Objectives To test a simple method for genotyping of C.trachomatis isolates and to investigate the clinical significance of the genotypes. Methods A part of the chlamydial genome encoding the major outer membrane protein(omp1) was amplified by polymerase chain reaction (PCR). The products were digested by endonucleases to see the characteristic patterns, after silver staining on 10% polyacrylamide gels. Results The omp1 genes of 15 serovars of C. trachomatis were amplified by PCR,which generated an 871 base pair gene fragment. AluⅠ digestion of the product gave characteristic patterns for the 15 serovars,but group C presented closely similar patterns. A triple digestion with HpaⅡ, followed by HinfⅠ and EcoRⅠ, would allow the differentiation of serovars in group C. Analysis of 74 clinical isolates revealed serovars E, F, D, G as the most prevalent genital serovars in the studied populations. Serovars B, H, J were occasionally identified. A mixed infection with serovars F and D was seen in a clinical sample. No significant relationship was observed between clinical manifestations of urogenital chlamydial infections and serovars,however,serovar D was more often associated with high titer of anti chlamydial antibody than other serovars. Conclusion The omp1 genotyping technique seems to be promising for epidemiological studies.

Objective To investigate the type distribution of urogenital Chlamydia trachomatis(Ct)among STD clinic outpatients in Guangxi Zhuang Autonomous Region, and to estimate the prevalence of Ct infection among the patients during posttreatment follow?up. Methods Urethral and cervical swabs were collected from male and female outpatients with confirmed urogenital Ct infection, respectively, in Institute of Dermatology of Guangxi Zhuang Autonomous Region. The patients with positive results in preliminary screening tests were followed up after treatment, and specimens were collected at follow?up visits. General and clinical information was also obtained from these patients. DNA was extracted from these samples by using the QIAxtractor instrument. Nested PCR was performed to amplify the major outer membrane protein A(ompA)gene for ompA typing, and to amplify CT046(hctB), CT058, CT144, CT172 and CT682 (pbpB) genes for high?resolution multilocus sequence typing (hr?MLST). Then, PCR products were sequenced, and ompA and MLST types of Ct were determined by sequence alignment and MLST analysis, respectively. The obtained MLST sequence types (STs) were compared with those from an Italian population by using the BioNumerics7 software, and a minimum spanning tree(MST)was generated. Results Totally, 44 and 6 Ct?positive specimens were collected at first visits and follow?up visits respectively. Among the 50 specimens, 42 underwent successful ompA typing and hr?MLST, and 7 ompA genotypes and 15 hr?MLST STs were identified, including 3 first reported STs. The distribution of STs of Ct isolates from Guangxi Zhuang Autonomous Region was significantly different from that from the Italian population. Among the 6 followed patients with posttreatment Ct infection, 3 were confirmed to be reinfected with Ct, and the other 3 failed to be diagnosed because of unsuccessful genotyping. Conclusion The genotypes of Ct strains isolated from STD clinic outpatients in Guangxi Autonomous Region were characteristic, and Ct reinfection occurred in some patients during follow?up.

Objective To compare the effects of medroxyprogesterone acetate (MPA) and compound norethisterone enanthate (CNE) on the susceptibility of BABL/c mice to lower reproductive tract infection with chlamydia trachomatis (Ct). Methods A total of 60 BALB/c mice were randomly and equally divided into 6 groups:MPA-pretreated control group and CNE-pretreated control group inoculated with MyCoy cell suspensions in the vagina on the 5th day after single treatment with MPA and CNE respectively, blank control group receiving no treatment, MPA-pretreated infected group and CNE-pretreated infected group inoculated with 1 × 107 inclusion-forming units(IFU)of Ct serovar E in the vagina on the 5th day after single treatment with MPA and CNE respectively, control infected group inoculated with the same quantity of IFU of Ct serovar E in the vagina but receiving no pretreatment. On day 4, 7 and 14 after inoculation, vaginal irrigation fluid was obtained from all the mice for cell culture of Ct. Three mice were randomly selected from each of these groups at the above three time points and sacrificed, and vaginal and uterine tissue specimens were obtained for hematoxylin-eosin(HE)staining and microscopic examination. Chi-square test and Fisher's exact test were conducted to compare infection rate among different groups. Results No growth of Ct was observed in the three control groups at the above time points. The culture-positive rate of Ct was 1/10 on day 4 but 0 on day 7 and 14 in both the CNE-pretreated infected group and control infected group, 7/10 on day 4, 2/7 on day 7 but 0 on day 14 in the MPA-pretreated infected group. Fisher's exact test revealed that the culture-positive rate of Ct was significantly higher in the MPA-pretreated infected group than in the control infected group and CNE-pretreated infected group on day 4 (both P =0.03), but similar among the three infected groups on day 7 (P = 0.23). Both the MPA-pretreated control group and infected group showed an increase in endovaginal mucus, thinning of vaginal stratified squamous epithelium, mucification of vaginal epithelium, presence of secretions in vaginal lumen and submucosal infiltration of a few inflammatory cells on day 4, 7 and 14, as well as appearance of pathological changes (including the presence of large quantities of purulent secretions in lumen, mild tissue edema and submucosal infiltration of a few inflammatory cells) in the vagina on day 4. Vaginal tissues were normal in both the CNE-pretreated infected group and control group at the above three time points, but mild tissue edema, lumen expansion, secretion retention and infiltration of scattered inflammatory cells were observed in the uterus on day 4 after inoculation. Conclusions MPA can arrest the estrous cycle of mice at diestrus with the mucification of vaginal epithelium, which may increase the susceptibility to Ct vaginal infection in mice. In contrast, CNE has no obvious effect on the estrous cycle and susceptibility to Ct vaginal infection despite of the appearance of pathological changes in the uterus.

