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.

Objective To make a nationwide external quality assessment for drug sensitivity testing of Neisseria gonorrhoeae, analyze the problems in and factors associated with the drug sensitivity testing, and to enhance the quality of drug sensitivity testing of N. gonorrhoeae at different monitoring sites. Methods Samples were uniformly delivered to monitoring sites by express mail service. Test results were analyzed in the National Center for STD Control, and the evaluation results were fed back to these monitoring sites. Results A total of 105 quality control samples were delivered from 2007 to 2009, with a response rate of 88.57% (93/105). Thirteen monitoring sites were enrolled in the external quality assessment, including 9 laboratories in 2007, 9 in 2008 and 13 in 2009. The total percentage amounted to 77.42% (24/31) for qualified laboratories during the 3 years, including 6 laboratories in 2007, 7 in 2008 and 11 in 2009. The coincidence rate increased for the detection of penicillinase-producing N. gonorrhoeae (PPNG), N. gonorrhoeae with chromosome-mediated ciprofloxacin resistance, and N. gonorrhoeae with chromosome-mediated spectinomycin resistance, and declined for the detection of N. gonorrhoeae with plasmid-mediated high level tetracycline-resistance (TRNG) and N. gonorrhoeae with chromosome-mediated ceftriaxone resistance. Conclusions The 3-year external quality assessment reveals an improvement in the overall quality of drug sensitivity testing of N. gonorrhoeae at national monitoring sites; the accuracy is improved markedly for the detection of PPNG, N. gonorrhoeae with resistance to spectinomycin and ciprofloxacin, but is needed to increase for the detection of ceftriaxone-resis- tant N. gonorrhoeae and TRNG.

Objective To make a nationwide external quality assessment of tests for isolation and identification of N.gonorrhoeae in medical and healthcare facilities at different levels,and to analyze current problems.Methods Lyophilized quality control samples were uniformly delivered to 252 medical and healthcare facilities providing sexually transmitted disease (STD) services at different levels.Test results were analyzed by the National Center for STD Control,China Center for Disease Control and Prevention,and evaluation results were fed back to participating laboratories.Results Finally,test results were received from 203 (80.56％) facilities.The comprehensive score averaged at 87.14,and facilities achieving a comprehensive score of 80 or greater amounted to 80.30％ (163/203).The coincidence rate was 53.69％ (109/203) for all of the 5 quality control samples,82.76％ (168/203) and 87.68％ (178/203) respectively for two quality control samples containing only N.gonorrhoeae,86.21％ (175/203) and 96.06％ (195/203) respectively for a sample containing Neisseria sicca and a sample containing Enterococcus faecalis,69.46％ (141/203)for a sample containing different species of Neisseria.Conclusion The external quality assessment reveals a disparity in the capability to isolate and identify Neisseria among medical and healthcare facilities providing STD services at different levels.

Objective To explore whether there were differences in genotypes between strains of M.leprae in China or not. Methods Polymerase chain reaction (PCR) was used to detect the genotypes of M.leprae in samples from patients with leprosy in different parts of China. Results Out of 16 samples from 9 provinces, 91 bp DNA fragments were present in 7 samples and 97 bp DNA fragments in 9 samples. Conclusion Above- mentioned results suggest the genotyping differences in strains of M.leprae with different distribution in areas of China.

This paper introduces the technical parameters,basic principle and investigation method of the multiparameter supervisor.The functions and implementation methods of its components are also presented.The supervisor can provide much physiological information of the patients to medical staffs.Thus,the level of medical supervision and analysis can be enhanced.The multiparameter supervision can be realized when the microcomputer system with PC-104construction is adopted to analyze the received signals and filter the interfering signals resulting from the random variations of frequency and amplitude.

Objective To observe clinical manifestations and immune responses in rabbit models of syphilis established by inoculation with Treponema pallidum through different routes. Methods A total of 20 rabbits were randomly and equally divided into two groups: testis-inoculated group injected with Treponema pallidum (Tp) suspensions into the testes, back-inoculated group intracutaneously injected with Tp suspensions at six sites on the shaved back. All the rabbits received a single injection with the total injection amount of Tp (Nichols strains)being 2 × 107 in both groups. Clinical symptoms and immune responses were evaluated in both groups every other day. Statistical analysis was carried out by a two-sample t-test and rank sum test with the SPSS 20.0 software. Results On day 8 after the inoculation, the testes of rabbits in the testis-inoculated group started to swell with induration, and the swelling was most severe during days 13 - 16. Afterwards, the testes gradually diminished, and returned to normal in size without swelling or hardening on day 28. No skin lesions occurred in the other sites in the testis-inoculated group within 2 months after the inoculation. Erythematous swelling occurred at inoculation sites on day 7 after inoculation in the back-inoculated group, which was most obvious between days 15 and 45. Moreover, cartilage-like indurated ulcers were observed at 12 inoculation sites in 3 rabbits in the back-inoculated group, and dark-field examination of the ulcers showed the presence of Tp. There was a significant difference in the time required for the severity of lesions to peak between the testis-inoculated group and back-inoculated group (14.50 ± 1.08 days vs. 29.00 ± 10.30 days, t=5.02, P<0.05). Rank sum test showed significant differences in the distribution of highest RPR titers between the two groups(P<0.05), and RPR titers were higher in the testis-inoculated group than in the back-inoculated group. Conclusions The injection of Tp suspensions at different sites can cause different clinical symptoms and immune responses in rabbits. RPR turned positive earlier with a higher titer in the testis-inoculated group compared with the back-inoculated group. The clinical manifestations in the back-inoculated group were similar to those of chancre in primary syphilis in human.

