Objective To establish an in vitro model for malignant melanoma with a malignant melanoma cell line MV3 on de-epidermized dermis (DED) and to study the invasion mode of melanoma cells. Methods A human de-epidermized dermis was prepared with some elements of basal membrane (BM). Then, the reconstructed BM was identified by periodic acid schiff (PAS) staining and immunochemical staining for collagen Ⅳ. MV3 cells were seeded onto the prepared acellular dermis and maintained at the air-liquid interface for 13-15 days after 3-day submerged culture. Subsequently, the reconstructed malignant melanoma tissue was examined with hematoxylin and eosin (HE) staining and immunohistochemical staining with antibodies to S-100 protein and HMB45. Results No obvious changes were observed by naked eye in DED after the inoculation with MV3 cells. PAS staining and immunochemical staining for collagen Ⅳ confirmed the presence of BM component on the surface of DED and in the cavity of skin appendages in DED. Histological examination and immunochemical staining revealed that on the BM zone, MV3 cells grew into irregularly sized clusters; in the .cavity of skin appendages, they attached onto the BM and aggregated into circular or bandlike shape; and at the lateral side of DED, they invasively and diffusely grew, broke through the BM and intruded into the surrounding tissues of DED. The reconstructed tissue was positive for S-100 protein and weakly positive for HMB45. Conclusions The in vitro model of malignant melanoma could be reconstructed by skin organ culture system. And, the experiment suggests that BM could affect the invasive growth pattern of malignant melanoma cells.

OBJECTIVE To understand drug susceptibility of Ureaplasma urealyticum(Uu) in the local region,and provide the scientific basement for the clinical use of antibiotic.METHODS The clinical specimens of 1108 cases were detected by bioM?rieux Mycoplasma IST2 kit.RESULTS The total positive detection rate of Uu was 54.24%(61.17% in female and 23.53% in male).Antibiotics susceptibility was as followed: josamycin(JOS) 99.00%,doxycycline(DOT) 98.17%,ofloxacin(OFL) 20.63%,erythromycin(ERY) 76.20%,tetracycline(TET) 96.84%,ciprofloxacin(CIP) 3.49%,azithromycin(AZI) 76.87%,clarithromycin(CLA) 87.35%,and pristinamycin(PRI) 99.67%.CONCLUSIONS The infection rate of Uu in the local region is higher than that in the other region of the country.And infection rate in female is significantly higher than that in male.

Objective To investigate the expression level of miR-133b in cancer tissues of patients with prostate cancer and its effect on the proliferation of prostate cancer cells.Methods The total RNAs in resected prostate cancer tissues and adjacent tissues from 30 patients with prostate cancer were extracted and reversely transcripted into cDNA,and then the expression levels of miR-133b were detected by real-time quantitative PCR.The correlations between the expression levels of miR-133b and the patients' clinicopathological features were analyzed.The expression of positive regulatory domain I-binding factor 16 (PRDM16) and proliferation of PC-3 cells transfected with miR-133b mimics by LipofectamineTM 3000 were determined by real-time quantitative PCR and the CCK8 method,respectively.Results The expression levels of miR-133b in prostate cancer tissues [(16.85 ± 0.94) × 10-4] was significantly lower than that in adjacent tissues [(22.95 ± 1.567) × 10-4,t =3.335,P ＜ 0.01].The expression levels of PRDM16 in PC-3 cells transfected with miR-133b mimics were significantly lower than that in the control group (0.371 ±0.031 vs 1.000 ±0.022,t =12.53,P ＜ 0.01).The proliferation ability of PC3 cells transfected with miR-133b mimics for 72 hours was significantly lower than that in the control group (t =6.811,P ＜ 0.01).Similarly,the proliferation ability of PC-3 cells transfected with PRDM16 inhibitor for 72 hours was also significantly lower than that in the control group (t =9.048,P ＜0.01).Conclusion The expression levels of miR-133b in prostate cancer tissues are significantly down-regulated,which regulate the proliferation of prostate cancer cells possibly through PRDM16.

Objective: To observe the therapeutic effect of treating senile intervertebral disc Herniation with acupuncture, tuina and traction and with physiotherapy and traction. Methods: Altogether 150 cases above 65 years old were randomly allocated into treatment group and control group, each containing 75 cases. Traction, acupuncture and tuina therapy were adopted in the treatment group, while traction and physiotherapy were adopted in the control group. Results: After one-month treatment, the recovery rate of the treatment group were 70.7%, which was significantly better than 53.3% of the control group (P＜ 0.05). Conclusion:Combination of acupuncture, tuina and traction has very good efficacy in treating senile intervertebral disc Herniation.

