ABSTRACT
Objective To investigate isolation of pathogens from bile and clinical characteristics of patients with hepatobiliary diseases.Methods Bacterial culture result of bile and related clinical data of patients with hepatobiliary diseases in a hospital were collected and analyzed by retrospective survey.Results A total of 406 bile specimens from patients with hepatobiliary diseases were collected,the positive rate of culture was 64.53％.Of 262 positive specimens,62.21％ (n =163),32.83％ (n =86),and 4.96％ (n =13) were isolated single pathogen,2 kinds of pathogens,and 3 kinds of pathogens respectively.374 pathogenic strains were isolated,242 (64.71％),131 (35.02 ％),and 1 (0.27 ％) were gram-negative bacteria,gram-positive bacteria,and fungus respectively.Patients with cirrhosis of liver,history of hepatobiliary surgery,and cholelithiasis had higher isolation rates of pathogens from bile than control group(all P＜0.05),isolation rates of pathogens from bile in patients with cholelithiasis of different sites were varied;but there was no significant differences among patients of different age,gender,and whether or not with hepatobiliary tumors(all P＞0.05).There were no statistical difference in constitute of pathogenic species from bile between patients with and without gallstones,as well as with and without history of hepatobiliary surgery(both P＞0.05);while constitute of pathogenic species from bile between patients with and without cirrhosis of liver was statistically different(x2 =14.058,P =0.001).Conclusion Pathogens from bile in patients with hepatobiliary diseases are mainly Enterobacteriaceae and Enterococcus spp.which caused single infection.Positive culture rate of pathogens from bile is higher in patients with cholelithiasis,history of hepatobiliary surgery,and liver cirrhosis.
ABSTRACT
Objective To construct the Neisseria surface protein A (NspA) DNA vaccine of Neisseria gonorrhoeae and evaluate the humoral and cellular immune responses induced by this vaccine in mice model.Methods The recombinant expression vector pcDNA3.1 (+)/NspA was constructed by inserting NspA gene into the eukaryotic expression vector pcDNA3.1 (+) and confirmed by poly merase chain reaction (PCR),restriction enzymes HindⅢ,XbaⅠand DNA sequencing.NspA mR- NA in transfected RAW264.7 cells and NspA protein expression in transfected COS-7 cells were de- tected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical stai- ning,respectively.Forty-five male BALB/c mice were immunized with pcDNA3.1 (+)/NspA recom binant plasmid.The level of serum anti-Neisseria gonorrhoeae antibody of the immunized mice was detected by tube agglutination test,and the level of interieron (IFN)-?was assayed by enzyme-linked immunosorbent assay (ELISA).The proliferation of splenocytes was determined by methyl thiazolyl tetrazolium (MTT) colormetry.The NspA gene in BALB/c mice was identified by PCR with the total DNA extracted from quadriceps femoris in immunized sites.Results Restriction enzymes digestion a- nalysis and DNA sequencing results revealed that the pcDNA3.1 (+)/NspA had been constructed successfully.NspA gene had been transcripted and expressed in mammalian cells.The peak titer of specific antibody was 1:640 in pcDNA3.1(+)/NspA immunized group and there was no specific an- tibody detected in both pcDNA3.1 (+) immunized group and PBS group.The IFN-?level in pcD NA3.1 (+) immunized group was (23.79?11.85)pg/mL and that in pcDNA3.1 (+)/NspA immu- nized group was(169.71?30.52)pg/mL (P