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:This study has two objectives. One is to detect the methylation status of reversion-inducing cysteine-rich protein with Kazal motifs (RECK, a new tumor suppressor gene) gene promoter in primary laryngeal squamous cell carcinoma and nor-mal laryngeal mucosa. The other is to analyze the correlation between RECK gene methylation status and radiosensitivity in laryngeal squamous cell carcinoma. Methods:Methylation-specific polymerase chain reaction was used to detect the RECK gene methylation of 70 specimens of laryngeal squamous cell carcinoma and 15 normal tissues of laryngeal mucosa. The patients underwent six cycles of ra-diotherapy and were followed-up for 5 years. The correlation between RECK gene methylation status and radiosensitivity in laryngeal squamous cell carcinoma was analyzed. Results: After six cycles of radiotherapy, 47 patients (67.14%) showed sensitivity and 23 (32.86%) showed tolerance to radiotherapy. The methylation level of the RECK gene was lower in the radiation-sensitive group than in the nonradiation-sensitive group (P<0.05). The methylation level of the RECK gene was lower in the remission group than in the non-remission group. RECK gene methylation could increase the risk of cancer by approximately 5.010 times (OR=5.010, 95%CI:1.616-15.533). Conclusion:RECK gene promoter methylation in human laryngeal squamous cell carcinoma is an early event that is correlated with the patient's sensitivity to radiotherapy. Thus, the patient's sensitivity to radiation can be predicted by detecting the meth-ylation status of the RECK gene promoter.

Objective: To develop price framework of medical service price project and introduce the weights calculation method of the new edition medical service price project. Methods: According to the new instruction manuals of medical service price and the research of the resource costs based on the relative value(RBRVS)of the U.S. Health Care Financing Administration (HCFA) and the American Medical Association ( AMA ) , an analysis framework and dimension of medical service price was raised and authoritative international medical service project price methods w ere summarized. Results: Based on the percentages of constituted factors and the professional features, it found the weights calculation model could be constructed with the weights model, and it could calculate the weights of the elements. Conclusion: Medical service price can be measured and comprehensively researched from the different dimensions of the medical service price, so as to carry out the pricing work of medical service price project and provide better service for pharmaceutical reform.

Objective To investigate the subjective life quality of hospitalized children and analyze its influencing factors for improving their life quality.Methods 48 hospitalized children who were admitted between May to August 2011 were investigated in convenience using inventory of subjective life quality (ISLQ).Results Patients investigated had a higher score than the national norm on Family Life Dimension.The satisfaction level of 20％ ～ 30％ children were below the average on Family Life and other eight dimensions,the cognitive components of 16 cases (33.3％) and emotional component of 11 cases (22.9％) were below the average.15 cases (31.3％) were not satisfied with their subjective quality of life.Female patients had higher scores than male patients on peer exchanges,body feelings and subjective quality of life satisfaction; the satisfaction level of experience depression for rural children was lower than urban children.Conclusions The subjective life quality of vast majority of hospitalized children is about the general level,but some patients still experience serious anxiety,depression and other negative psychological emotion.Nurses should guide the patients positively and specially,considering the influence of their gender and living places while providing caring.Based on the health education of patients and their family,we promote them to face the disease.

Objective To evaluate plasma BNP level change in acute pulmonary embolism and its clinical value.Methods 33 cases were diagnosed by CT pulmonary angiography or DSA.they were divided into two groups:the group with right ventricular(RV)dysfunction group and the group without right ventricular(RV)dysfunction.Plasma BNP was detected by Triage BNP test.otherwise,we observed severe clinical presentation、 death rate and mechanical ventilation in two groups.Results Plasma BNP level is higher in the group with RV dysfunction(288?102pg/ml)than that in the group without RV dysfunction(61?32pg/ml)(P

Objective:To evaluate the effect of Helicobacter pylori infection on hyperdynamic circulatory state and portal hypertensive gastrropathy in liver cirrhotic patients.Methods:Helicobacter pylori infection,IL-1,IL-8,TNF-?,Endotoxin,NO,ET,anti-VacA and anti-CagA levels were determined in cirrhotic patients by using rapid urease test,()~(14)C-urea expiratory test,immunoimpression test,nitrite colorimetric test and ELISA.The parameters of portal venous system flow were determined by doppler ultrasonography.Results:HP prevalence in patients with liver cirrhosis was 77.1%,anti-VacA and antiCagA positive rate was 60.4% in common.IL-1,IL-8,TNF-?,Endotoxin,NO and ET levels in anti-VacA and anti-CagA positive group were significantly higher than in anti-VacA and anti-CagA negtive group.The amount of splenic venous flow(SVF) and superior mesenteric venous flow(SMVF)in anti-VacA and anti-CagA positive group was greater and the amount of portal venous flow(PVF) was smaller than those in negtive group.Conclusion:There was the effect of Helicobacter pylori infection on hyperdynamic circulatory state and portal hypertensive gastrropathy in liver cirrhotic patients.

