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.

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.

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.

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 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.

Objective To investigate the effect of JUC on the incisions for episiotomy.Methods Three hundred primiparas to undergo episiotomy in our hospital were divided into two groups in equal number.The experiment group was given JUC Spray before suturing and the control group did not use any solution.In the two groups,antibiotics were not used after the operation,and the incisions were only cleaned with 0.5%povidone-iodine 2 times a day.Result There were significant differences between the two groups in terms of postoperative pains,inflammation and healing in the wounds,and hospital stay(P<0.05).Conclusions Application of JUC after episiotomy could be long-acting in antibacteria.It can reduce wound pain,improve wound healing rate, decrease the medical cost and shorten the hospital stay.

Objective:To investigate the inhibitory effects of halofuginone on radiation-induced pulmonary injury and to explore the therapeutic mechanism of this drug. Methods:A total of 72 healthy female C57BL/6 mice were randomized into 4 groups, namely, control, irradiation, halofuginone, and irradiation plus halofuginone groups, with 18 mice in each group. No treatment was performed in the control group. In the halofuginone group, the halofuginone lavage was conducted once a day, with a continuous course treatment for a month or until sacrifice of the mice. In the therapeutic alliance group, the treatment mode was the same as that in the halofuginone group. Then, a 6MV-X ray single fraction irradiation was performed after the completion of a 15-day intragastric administration. At 24 h, 1 week, 2 weeks, 4 weeks, 12 weeks, and 20 weeks after the irradiation, 3 mice from each group were randomly sacrificed, and total lung tissues were harvested. The lung was dissected to prepare pathological sections. The sections were stained with hematoxylin and eosin staining (H&E) to explore morphologic changes. The protein and mRNA expression levels of TGF-β1 were analyzed by a combi-nation of immunohistochemistry and polymerase chain reaction. The level of hydroxyproline was also measured. Results: The out-comes of H&E staining showed that halofuginone markedly ameliorated the acute pulmonary inflammation and fibrosis induced by irra-diation. The combination group had a lower level of hydroxyproline than the irradiation group, with statistically significant differences at 20 weeks after irradiation (P=0.037). The protein and mRNA expression levels of TGF-β1 were higher in the irradiation and combi-nation groups than in the control group and (or) halofuginone group at different time points (P<0.05). The combination group had lower TGF-β1 protein expression than the irradiation group at different time points, with statistically significant differences at 2, 4, 12, and 20 weeks after the irradiation (P<0.05). Meanwhile, TGF-β1 mRNA level was lower in the combination group than in the irradiation group only at 4 and 12 weeks after the irradiation (P<0.05). Conclusion:Halofuginone can ameliorate the irradiation-induced lung inflamma-tion and fibrosis probably by inhibiting the radiation-induced TGF-β1 expression. Therefore, halofugione is expected to be a therapeu-tic drug for preventing irradiation injury of the lung.

This study was aimed to explore pharmacological effects of epimedium pubescen flavonoid (EPF) on biomechanical properties among ovariectomized rats. Sixty female Sprague-Dawley (SD) rats (aged 2-month-old) were randomly divided into six groups (n = 10), which were the sham control group (Group A), the model group (GroupB), the standard group (Group C), the treated 1 group (Group D), the treated 2 group (Group E), and the treated 3 group (Group F). Except the sham control group (Group A), rats in other groups had been ovariectomized. All rats were given the same feedstuff. Meanwhile, Group C was given calcium 75 mg·kg-1 combined with VitD3 21 IU·kg-1 by gastrogavage every day for 4 months; Group D was given EPF 75 mg·kg-1; Group E was given EPF 150 mg·kg-1;Group F was given EPF 300 mg·kg-1. At the end of the 4th month, all rats were sacrificed. Bones, which included tibia, femur and humerus of both sides and all lumbar vertebra bodies, had been taken out. Measurement was made on the elastic modulus, maximum loading capability, maximum stress, potential energy of deformation, and structural rigidity of biomechanical properties of the fourth lumbar vertebra body (LV4); the maximum loading capability, bone break load, potential energy of deformation, structural rigidity of the structural dynamics properties of the femur com-pact bone; the elastic modulus, maximum stress, maximum inherent strain, bone break stress, and bone break strain of the mechanical properties of a material of the femur compact bone in the experimental rats. The results showed that compared with Group B, the elastic modulus, maximum loading capability, maximum stress, potential energy of deformation, and structural rigidity of LV4; the maximum loading capability, bone break load, potential energy of de-formation, structural rigidity of the structural dynamics properties of the femur compact bone; the elastic modulus, maximum stress, maximum inherent strain, and bone break strain of the mechanical properties of a material of the fe-mur compact bone were obviously increased in Group A, D, E and F (P< 0.05). Group C had increasing tendency. There were no statistical differences among Group A, C, D, E and F. Group D, E, and F had increased with EPF dose-dependently. However, there were no statistical differences among them. There were no statistical differences on bone break strain of the mechanical properties of a material of the femur compact bone among Group A, C, D, E, and F. It showed that ovariectomization reduced the biomechanical properties of vertebra bodies, structural dynamics properties of the femur compact bone, and the mechanical properties of a material of the femur compact bone. The application of EPF can effectively prevent and treat the decreasing of biomechanical properties of ovariectomized rats, so as to keep them in a relatively higher level.