Background and purpose:To study the expression of PTEN and P53 and analyse their roles in HGC oncogenesis,and to discover a good marker for diagnosis and prognosis of HGC.Methods:The expression of PTEN mRNA,PTEN protein and P53 protein were detected by in situ hybridization(DNA-RNA) and immunohistochemistry(SP method) in 64 cases of HGC.We took 22 cases of gallbladder adenoma and 10 cases of chronic cholecystitis as controls.Results:① The positive expression rates of PTEN mRNA and protein in HGC(56.3%,59.4%) were significantly lower than that in gallbladder adenoma(81.8%,90.8%) and chronic cholecystitis(100%,100%)(P

Objective To study the intercellular adhesion molecule-1(ICAM-1)and fat metabolism in the contribution of the incidence of steroid avascular necrosis of femoral head(ANFH).Methods 32 New Zealand rabbits were randomly divided into two groups:animals in experiment group(n=24)were given an injection of hydrocortisone acetate(8.0mg/kg)two times per week,and animals in control group(n=8)were given an injection with the same volume of normal saline.Serum total cholesterol(TC),triglyceride(TG)and ICAM-1 were quantitated before and 2,4,6,8 weeks after the treatment.2,4,6 and 8 weeks after treatment,6 animals of the experiment group and 2 of the control group were randomly chosen and sacrificed.ICAM-1 expression in soft tissue around hip was determined immunohistochemically,and pathohistological changes in femoral head were observed by light microscopy.Results Serum TC and TG levels of the animals in experimental group were significantly higher than those before treatment and those in control group in the 6th week after treatment(P

Objective To establish an HPLC method to determine the contents of dauricine in Menispermi Rhizoma from different producing areas. Methods C18 was set as chromatographic column filler, with acetonitrile-water-triethylamine (45:55:0.1) as the mobile phase, 284 nm as the ultraviolet wavelength detection, 1 mL/min as the flow rate, 30 ℃ as the column temperature. HPLC chromatograms of eight different batches of Menispermi Rhizoma were established. Results HPLC testing conditions of Menispermi Rhizoma was established. Within 20-100 μg/mL, there was a good linear relationship between the injection volume of the reference substance and the peak area (r=0.9995). The average recovery of dauricine was 100.30%, RSD=1.000%. The contents of dauricine in Menispermi Rhizoma from different producing areas were different. Conclusion The HPLC method is with sensitivity, accuracy, precision, good reproducibility and simple operation, which can be used as detection method to determine the content of dauricine in Menispermi Rhizoma.

Objective:To develop and optimize a novel assay for determination of cytotoxicity based Calcein -AM release.Methods:The target cells stained by Calcein-AM dye,then effectors and targets were incubated at E/T ratios from 30∶1-1∶1 for 4 h at 37℃,and the supernatant of reactions were detected by Fluorescence-Measurement to analyze specific cytotoxity.Results:The optimal excitation and emission wave lengths of Calcein were 485 nm and 515 nm.Dilutions of target cells stained by Calcein-AM had a linear relationship with measured fluorescence values.The Calcein-AM dye used to stain the living cells was shown to have a low spontaneous leakage rate-less than 15% in 4 hours at 37℃.Cytotoxicity activity of CIK showed a significant and positive correlation with E/T ratio when incubated at 4 h.Conclusion:The developed cytotoxicity test by Calcein-AM release is accurate and can avoid the application of radioactive reagents.

