Objective To evaluate whether different DNA polymerases have an impact on the results of research on gut microbial diversity on the basis of full-length 16S rRNA gene as the target of sequencing.Methods Bacterial 16S rRNA gene sequences were amplified from the genomic DNA of three canine fecal samples with KAPA HiFi~(TM) HotStart DNA polymerase and PCRBIO HotStart DNA polymerase,respectively.The full-length 16S rRNA genes were sequenced using the PacBio single molecule real-time sequencing technology to profile gut microbial communities at the species level.Results Based on t test in a pairwise manner,the results showed there were no significant differences in the levels of phylum,genus and species between the two different DNA polymerases(P0.05),but in amplification efficiency when it came to some of relatively less operating groups(OTUs).UPGMA cluster analysis of unweighted UniFrac distance and PERMANOVA analysis of weighted UniFrac distance all showed that there was no significant difference in the diversi
Objective To analyze the distribution and drug sensitivity of the pathogenic bacteria in ear canal secretion of patients with chronic suppurative otitis media(CSOM).Methods Specimens of ear canal secretion collected from 100 patients(116 ears)with CSOM were collected for routine pathogenic bacteria isolation,identification and drug sensitivity test.Results In the 116 ears,pathogenic bacteria were detected in 97 ears(83.6%).A total of 103 strains of pathogenic bacteria were detected,including 53.4%(55/103)of Gram positive bacteria,41.7%(43/103)of Gram negative bacteria and 4.9%(5/103)of fungi.Staphylococcus aureus and Staphylococcus epidermidis had higher resistance to Penicillin and Sulfamethoxazole/Trimethoprim,and higher sensitivity to Rifampin and Moxifloxacin;and they were completely sensitive to Furazolidone and Vancomycin.Pseudomonas aeruginosa and Klebsiella pneumoniae had higher resistance to Cefazolin,Ampicillin,Cefepime,Ceftriaxone and Piperacillin/Sulbactam,and higher sensitivity to Tobramycin,Aztr
Objective To analyze the clinical distribution and drug resistance of K.pneumoniae from bloodstream infection(BSI),so as to provide evidence for clinical rational use of antibiotics and effective control of the infection.Methods 289 strains of K.pneumoniaeisolated from BSI patients in our hospital from 2014 to 2016 were collected for bacterial identification,antimicrobial susceptibility test and ESBLs phenotype confirmation.The clinical distribution and antibiotic resistance of K.pneumoniaefrom bloodstream infection were analyzed.Results Patients with K.pneumoniae BSI mainly distributed in the emergency department(72/289)and intensive care unit(69/289).During these 3 years,K.pneumoniaeshowed low resistance rates to Carbapenems and Amikacin,at around 20.00%.A total of 135(46.71%)ESBL producing strains were identified;their resistance rates to commonly used antibiotics were significantly higher than those of non-ESBL producing strains(P0.01).The resistance rates of Carbapenem resistant strains to common antibio
Tuberculosis is caused by Mycobacterium tuberculosis(M.tuberculosis).CD_4~+ T cells,CD_8~+ T cells and Th17 cells play the most important roles in the immune responses against M.tuberculosis.Besides,specific glycosylated IgG in the humoral immunity contributes to the eradication of pathogens.The current vaccine,Bacillus Calmette-Guérin(BCG),protects infants efficiently against severe forms of tuberculosis,but does not prevent adult pulmonary tuberculosis satisfactorily.Therefore,it is necessary to develop new vaccines.At present,several candidates are being studied in clinical trials.In this review,the immune mechanisms,as well as the knowledge of BCG,are clarified,and the development of new vaccines is summarized.
