A mouse model of bacterial vaginosis established by infecting estrogen-treated mice with Prevotella bivia / 中华微生物学和免疫学杂志

Objective To establish a mouse model of bacterial vaginosis ( BV) by infecting estro-gen-treated mice with Prevotella bivia ( P. bivia) . Methods The mice were intraperitoneally injected with beta-estradiol 17 valerate which was suspended in sesame oil and then inoculated with different doses of P. bivia strains at the logarithmic phase. Samples of vaginal flushing fluid were collected at different time points after inoculation and used for the isolation of P. bivia strains and the detection of sialidase activities. Altogether 30 mice treated with estrogen and high dose of P. bivia were killed on days 2, 7 and 21 (n=10). Samples of cornua uteri, bladder and kidney were collected from those mice for P. bivia strains isolation. Re-sults Injection the estrogen-treated mice with P. bivia via vagina could cause the P. bivia infection for more than 14 days. The numbers of P. bivia strains isolated on day 21 decreased significantly. Enhanced sialidase activities and clue cells were observed in vaginal secretions of mice with P. bivia infection. Injection of mice with the high dose of P. bivia could spread the infection to cornua uteri. Conclusion Estrogen-treated mice could be used as an animal model for researches on BV.

The coexistence expression of TRPV1 and μ-opoid receptor in spinal dorsal root ganglion projected to stomach and its clinical significance / 实用医学杂志

Objective To observe the coexistence expression of TRPV1 and μ-opoid receptorin spinal dorsal root ganglion (DRG) projected to stomach , and to investigate the relationship between TRPV1 andμ-opoid receptorand its clinical significance in rats with acute gastric mucosal lesion induced by water immersion and restraint stress. Methods FortyWistar rats were randomly assigned to 3 groups, including normal control group(group NC, n = 10), WIRS group (group WIRS, n = 20), and sufentanil pretreatment group (group SP, n = 10). A rat model of gsatric mucosal lesion was induced by WIRS. 6 hours after WIRS treatment, gastric tissues were excised and microscopically observed; ulcer index (GI) was noted. The coexistence expression of TRPV1 and μ-opioid receptor in DRG neurons was detected by immunofluorescence assay, and the levels of CGRP was measured by ELASA. Results As compared withgroup WIRS, the degree of gastric injury was obviously relieved in group SF. Coexistence of TRPV1 and μ-opioid receptor was detected in thoracic DRG neurons projected to stomach; the CGRP level was higher in group WIRS than in group NC. ConclusionsTRPV1 isinvolved in protection of acute gastric mucosal lesion. Activation of μ-opioid receptor can induce TRPV1 to release CGRP, resulting in protection of gastric mucosa.

Construction of a recombinant Neisseria gonorrhoeae strain with rmp gene deletion and study on the bactericidal activities of its antibodies / 中华微生物学和免疫学杂志

Objective To study the role of the outer membrane protein Rmp of Neisseria gonor-rhoeae strain in immunosuppression and the strategy of eliminating it .Methods The rmp gene of Neisseria gonorrhoeae strain was amplified by PCR and inserted into pMD 19-T vector .The recombinant vector pMD 19△rmp∷Kan containing Kan and the 5′-and 3′-flanking regions of rmp (△rmp∷Kan) was constructed by replacing 200 nucleotide residues of pMD 19-rmp with kanamycin resistance gene Kan and transformed into Neisseria gonorrhoeae WHO-A strain.PCR and Western blot assay were used to screen and identify the re-combinant mutant strains that could not express Rmp .Mice were immunized with mutant strains and bacteri-cidal activities of the immune sera were detected by antibody-mediated complement-dependent cytotoxicity assay.Results The mutant strains that could not encode Rmp were successfully constructed .Antibodies in-duced by mutant strains showed stronger bactericidal activity against Neisseria gonorrhoeae in comparison with those induced by wild strains .Conclusion The recombinant Neisseria gonorrhoeae strain with rmp gene de-letion might eliminate the immunosuppressive effects of Rmp expressed in wild gonococcal strains , which provides a reference for further development of novel live attenuated whole-cell vaccines of Neisseria gonor-rhoeae.

Prokaryotic expression of Neisseria gonorrhoeae NspA and its antigenicity / 中华微生物学和免疫学杂志

Objective To develop fast detection techniques for the diagnosis of gonococcal infections.Methods Prokaryotic expression vector for Neisserial surface protein A (NspA) was constructed using the NspA gene cloned by PCR.Mice were immunized with the renatured recombinant NspA (rrNspA)to prepare antibodies against NspA.Western blot and ELISA was used to analyze the binding of NspA antibodies to lysate of gonococcal cells or to intact gonococci.Results NspA antibodies that were prepared by the rrNspA expressed in E.coli could bind to rrNspA,the natural NspA existing in gonococcal cells,or intact gonococci.Conclusion RrNspA and its antibodies have potential value in developing fast diagnostic kits for gonococcal diseases.

