The chemical composition, separation and identification of the components as well as the tumor photobiological activities of the major components of the new tumor-photochemodiagnostic and photochemotherapeutic agent photocarcinorin (PsD-007) were represented in this paper. It has been shown by the results of HPLC analysis in combination with spectroscopic determinations that PsD-007 is composed of 7 different porphyrins: MHD, DMD, MVD, AHD, HVD, Hp and Pp. The experimental results show that MHD, DMD and MVD are the major tumor photobiologically active components of PsD-007.

Objective To establish a molecular typing method (opa-typing) for Neisseria gonorrhoeae, and to evaluate its performance based on epidemiological data. Methods Twenty-six gonorrhea patients were recruited from March to April 2006 at two sites, including 17 cases from the STD Clinic of Institute of Dermatology, Chinese Academy of Medical Sciences and Peking Union Medical College and 9 cases from the STD Clinic of Shandong Provincial Institute of Dermatology. Of the 26 patients, 6 were from three known sexual links, while the remaining 20 patients did not have any sexual contact with each other. The opa gene was amplified by using PCR from Neisseria gonorrhoeae isolates from these patients followed by overnight digestion with restriction enzymes Taq Ⅰ and Hpa Ⅱ. The enzyme digestion band patterns were analyzed using the Cel-Compar program. Results The opa gene was successfully amplified from all the 26 Neisseria gonorrhoeae strains, and restrictedly digested by endonucleases Taq Ⅰ and Hpa Ⅱ . Identical band patterns were observed between patients with sexual links, but not among the remaining 20 patients. Conclusions The results of opatyping of Neisseria gonorrhoeae coincide with the information on sexual behaviour provided by patients. Opatyping may serve as a reliable tool in sexual network analysis.

To investigate the clinical improvement of limping gait in patients with ankylotic hips and limb length discrepancy.Methods:From 1996 to 2005,12 patients with ankylotic hips and limb length discrepancy were treated by distraction osteogenesis with a mono-lateral external fixator and an intramedullary nail.The limb length discrepancy was 6.20-12.50 (median 8.45) cm.Limping gait was classified according to the recommendations of the American Academy of Orthopedic Surgeons/Hip Society and scored according to Harris:no limping scored 11 points,mild limping scored 8 points,moderate limping scored 5 points,while severe limping scored 0 points.Limping gait was severe in all patients pre-operatively and the total score was 0.Results:All patients were followed up for 30.00-46.00 (median 38.55) months,and all reported improvement in limping gait.The gain in length was 6.00-12.50 (median 8.20) cm,and the mean residual limb length discrepancy was 0-0.50 (median 0.20) cm.The total treatment time was 41.00-82.00 (median 61.50) weeks,the lengthening time was 14.00-38.00 (median 29.55) weeks.At the last follow-up,10patients had mild limping gait and 2 had moderate limping gait; the total score was 90.00.The median score was 7.50 (P25 was 8.00,P75 was 8.00).According to Wilcoxon signed rank test,the post-operative limping gait scores were significantly higher than pre-operative (P=0.001 ).Conclusion:Femoral lengthening can improve the limping gait significantly in ankylotic hips and limb length discrepancy.

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 develop a gene chip for the detection of pathogens causing genital ulcer diseases (GUDs). Methods Specific probes of 4 different pathogens were designed and synthesized. Gene chip was prepared by blotting the probes onto specially treated glass slides with the use of a robotics. Target genes of standard strains for the 4 different pathogens and the clinical specimens were amplified by PCR with Cy5 fluorescence labeled primers. The labeled amplicons were hybridized with gene chips, and then scanned and analyzed using computer software. Results The fluorescence signal for specific pathogen could be found in the gene chip, illustrating that one specific fluorescence signal denoted a single pathogen, and the combination of different signals denoted the corresponding co-existence of pathogens. Examination of 40 clinical specimens obtained from 40 patients with genital ulcers with gene chip was in good concordance with dark-field microscopy plus PCR or HSV culture plus PCR, showing Kappa values of 0.882 and 0.947, respectively. In addition, mixed infections were detected in 2 specimens. Conclusion Gene chip is a sensitive method with a reliable result and it can detect multiple infections simultaneously.

