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 investigate the characteristics and clinical application value of anti-double stranded DNA antibody de-tected by Crithidia indirect immunofluorescence assay method and enzyme linked immunosorbent assay method.Methods Eighty-five patients with systemic lupus erythematosus,20 disease controls and 75 healthy controls were selected.The serum anti-double stranded DNA antibody was detected simultaneously by the methods of Crithidia indirect immunofluorescence assay and enzyme linked immunosorbent assay and their diagnostic efficacies for detection were compared.Results For each method the positive rate in the systemic lupus erythematosus group was significantly higher than that in the disease control group and healthy control group. The difference had statistical significance (P <0.05).The positive rates of Crithidia indirect immunofluorescence assay and enzyme linked immunosorbent assay in the systemic lupus erythematosus group were 72.94% and 88.24% respectively,and the positive predictive value of enzyme linked immunosorbent assay is lower(P <0.05).Meanwhile the anti-double stranded DNA antibody con-centrations detected by enzyme linked immunosorbent assay method showed statistically significant difference among the active sys-temic lupus erythematosus group,the stable systemic lupus erythematosus group and the control group (P <0.05 )and presented linear trend.Conclusion Using Crithidia indirect immunofluorescence assay method to detect anti-double stranded DNA antibody for the systemic lupus erythematosus group has high specificity and is helpful for the diagnosis of systemic lupus erythematosus. Enzyme linked immunosorbent assay can be used to detect anti-double stranded DNA antibody concentration quantitatively,which is linearly related with systemic lupus erythematosus activity and the method is of high sensitivity,which can effectively screen the pa-tients with systemic lupus erythematosus.

Objective To investigate the possible regulating effect of integrin-linked kinase (ILK) towards matrix metalloproteinase-9/tissue inhibitor of metalloproteinase-1 (MMP-9/TIMP-1) ratio in the process of transforming growth factor-β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) in human kidney proximal tubular epithelial (HK-2) cells.Methods HK-2 cells were cultured and stimulated with 10 ng/ml TGF-β1.Specific ILK-small interfering RNA (ILK-siRNA) was used to inhibit ILK expression.The characteristic epithelial marker (E-cadherin) and mesenchymal marker (α-SMA) of EMT were examined by Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) and Western blot.The expressions of ILK,MMP-9,and TIMP-1 were also examined,to determine the regulating effect of ILK towards MMP-9/TIMP-1 ratio.Results In the HK-2 cells cultured with TGF-β1,the expression of E-cadherin decreased,and α-SMA expression increased;overexpression of ILK and an abnormal changing of MMP-9/TIMP-1 ratio were observed.ILK inhibition by ILK-siRNA could adjust MMP-9/TIMP-1 ratio to near normal.Meanwhile,the overexpressed ILK and α-SMA were decreased.Conclusions Our data indicates that ILK-siRNA successfully inhibits ILK expression,which regulates the MMP-9/TIMP-1 ratio in HK-2 cells.The inhibition of ILK expression suppresses TGF-β1-induced EMT partially.

Humans are exposed to the ubiquitous radiofrequency (RF, 100 kHz-300 GHz) electromagnetic fields because of the mushroom development of wireless communications,raising concerns over the possible hazards of RF radiations.Epidemiological investigation has showed that chronic use of cellphones increases the risk of brain tumors.Since genetic damage is closely related to tumors, researchers have been trying to find out whether cellphones and other RF devices are genotoxic.However, the investigations have yielded both negative and positive results.This review summarized the recent in vitro and in vivo researches about genotoxicity of RF radiations and proposed a possible mechanism by which of RF radiations cause genetic damage.

Objective:To compare color level among Vita,Shofu and Dentsply shade guide. Methods:Vita,Shofu and Dentsply shade guide were scanned into a computer and saved as BMP pictures. L *, a * and b * values of the color of the images were measured by Photoshop. Results:There was same trend of variation in color of the shade guides, but the maximum and minimum of L *, a * and b * in those shade guides were different. Conclusion: For matching color accurately, one shade guide can not be used to replace the other one in clinical application.

Objective To discuss the allocation of oral medical resource in west area of Chongqing,and investigate the caries prevalence in these population.Methods Study samples was raised with the multistage stratified random cluster sampling method,then data was analyzed to compare the allocation of oral medical.Resource,awareness degree on oral health and caries prevalence in each age grade between urban and rural area.Results In west area of Chongqing,the allocation of oral medical resource and awareness degree on oral health were better in urban area than those in rural area.In each age grade,the caries prevalence is higher in rural area,when compared with that in urban area.In addition,the caries prevalence of 5 years old group and 12 years old group is the same between male and female in both urban area and rural area (P＞0.05).Nevertheless,in 35-44 years old and 65-74 years old group,the caries prevalence was higher in female compared with male in both urban area and rural area (P＜0.05).Conclusion In west area of chongqing,the rural allocation of oral medical resource is bad and needs improvement,the awareness of oral hygiene is weak among population of rural area.In west area of Chongqing,women in 35-44 years and 65-74 years old should pay more attention to caries prevention and treatment.

