Objective To study the role of Th1/Th2 cytokine profile in the pathogenesis of recurrent genital herpes (RGH), and to find out the relationship between Th1/Th2 cytokines. Methods A two－ colour immunofluorescent staining of cell surface antigen and intracellular cytokines (IL－ 2、 IL－ 10、 IL－ 12、 IFN－ γ and TNF－ α ) in CD3＋ T－ lymphocytes of 20 patients with RGH with flow cytometric analysis were performed. Results Compared to controls, patients with RGH showed a decreased number of CD3＋ T cells（ P0.05） . Conclusions RGH patients seem to have a predominance of Th2 cytokine profile, which may play an important role in the pathogenesis of RGH.

Objective To investigate the antimicrobial susceptibility of Neisseria gonorrhoeae isolated from 10 cities of China, and to provide reference for the formulation of treatment guideline and making control strategy. Methods Agar dilution was used to detect antimicrobial susceptibility and acidometric method for PPNG testing. Results A total of 3186 gonococcal isolates were tested from 1993 to 1998. The resistant rate for penicillin was 66.70％ . PPNG isolates accounted for 8.14％ . Tetracycline－ resistant isolates accounted for 93.02％ , and high level tetracycline－ resistant isolates (TRNG) accounted for 4.65％ . The resistant rate for ciprofloxacin was also high (34.25％ ). The resistant rates for spectinomycin and ceftriaxone were 0.44％ and 0.57％ respectively. Conclusions The gonococcal isolates are highly resistant to penicillin, tetracycline, and ciprofloxacin, while still sensitive to spectinomycin and ceftriaxone. These results suggest that the present treatment for gonorrhea can still be used.

Objective To investigate whether baicalein inhibits the proliferation, cell cycle of and pseudopod formation in A431, a skin squamous cell carcinoma cell line, by suppressing the expression of ezrin protein. Methods A431 cells were grouped to be transfected with ezrin-targeting siRNA (siRNA group), treated with baicalein of 5, 10, 20, 40 μmol/L, respectively (baicalein group), or remain untreated (control group). After additional culture, wound healing assay and Transwell assay were performed to observe the migration and invasion of A431 cells, RT-PCR to detect the mRNA expression of ezrin in A431 cells, Western blot and immunoflu-orescence to measure the expression of ezrin protein and its phosphorylation. The pseudopod formation in A431 cells was observed by using scanning electron microscopy. Results After 24-hour culture, wound healing assay displayed that the percent wound closure was 13.3 ± 1.7, 7.6 ±1.6 and 5.9 ± 1.3, respectively, in A431 cells treated with baicalein of 5, 10, 20μmol/L, significantly lower than that in untreated A431 cells (16.3 ± 2.3, all P < 0.01), and the inhibition of baicalein on the migration of A431 cells was concentration-dependent. In the Transwell assay, a significant decrease was observed in the number of A431 cells per high power field permeating through the artificial basement membrane in the groups treated with baicalein of 5, 10, 20 μmol/L for 48 hours compared with the control group (46.5 ± 3.8, 34.3 ± 3.4, 25.3 ± 2.3 vs 56.3 ± 3.8, all P < 0.01), whereas no significant difference was noted between these baicalein-treated groups and siRNA-transfected group (28.3 ± 2.1, all P > 0.05). RT-PCR analysis showed that the mRNA expression of ezrin in baicalein-treated A431 cells significantly decreased compared with that in untreated cells (all P< 0.01), but showed no difference from that in siRNA group (P > 0.05). A statistical difference was also observed in the expression of ezrin and phosphorylated ezrin protein between baicalein-treated A431 cells and untreated cells (all P< 0.05), but not between 40 μmol/L baicalein-treated A431 cells and siRNA-transfected cells (P> 0.05). Furthermore, the suppression of baicalein on ezrin protein and mRNA expression was concentration dependent. The number of pseudopod per cell was significantly lower in 20 μmol/L baicalein-treated A431 cells and siRNA-transfected cells than that in untreated A431 cells (5.3 ± 1.9, 4.5 ± 2.8 vs 22.6 ± 2.8, both P < 0.01), while no significant difference was observed between the former two groups of cells (P > 0.05); the length of pseudopodia also reduced in baicalein-treated cells. Conclusions Baicalein may inhibit the proliferation and invasion of A431 cells by directly or indirectly suppressing the expression of ezrin and phosphorylated ezrin, which in turn contributes to the effect of baicalein against tumor proliferation and metastasis.

