Objective:To study the effect of Hypoxia-inducible factor-1 alpha(HIF-1?)on the radiosensitivity of rat cervical carcinoma and discuss the possible mechanism.Methods:The transplantation tumor model of cervical carcinoma was established. 77 rats were randomly divided into five groups:control group,the large fraction irradiation group,the small fraction irradiation group,the large fraction irradiation combining HBO group and the low dose irradiation combining HBO group.Expressions of HIF-1? and bcl-2 in transplantation tumor after treatment were detected by immunohistochemical staining,and the tumor inhibitory rate by the change of tumor volume.The correlation between expressions of HIF-1 ? and bcl-2(and tumor inhibitory rate)was also observed.Results:Expression of HIF-1 ? and carcinoma growth inhibitory rate showed a reverse correlation(r=-5.147,P

Objective:To investigate the radiosensitivity enhancement effect of COX-2 inhibitor-Nimesulide on breast cancer cell line(MCF-7) in vitro and to initially disclosure its mechanism.Methods:The clonogenic assay was performed to determine the radiosensitizing effect of Nimesulide on MCF-7 cell line.The morphological changes of MCF-7 were observed by electron microscopy.The apoptosis and expressions of Bcl-2 and Bax were measured by flow cytometry(FCM).Results: Nimesulide at different concentration significantly enhanced the radiosensitivity of MCF-7 cells.in a dose-dependent manner,under the combined treatment of Nimesulide and radial the protein expression of Bax was up-regulated,while that of Bcl-2 was inhibited and the apoptpsis increased significantly.Conclusion:Cox-2 inhibitor could obviously enhance the radiosentivity of human breast cancer cell line MCF-7.

Objective:To investigate the inhibitory effect of cyclooxygenase inhibitor acetylsalicylic acid(ASA),and a selective COX-2 inhibitor(Nimesulide) on the growth of human breast cancer cell line MDA-MB-231 in vitro.Methods:The inhibitory effect was detected by MTT assay.Immunohistochemical assay was performed to examine the expressions of COX-2 and c-erbB-2 in SKOV3 cell line treated by NSAIDs.Results:A decrease in cell number compared with controls was observed in all of the cell line treated with ASA and Nimesulide.It was a dose-dependent inhibition of cell proliferation.COX-2 was expressed positive in MDA-MB-231 and the expression of cerbB-2 was found to decrease by immunohistochemical assay.Conclusion:NSAIDs can inhibit the growth of human breast cancer cell line MDA-MB-231 in vitro.

BACKGROUND: Leptin is a hormone produced predominantly by adipocytes and has a variety of physiological functions. It has been a hot spot in energy metabolism research. However, leptin presently used is usually produced by E coli, in which leptin cDNA is expressed and is in the form of insoluble inclusion body. Therefore, extremely complicated reactivating procedure is needed to obtain biologically activated human leptin.OBJECTIVE: To explore the condition for purification of leptin in non-affinity chromatography in order to obtain soluble human leptin.DESIGN: An observational experiment.SETTING: Molecular laboratory of biochemical department in a medical college.MATERIALS: The strong anion exchanger sepharose Q and hydrophobic phenyl sepharose 6 were used in different conditions for removal of as many contaminants as possible.METHODS: The supernatant of pichia pastoris yeast culture solution was first purified through Q column chromatography, the protein was collected and was further purified through hydrophobic support with 1 mol/ L (NH4) 2SO4.MAIN OUTCOME MEASURES: Gel scanning revealed that the purity of human leptin was 42. 3% before purification, 89.6% after Q column chromatography and 96.2% after hydrophobic interaction chromatography.RESULTS: The post-purification product presented a s. ingle band in SDS-PAGE. Gel scanning revealed that the purity of human leptin was 42.3% before purification, 89.6% after Q column chromatography and 96.2% after hydrophobic interaction chromatography.CONCLUSION: The combined use of strong anion exchange chromatography and hydrophobic interaction chromatography can effectively purify leptin expressed by pichia pastoris yeast and the purity is identical to that of nickel affinity column chromatography. It provides reliable evidence and method for possible manufacture of human leptin and lays experimental basis for leptin-related research.