Objective To clone and express immunodominant fragment of glycoprotein G of HSV-2 (FgG-2). Methods The target gene was amplified by polymerase chain reaction (PCR). The PCR products were ligated into directional TOPO expression vector. After identification, the recombinant expression vector was transferred into BL21 StarTM cell for expression. Finally, recombinant protein of FgG-2 (rFgG-2) was detected by Western Blot (WB). Results A 616 bp DNA fragment was obtained with PCR and then confirmed in recombinant vector by PCR and sequencing, bearing 99.5% consistent sequence with target gene. Highest recombinant protein production was obtained at the time point of 3 hours. Expression of target protein was confirmed by WB with anti-gG monoclonal antibody. Conclusions The immunodominant fragment of gG-2 has been successfully cloned and expressed in E.coli, which might be used for the development of serum diagnostics assay kits for HSV-2 infection.

Objective To study the current status of antimicrobial resistance of clinical Neisseria gonorrhoeae isolates in China by analyzing the surveillance results in 2008.Methods N. gonorrhoeae strains were collected from 951 eligible patients at national monitoring sites for resistance of N. gonorrheae,including 156 patients from Jiangsu province,71 from Zhejiang province,102 from Fujian province,207 from Guangdong province,77 from Guangxi province,43 from Hainan province,80 from Sichuan province,44 from Chongqing,45 from Xinjiang Uygur Autonomous Region,72 from Shaanxi province,and 54 from Tianjin.The production of β-lactamase was detected by paper acidometric testing,and minimum inhibitory concentrations(MICs)were determined by agar dilution method for spectinomycin,ciprofloxacin,cefiriaxone,tetracycline,respectively.Results Among the 951 N. gonorrhoeae isolates,2(0.21%)were resistant to spectinomycin,451(47.42%)showed reduced sensitivity to ceflriaxone,928(97.58%)were resistant to ciprofloxacin.Penicillinase-producing N.gonorrhoeae (PPNG) and plasmid mediated tetracycline-resistant N. gonorrhoeae (TRNG) accounted for 34.91%(332/951)and 51.21%(487/951) of these isolates respectively.Kendall rank correlation analysis showed that there was a positive correlation between the positivity rate of TRNG and PPNG(r=0.20,P<0.01),but a negative correlation between the susceptibility to cefiriaxone in N.gonorrhoeae and positivity rate of PPNG(r=-0.09,P<0.01).No correlation was observed between the susceptibility to cefiriaxone and susceptibility to ciprofloxacin or the positivity rate of TRNG,or between the susceptibility to ciprofloxacin and positivity rate of PPNG or TRNG.Chi-square analysis showed a marked increase in the percentage of N.gonorrhoeae isolates with reduced susceptibility to ceftriaxone in Guangxi province,Hainan province,Xinjiang Uygur Autonomous Region and Shaanxi province,the percentage of N.gonorrhoeae isolates with resisitance to spectinomycin in Shaanxi province,prevelance of TRNG in Guangdong province,and prevelance of PPNG in Sichuan and Zhejiang provinces compared with the average level (all P<0.05).Conclusions Thero is a significant diffefence in antimicrobial susceptibility of N.gonorrhoeae from difierent areas of China.A significant elevation is observed in the percentage of N.gonorrhoeae with reduced susceptibihty to cefifiaxone and resistance to spectinomycin in Shaanxi province.to which close attention should be paid.