BACKGROUND:There is no effective drug for idiopathic pulmonary fibrosis (IPF), because of a lack of the animal model imitating the complete pathogenesis of human IPF. Therefore, it is critical to establish an ideal animal IPF model used for investigating the underlying pathogenesis and developing a kind of effective drug. OBJECTIVE:To establish an animal model that can mimic more characters of human IPF. METHODS:Seventy male C57BL/6 mice were randomly divided into two groups, fol owed by subjected to the intraperitoneal injection of bleomycin (35 mg/kg) on days 1, 4, 8, 11, 15, 18, 22, and 25, twice (group A) or once (group B) a week. Mice were sacrificed at 2, 4, 6, 8, and 10 weeks after the eighth injection, and the lung tissues were moved used for hematoxylin-eosin, Masson and immunohistochemical stainings. RESULTS AND CONCLUSION:There were various degrees of alveolitis and pulmonary fibrosis in the two groups at different time points after the last injection. The scores of alveolitis and pulmonary fibrosis in the group A began to gradual y increase from the 2nd week and reached the highest level at the 6th-8th weeks until the 10th week. In contrast, the scores of alveolitis and pulmonary fibrosis in the group B peaked at the 2nd week, then fluctuately decreased, and were significantly lower than those in the group A at the 6th week (P<0.05). Immunohistochemistry showed that type I col agen deposition was mainly distributed in the subpleural region, peri-vascular region and alveolar septa, which was consistent with Masson staining findings. The expression levels of transforming growth factorβ1 (TGF-β1) andα-smooth muscle actin (α-SMA) in the regions developing alveolitis and pulmonary fibrosis were significantly increased. In the group A, the expression levels of type I col agen, TGF-β1,α-SMA, and the hydroxyproline content in the lung tissues reached the peak level at 6-8 weeks. However, in the group B, al above indicators reached the highest level at the 2nd week, but gradual y decreased thereafter. At the 4th week, the expression Levels of TGF-β1 andα-SMA in the group B were significantly lower than those in the group A (P<0.05). At the 6th week, the hydroxyproline and type I col agen levels in the group B were significantly lower than those in the group A (P<0.05). In conclusion, the mouse model of pulmonary fibrosis induced by intraperitoneal injection of 35 mg/kg bleomycin twice weekly can be used to mimic the repetitive wound healing process, pathological morphology and cytokine changes of human IPF, which is prone to administration, with better stability and repeatability. This model is of great significance for the study on IPF. Subject headings:Disease Models, Animal;Pulmonary Fibrosis;Bleomycin

BACKGROUND:It is particularly important to establish an ideal animal model of pulmonary fibrosis to investigate the underlying pathogenesis and screen effective drugs to prevent and control pulmonary fibrosis. OBJECTIVE: To establish a modified scheme of establishing mouse models that can reflect pulmonary fibrosis formation in humans. METHODS: Fifty-six male C57BL/6 mice were randomly divided into two groups: A (a single large-dose injection) and B (multiple smal-dose injections). Mice in group A were subjected to a single intravenous injection of bleomycin 200 mg/kgviathe tail vein; and mice in group B received intravenous injections of bleomycin 50 mg/kg via the tail vein per week, totaly for 6 weeks. 

p53, as a transcription factor, is an important tumor suppressor gene and plays the key role in the p53-dependent gene regulatory network. Therefore, it is important to understand its biological function at the level of the whole system. In this paper, based on KEGG database and related literatures in English and Chinese, the interaction mode and quantitative relationship of the related molecules involved in p53 signaling pathway were extracted. By using S-system equations and 'Simulink' toolbox of Matlab7.0, a dynamic model of p53 signaling pathway was developed, and the dynamic regulatory characteristics of p53 signaling pathway were analyzed on model simulation. The results were in accord with the literatures and could reflect quantitatively the complex regulatory relationship between the interacting molecules involved in p53 signaling pathway. In addition, model simulation helped us find and identify the key molecules in this signaling pathway. Thus, this model can be used as a basis for the follow-up study of the relationship by precise and quantitative assessment.
Algorithms ; Computer Simulation ; Gene Expression Regulation ; Humans ; Models, Biological ; Signal Transduction ; physiology ; Transcription Factors ; genetics ; metabolism ; Tumor Suppressor Protein p53 ; genetics ; physiology