Objective To investigate the role of stem cell factor (SCF) and mast cells (MC) in the pathogenesis and progression of dermal lesions caused by chronic renal failure.Methods Thirty-six Wistar rats were randomly divided into model group (adenine lavage at a dose of 150 mg·kg-1·d-1) and control group (physiological saline lavage at equal volume).Six rats from each group were sacrificed respectively at week 4,8 and 12.The intensity of MC infiltration was examined by toluidine blue staining. The expression of SCF was detected by immunohistochemistry and real-time fluorescence quantitative PCR. Results Compared with control group,the intensity of MC and the expression of SCF were significantly higher in dermal tissue of model group (P＜0.O1,respectively),and they were increased with time.In the model group,the number of MC infiltration was positively correlated with both the protein expression of SCF (r=0.81,P＜O.01) and the level of SCF mRNA (r=0.65,P＜0.01). Conclusion The increased SCF and MC may participate in the pathogenesis and progression of dermal lesions caused by chronic renal failure.

BACKGROUND: It demonstrated that combined application of thymosin α 1 (TM-α1) and interferon (IFN) can enhance the antiviral activity of IFN.OBJECTIVE: To obtain recombinant fusion protein of TM- α1 and consensus IFN α (IFN α -con) with double activity of antiviral activity and immunity enhancement.DESIGN, TIME AND SETTING: The in vitro contrast experiment was conducted in the Biochemical Laboratory of Research Institute of Medicine, Chongqing K.E.W Pharmaceutical Co., Ltd. from March 2003 to December 2004.MATERIALS: The fusion gene fragment (TM- α 1+IFN α -con) was synthesized by Shanghai Sangon Biological Engineering Technology And Service Co., Ltd, WISH cell, and vesicular stomatitis virus (VSV) was purchased from Institute of Biochemistry and Cell Biology, commercial products of IFN α 1b, IFN α 2a and TM- α 1 was served as reference substance.METHODS: The preference for E. coli of fusion sequence coding TM- α 1 and IFN α -con were cloned into expression vector of pET-22b(+) and expressed in BL21(DE3)-Codon plus-RP-X, which was purified by precipitation of (NH4)2SO4, hydrophobic interaction chromatography, anion-exchange chromatography, cation-exchange chromatography and molecular exclusion chromatography. The antiviral activity of fusion protein was tested by cytopathic-effect inhibition assay, and effect of fusion protein on lymphocyte proliferation was tested by cell proliferative assay.MAIN OUTCOME MEASURES: The specific activity of fusion protein and its biological activity in promoting lymphocyte proliferation.RESULTS: The fusion protein was expressed as a soluble form, accounting for over 20% of the total cell protein in E. coli, which approached 96% after purification. The antiviral activity of fusion protein was superior to IFN α 1b and IFN α 2a. However, the activity of fusion protein for promoting lymphocyte proliferation was similar to TM- α 1.CONCLUSION: The fusion protein of TM- α 1 and IFN α -con expressed in E. coli has both effects on anti-virus and promoting lymphocyte proliferation.