ObjectiveTo establish a rapid,sensitive and accurate method to detect Mycoplasma genitalium,and to evaluate the prevalence of M.genitalium among unlicensed prostitutes from Hezhou city in Guangxi Zhuang Autonomous Region.MethodsA pair of primers and Taqman MGB probe were designed and synthesized for the Pa gene of M.genitalium.Standard samples were prepared with the M.genitalium type strain G37.The established Taqman MGB real time fluorescence-based PCR assay was used to detect M.genitalium in the standard samples and cervical swab specimens collected from unlicensed prostitutes in Hezhou city of Guangxi Zhuang Autonomous Region.ResultsThe established Taqman MGB real time PCR exhibited a wide linear range( 1 × 10 copies/μl to 1 × 106 copies/μl,R2 =0.993),good repeatability(intra-assay variation;0.7％,inter-assay variation:1.09％) and hign sensitivity with the limit of detection being 10 copies/μl and limit of quantification being 50 copies/μl.As the assay showed,12.1％ of the 404 cervical swab samples were positive for M.genitalium.ConculsionThe Taqman MGB real time fluorescence-based PCR is a rapid and sensitive method for the quantitative and qualitative detection of M.genitalium.

Objective To test the ceftriaxone susceptibility of Neisseria gonorrhoeae (NG) isolates from Nanjing city,and to assess their genotypes by using the NG multi-antigen sequence typing (NG-MAST) method.Methods A total of 204 NG strains isolated in 2007 and 81 in 2012 from Nanjing city were included in this study.The minimum inhibitory concentration (MIC) of ceftriaxone was determined for these strains using an agar dilution method.DNA was extracted by the Qiagen commercial kit from these strains followed by NG-MAST.Results All the isolates were susceptible to ceftriaxone (MIC,≤ 0.25 μg/ml).The MIC of ceftriaxone was ≥ 0.06 μg/ml for 63.2％ of all the NG strains,70.6％ of those isolated in 2007 and 44.4％ of those in 2012,and ≥ 0.125 μg/ml for 31.6 ％ of all the NG strains,39.7％ of those isolated in 2007,11.1％ of those in 2012.Totally,166 genotypes were identified among the 285 isolates,of which,73 had been reported,and 93 were previously unreported.The most prevalent genotype was ST568 (n =13) in NG strains isolated in 2007,followed by ST270 (n =9),ST421 (n =7),ST2288 (n =5),ST1731 (n =4),ST1766 (n =4),ST1866 (n =4),ST1870 (n =4),while ST2318 (n =5),ST1053 (n =4),ST5990 (n =4),ST8726 (n =4) were the common genotypes in 2012.Those isolates with identical or similar genotypes tended to display similar MICs for ceftriaxone.Conclusions The prevalent genotypes of NG are markedly different between 2007 and 2012 in Nanjing region,and there is a strong association between the genotypes and ceftriaxone susceptibility of NG.NG-MAST results may serve as a genetic marker in the surveillance of antibiotic susceptibility in NG.

Objective To compare the validity of four commercially available ELISA kits for detecting HSV-2 type-specific IgG antibodies. Methods A total of 125 serum specimens were collected from 105 patients with genital ulcers and 20 normal individuals without history of STDs. Four ELISA kits which are commercially available in China for the detection of HSV-2 type-specific IgG antibodies were selected for the evaluation. Western blot assay was used as the gold standard. Results Based on the results of detection by Western blot assay, the sensitivity and specificity of these ELISA kits including home-made 1, home-made 2, improted 1 and improted 2 were 13.1% and 98.4%, 7.5% and 100%, 100% and 11.1%, 87.7% and 96.7%, respectively. The areas under the ROC curve of three kits including home-made 1, imported 1 and imported 2 were 0.885 (0.822 - 0.948), 0.852 (0.747 - 0.902), 0.947 (0.950 - 0.998), respectively. Conclusions The results of imported 2 are well consistent with those of Western blot, while the results of other 3 kits are poorly consistent with those of Western blot. It is also indicated that the commercially available ELISA kits for detecting HSV-2 type-specific antibodies should be re-evaluated in terms of their validity befor being applied for the clinical diagnosis as well as laboratory research.