BACKGROUND:The mechanical valves used in the adult Luo-Ye pump have a large size and great destruction to blood, which are not suitable for infant ventricular assist pump. Therefore, designing and producing a high molecular valve with smal size and low incidence of thrombosis is a research hotspot. OBJECTIVE:To design and produce a valve of 20 mL infant Luo-Ye pump, and to test its basic functions and fatigue properties. METHODS:The size and shape of valve was designed with MASTERCAM software, polyurethane valve was obtained through producing the valve model and plastic injection;the static leakage, pressure drop and fatigue resistance of polyurethane valve were tested according to the ISO5840 requirements. RESULTS AND CONCLUSION:The polyurethane trefoil valve was produced, but the failure rate of plastic injection was high;the basic function of the trefoil valve met the ISO5840 requirements bascial y;after continuously operated 1.0×107 times, stroke volume of 20 mL Luo-Ye pump was changed 5.2%, and two polyurethane valves and valve leaflets did not change and damage. Polyurethane trefoil valve was designed and produced successful y;polyurethane valves could meet the needs of 20 mL Luo-Ye pump, which already have the ability to clinical trials.

Objective To research the effect to the semen quality of the three operation methods of superiority of highly selective varicocele high ligation,transinguinal canal and laparoseopic of renovated polomo management of varicocele.Methods 561 patients in our hospital who charged by infertility had the operation were analyzed retrospectively and were recruited with primary varicocele above grade II for this study.These patients were divided into three group according to three kinds of operation methods of varicocele :group A was treated with highly selective varicocele high ligation and had 300 patients;group B was treated with transinguinal canal operation and had 181 patients; and group C was treated by laparescopic of renovated polomo operation with 80 patients.Through the comparing of the fertility ability of the three operation methods,to evaluate the therapeutic effect of the three methods.Results The increasing rates of the quality of semen in group B was higher than other groups if patients' ages were lower than 30.The increasing rates of the quality of semen in every group was also higher if patients' ages were lower than 30.No difference was found between three groups in natural conception rate (P＞0.05).Conclusion The methods of highly selective varieecele high ligation would be a better choice in the management of patients with primary varicecele because of higher quality of semen.

Objective:This study aims to evaluate the correlation of combined preoperative plasma levels of fibrinogen (Fbg) and lym-phocyte to monocyte ratio (LMR) (F-LMR) with the prognosis of patients with non-small cell lung cancer (NSCLC) after complete resec-tion. Methods:The clinical data of 589 patients with NSCLC who underwent complete resection in our hospital were retrospectively analyzed. Receiver operating characteristic curve (ROC) analysis was used to select the cut-off values of Fbg and LMR. Based on the cri-teria of F-LMR, we divided the patients into three groups:F-LMR 0 score, F-LMR 1 score, and F-LMR 2 score. The association between F-LMR and the clinicopathological characteristics was analyzed by theχ2 test. Kaplan-Meier analysis was used to analyze the prognostic factors, and the log-rank test was used to determine the differences in survival rates. Prognostic factors were assessed by univariate and multivariate analyses (Cox's proportional hazards regression model). Results:According to the ROC curve, the cut-off values of Fbg and LMR were 3.48 g/L and 3.23, respectively. F-LMR 0 score had n=215, F-LMR 1 score had n=228, and F-LMR 2 score had n=146. Pre-operative F-LMR was closely related to age, gender, smoking history, tumor location, surgical type, pathological stage, pathological type, and tumor size (P<0.05). Univariate analysis showed that tumor location, surgical type, pathological stage, tumor size, F-LMR score, LMR, and Fbg were associated with survival (P<0.05). Multivariate analysis showed that the pathological stage [disease-free sur-vival (DFS): hazard ratio (HR)=1.700, 95%confidence interval (CI)=1.483-1.950, P<0.001;overall survival (OS):HR=1.703, 95%CI=1.486-1.952, P<0.001] and F-LMR score (DFS:HR=1.264, 95%CI=1.077-1.484, P=0.004;OS:HR=1.301, 95%CI=1.107-1.528, P=0.001) were the independent prognostic factors of NSCLC patients. Conclusion:The preoperative F-LMR score may be a useful blood marker for predicting the prognosis of patients with NSCLC with radical resection.