BACKGROUND:As a by-product in sugar industry, sugar cane wax has been widely used in non-medical field. Some researches indicate that sugar cane wax plays a great role in reducing blood cholesterol; however, the therapeutical effect and clinical application should be studied further.OBJECTIVE: To separate the high-class eicosanoic acid and the high-class alkanols, which are suitable for medical application, and further to observe the effect of them on reducing blood cholesterol of model rets with hyperlipemia.DESIGN: Randomized control animal study.SETTING: Pharmacological Institute, Chongqing Kangerwei Pharmaceutical Co., Ltd.MATERIALS.: The experiment was carried out in the Pharmacological Institute of Chongqing K.E.W Pharmaceutical Go.,Ltd. From April 2005 to January 2006. A total of 65 adult female Wistar rats, aged at 3-6 months, weighing 180-220 g, of SPF grade, were provided by Experimental Animal Center of Chongqing Institute of Traditional Chinese Medicine. Raw sugar cane wax was provided by Beijing Jiade Hongsheng Biochemical Technology Co., Ltd. High-class alkanols C26,C28,C30, C32 and high-class eicosanoic acid C28, C30, C32, C34 were provided by Sigma Company (standard materials of gas phase chromatography), and other reagents were national analytical pure.METHODS: ① Sugar cane wax was extracted from raw sugar cane wax with ethanol and other organic solution and separated from the mixture of high-class eicosanoic acid and the mixture of high-class alkanols with saponification and calcification. Main components were analyzed with gas phase chromatography. The main components of high-class alkanols were C26, C28, C30 and C32 and the main components of high-class eicosanoic acid were C28, C30, C32 and C34, ② Based on references, rats were fed in 3 days and randomly divided into blank group (n =10) and experimental group (n =55).And then, all rats were cut off their tails to collect blood and the triacylglyoerol (TG), total cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) were measured with automatic biochemistry analyzer. Rats were fed with common granule feeds in blank group or with high-lipid feeds (containing 0.1 mass fraction of oiliness, 0.1 mass fraction of yolk powder, 0.01 mass fraction of cholesterol, 0.002 mass fraction of pig's gall salt, 0.788 mass fraction of common feeds) in experimental group. All rats ate and drank freely. Seven days later, blood was collected again from tail tip to measure the contents of TG, TC and HDL-C. Based on level of serum TG, rats in the experimental group were randomly divided into 5subgroups (n =11): negative control group, low-dosage high-class alkanols group, high-dosage high-class alkanols group,Iowdosage high-class eicosanoic acid and high-dosage high-class eicosanoic acid group. Rats in low-dosage and high-dosage high-class alkanols groups were perfused with 5 and 50 mg/(kg·d) high-class alkanols; meanwhile, rats in low-dosage and high-dosage high-class eicosanoic acid groups were perfused with 20 and 200 mg/ (kg·d) high-class eicosanoic acid. Rats in negative control group and blank group were perfused with the same volume of 0.3% carboxymethylcellulose sodium and distilled water, respectively, once a day for successive 30 days. At 16 hours after the last administration, rats were anesthetized to collect blood from heart to measure contents of TG, TC and HDL-C in serum.MAIN OUTCOME MEASURES: ① Percentage of main component in separated mixtures of high-class eicosanoic acid high-class alkanols; ② levels of serum cholesterol, HDL and TG.RESULTS: A total of 65 experimental rats were involved in the final analysis. ① Gas phase chromatography suggested that the content of C28 high-class alkanols was the most (73.6%), and other three kinds of high-class alkanols were counted for 5.3% (C26), 6.2% (C30) and 5.1% (C32), respectively. The total quantity was 90.2%. In the mixture of high-class eicosanoic acid, content of C28 high-class eicosanoic acid was the most (46.6%) and the other three kinds of high-class eicosanoic acid were counted for 16.7% (C30), 6.8% (C32) and 9.3% (C34), respectively. The total quantity was 79.3%. ②Levels of serum TC were (1.46±0.27), (1.66±0.33), (1.44±0.25) and (2.16±0.52) mmol/L in high-dosage and Iow-dosage high-class alkanols groups and high-dosage and Iow-dosage high-class eicosanoic acid groups, respectively, which were lower than those in negative control group [(2.52±0.83) mmol/L, P＜0.01]. Levels of HDL-C were (0.73±0.09), (0.71±0.07), (0.79±0.10) and (0.70±0.08) mmol/L in the four treatment groups, respectively, which were higher than those in negative control group [(0.58±0.13) mmol/L, P＜0.05-0.01].CONCLUSION: The high-class alkanols and the high-class eicosanoic acids separated from sugar cane wax made in China significantly have the activity of reducing blood cholesterol; however, the effect on decreasing TG is not obvious.

Objective To study the effect of repeated rectal exposure of low -dose simian immunodeficiency virus on the systemic cellular immunity in monkeys .Methods Eight 3-to 4-year old rhesus macaques ( Macaca mulatta) (male:female 1:1) were used in this study.The monkeys were inoculated with 10 TCID50 SIVmac239 virus through rectum twice a week for consecutive 6 weeks to establish a multiple rectal exposure model of SIVmac 239 virus infection.Then, plasma viral load, CD4+ T cell count, T cell subsets and IFN-γsecretion of the experiment monkeys were determined . Results Low-dose SIVmac239 virus induced some changes in the immune system through the rectal mucosa , but didn’t induce typical infection.Repeated rectal mucosal low-dose virus exposure can activate the cellular immune system . Conclusions This study defines the effect of repeated low -dose simian immunodeficiency virus exposure on the systemic cellular immunity, and provided basic information for HIV-1 vaccine research.