Objective To investigate colloidal gold immune chromatography (GICA ) and enzyme linked immunosorbent assay (ELISA) detection pepsinogen (PG) consistency of results ,so as to provide experimental evidence for the use of colloidal gold im‐munochromatography assay PG and briefly analysis of its clinical utility .Methods Totally 40 cases in our hospital digested samples and medical centers 60 healthy population sample in Endoscopy Center by endoscopy diagnosed with inflammation of the stomach disease ,respectively ,with colloidal gold immune chromatography and enzyme‐linked immunosorbent assay PG ,colloidal gold immu‐noassay chromatography method to be evaluated ,as the reference ELISA method .The results were compared to the method and bias analysis .Results GICA method with a correlation coefficient ELISA test results were greater than 0 .95 ,indicating a good correla‐tion between the two methods and there is no overall bias;sensitivity and specificity of the two methods were (0 .775 ,0 .85) and (0 .782 ,0 .823) .Conclusion Both GICA and ELISA can be used for screening or diagnosis of stomach illnesses .

Objective To investigat the changes of hemodynamics and arterial blood gas during the establishment of ECMO model and provide an ideal platform for the study of the mechanism of DCD protection using ECMO, a porcine model of controlled donors of cardiac death (CDCD) with extracorporeal support was established in the paper. Methods Eight domestic crossbred pigs were anes-thetized, atracurium (1mg/kg) and heparin (150~200 U/kg) was administered and the ventilator was discontinued while the animal was under deep anesthesia to kill the animal.Meanwhile, blood was drained from the left jugular vein catheter into the ECMO circuit and re-turned into the left femoral artery to establish the porcine model of CDCD with extracorporeal support . The average blood press and arterial blood gas were ministered at normal, cardiac arrest and ECMO supporting 1 h, 3 h and 5 h. Results The model of CDCD with extra-corporeal support was established successfully.Two animals restored the beating of the heart and the high doses of potassium chloride were given untill cardiac arrest during extracorporeal support.Blood gases throughout the apneic time showed a severe hypoxemia and acidosis, pH and PaO2 reduced to 7.09 ±0.24 and 5.3 ±0.93mmHg respective-ly, which can be improved during extracorporeal support.MAP and PaO2 were maintained over 300mmHg and 65mmHg respectively. Conclusion A new porcine model of controlled donors of cardiacdeath with extracorporeal support has been successfully established and showed to be feasible for related pathophysiological research work.

Objective To use the colloidal gold immunochromatography (GICA ) method to conduct the methodological prelimi-nary evaluation on pepsinogen Ⅰ (PGI) reagent kit .Methods The method provide by the Preliminary Evaluation of Clinical Quan-titative Experimental Methods :Approval Guide Second Edition (EP10-A2) formulated according to the Clinical Laboratory Stand-ards Institute (CLSI) was used to continuously detect the low ,moderate and high concentrations of serum for 5 d ,Then the related data were collected for analyzing the dispersion degree ,linearity ,offset ,precision and so on .Results The PGⅠ quality control ser-um (concentration 25 ,50 ,100 μg/L) was detected at low ,medium and high concentration levels ,the linear regression equation ob-tained by analysis was Y=0 .9939X+0 .7433 ,correlation coefficient (R2 )=0 .9992 ;the offsets were 0 .37 ,0 .77 ,0 .78 μg/L re-spectively ,total imprecision was 3 .04% ,1 .17% and 1 .08% respectively .Conclusion The GICA related technical indicators of PGⅠreagent kit reach the standards of EP10-A2 document ,the detection results are accurate with high sensitivity and good stability , and conform to the requirements of clinical applications .

To observe the expressions of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in rats with pulmonary fibrosis (PF) induced by bleomycin, and to explore the mechanisms of Shenlong Decoction in preventing and treating PF.