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Objective To investigate the feasibility and prospects of nested real-time PCR(NR-PCR)technique for Treponema palladium(Tp)detection in various samples of different stages of syphilis from patients preliminarily diagnosed as syphilis. Methods Targeting the Tp polA gene, NR-PCR was performed to detect Tp DNA in various samples from the patients with various stages of syphilis at the first clinic visit, including skin tissue fluid swabs, serum, whole blood, cerebrospinal fluid(CSF)and earlobe blood. Data were analyzed with SPSS software version 13. Results A total of 368 clinical samples were collected from 200 patients with syphilis. With a detection limit of 2 Tp/ml, NR-PCR showed that the total positive rate for Tp DNA was 71.7%(264/368). The Tp DNA positive rate was highest in earlobe blood samples (92.0%, 23/25), followed by CSF samples(90.2%, 46/51), skin tissue fluid swabs(74.3%, 26/35), serum samples(66.9%, 99/148)and whole blood samples(64.2%, 70/109). There was good agreement between NR-PCR results and serologic test results, with a consistency rate of 76.0%(152/200). Furthermore, the Tp DNA positive rate did not differ between patients with primary(12/19)and secondary syphilis(14/16)in skin tissue fluid swabs(χ2 = 2.62, P > 0.05), and was slightly but insignificantly higher in patients with secondary syphilis than those with primary syphilis in the serum samples(χ2=3.6, P=0.06). The Tp DNA positive rate of whole blood samples was also higher in patients with secondary syphilis than those with any other types of syphilis. Among patients with neurosyphilis, no significant difference was observed in the Tp DNA positive rate between earlobe blood samples and CSF samples(P=0.06). Among patients with latent syphilis, the Tp DNA positive rate was significantly higher in serum samples with an RPR titer of ≥ 1:8 than those with an RPR titer of≤1:4. Conclusion NR-PCR is feasible for detecting Tp DNA in various kinds of samples, and the Tp DNA positive rate is influenced by stages of syphilis and types of samples, as well as RPR titers.
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Objective To profile the omp1 genotypes of Chlamydia trachomatis (Ct) in patients with nongonococcal urethritis (cervicitis) in Guangzhou region. Methods Swab samples were obtained from the urethra of males and cervix of females in clinical settings of venereology and gynecology as well as at outreach sites for the prevention and control of sexually transmitted diseases (STDs). DNA was extracted from the swabs and nested PCR was performed to amplify the variable domain (VD) 1 - 3 of omp1 gene of Ct followed by gene sequencing. The genotypes of Ct were determined based on the amino acid mutation in VD 1 - 2 of omp1 gene. Results Totally, 1208 swabs were collected. Of them, 132 were Ct positive, and 130 positive samples underwent genotyping. Ten ompl genotypes were determined in total, including serotype E (38, 29.23%), D (25, 19.23%), J(24, 18.46%), F(21, 16.15%), G(7, 5.38%), H(5, 3.85%), K(5, 3.85%), B(2, 1.54%), Ja (2, 1.54%), I (1, 0.77%). E, D, J and F were the dominant type of Ct in this region, and amounted to 83% of all the Ct isolates. Mutations were observed within VD 1 and 2 of omp1 gene in serotype D, B and K.Serotypes were undetermined for Ct in 2 patients with mixed infection. Conclusions In Guangzhou region, E,D, F and J are the predominant genotypes of Ct, and amount to 83% of all the Ct isolates. Ct serotype B is also observed in the urethra of males and cervix of females in this region.
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[Objective] Expressions of human epidermal growth factor receptor 2(HER-2),p53,estrogen receptor (ER),and progesterone receptor (PR) in tissues of breast invasive ductal carcinoma are not only applied to determine the therapeutic regimen,but they may also be related to the prognosis.We investigated the levels of these proteins among different clinical stages and the correlations.[Method] One hundred and thirty-eight tissues from cases with breast invasive ductal carcinoma were tested with immunohistochemistry.New scoring standards and rank test were applied.The indices were digitalized and semi-quantified.[Results] In the tissues from high clinical stage,the expression of HER2 was significantly increased,while expression of PR was markedly decreased.[Conclusion] Expression of HER2 and PR might be better markers for predicting clinical stages and prognosis.
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<p><b>OBJECTIVE</b>To study the distribution of poly(ADP-ribose) polymerase(PARP) pseudogene polymorphism and the association with susceptibility to lung cancer in Chinese people.</p><p><b>METHODS</b>The subjects of this study included 63 patients with lung cancer and 82 healthy controls matched in gender and age. Genome DNA was extracted from white blood cells. Products from PCR with a pair of specific primer were electrophoresized in agarose including EB. Under ultraviolet, observation and imaging were performed.</p><p><b>RESULTS</b>There was no significant difference in genotype between the cases and controls. The frequencies of B allele in cases and controls were 0.095 and 0.116 respectively. Whether there was B allele or not, smoking was a risk factor of lung cancer (P<0.05). As the genotype was AA and AB or BB, smoking OR was 2.28 and 4.83 respectively. Among non-smokers, the risk at lung cancer did not increase in AB or BB genotypes(P=0.202).</p><p><b>CONCLUSION</b>Frequency of B allele is relatively lower in Chinese people than in other races. In smokers, B allele may be a susceptible marker of lung cancer, and there is synergistic function between B allele and smoking.</p>