Objective Clone and mutagenesis analysis of nhaA gene in Vibrio cholerae O1 and O139. Methods 40 strains of Vibrio cholerae O1 and O139 were collected. A full length nhaA gene fragment was amplified with PCR and cloned into plasmid vector pcDNA3. Homology and mutagenesis of nhaA gene in Vibrio cholerae O1 and O139 were analyzed after sequencing the nhaA gene. Results nhaA gene of Vibrio cholerae O1 and O139 were successfully amplified and cloned. Sequence analysis manifested that nhaA gene of Vibrio cholerae O1 and O139 in China share a high homology with reference sequence of wide-type Vibrio cholerae O1 in GENEBANK(99% and 96% respectively). The amino acid mutagenesis rates of nhaA gene in Vibrio cholerae O1 and O139 were 2% and 11% respectively. The important residues(Asp133,Asp163,Asp164,His225,Leu73 and Gly338 )had no mutation. But in residues 203 and 221 of nhaA gene Vibrio cholerae O1 and O139 had common mutation. Conclusions Mutagenesis of nhaA gene and NhaA protein may be the result of Vibrio cholerae adaptation to survival environment.

Objective To investigate the pathogens inducing a posto perative nosocomial infection. Methods Specimens was collected fro m exudates or air for bacteria culture and identification. Re sults The postoperative infection was induced by staphylococcus x ylosus. Conclusions The relevant factors affecting the po stoperative nosocomial infection include incomplete sterilization of operative r oom and operative tools. Thus strict control measures must be put into effect.

Objective To evaluate the therapeutic effect of transplantation of autologous melanocytes cultured with individualized medium in vitiligo. Methods Donor skin was obtained by suction blisters from a normally pigmented area of the abdomen of 155 patients with vitiligo. The roof of the blisters was clipped and digested with trypsin, then the suspension of epidermal cells and melanocytes were cultured in Hu16 medium.The cell division time (DOT) and melanin content of cultured melanocytes were measured followed by the adjustment of concentration of fetal calf serum, cytokines and cAMP elevating agents based on the DOT,melanin content and morphology of melanocytes for the individualized culture of melanocytes. After 2 - 5 passages, melanocytes were harvested and inoculated into ultrapluse CO2 laser-denuded lesions. All patients were followed up for at least 6 months. Results One hundred and fifty-five vitiligo patients with 204 lesions were treated with transplantation of autologous melanocytes. Of the 155 patients, 119 received 1 session of transplantation, 36 received 2 to 4 session of transplantation. Cells were expanded by 50 - 80 times in vitro after individualized culture. Repigmentation was more than 50% in 84.8% of these lesions, more than 90% in 52.94% of the lesions. A homogeneous skin color was obtained in repigmented skin, and no scarring or other side effects were observed. No influence was noted on the outcome of transplantation for sex, age, course of disease or lesion size of patients. Segmental vitiligo showed better response than vitiligo vulgaris: the effective rate and cure rate were 93.62% and 65.96% respectively for segmental vitiligo, 82.16% and 49.04% respectively for vitiligo vulgaris. Lesions located on the arms and legs (not including elbows and knees) showed the best response, with a cure rate of 73.08%, whereas acral sites were the most difficult area to repigment, with a cure rate of just 25.93%. Conclusions Individualized culture can significantly increase the success rate of melanocyte culture and expanding times of melanocytes. Transplantation of cultured autologous melanocytes is an effective modality deserving clinical application in the treatment of stable vitiligo, with the advantage of treating large depigmented area with melanocytes from a small donor site.

Objective:To study the possibility of dengue virus E gene vaccine.Methods:The recombinant eukaryotic expression plasmid pcDNA3 E was first transfected into NIH3T3 cells by lipofectin SDS PAGE and Western blotting analyzed the expression of E gene Then the recombinant plasmid was intramuscularly injected to BALB/c mice,and the specific humoral and cellular immunity were tested Results:The recombinant plasmid DNA could induce specific immune reactions and the immune response could last a long time Conclusion:The dengue virus E gene vaccine could induce specific immune reaction,which might have provided some material and new experimential data for the further study of dengue vaccines

