.0％-43.3％. No imipenem and meropenem resistant strains were identified. Compared with cefoxitin-resistant strains,the resistance rates of non-cefoxitin resistant strains against piperacillin (82.9％ vs 28.6％),ceftazidime (63.4％ vs 9.5％),aztreonam (68.3％ vs 9.5％),amikacin (68.3％ vs 20.6％),ciprofloxacin (48.8％ vs 19.1％) and chloramphenicol (90.3％ vs 58.7％) were all lower (all P ＜ 0.05 ). Conclusions S. marcescens is one of the most common conditional pathogenic bacteria leading to nosocomial infections,which is resistant to many kinds of antimicrobial agents.The surveillance of antimicrobial resistance in S. marcescens should be strengthened for purpose of preventing the transmission of multidrug resistant strains.

ObjectiveTo investigate the variations and distributions of the plasmid-mediated quinolone resistance genes in clinical isolates of Shigella and their resistance to antimicrobial agents. Methodsqnr, aac(6')-Ib-cr and qepA genes were identified by polymerase chain reaction (PCR) in 137 clinical isolates of Shigella.DNA sequencing of gene-positive strains were analyzed and the conjugation experiment was performed. The minimal inhibitory concentrations (MIC) of Shigella isolates, recipient strains and transconjugants were tested by agar dilution method for quinolones and other antimicrobial agents. The genotype of transconjugants were determined by PCR and sequencing. ResultsFour (2.9％) strains of the 137 Shigella isolates were qnr gene positive, including 3 qnrS2 positive and 1 qnrB4 positive (GenBank accession numbers of the complete sequence were JF261185 and HQ917003, respectively).Furthermore,five (3.6％) aac ( 6')-Ib-cr gene-positive strains (GenBank accession number JF261186 ) and one (0.7％) qepA gene-positive strain were identified in all isolates. The conjugation experiments were successfully carried out in six out of ten PCR-positive isolates. The MIC of transconjugants against quinolones and other antimicrobial agents increased differently compared to recipient strains. Conclusions The plasmid-mediated quinolone resistance genes are lowly prevalent in clinical isolates of Shigella. However, these resistance genes have the characteristic of horizontal transfer, which indicates that more attention should be paid to this phenomenon.

Objective: To explore the impact of five-flavor Sophora flavescens enteric-coated capsules (FSEC) on the intestinal flora of rats with ulcerative colitis (UC), so as to provide the reference for the mechanism of FSEC in treating UC. Methods: Forty SPF-grade Wistar rats were randomly divided into the control group, the model group, the mesalazine group and the FSEC group. Except the control group (0.9% sodium chloride solution), the other 3 groups used 3% dextran sulfate sodium (DSS) for 7 days to establish UC model. After successful modeling, the control group and the model group were given 2 mL/kgbw of 0.9% sodium chloride solution by gavage for 2 weeks, while the mesalazine group and the FSEC group were given 2 mL/kgbw of mesalazine suspension (0.2 g/kg) and FSEC granule suspension (2.16 g/kg), respectively. Pathological changes of colon tissue were observed after hematoxylin-eosin (HE) staining. Rat fecal samples were collected, and 16S rDNA high-throughput sequencing and bioinformatics analysis were performed on intestinal flora. The α and β diversity of intestinal flora among the four groups were compared, and the dominant flora was screened using LEfSe analysis. Results: Compared with the control group, the model group showed a significant loss of colonic crypts and a large infiltration of inflammatory cells. Compared with the model group, the mesalazine group and the FSEC group exhibited a slight loss of colonic crypts, a small amount or an absence inflammatory cell infiltration, and improved tissue damage. The α-diversity analysis showed that compared with the control group, the Chao1 and Shannon indices in the model group increased, while the Simpson index decreased; compared with the model group, the Chao1 and Shannon indices in the mesalazine group and the FSEC group decreased, and the Simpson index increased（all P<0.05）. The β-diversity analysis showed that the sample distance between the FSEC group and the control group were more closer than that between the model group and the control group. LEfSe analysis results showed that the dominant bacteria in the model group were mainly from the Alistipes and Oscillospira. In the FSEC group, the dominant bacteria were from the Ruminococcus and Prevotella. Conclusion FSEC can improve the structures of intestinal flora, increase the abundance of beneficial bacteria such as Ruminococcus and Prevotella, reduce the abundance of pathogenic bacteria such as Alistipes, and alleviate the inflammatory response in UC rats.