85%. The concentrated MCS in different amount was added to the IFN-?(100 pg/mL) and LPS (10 ng/mL) enriched culture media. The IL-12 production by monocytes was determined by the enzyme- linked immunosorbent assay(ELISA).The expression of CD14 and CD1a was analyzed by flow cytometry 5 days after the monocytes were co-cultured with MCS. Results The production of monocytic IL-12 was down-regulated by MCS in a dose dependent manner. The amount of IL-12 from monocytes decreased along with an increased dose (25-100?L) of MCS applied in the reaction. It was also observed that the differentiation from CD14 expressing monocytes to CD1a dendritic cells was impaired by MCS. The ability of MCS to inhibit the production of IL-12 by monocytes and to suppress the differentiation of monocytes to dendritic cells in vitro could be disrupted by PD98059,an ERK specific inhibitor. Conclusions MCS appears to inhibit IL-12p40 production by monocytes and inhibit differentiation of monocytes in vitro via secretion of ERK stimulating factor. The inhibitory factors in MCS and their chemical natures need further research.

Objective To make a nationwide external quality assessment of serologic tests for syphilis in China,in hope to increase the quality of syphilis serology in laboratories at different levels.Methods From 2006 to 2008,a nationwide external quality assessment scheme was conducted for serologic tests for syphilis in laboratories of some medical and healthcare facilities each year by the Reference Laboratory,National Center for Sexually Transmitted Disease Control,China Center for Disease Control and Prevention.Five quality control samples and corresponding questionnaires were sent to the participating laboratories.Tests were conducted and test results were reported within stipulated time.Subsequently,the test results were statistically analyzed by the Reference Laboratory,and the final results were fed back to all of these participants.Results From 2006 to 2008,the number of participating provinces increased from 17 to 31,and the number of participating laboratories from 23 to 145.Laboratories achieving a full score amounted to 79.9％,36.8％ and 57.6％,and those gaining a score of 80 or greater amounted to 95.7％,88.2％ and 89.7％,respectively,in 2006,2007 and 2008.Conclusion The external quality assessment scheme has enhanced the capacity of participating laboratories for syphilis serology to a certain extent from 2006 to 2008.

Objective To investigate the in vitro effects of various antibiotics (spectinomycin, ceftriaxone, erythromycin, ofloxacin and doxycycline) against 12 isolates of C. trachomatis. Methods Minimal inhibitory concentrations (MICs ) and fractional inhibitory concentrations (FICs) of the antimicrobials against all C. trachomatis were calculated. Checkerboard method was used for the determination of FICs and Ridit test for the comparison of the interactions among the various combinations. Results No difference was observed in most of the combinations. No antagonism was found in all except for ceftriaxone-doxycycline combination. Synergism was observed in 42% (5 of 12) and 50% (6 of 12) of the chlamydial isolates for erythromycin-spectinomycin and doxycycline-spectinomycin combination, respectively. No significant difference was observed among triple combinations with spectinomycin or with ceftriaxone. When interactions of erythromycin, ofloxacin and doxycycline with spectinomycin were compared to those with ceftriaxone respectively, both interactions of erythromycin (U = 2.46, P = 0.014) and doxycycline (U = 2.83, P = 0.002) were more synergistic with spectinomycin than those with ceftriaxone. Conclusions This study indicates that the combination of spectinomycin with erythromycin or doxycycline is more effective against C. trachomatis than that of ceftriaxone. Therefore, spectinomycin rather than ceftriaxone might be recommended in the dual therapy against C. trachomatis and N. gonorrhoeae.