BACKGROUND:Alcohol has become pathogenic factors of avascular necrosis, and the alcohol induced
abnormal lipid metabolism in bone marrow may be the important reason for the onset of avascular necrosis, but the mechanism is not clear yet.
OBJECTIVE:To observe the changes of structure and function of fat cel s under the action of alcohol, in order to analyze the pathogenesis of alcoholic femoral head necrosis.
METHODS:Primary adipocytes in vitro culture technique was used to obtain rabbit femoral head intramedul ary adipose tissue, and then the fat cel s were separated, and the phenotype was identified with oil red O staining. The passaged stable intramedul ary fat cel s were col ected. Coverslip was cut into 1 cm × 1 cm in size, and placed in the 24-wel culture plate before planting. The cel s were randomly divided into alcohol group and control group, 24 holes (each hole for a sample) in each group. The control group was without alcohol, while the alcohol group was added with 0.15 mol/L alcohol. At 4, 6, 8 and 10 days, the culture medium was replaced. Medium was changed and no longer adding alcohol, and then cultured for 10 days. When the culture terminated, the coverslip was removed for oil red O staining. Final y, the morphology and the number of the fat cel s were observed under light
microscope.
RESUTLS AND CONCLUSION:With time prolonging, the number of fat cel s in the alcohol group was significantly more than that in the control group (P<0.001). The lipid droplets in the two groups were gradual y increased and enlarged, but more significant in the alcohol group. The number of intramedul ary fat cel s in the alcohol group after cultured for 4, 6, 8 and 10 days was respectively (200.90±24.60), (1 102.30±76.73), (1 160.30±28.37) and (1 199.70±44.74)/cm2;the
number of intramedul ary fat cel s in the control group was respectively (99.80±10.82), (0.40±94.71), (1 000.20± 41.85) and (1 059.80±26.79)/cm2, the number of fat cel s increased with the time of alcohol influence. Alcohol can promote the intramedul ary fat cel s to increase and enlarge, and this may be the main reason for femoral head necrosis, as long-term alcoholism can lead to bone marrow fat tissue increasing, intraosseous pressure increasing and perfusion reducing, thus resulting ischemia.

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 screen primers used in polymerase chain reaction (PCR) for detecting Trichomonas vaginalis. Methods Three pairs of PCR primer reported in the literatures (TVA5-TVA6, TV1-TV2 and TVK3-TVK7) were screened. For each PCR, four components, including primers, Mg2+, dNTPs and Taq polymerase, were optimized using Taguchi methods to determine the optimal PCR conditions. With the optimal conditions, the sensitivities of three PCR were compared. Vaginal swabs were collected to detect Trichomonas vaginalis by culture and PCR, and the PCR with highest sensitivity was evaluated. Results All three PCR were of high specificity, and the PCR with primers of TVK3-TVK7 had the highest sensitivity. Of 25 clinical vaginal swabs, T. vaginalis was detected in 7 samples by the culture, however, it was detected in 8 samples by the PCR. All culture-positive samples were also positive by PCR. Conclusions The PCR with the primers of TVK3-TVK7 is highly sensitive and specific, which could be useful to detect T. vaginalis in vaginal swab samples.

Objective To genotype the clinically isolated strains of M.leprae by genome sequencing analysis. Methods PCR amplification was used to produce the 200 bp partial rpoT gene fragments from 2 standard strains and the isolated strains of M. leprae isolated from clinical specimens in 7 areas of China. The fragments were sequenced by BigDye Terminator Cycle Sequencing Reaction kit. Results① 12 rpoT－ 91/97bp DNA fragments and 2 rpoT－ 194/200 bp DNA fragments were obtained from 13 clinically isolated strains of M. leprae by PCR amplification;② based upon genome sequencing analysis, a component of 3－ copy or 4－ copy“ GACATC” repeat sequence was found in the nucleotide sequence of rpoT－ 194/200 bp DNA fragment. Conclusion The genome sequencing analysis can be used to objectively and accurately genotype M.leprae, and it is a useful tool for epidemiological study on transmission and infection of leprosy. However, the longer DNA fragments are necessary when sequencing analysis is conducted. Therefore this method needs further improvement because only the shorter DNA fragments amplified from paraffin－ embedded tissue.