Objective To construct a recombinant Mycobacterium tuberculosis Rv1776c gene,and identify its activity in recombinant Mycobacterium smegmatis.Methods PCR method was used to clone Mycobacterium tuberculosis Rv1776c gene.Escherichia coli-mycobacteria shuttle expression plasmid pMV-Rv1776c was constructed,and confirmed by enzyme digestion and sequencing.The recombinant plasmid was transfected into Mycobacterium smegmatis mc~2 155 with electroporation method.The expression of Rv1776c protein in recombinant Mycobacterium smegmatis was confirmed by using SDS-PAGE and Western blot.Results The recombinant Mycobacterium smegmatis was constructed successfully.The growth curve showed that the recombinant plasmid did not affect the growth of Mycobacterium smegmatis in vitro.SDS-PAGE and Western blot confirmed that the Rv1776c expressed in the Mycobacterium smegmatis had a relative molecular weight of about 42 kD.Conclusion The shuttle plasmid pMV-Rv1776c of Rv1776c gene was successfully constructed,and its biological act
Objective To learn the drug resistance of Acinetobacter baumannii and detect the resistant gene of carbapenem-resistant Acinetobacter baumannii,so as to provide theoretical basis for clinical rational use of drugs and control of nosocomial infections.Methods The drug resistance of 45 strains of Acinetobacter baumannii clinical isolates were detected by using K-B method.The phenotype of carbapenemase was analyzed through modified Hodge test,Carba NP test and EDTA co-trial.The OXA-23 and NDM-1 genes in Acinetobacter baumannii were detected with PCR.Results A total of 42 multidrug-resistant strains were detected.36 strains of carbapenemase positive strains were detected by using modified Hodge test and Carba NP test.OXA-23 gene was amplified by using PCR while no NDM-1 gene was found.Conclusion The drug resistance of Acinetobacter baumannii is very high,and most of the strains carry OXA-23 gene.Antibacterial drugs should be used reasonably based on the results of drug susceptibility test.
Objective To investigate the genetic variation of H3N2 subtype influenza virus and the match between the epidemic strain and vaccine strain in Liaoning during 2016 and 2017.Methods Reverse transcriptase polymerase chain reaction(RT-PCR)was performed to amplify the HA1 genes of the isolated influenza H3N2 strains,and the amplified fragment was sequenced and blasted with Northern Hemisphere vaccine strain recommended by WHO in recent years.Results HA1 phylogenetic analysis showed that during 2016-2017 the H3N2 subtype influenza virus strain and the vaccine strain in latest three years were not in the same branch.Genetic characteristics analysis revealed more than two variations occurred in the antigenic determinants A and B of all the viruses.19 strains of viruses had a new mutation at the receptor binding site of 131 amino acid.1 out of the 20 strains of viruses produced a new mutation of cysteine,suggesting possible occurrence of new disulfide bond.No new mutations were detected at glycosylation sites.Conclus
Liver cirrhosis is a kind of common diseases in China.Nowadays,more and more researches had demonstrated that the gut microbiota dysbiosis was associated closely with liver cirrhosis and its complications.In this article,we described the features of the complex interaction between gut microbiota and liver,i.e.the so called "gut-liver axis".In cirrhosis,the overgrowth of bacteria and the dysfunction of intestinal mucosa and immune system result in bacteria translocation and gut microbiota imbalance.On the contrary,the gut microbiota dysbiosis deteriorates the function of liver and results in the occurrence of the complications.This review aims to summarize the researches on the role of bacterial and fungal microbiota in liver cirrhosis and its complications.
Objective To discuss the influence of Trimebutine combined with Bifid Triple Viable Capsules on intestinal microecology of patients with functional dyspepsia(FD)and diarrhea-predominant pattern irritable bowel syndrome(IBS-D).Methods 90 patients with FD and IBS-D were randomly divided into observation group(n=45)or control group(n=45).Both groups were given Trimebutine tablets 100 mg tid,while the observation group was additionally given Bifid Triple Viable Capsules 630 mg bid,for 8 weeks.The changes of intestinal flora in both groups were observed,and the clinical efficacy and adverse reactions were compared between groups.Results After treatment,the quantities of Bifidobacteriaand Lactobacilli obviously increased,while the quantities of Enterobacters,Enterococci and Saccharomycetes were decreased in observation group(P0.05).No obvious changes in the quantities of the 5 bacteria were observed in control group(P0.05).The total clinical efficiency rate in observation group was higher than that in control group
Objective To analyze the effect of colonscopic probiotics administration on the symptoms of irritable bowel syndrome(IBS)and potential adverse events.Methods The patients with IBS in our hospital were included according to IBS Roma IV diagnostic criteria after obtaining their informed consent.Suspension of Bifidobacterium,Lactobacillus acidophilus and Enterococcus faecalis(100 mL,33.6 mg/mL)was sprayed into the right hemicolon during colonscopy.The Bristol stool scale,abdominal visual analogue score(VAS),colon symptom score and IBS severity scoring system(IBS-SSS)were used at 0,2 and 4 weeks,while Hospital anxiety and depression scale(HADS)was used at 0 and 4 weeks to evaluate the symptom improvement.Results 5 cases of constipation predominant IBS(IBS-C)and 13 cases of diarrhea predominant IBS(IBS-D)were enrolled.The average age were(49.4±10.0)and(36.1±8.9)respectively,and the male/female ratios were 1∶0.66 and 3.3∶1,respectively.No improvements were observed in IBS-C patients.In IBS-D group,the composite eff