Aim To explore the growth inhibition effects of jasmonates on human neuroblastoma SH-SY5Y cell line,and to investigate its mechanisms.Methods After administration of 0.5~2.5 mmol?L-1 jasmonates for 6~24 hrs, the growth inhibition rates of SH-SY5Y cells were studied by MTT colorimetry.Cell cycle phases were assayed by propidium iodide staining flow cytometry. Cellular apoptosis was inspected by Hoechst 33258 fluorescent staining and Annexin V-FITC and propidium iodide staining flow cytometry.Gene expressions of PCNA, cyclin D1 and N-myc were determined by reverse transcription polymerase chain reaction.Results Jasmonates inhibited the growth of SH-SY5Y cells in a dose-and time-dependent manner,while the methyl jasmonate was the most efficient. After administration of 0.5 to 2.5 mmol?L-1 of methyl jasmonate for 24 hrs,the growth inhibition rates of cells reached 5.75%~88.7%(P

Objective To explore the safety and feasibility of application of nalbuphine in uterine artery embolization. Methods We selected 70 cases of gynecologic fibroids receiving interventional surgery from January 2015 to October 2016 in our hospital.According to random number table,they were divided into either experimental group or control group,with 35 cases in each group.After selective uterine artery embolization,conventional intravenous analgesia pump was used to relieve pain.The experimental group was given nalbuphine 1 mg/kg and flurbiprofen 100 mg,and the control group was given sufentanil 2 μg/kg and flurbiprofen 100 mg.The visual analogue scale(VAS), nausea and vomiting, respiratory depression and other complications were observed in both groups after operation at 2,6,12,24 and 48 h. Results There were no significant differences between two groups at each time point after surgery in VAS scores[2 h:(4.2 ±0.7)points vs.(4.0 ±0.5)points,t=1.375,P=0.174;6 h:(3.5 ±0.4)points vs.(3.3 ±0.6)points,t=1.641,P=0.105;12 h:(3.0 ±0.7)points vs.(2.8 ±0.5)points, t=1.375, P=0.174;24 h:(2.8 ±0.5)points vs.(2.6 ±0.6)points,t=1.515,P=0.134].At 48 h after operation,the VAS score was significantly higher in the control group than that in the experimental group[(2.3 ±0.3)points vs.(2.1 ±0.4)points, t=2.366, P=0.021].The incidence of nausea in the experimental group was lower than that in the control group(22.9%vs.2.9%,χ2=4.590,P=0.032). Conclusion Use of nalbuphine for analgesia after uterine artery embolization has good analgesic results and reduces the incidence of nausea and vomiting after operation.

OBJECTIVETo investigate the correlation between intrahepatic eovalently closed circular (ccc)DNA of hepatitis B virus (HBV) and pathogen-and patient-related parameters.

METHODSUltrasound-guided liver biopsies were obtained from 60 patients with chronic HBV infection. Levels of intrahepatic HBV cccDNA and serum HBV DNA were measured by quantitative fluorescence PCR. Level of serum hepatitis B surface antigen (HBsAg) was measured by chemiluminescence immunoassay. Clinical parameters, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alkaline phosphatase (AKP), albumin, globulin (GLO), white blood cell, platelet, prothrombin-international normalized ratio, were measured by standard assay. Demographic information was recorded.The correlation between intrahepatic HBV cccDNA and pathogen-and patient-related parameters was assessed.

RESULTSIntrahepatic HBV cccDNA level was negatively correlated with age, GLO, ALT and grade of necroinflammation. Patients with age of 30 years or more showed significantly higher level of HBV cccDNA level than patients under 30 years-old (7.44±0.58 and 5.66±1.35; t=7.157, P less than 0.001). Intrahepatic HBV cccDNA level was positively correlated with serum HBV DNA level (r=0.916, P less than 0.001) and serum HBsAg level (r=0.727, P less than 0.001). The median ratio of HBV cccDNA to HBV DNA was 1.18, and of HBV cccDNA to HBsAg was 1.67.

CONCLUSIONIntrahepatic HBV cccDNA levels decrease with age, level of ALT, level of GLO and grade of liver necroinflammation, but increase with level of serum HBV DNA and level of serum HBsAg. To a certain extent, serum HBV DNA and serum HBsAg levels may be a sufficient marker of intrahepatic HBV cccDNA levels.

Age Distribution ; Alanine Transaminase ; Aspartate Aminotransferases ; Biomarkers ; DNA, Circular ; DNA, Viral ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Real-Time Polymerase Chain Reaction ; Serologic Tests