Objective To explore the application value of endoscopic ultrasonography (EUS) in the diagnosis of suspected obstructive jaundice in acute biliary pancreatitis and its effect on treatment outcomes. Methods Clinical data were retrospectively collected in 96 patients with acute biliary pancreatitis (ABP) who were suspected obstructive jaundice. Patients were divided into early EUS scanning group (EES) and delay EUS scanning group (DES). Clinical treatment results and test results were compared between the two groups. Results The diagnosis sensitivity, specificity, accuracy, positive predictive value and negative predictive value of EUS were 96.43 %, 82.5 %, 90.63 %, 88.52 % and 96.29 %. Early EUS scan with 72 h could identify the etiology of ABP and subsequent treatment based on the EUS can easily decrease the white blood cell count, percentage of neutrophils, total bilirubin and serum amylase (P < 0.05). The pain relief rate in EEI group after 1 week treatment was significantly higher than that in the DES group (87.50 % vs 66.67 %, P = 0.027) and average length of hospitalization was shorter [(12.70 ±2.10) d vs (14.10 ± 3.00) d, P = 0.006]. Delay EUS scan were likely to have higher white blood cell count and total bilirubin, meanwhile seemed to increase the organ failure rate and necrotic infection of the pancreas. Conclusion ABP with suspected obstructive jaundice should be considered to have endoscopy intervention. Results of EUS are the basis for treatment decision, which can avoid the unnecessary endoscopy treatment.

Objective To investigate the strategies of nutritional therapy for severe acute pancreatitis (SAP) patients.Methods Two hundred and eight patients with SAP were randomly divided into early enteral nutrition (EEN) group,late enteral nutrition (LEN) group and total enteral nutrition (TPN) group.EEN group received enteral nutrition through nasojejunal tube feeding within 72 hours in the course of disease,LEN group received the same treatment after 6 d in the course of disease,TPN group received total enteral nutrition.The index of nutrition,incidence of complication were recorded,efficacy and safety were analyzed.Results EEN group included 67 cases,while 72 cases in LEN group and 69 cases in TPN group.The incidence rate of malnutrition after 14 d of treatment in EEN group and LEN group was significantly lower than that in TPN group [59.7％ (40/67),58.3％ (42/72) vs.84.1％ (58/69),P ＜0.05],but the difference between EEN group and LEN group was not statistically significant (P ＞ 0.05).There was no statistically significant difference in the incidence of hyperlipidemia and hyperglycemia among three groups (P ＞ 0.05).The expression level of albumin,prealbumin and retinol-binding protein after 14 d of treatment in three groups compared with those before treatment and the differences were statistically significant (P ＜ 0.05).The expression level of albumin,prealbumin and retinol-binding protein after 14 d of treatment in EEN group and LEN group compared with those in TPN group and the differences were statistically significant (P＜ 0.05),there was no statistically significant difference between EEN group and LEN group (P＞ 0.05).The incidence rate of total infections,abdominal infections,bloodstream infections,secondary superinfections in TPN group were significantly lower than those in EEN group and LEN group [62.3％(43/69) vs.25.4％ (17/67),19.4％(14/72) ;21.7％(15/69) vs.4.5％(3/67),6.9％ (5/72);39.1％(27/69) vs.14.9％(10/67),11.1％(8/72) ;33.3％ (23/69) vs.9.0％ (6/67),9.7％(7/72),P ＜ 0.05],there was no statistically significant difference in the incidence of peripancreatic infections,lung infections among three groups (P ＞ 0.05).There was no statistically significant difference in the incidence of total infections,different infections between EEN group and LEN group.There was no statistically significant difference in the incidence of noninfections among three groups (P ＞ 0.05).The rate of recovery in EEN group and LEN group was significanby higher than that in TPN group [91.0％ (61/67),94.4％ (68/72) vs.81.2％ (56/69),P ＜ 0.05],but the difference between EEN group and LEN group was not statistically significant (P ＞ 0.05).Conclusion Early enteral nutrition therapy for SAP patients is safe and effective,and could significantly improve the prognosis of patients.