RESUMO
Gardnerella vaginal is one of the main pathogenic floras of female bacterial vaginosis. Sexually transmitted, it causes fallopian pregnancy, premature rupture of the foetal membrane, neonate premature delivery and other bad pregnancy outcome. Gardnerella vaginal is also involved in male urogenital tract infection. This review summarizes the recent advances in the studies of Gardnerella vaginal in such aspects as its categorization, biocharacteristics, biogrouping, epidemiology, pathogenesis, diagnostic criteria, detection, treatment and clinical significance.
Assuntos
Gardnerella vaginalis , Feminino , Humanos , MasculinoRESUMO
AIM: To investigate the possible functions of human sperm membrane protein (hSMP-1) in the process of fertilization. METHODS: A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 (anti-rhSMP-1) antibodies. Sperm acrosome reaction and sperm-zona pellucida (ZP) binding assay were carried out in 10-week-old BALB/c mice. RESULTS: Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 (P < 0.05). CONCLUSION: The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.
Assuntos
Reação Acrossômica/fisiologia , Anticorpos/farmacologia , Antígenos de Superfície/genética , Proteínas de Membrana/genética , Reação Acrossômica/efeitos dos fármacos , Reação Acrossômica/imunologia , Animais , Clonagem Molecular , Primers do DNA , Escherichia coli/genética , Feminino , Fertilização , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Zona Pelúcida/imunologia , Zona Pelúcida/fisiologiaRESUMO
OBJECTIVE: To acquire the purified human nuclear autoantigenic sperm protein (hNASP) and its polyclonal antibody for investigating the possible functions of hNASP involved in fertilization. METHODS: The coding sequence of hNASP gene was amplified from human testis RNA with specific primers, and the PCR product was cloned first into pMD-18T and then into pET-28a ( + ) after restriction digestion with BamH I and Hind III. The fusion protein was expressed in E. coli BL21 (DE3) after induction with IPTG. The recombinant protein NASP was purified from the supernatant with Ni2 -NTA His-bind resin under native conditions. RESULTS: The results of DNA sequencing and SDS-PAGE analysis showed the protein to be what we had hoped to acquire. ELISA showed that we had acquired rabbit anti-hNASP serum with high titer. CONCLUSION: High purity recombinant hNASP protein could be obtained with the above-mentioned prokaryotic expression method, and so could the rabbit anti-hNASP serum with high titer and high specificity.