Objective To profile the phenotype and pathogens of cutaneous and mucous mycoses in a dermatology outpatient clinic in Nanchang region. Methods A review was performed to assess cutaneous and mucous mycoses diagnosed in the dermatology outpatient clinic of Dermatology Hospital of Jiangxi Province from 2006 to 2008. The relationship of clinical phenotype and pathogens to season, patients' age and gender was analyzed. Results A total of 7251 cases were collected, and the ratio of male to female patients was 2.3: 1. The most prevalent mycoses included tinea cruris (2702, 37.1%), pityriasis versicolor (1505, 20.8%) and tinea manus (727, 10.0%). In total, 4953 fungal strains were isolated from all the patients except for those with pityriasis versicolor, of them, Trichophyton rubrum accounted to 69.9%, Candida to 20.4%, and Trichophyton violaceum to 4.5%. Season, patients' age and gender were found to be associated with clinical phenotypes and pathogens of mycoses. Conclusions In the dermatology outpatient clinic of Nanchang region, tinea cruris is the most common superficial fungal disease, with the predominant pathogen being Trichophyton rubrum. Trichophyton violaceum is the primary pathogen of tinea capitis, which is different from other reports.

Objective To retrospectively assess the clinical manifestations and treatment of secondary syphilis with ocular impairment as the initial symptom. Methods Clinical data were retrospectively analyzed on secondary syphilis with ocular impairment as the initial symptom collected from September 1998 to October 2008. Results There were 11 syphilitic patients presenting acute ocular impairment as their initial manifestation. Skin eruptions developed simultaneously with ocular impairment in 2 patients, following ocular impairment in 9 patients. All patients were positive for rapid plasma reagent test (RPR) and treponema pallidum haemagglutination assay (TPHA), but negative for HIV. Of these patients, 9 suffered from uveitis (iridocyclitis, choroiditis or panuveitis), 2 from optic neuritis; 3 had unilateral ocular involvement, 8 had bilateral ocular involvement. After treatment with injected penicillin or ceftriaxone sodium, 9 patients experienced complete visual recovery, 2 partial visual recovery. Conclusions Ocular impairment occurs in patients with secondary syphilis at a low incidence, with no characteristic clinical manifestations. For patients who have no response to conventional ocular therapy, ocular syphilis should be considered and serological examination for syphilis is recommended.

Objective To study the effect of down-regulation of PTTG on the proliferation and migration of cutaneous squamous cell carcinoma cell line SCL-1 and its related mechanism. Methods SCL-1 cells were transfected with control siRNA or PTTG-targeting siRNA (PTTG-siRNA), or remained untransfected. After additional culture, the proliferation of SCL-1 cells as observed with cell counting kit-8 (CCK-8), and cell migration with Boyden chamber. Real-time PCR and Western blot were performed to detect the expression of matrix metalloproteinase 2 (MMP-2), MMP-9 and PTTG. Results The proliferation of SCL-1 cells transfected with PTTG-siRNA was markedly deccelarated in comparision with that of untransfected cells and those transfected with control siRNA (both P< 0.05). Real-time PCR and Western blot disclosed a significant decrease in the mRNA and protein expression of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells compared with the other two groups of cells. As real-time PCR showed, the expressions of MMP-2, MMP-9 and PTTG in PTTG-siRNA-transfected SCL-1 cells were 0.8%, 23.2% and 21.3% of those in untransfected cells, respectively. Further more, the number of SCL-1 cells migrating through microporous membrane in the Boyden chamber was significantly smaller in PTTG-siRNA-transfected group than in untransfected group and control siRNA-trans-fected group (51.38 ± 4.71 vs 131.33 ± 6.12 and 127.72 ± 5.20, both P< 0.05). Conclusion The down-regulation of PTTG may deccelarate the proliferation and migration of SCL-1 cells and inhibit the expression of MMP-2 and MMP-9 in SCL-1 cells.