Objective:To study the relationship between pre-pregnancy body mass index (BMI)and preterm birth.Methods:A case-control study was conducted in Haidian Maternal and Child Health Hos-pital in Beijing from January to April in 201 3.This study contained 1 74 preterm births in the case group and 382 term deliveries in the control group.The height,pre-pregnancy body weight,body weight before delivery,gestational weeks,history of diseases,family history of diseases,and complications during pregnancy of the subjects were collected.Multivariate Logistic regression was used to estimate the odds ratio and 95% confidence intervals (CI)after adjustment by maternal age,education,smoke during pregnancy,primiparous,mean income,and mean family living space.Results:After analyzing the rele-vant risk factors of preterm birth,the multivariate Logistic regression showed that pre-pregnancy obesity was a risk factor for preterm birth,the adjusted odds ratio was 2.461 (95% CI:1 .1 74 -5.1 59,P =0.01 7).The associations between pre-pregnancy overweight and preterm birth or pre-pregnancy under-weight and preterm birth were not found.The gestational diabetes mellitus,pregnancy-induced hyperten-sion,and family history of preterm birth were risk factors for preterm birth,the adjusted odds ratios were 1 .781 (95% CI:1 .025 -3.095,P =0.040),3.831 (95% CI:2.044 -7.1 80,P <0.001 ),and 3.675 (95% CI:1 .358 -9.942,P =0.01 0),respectively.Conclusion:Pre-pregnancy obesity ap-peared to be a risk factor for preterm birth.To decrease the incidence of preterm birth,women should improve preconception care and keep their BMI in a normal range before pregnancy.

BACKGROUND: Transformation growth factor β1 (TGF-β1) can promote bone marrow mesenchymal stem cells (BMSCs) migration and proliferation, but the underlying mechanisms remain unclear.OBJECTIVE: To observe the invasiveness of TGF-β1 on BMSCs cultured in vitro, and to investigate regulatory effect on Snail and matrix metalloproteinase 2 (MMP-2) expression.METHODS: Rat BMSCs were isolated and cultured with density gradient centrifugalization and adherence method. The influence of different concentrations of TGF-β1 on the BMSC migration was detected using the modified Transwell chambers. Small interfering RNA for Snail gene was synthesized and transfected into BMSCs by liposomel before TGF-β1 was treated, and the expression of Snail and MMP-2 before and after transfection were measured by western blot assay.RESULTS AND CONCLUSION: The exogenous TGF-β1 can induce a dose-dependent increase in cell migration, which peaked at 2 μg/L. The expression levels of Snail mRNA and MMP-2 mRNA were significantly increased after 2 μg/L TGF-β1 treatment. Snail gene can effectively inhibit the expression of MMP-2 promoted by TGF-β1. Experimental findings indicate that TGF-β1 could increase the MMP-2 expression and then promote the BMSCs migration through the upregulation of the Snail expression.

ObjectiveTo investigate the methods of treatment and effect in suppurative perichondritis of auricle.Methods27 cases with suppurative perichondritis of auricle were mentioned with sensitive antibiotic treatment in the whole and partly of the body.Carve and extradite the cases of abscess till entire debridement.ResultsAfter follow-up for 6 to 12 months,all 27 cases of auricles were preserved without feeling of causalgic.The efficient rate was 100％.Although 5 cases of auricle's shape were shrinked,the others preserved a good shape of auricle.The relief rate was 81.5％.ConclusionThe effective way of treating suppurative perichondritis of auricle was using sensitive antibiotic and entire debridement.