Human carcinoembryonic antigen-related cellular adhesion molecules 1 mediated susceptibility of Neisseria gonorrhoeae to mouse / 中华微生物学和免疫学杂志

ObjectiveTo develop a transgenic mouse model for N.gonorrhoeae researches.Methods Human carcinoembryonic antigen-related cellular adhesion molecules 1 (hCEACAM1) eukaryotic expression vector,pCDPGICAM1,was used to generate transgenic mice by microinjection.The funder mice were screened by PCR,sequence analysis,Western blot and fluorescence-activated cell sorting analysis,respectively.The transgenic mice expressing hCEACAM1 were inoculated with N.gonorrhoeae intravaginally.Adhesion and infection of gonococci to mice were analyzed by bacteria culture and microscopy.Results Four (lines 50,53,54,and 59) of the 22 F0 generation transgenic mice were found to carry the transgene.The hCEACAM1 protein was expressed on the cell membrane of various tissues in the line 53 transgenic mouse.Compared with normal mice,N.gonorrhoeae can successfully infect and cause inflammation in the transgenic mice.Conclusion The hCEACAM1 transgenic mouse can be used as an animal model for gonococcal infections.

Effects of isoflurane on spatial reference memory and changes of Caspase-3 expression and apoptosis in brain of aged mice / 中华行为医学与脑科学杂志

Objective To investigate the effect of isoflurane on the spatial learning and memory in aged mice,and whether this is associated with the changes of Caspase-3 expression and apoptosis in brain.Methods Twenty-four CO57BL/6 aged mice(16 months)were randomly divided into isoflurane treatment group(Iso Group,n=12) and sham control group (Con Group,n=12).Mice in Iso group were exposed to 1% isoflurane in carrying gas of 30% oxygen,balance nitrogen in a warmed,humidified chamber for4 h per day for2 days.For Con group,animals were treated at the same condition with only carrying gas.After anesthetic exposures,behavioral testing was performed using the Morris water maze(MWM),and then changes of Caspase-3 expression and apoptosis in hippocampus CAI,dentate gyrus(DG) and cortex(CX) in brain were determined by using immunofluorecence staining and TUNEL staining.Results In hidden-platform training of MWM,the mean escape latency to platform showed no significant difference between the two groups (F=0.007,P=1.235),but the mice in Iso group showed obviously impaired retention of memory by spending more percentage of time swimming in the probe quadrant as compared to the control animals in the probe test((34.5±5.0)%vs(45.1±4.9)%.P<0.01).There was no significant difference in average swimming speed during the MWM testing trials between the two groups (F=1.537,P=0.241).A few Caspase-3 and apoptosis positive cells were found in hippocampus Cal,DG and CX regious,while no difference was found in the density of positive cells between the two groups(P>0.05).Conclusion 1% isoflurane repeatedly exposure significantly impaires the spatial reference memory in aged mouses,however does not significantly change the expression of easpase-3 and apoptosis in brain.

The adhesion of Neisseria gonorrhoeae to COS-1 cells mediated by human CEACAM1 / 中华微生物学和免疫学杂志

Objective To study the role of human carcinoembryonic antigen-related cellular adhesion molecule 1 (hCEACAM1)in mediating the specific adhesion of N. gonorrhoeae to its human host cells.Methods A recombinant eukaryotic expression vector pCDPGICEA1 was constructed by putting hCEACAM1 cDNA behind both hCD46 promoter and rabbit β-globulin intron 2,and with which,the COS-1 cells were transfected. Following G418 selection, the COS-1 cells expressing hCEACAM1 were sorted out with flow cytometry. The adhesion of N. gonorrhoeae to the gene transfected COS-1 cells was analyzed with bacterial binding assay. Results hCEACAM1 cDNA could be expressed effectively under the direction of hCD46 promoter and rabbit β-globulin intron 2,and N. gonorrhoeae could adhere to COS-1 cells expressing hCEACAM1. Conclusion hCEACAM1 can mediate the adhesion of N. gonorrhoeae to animal originated COS-1 cells. thus its transgenic mice may be used as a novel animal model for studying N. gonorrhoeae infection.

THE EARLY STAGE DIGNOSIS OF PRIMARY GALLBLADDER CARCINOMA:ANALYSIS OF 679 CASES / 中国现代普通外科进展

Objective:In order to enhance the early dignosis level and strengthen the understanding of pathogeny、pathology by stage and prognosis of primary gallbladder car-cinoma(PGC). Methods : The results of 679 patients of PGC from 1956 to 1998 in first affiliated hospital of Xi'an medical university were retrospectively analyzed. Results: Most of the patients belong to advanced stage when dignosised. The PGC were usually found among the old women patients. The sex ratio(female/male) was 3:1. There is a intimate relatioship between gallstone and PGC. About sixty percents of PGC might be found gallstone.The dignostic accordance rate was low between preoperation and postopera-tion.Most patients with PGC were found unexpectedly in operation due to gallstone or a-cute cholecystitis. To worse more, many of patiens with PGC were missed the opportunities of dignosis and treatment because of only paying attention to gallstone. Most of the pathological types were adenocarcinoma. The development of imaging examination is hoped to find early stage cases and improve its prognosis. Conclusion : It is a basic way for early dignosis and improving prognosis to strengthen the understanding of pathogeny, pathology by stage and prognosis and rationalize application of all kinds of exzamination means in PGC.

Relationship between expression of CD44v6 and matrix metalloproteinase-2 and the invasion and matastasis of gallbladder carcinoma / 中国普通外科杂志

Objective To evaluate the role of CD44v6 and matrix metalloproteinase-2(MMP-2) in the invasion and metastasis of gallbladder carcinoma (GC). Methods Immunohistochemical techniques were adopted to detect the expression of CD44v6 and MMP-2 in 42 cases of GC and 10 cases of cholecystolithiasis. Results (1)The expression of CD44v6 in the GC group (64.3%, 27/42) was significantly higher than that in the cholecystolithiasis group (0%,0/10). The expression of CD44v6 was correlated with GC histologic grade (P