Heilongjiang Provincial Hospital has scored an initial success in dealing with medical disputes since the medical liability insurance and third-party mediation mechanism were introduced into the hospital in 2013.The paper identified problems found in the practice,and recommended the following:rationalizing the cost of insurance coverage,expanding scope of third-party mediation properly,enhancing professional authority in assessment of medical dispute cases,simplifying insurance compensation procedure,and consolidating the legal status of the medical dispute mediation institutions,for better resolution of such disputes.

ObjectiveTo investigate the therapeutic effect of human umbilical cord mesenchymal stem cell-paracrine substance on fulminant hepatic failure (FHF) rat, and to study the effect on liver function and hepatocyte proliferation. MethodsMesenchymal stem cells(MSCs)were separated from human umbilical cord, and surface makers of cells were detected by flow cytometry. Human umbilical cord mesenchymal stem cells-conditioned medium(MSC-CM) was prepared. FHF rat model was induced by intraperitoneal injection of D-galactosamine and they were randomly diveded into three groups: MSC-CM group, NS group, PHGF group. 24 h later, 1 ml MSC-CM, 1 ml 0. 9% NaCl solution and lml PHGF solution was injected into the tail vein of MSC-CM, NS, and PHGF rats, respectively. In each group (n=8 per group), blood samples were collected at 12, 24, 36, and 60 h after treatment from inner canthus for analysis of blood ALT and TBIL levels. We used five rats per group for tissue collection after sacrifice at 36 h after treatment and 10 animals per group for survival analysis. PCNA immunohistochemical staining was used in the sections of liver tissue to detect hepatocyte proliferation. Results24 h after treatment, the levels of ALT and TBIL in the MSC-CM and PHGF groups were lower than those in the NS group(P＜0. 05), but there was no significant difference between the MSC-CM and PHGF groups. There were more PCNA-positive hepatocytes in the MSC-CM and PHGF groups than in the NS group(P＜0.01), but there was no significant difference between MSC-CM and PHGF group. Survival analysis found that the survival rate of rats in the MSC-CM and PHGF groups was higher than that of rats in the NS group (P=0. 049), but there was no significant difference between the MSC-CM and PHGF group. ConclusionsThe paracrine substance of human umbilical cord mesenchymal stem cells can stimulate hepatocyte proliferation and improve liver function of FHF rats, potentially creating a new avenue for the treatment of FHF.

OBJECTIVE: Panel reactive antibody (PRA) can induce acute rejection following kidney transplantation, however, it is poorly understood which PRA is more associated with rejection. Therefore, the aim of this study is to analyze the correlation between PRA and rejection by observing the change of PRA Ⅰ and PRA Ⅱ prior to and after the kidney transplantation. METHODS: Levels of PRA Ⅰ and PRA Ⅱ were observed in 100 patients received kidney transplantation at the Department of Urology, Beijing Chaoyang Hospital Affiliated to the Capital Medical University. During these 100 patients, 18 patients had PRA changes after operation. The relationship between PRA changes after kidney transplantation and acute rejection were analyzed. RESULTS: Totally 18 patients were included in the final analysis. Nine of them occurred acute rejection with obviously increased PRA Ⅱ (P=0.040), however, the PRA Ⅰ had no significant changes (P=0.707). The changes of PRA Ⅰ and PRA Ⅱ had no significance in the remaining 9 patients prior to and after kidney transplantation. The overall level of PRA increased in 7 patients, in 5 patients with increased PRA Ⅱ, 4 patients suffered acute rejection, 1 of which was renal allogreft failure; 2 cases with PRA Ⅰ increasing did not occur acute rejection. The overall level of PRA declined in 11 patients, including 5 patients with PRA Ⅱ decreased, 1 patient occurred acute rejection; 4 patients in 6 patients with PRA Ⅰ declined suffered acute rejection. CONCLUSIONS: The increased PRA Ⅱ after transplantation easily result in acute rejection, which has definite correlation to acute rejection, however, the PRA Ⅰ changes has no impact on acute rejection.

Objective To construct tissue-engineered skin via in vitro inoculation of epidermal stem cells(ESCS)onto de-epidermized dermis.Methods Skin tissue was obtained from the foreskin of a healthy 6-year-old child.and keratinocytes were isolated by two-step trypsinization method followed by the collection of ESCS via rapid adhesion by collagen Ⅳ.The ESCS were identified by morphological observation and immunohistochemical staining with K19 and integrin β1.To construct tissue-engineered skin,selected ESCS were seeded onto the surface of de-epidermized dermis followed by a one-week culture immersed in the medium and a subsequent 4-week culture at the air-medium interface.The tissue-engqneered skin was evaluated with haematoxylin & eosin(HE)staining as well as keratin immunohistochemistry.Results Micro scopically,cultured ESCs showed a paving stone-like appearance and grew into colonies.Immunohistochemistry revealed that the ESCs were positive for integrin-β1 and keratin 19.After 5 weeks of culture,3-6 layers of epidermal cell were observed on the dermis with the formation of stratum corneum.Keratin protein was observed in the artificial epidermal skin.Conclusion Tissue-engineered skin is successfully constructed with epidermal stem cells and de-epidermized dermis in vitro.