ObjectiveToevaluateourrevisedsyndromicalgorithmforthemanagementinpatientswithvaginaldischargeanddetermineitssensitivity,specificity,andpositivepredictivevalue(PPV).MethodsPatientswithvaginaldischargesyndromewereselectedintheirfirstvisitstotwoSTDclinicsinShanghaiandSichuan.Theyweremanagedaccordingtorevisedsyndromicflowcharts.Theetiologyofthesyndromewasdetectedbylaboratorytesting.ThedatawereanalyzedusingEPIINFOV5.0software.ResultsTherewere27(8.1%)patientswithgonorrhea,57(17.1%)withchlamydialinfection,and18(5.4%)withbothinfectionsin334patientswithvaginaldischarge.Thesensitivitywas70.6%,specificity54.7%,PPV40.7%,andnegativepredictivevalue(NPV)80.9%forthediagnosisofgonorrheaand/orchlamydialinfectionbysyndromicapproach.ConclusionThespecificityandPPVforsyndromicmanagementofvaginaldischargearenotsatisfied.Furthervalidationandrevisionareneededforsyndromicapproachesofvaginaldischarge.

Objective To estimate the prevalence of hepatitis C virus (HCV) infection among sexually transmitted disease (STD) clinic attendees infected with human immunodeficiency virus type 1 (HIV-1) in Guangxi Zhuang Autonomous Region.Methods Totally,11 553 blood plasma samples were collected from STD clinic attendees in Guangxi Zhuang Autonomous Region,and subjected to HIV-1 antibody screening and confirmatory testing.Enzyme linked immunosorbent assay (ELISA) was performed to detect anti-HCV antibodies in 140 anti-HIV-1 antibody-positive samples and 282 anti-HIV-1 antibody-negative samples from age-and marital status-matched attendees.Chi-square test was performed to assess the differences in the prevalence rate of HCV infection between anti-HIV-1-negative and-positive samples,and Logistic regression analysis to evaluate the risk factors for HCV and HIV co-infection.Results The positivity rate of anti-HCV antibodies was 33.57％ (47/140)among anti-HIV-1-positive samples,significantly higher than that in anti-HIV-1-negative samples (1.06％ (3/282),x2 =94.66,P ＜ 0.05).Logistic regression analysis showed a statistical increase in the prevalence of HCV/HIV co-infection in individuals reporting more than one sexual partners compared with those reporting only one sexual partner (OR =2.4,95％ CI (1.0-5.6),P =0.05),and in intravenous drug users compared with non-intravenous drug users (OR =20.8,95％ CI(5.7-76.5),P ＜ 0.05).Conclusions HCV infection appears to be associated with HIV-1 infection,and comprehensive intervention on HIV-1-infected patients may slow down HCV transmission.

Objective To perform a nationwide external quality assessment for detection of Chlamydia trachomatis, and to improve the performance of laboratories in the detection of Chlamydia trachomatis. Methods Totally, 419 quality control samples were sent to tested laboratories, including 76 samples in 2007, 168 samples in 2008 and 175 samples in 2009. The laboratories were required to test the samples and report test results, within stipulated time, to the reference laboratory in National Center for Sexually Transmitted Disease (STD) Control, Chinese Center for Disease Control and Prevention. The reported results were statistically analyzed by the National Center for STD Control, who finally fed back the statistical results to all of the participants. Results The percentage increased from 84.93% in 2007 to 92.14% in 2009 for laboratories showing an 80% or more consistency with the reference laboratory in the detection of Chlamydia trachomatis from quality control samples (qualified), from 47.95% in 2007 to 70% in 2009 for those showing a 100% consistency (excellent), and dereased from 5.48% in 2007 to 0.71% in 2009 for those showing a consistency of lower than 60% (unqualified). The centralabs of provincial CDC and volunteer laboratories exhibited a satisfactory performance for the detection of Chlamydia trachomatis, while the performance of a small number of national STD surveillance sites needed to be increase. Conclusion The external quality assessment reveals a continuous improvement in the capability of detecting Chlamydia trachomatis in STD laboratories at different levels in China.

Objective To study the biological changes and the collagen synthesis of the primary cultured skin fibroblast treated with Cigarette Smoke Extract (CSE). Methods The morphological changes of fibroblasts after 24 hours' treatment with CSE were observed with invert microscope. The inhibitory effect at different concentrations of CSE on fibroblast activities was determined by the tetrazolium dye colorimetric test (MTT Test). The growth curves of fibroblasts treated with CSE were drawn with MTT method. Cell aging was observed with β-galactosidase, which was the biological marker of senescence. Flow cytometry (FCM) was used to estimate cell cycle phases after the fibroblasts were treated at different concentrations of CSE and different time. The mRNA expression of type Ⅰ procollagen was detected by RT-PCR. Results After the treatment, the fibroblasts displayed morphological changes and the growth of fibroblasts was apparently slowed down by CSE. The positive β-galactosidase staining was observed in the treated fibroblasts, which were affected by CSE for 5 passages. FCM analysis demonstrated that CSE decreased the cells in S phase and increased the cells in G1 and G2 phase. The result of RT-PCR showed that type Ⅰ procollagen was decreased after the treatment with CSE. Conclusion CSE can not only inhibit the growth and proliferation of the skin fibroblasts, but also decrease collagen synthesis of dermal fibroblast which is very important to the skin health.