1.Regulation of apoptosis of host cells by in vitro azithromycin-induced persistent Chlamydia trachomatis infection
Wentao CHEN ; Yaohua XUE ; Jinmei HUANG ; Jieyi YANG ; Yunhu ZHAO ; Yinyuan LAN ; Mingheng FANG ; Biying ZHENG ; Heping ZHENG
Chinese Journal of Dermatology 2018;51(5):347-351
Objective To evaluate the regulatory role of azithromycin-induced persistent Chlamydia trachomatis (Ct) infection in the apoptosis of Hela229 cells.Methods Hela229 cells were firstly co-cultured with Ct for 22 hours,and then cultured with Dulbecco's modified Eagle's medium (DMEM) containing 0.08 mg/L azithromycin for 26 hours to establish a cell model of persistent Ct infection (persistent infection group).These infected Hela229 cells cultured with azithromycin-free DMEM served as a cell model of acute Ct infection (acute infection group).After 48-hour infection with Ct,azithromycin was removed,and infected Hela229 cells in the above 2 groups were successively cultured with DMEM for the resurgence of Ct.Immunofluorescence assay and electron microscopy were performed to verify the persistent Ct infection model.The Hela229 cells in the persistent infection group and acute infection group as well as uninfected Hela229 cells (control group) were treated with staurosporine (STS) for 4 hours to induce the apoptosis,and then cell apoptosis was detected by Hoechst 33258 staining,annexin V/propidium iodide staining and flow cytometry.Results After the treatment with azithromycin,atypical inclusions with aberrant reticulate bodies appeared in the Ct-infected cells.After removing azithromycin,cells were cultured until 96 hours after infection,and infectious elementary bodies reappeared in the Ct inclusions.After the treatment with STS,Hoechst staining showed that there was loose chromatin in the persistently infected cells,while chromatin condensation was observed in the uninfected cells.After 24-hour infection with Ct and 4-hour induction with STS,the apoptosis rate was significantly higher in the persistent infection group (45.567% ± 2.631%) than in the acute infection group (38.567% ± 1.701%,t =2.686,P =0.028),but significantly lower in the persistent infection group than in the uninfected group (69.800% ± 2.835%,t =8.187,P < 0.001).After 48-hour infection with Ct and 4-hour induction with STS,there was a significant difference in the apoptosis rate between the persistent infection group (46.700% ± 5.257%) and acute infection group (61.767% ± 1.815%,t =5.781,P < 0.001),as well as between the persistent infection group and the uninfected group (68.667% ± 3.156%,t =7.421,P < 0.001).Conclusion This study showed that azithromycin-induced persistent Ct infection regulated the apoptosis of host cells,and this effect lasted 48 hours.
2.Genital Chlamydia trachomatis infection and associated risk factors in male clients attending sexually transmitted disease clinics in 9 cities in Guangdong province
Hongcheng SHEN ; Shujie HUANG ; Xiaolin QIN ; Peizhen ZHAO ; Yinyuan LAN ; Huachun ZOU ; Jiangli OU ; Lei CHEN ; Xiaomin LUO ; Heping ZHENG ; Yan LI ; Bin YANG
Chinese Journal of Epidemiology 2017;38(3):364-368
Objective To investigate the prevalence of genital Chlamydia trachomatis (GCT) infection and associated risk factors in male clients attending sexually transmitted disease (STD) clinics in Guangdong and provide integrated intervention strategy for this group.Methods Convenient sampling was used to recruit participants from April to June in 2015 in Guangdong province.The information about their socio-demographic characteristics and sexual behaviors were collected by using a questionnaire,and blood samples were taken from them to test the antibodies against HIV,syphilis and HCV.First pass urine was taken to test GCT and gonorrhea.Results A total of 1 749 participants with the average age of 39.53 years were recruited.The majority of them were married (73.87%,1 292/1 749),residents of Guangdong (92.28%,1 614/1 749) and in Han ethnic group (99.49%,1 740/1 749).The positive rates for GCT,HIV,syphilis,HCV,Neisseria gonorrhea,and WBC in urinalysis were 6.06% (106/1 749),0.46% (8/1 749),3.43% (60/1 749),0.45% (7/1 550),2.74% (48/1 749),7.89% (138/1 749) respectively.Multivariate analysis showed that risk factors for GCT infection include IDUs (OR=13.98,95%CI:3.35-58.38),anal sex with men (OR=3.11,95% CI:1.45-6.71),Neisseria gonorrhea positive (OR =9.64,95% CI:5.09-18.24),and WBC positive (OR =1.96,95% CI:1.08-3.55).Conclusions This study demonstrated the high prevalence of GCT infection in male clients attending STD clinics in Guangdong.Therefore precision intervention should target this population at high-risk.