ObjectiveTo investigate the action mechanism of Th17 cells in immunoinflammatory response in systemic lupus erythematosus(SLE).MethodsReverse-transcription PCR was performed to measure the mRNA expre ssion of the retinoic acid receptor-related orphan nuclear receptor RORγt in peripheral blood mononuclear cells (PBMCs) from 12 patients with active SLE,9 patients with inactive SLE and 12 normal human controls.Data were statistically analyzed by approximate F test(Welch test) and Dunnett's T3 multiple comparison test (corrected).ResultsIn the case of RORγt mRNA expression in PBMCs,significant differences existed among the 3 groups(F =23.286,P ＜ 0.01 ); in detail,the patients with active SLE were significantly higher than patients with inactive SLE and normal controls ( 1.06 ± 0.44 vs.0.65 ± 0.25,F =2.453,P ＜ 0.05;1.06 ± 0.44 vs.0.22 ± 0.08,F =6.504,P ＜ 0.05),and the patients with inactive SLE were significantly increased compared with the normal controls (F =3.343,P ＜ 0.05).The expression level of RORγt mRN A was significantly positively correlated with SLE disease activity index (rp =0.623,P ＜ 0.01 ).ConclusionsThere is a polarization of Th17 cells in patients with SLE.To antagonize the transcription factor RORγt,which plays an essential role in the regulation of Th17 cell differentiation,may facilitate the control of SLE via attenuating the immunoinflammatory response.

Objective To establish a method for the determination of peoniflorin and glycyrrhizic acid in modified Shaoyao Gancao Granules.Methods Reversed-phase high performance liquid chromatography(HPLC) was used.The chromatographic condition was as follows: Diamonsil C18 column(4.6mm ?250mm,5?m),column temperature being room temperature,mobile phase consisting of acetonitrile(A)-10g/L acetic acid(B) with adjusting pH value being 3.5,flowing rate at 1.0mL/min,detective wavelength at 250nm,and the injection volume being 20?L.The gradient elution procedure was as follows: elution with 18% A for 0～5 min,elution with 19%～29% A for 6～10 min,elution with 30%～45% A for 11～25min,and A+B=100%.Results Peoniflorin at the concentration of 1.916～122.6?g/L and glycyrrhizic acid at the concentration of 2.625～168?g/L showed a good linearity.The mean recovery was 101.12% for peoniflorin and 99.68% for glycyrrhizic acid.Conclusion This method is simple and reproducible,and can be used for the determination of peoniflorin and glycyrrhizic acid in modified Shaoyao Gancao Granules.

Wet age-related macular degeneration is the leading cause of the severe, irreversible vision loss in individuals over the age of 65 years. Anti-VEGF therapy has been shown to play a key role in the pathogenesis of wAMD, which is the front-line therapy admittedly. lt has no clear steady curative effect for some patients even if they accepted repeating Anti-VEGF therapy. For the purpose of visual acuity improved more, a steady flow of new therapy has emerged, such as function towards the same or different targets of antiangiogenesis, consolidating the effect by combination therapy, improving or simplifying the mode of administration, etc. This paper gives a brief review of the progress of anti-VEGF for the therapy in wet age-related macular degeneration.

26 heterotrophic bacteria strains were isolated from grown abalone digestion guts and their fanning waters in Jiansheng Abalone Farm in Shangwei, Guangdong province. Analyses of extra-cellular pathogenic factors were performed and API 20 E strips were employed to identify all the isolates. Results indicated that isolates from digestion guts displayed greater ability of producing protease , amylase, gelatinase and/or hemolysis than those from farming waters, while their ability of producing lipase and phospholipase were lower than the later. Regardless of their source of origins, there were some isolates which had great abilities of producing extra-cellular products and most of them were Sphingomonas paucimobilis. Therefore, Sphingomonas paucimobilis should be considered as an opportunistic pathogen in the abalone fanning environments, while the digestion guts and fanning waters should be both regarded as the sources of harboring potential pathogens. In addition, apart from predominant strains, the roles of extra-cellular products of the bacteria community as a whole should be taken into consideration when dealing with fish diseases.

Child, Preschool ; Humans ; Male ; Salmonella Infections ; Salmonella enterica

Objective To explore the optimal semi-quantitative uptake ratio for differentiation between benign and malignant lung lesions with 18F-FDG coincidence imaging.Methods One hundred and thirty-seven patients (89 males,48 females,age range:33-78 years) with lung diseases underwent 18 FFDG coincidence imaging.The maximum radioactivity counts of the lung lesions (T),normal chest wall soft tissues (NT1) and the contralateral lung tissue (NT2) were measured.The ratios of R1 (T/NT1) and R2(T/NT2) were calculated.The optimal threshold values of R1(cutoff)and R2(cutoff) were identified by ROC curve analysis.Using the optimal threshold,the sensitivity,specificity and accuracy were calculated.Results The optimal threshold values R1(cutoff)and R2(cutoff) were identified as 3.58 and 4.40.The sensitivity,specificity and accuracy were 90.0％(90/100),89.2％(33/37),89.8％(123/137) according to R1(cutoff) and 90.0％ (90/100),78.4％(29/37),86.9％(119/137) according to R2(cutoff) Conclusion Based on which the chest wall soft tissue is taken as a reference point,the optimal threshold value of T/NT1 is 3.58 in differentiation between benign and malignant lung lesions with 18F-FDG coincidence imaging.