ObjectiveTo establish an individualized culture system for melanocytes, and to estimate its efficacy for the treatment of large-area vitiligo. MethodsHu 16 medium was used for in vitro primary culture of melanocytes isolated from patients with stable segmental vitiligo.Doubling time(DOT), melanin content (M), melanin production(MP) and number of dendrites were examined to evaluate the biological activity of melanocytes. To obtain melanocytes with better biological activity, the components of Hu16 culture medium were adjusted. Ultra pulse CO2 laser was utilized to shave the vitiligous lesions and remove the epidermis followed by autologous transplantation. Follow-up was carried out. ResultsMelanocytes were obtained from 10 patients with stable segmental vitiligo and cultured. The melanocytes from 6 patients showed relatively short DOT, stable M and MP during the first and seventh passage, and were considered to be at initial or growth stage and applicable to transplantation. The remaining melanocytes from the other 4 patients had displayed long DOT, instable M, MP and dendrite quantity since the third passage; by adjusting the components of culture medium, these cells were induced into growth stage and finally applied to transplantation. A 12-month follow-up revealed that the repigmentation rate was higher than 90％ in 7 patients, ranged between 70％ and 80％ in the remaining 3 patients, with the transplantation area being 116.8 + 75.6 cm2. ConclusionsThe individualized culture system with adjusted components in culture medium yields melanocytes with satisfying biological activity, which are proved to be effective for the treatment of large-area, segmental and stable vitiligo.

Objective To evaluate the relationship between cell seeding density and clinical efficacy of autologous cultured melanocyte transplantation in the treatment of vitiligo.Methods A total of 632 patients with vitiligo were enrolled in this study,and randomly classified into 4 groups to be treated with transplantation of autologous cultured melanocytes at 4 different seeding densities respectively,i.e.,(3.0-4.9)× 104/cm2 (n =201),(5.0-7.9) × 104/cm2 (n =303),(8.0-9.9) × 104/m2 (n =82),(10.0-12.0) × 104/cm2 (n =46).Epidermal sheets were obtained by suction blister biopsy from the normal skin of the vitiligo patients,and subjected to the isolation and culture of melanocytes.After 2 to 5 passages,the cultured autologous melanocytes were transplanted at different seeding densities to vitiligous lesions,which were abraded previously by ultra-pulsed CO2 laser,of these patients.All the patients were followed for 6-12 months.Results At 6 months after the transplantation,52.85％of these patients achieved more than 90％ repigmentation,and 82.28％ more than 50％ repigmentation,with no differences in the cure rate and response rate between the 4 groups (both P ＜ 0.05).The percentage of patients obtaining excellent color matching was significantly higher in the group treated with transplantation of melanocytes at a seeding density of (5.0-7.9) × 104/cm2 than in the other 3 groups at 6,12 and 24 months after treatment (all P ＜ 0.05),and higher in all the 4 groups at 12-and 24-month points compared with the 6-month point (all P ＜ 0.05),but no statistical difference was observed between the 12-and 24-month point in any of these groups (all P ＞ 0.05).Conclusions The transplantation of autologous cultured pure melanocytes is effective for the treatment of stable vitiligo with the optimal cell seeding density of melanocytes being (5.0-7.9) × 104/cm2,and the color matching appears to improve with time.

Objective To evaluate the therapeutic efficacy of autologous melanocyte transplantation for the treatment of vitiligo in patients with abnormal thyroid function.Methods A total of 60 patients with vitiligo were enrolled in this study,including 30 with abnormal thyroid function and 30 without.Epidermal sheets were obtained by suction blister biopsy from the normal skin of all the patients followed by melanocyte isolation and culture.After 2-5 passages of subculture,the melanocytes were transplanted onto vitiliginous lesions,which were abraded previously by ultra-pulsed CO2 laser,in the corresponding patients.All the patients were followed for 6-12 months.Results Of the 30 patients with abnormal thyroid function,7 patients achieved more than 90％ repigmentation,9 patients 50％-89％ repigmentation,53.3％ more than 50％ repigmentation,with the average repigmentation rate being 47％ within 6 months after the transplantation.Meanwhile,13 out of the 30 patients without abnormal thyroid function showed more than 90％ repigmentation,11 showed 50％-89％ repigmentation,with the average repigmentation rate being 75％.Both the cure rate and response rate were significantly higher in the patients without abnormal thyroid function than in those with (cure rate,43.3％ vs.23.3％,P＜ 0.05; response rate,80％ vs.53.3％,P＜ 0.05).Significant differences were also found in the response rate for lesions on the face or neck and for those sized more than 20 cm2 between the two groups of patients (both P ＜ 0.05).The lesions transplanted with epidermal melanocytes from the waist exhibited the lowest cure rate and response rate.Conclusion Clinical or subclinical thyroid dysfunction may have a negative impact on the efficacy of autologous melanocyte transplantation in vitiligo.