Objective To investigate the relationship between the methylation of CpG island of RUNX3 gene promoter and its expression in a human cutaneous malignant melanoma cell line A375, and to assess the role of RUNX3 gene methylation in the pathogenesis of human cutaneous malignant melanoma. Methods Cultured A375 cells were treated with various concentrations (0, 1, 5, 10, 20 μmol/l) of 5-azacyti-dine for 24 or 72 hours followed by another 5 days of culture. Then, methylation-specific PCR (MSP) was performed to evaluate the methylation status of RUNX3 promoter region, and Western-blot analysis to detect the protein expression of RUNX3 in A375 cells. Results The RUNX3 gene promoter region was hypermethylated in untreated A375 cells, along with the absence of protein expression of RUNX3. However, after the treatment with 5-azacytidine, the promoter region of RUNX3 gene was demethylated partly, and the expression of RUNX3 protein was restored in A375 cells. Further, the expression intensity was directly correlated with the concentration of 5-azacytidine. Conclusions The promoter hypermethylation of RUNX3 gene may be related to the silencing of RUNX3 gene expression in A375 cells, whereas 5-azacytidine can cause the demethylation of RUNX3 gene, reactivate the gene which has been inactivated by the promoter hypermethylation, and finally induce the re-expression of RUNX3 protein.

Objective To screen, analyze and predict microRNAs (miRNAs) related to systemic scle-roderma (SSc). Methods Differentially expressed miRNAs between tissue samples from 3 patients with SSc and 3 normal human controls were screened with a gene chip including 924 miRNAs. Target genes regulated by differentially expressed miRNAs were searched with bioinformatics method. Finally, miRNAs related to SSc were predicted. Results There were 24 miRNAs differentially expressed between tissue samples of SSc and normal controls, including 9 up-regulated miRNAs and 15 down-regulated miRNAs. Literature review disclosed that SSc was associated with target genes regulated by hsa-miR-206, has-let-7g, hsa-miR-133a, hsa-miR-125b, hsa-miR-40-5p and hsa-miR-23b. In particular, 15 target genes regulated by hsa-miR-206 were closely correlated with the pathogenesis of SSc. Conclusions In lesions of SSc, there is an expression of miRNAs related to the pathogenesis of SSc, which may include hsa-miR-206 as well as 5 other miRNAs.

Objective To measure the expressions of IL-10, IL-23 and CD86 in lesions of epidermodysplasia verruciformis (EV), and to explore the relationship between cellular immune abnormality and EV pathogenesis. Methods Immunohistochemistry was used to measure the expressions of IL-10, IL-23 and CD86 in tissue samples from 10 patients with EV and 10 normal human controls. Results Three cytokines were observed in all the samples of EV, with the expression score ranging from 3 to 6 and expression intensity from moderate to high. However, of the control specimens, only 1 was positive for IL-10 with the expression score being 3, and expression intensity being moderate. Conclusion The pathogenesis of epidermodysplasia verruciformis may be correlated with the expression abnormality of some cytokines secreted by keratinocytes.

Objective To study the survival and melanogenic potential of human melanocytes reversibly immortalized via SV40T antigen gene and Cre/loxP system in Guinea pigs. Methods The supernatants of retrovirus vector Cre-ERT2 were used to infect melanocytes which had been successfully transfected by SV40TAg gene (MCT), then the expression of Cre recombinase was induced with tamoxifen in infected cells; subsequently, the surviving cells, which were named as MCTC, were subjected to expansion culture. Guinea pigs were utilized to establish animal models of vitiligo, then MCTC and primary melanocytes were transplanted respectively into the animal models. The repigmentation at the transplanted area was observed with naked eyes successively until 3 months after the transplantation when tissue samples were obtained from implanted area and nonimplanted area of guinea pigs and subjected to Masson-Fontana silver stain and Hematoxylin-eosin stain for the analysis of melanocyte distribution and melanin deposition in epidermis. Results Repigmentation started 4 weeks after the transplantation, and dark or brown patches, which ranged in size from 0.5 to 1 cm, were observed in the implanted area 3 months after the transplantation. The repigmentation rate was of no significant difference between pigs transplanted with MCTC and those with primary melanocytes (82.5% vs 76.7%, P > 0.05). Pathological examination revealed melanin deposition in the basal layer of epidermis and some hair follicles in transplanted area. Conclusions SV40T antigen gene combined with Cre/loxP site-specific recombinase system can induce the reversible immortalization of human melanocytes, and the immortalized melanocytes have a favorable profile of biological safety and similarity in survival rate and melanogenic potential to primary melanocytes.