Animals ; Carboxylic Ester Hydrolases ; metabolism ; Chickens ; Drosophila ; Enzyme Activation ; Humans ; Mice ; Nervous System Diseases ; chemically induced ; enzymology ; physiopathology ; Organophosphate Poisoning ; Phosphorylation

In the light of requirements of medical equipment simulation system, the overall framework of joint control system of medical equipment integrated support in the theater is designed in this paper. Such modules are developed with Delphi as communication module, module for equipment basic information, module for equipment video information and module for medical faculty information. A joint control system suiting integrated support simulation sand table for medical equipment is complteted,which provides a multidimensional informational platform for integrated demonstration of medical equipment.

Objective To express and purify mammalian glycosylation modified trimeric Ebola virus trimeric glycoprotein (EBOV-GP) using novel glycoengineered Pichia pastoris.Methods The EBOV-GP and EBOV-GPΔMLDΔTM genes were cloned into the pPICZ-αA vector,electrochemically converted to glycoengineered Pichia pastoris,and compared with EBOV-GP expressed in HEK-293T cells.Glycosylation was analyzed by PNGaseF and EndoH digestion,while the target protein was purified by affinity chromatography and ion exchange chromatography.N-terminal sequencing was used to determine whether the signal peptide was correctly cleaved during protein translation and gel column analysis was used to find out whether the trimeric structure was formed.Results The results of PNGaseF showed that EBOV-GP expressed by glycoengineered Pichia pastoris and HEK-293T cells had the same relative molecular mass and N-glycosylation degree.EndoH digestion showed that the N-glycosylation modification of EBOV-GPΔMLDΔTM was in a non-high mannose form.N-terminal sequencing showed that the signal peptide of the GP protein itself was correctly excised.Gel column analysis showed that the purified protein was in a trimeric form.Conclusion An EBOV-GP is obtained with complex glycosylation modification based on Glycoengineered Pichia pastoris.

Animals ; Flunarizine ; pharmacology ; Hippocampus ; drug effects ; physiopathology ; Male ; Neurons ; drug effects ; physiology ; Penicillins ; adverse effects ; Rats ; Rats, Wistar ; Seizures ; chemically induced ; physiopathology

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

Objective To study the relationship between serum tumor marker level and the apoptosis regulation gene of tumor tissue in the patients with primary hepatocarcinoma .Methods 40 cases of primary hepatocarcinoma and 40 healthy people were in‐cluded into the observation group and control group .Then the levels of tumor marker GP73 ,TK1 ,DKK1 in serum and the expres‐sion of apoptosis regulation gene in tumor tissue were detected in the two groups .Results The serum GP73 ,TK1 and DKK1 levels of the observation group were significantly higher than those of the control group ,the difference was statistically significant (P<0 .05) .The apoptosis inhibiting gene Plk1 ,Livin and Xiap levels in the hepatocellular carcinoma tissue were higher than those in the adjacent normal tissues ,while the pro‐apoptotic gene M TS1 ,Caspase‐3 and Caspase‐8 levels were lower than those in the adjacent normal tissues ,the difference had statistical significance (P< 0 .05) ;serum GP73 ,TK1 and DKK1 levels were positively correlated with Plk1 ,Livin and Xiap levels and negatively correlated with M TS1 ,Caspase‐3 and Caspase‐8 levels .Conclusion The levels of se‐rum GP73 ,TK1 and DKK1 in the patients with primary hepatocarcinoma are abnormally increased ,moreover which are closely cor‐related with the apoptosis regulating gene expression and the ideal indexes to evaluate the disease condition of primary hepatocarci ‐noma .

Objective To develop an interleukin-4(IL-4) antagonist named M5-IgG1Fc protein constructed by genetic engineering of antibody Fc fragment-cytokine mutein fusion protein which has a long half-life time in plasma.M5-IgG1 Fc protein binds to IL-4 receptor but cannot activate downstream signalling pathway , which provides a basis for drug develop-ment for allergic diseases .Methods The synthesized interleukin-4 mutant gene ( named M5 ) was cloned into the expres-sion vector pBV220 and transformed into E.coli DH5α.Chimeric gene M5-IgG1Fc obtained by overlap extension (SOE) method was transformed into glycoengineered Pichia pastoris GJK01 through expression vector pPICZαA .Then M5-IgGFc fusion protein was obtained by protein purification after being induced by methanol in 72 hours.The anti-IL-4 biologicial ac-tivity assay of M5 and M5-IgG1 Fc was performed with CTLL-2/IL-4R cells and detected with MTT colormetry .Finally,the half-life time of M5 and M5-IgG1 Fc protein in mice was compared by detecting the remaining amount in plasma with ELISA kit.Results The M5 protein expressed in E.coli and M5-IgG1 Fc fusion protein expressed in P.pastoris GJK01 both had IL-4 antagonistic bioactivity .The EC50 of both, which inhibited 5.6 ×10 -2 nmol/ml of IL-4, were 0.31 ±0.05 and 0.77 ± 0.03 nmol/ml,respectively.The maximum of M5 in plasma at 0.5 h was 5.8 ×10 -2 nmol/ml but the remaining amount was 2.8%of the maximum at 2 h.M5 protein could not be detected after administration at 8 h because of the detection line . The maximum of M5-IgG1 Fc fusion protein was 4.7 ×10 -2 nmol/ml,while fusion protein M5-IgG1 Fc decreased to 4.3%of its maximum at 120 h and could not be detected at 168 h.Conclusion M5 protein has IL-4 antagonistic bioactivity .M5-IgG1 Fc fusion protein expressed in glycoengineered P.pastoris GJK01 has IL-4 antagonistic bioactivity and long retention time in mice,which can be potentially used for treatment of allergic diseases .

In order to realize the sustained education concept in clinical medical English teaching,several measures were taken in the first clinical medical college of Nanjing University of Traditional Chinese Medicine,such as training the teaching staff,using original textbooks and redesigning the curriculum.Particularly the tutorial system was introduced to the education frame.The teaching and research section of clinical medical English explored the new teaching routes for postgraduates in traditional Chinese medicine university.

Aim To evaluate the effects of pantoprazole on various experimental acute ulcer inrats and mice. Methods The model of a gastric ulcer of rats or mice was caused bystree- induced ulcer and ligatel pylurus-induced ulcer. Results & Conclusions At adose of 5, 10, 20 mg? kg-1 of Pantoprazole can markedly decrease the ulcer index ofstree-induced ulcer. Pantoprazole(4, 8, 16 mg? kg -1 ) significantly decrease the areaof ligated pylorus-induced gastric ulcer. It was also found that pantoprazole caninhibit the output of basic gastric acid.

Objective To provide reference for CT performance/quality test. Methods Researches were made in Chinese documentations on CT performance/quality test issued from 1998-aug 2002.Results Approximately a quarter of CTs among the documents assembly released public were tested unqualified . Qualification rate of new CTs was higher Than second-handed ones. Part of items and data in the documentations were insufficient. Conclusion CT performance/quality test is a crucial part of system Engineering of QC(quality control),therefore it must be implemented seriously and relevant regulations should be optimized in order to benefit both patients and health-care communities.