The TSR1(thermosensitive rescued)gene associated with the secretion of proteins of the yeast Yarrowia lipolytica was site-directed mutagenesis,restricted and spliced to obtain a series of deletion mutants.These mutants would be useful for studing the function of different domains of TSR1 gene in yeast Yarrowia lipolytica

Adult ; Aged ; Aged, 80 and over ; Female ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; classification ; pathogenicity ; Humans ; Lymphatic Metastasis ; Male ; Microvessels ; pathology ; Middle Aged ; Neoplasm Invasiveness ; Neovascularization, Pathologic ; microbiology ; pathology ; Stomach Neoplasms ; blood supply ; metabolism ; microbiology ; Vascular Endothelial Growth Factors ; metabolism

A novel nucleic acid amplification method,termed loop-mediated isothermal amplification(LAMP),which amplifies DNA with high specificity,efficiency,and rapidity under isothermal conditions,may be a valuable tool for the rapid detection of infectious diseases.This method employs a DNA polymerase that have activity of strand displacement DNA synthesis and a set of four specially designed primers that recognize a total of six distinct sequences on the target DNA.LAMP can amplify a few copies of DNA to 109 in less than an hour.The final products are stem-loop DNA with several inverted repeats of the target and cauliflower-like structures with multiple loops.A positive reaction would be shown as a ladder-like pattern in a gel electrophoresis analysis.Because of the advantage,the LAMP method will be widely applied to research of nucleic acid,clinical diagnosis of infectious diseases and detection of genetically modified organisms etc.

Antimicrobial peptides are a class of small peptides with anti-extrogenous pathogen activities.They are derived from organism and possess antibacterial,antifungus,antiviruses and anticancer cell actions.In recent years,it’s found that some microbial pathogens are able to resist antimicrobial peptides.The constitutive and inducible mechanism of a pathogen resists a given peptide were reviewed in this paper.

Objective To explore the effect of chronic intermittent hypoxia (CIH) on hippocampus neuron and growth associated protein-43 (GAP-43) in rats. Methods 40 adult male Sprague-Dawley rats were randomly divided into control group, CIH 1 week group, CIH 3 weeks group and CIH 5 weeks group with 10 rats in each group. The CIH groups were exposed to intermittent hypoxia in designed cabin 8 hevery day for 1 week, 3 weeks and 5 weeks. The pathological changes of hippocampal neurons were observed by HE staining. The GAP-43 expression of hippocampus was detected by immunohistochemical staining. Results There was degeneration and necrosis of hippocampal neurons in the CIH rats, and it became worse gradually along with the hypoxia. Compared with the control group, the GAP-43 expression in hippocampus increased in CIH rats. The peak of GAP-43 expression appeared in the 1st week (P=0.038) and then it decreased in the 3rd week (P=0.382) and it was closed to the normal level in the 5th week (P=0.860). Conclusion Degeneration and necrosis of hippocampal neurons exacerbated gradually along with the hypoxia, while GAP-43 expression changed inconsistently.

Objective To investigate the diagnostic value and significance of myocardial injury markers in neonates.Methods A retrospective analysis of 838 cases of myocardial injury markers in neonates,the total positive rate of hs -cTnI and the positive rates of hs -cTnI(hypersensitivity cardiac troponin I)in different kinds of neonatal disease were calculated,the levels of abnormalities consistent rate in hs -cTnI and CK -MBmass(creatine kinase MB mass)were compared with hs -cTnI and MYO(Myoglobin).Results The total positive rate of hs -cTnI was 40.33% in the 838 neonates.The highest incidence of myocardial injury was neonatal sepsis (57.14%),followed by neonatal pulmonary hemorrhage (55.56%)and neonatal convulsions (54.55%).The abnormalities consistent rate of hs -cTnI and CK -MBmass was better than hs -cTnI and MYO (85.50% vs 28.11%,χ2 =226.9,P <0.05). Conclusion Neonatal hospitalized children often complicated by myocardial injury;As a biochemical myocardial injury marker,the hs -cTnI detection is important for early detection of myocardial injury,it should be recommend as routine test items;CK -MBmass has better correlation with hs -cTnI than MYO,which can provide guide for doctors to interpret the data of myocardial injury markers.

OBJECTIVETo explore the inhibition of Tangtong Recipe (TTR) on proliferation of vascular smooth muscle cells (VSMCs) in rat model of diabetes mellitus using seropharmacological and modern molecular biological technologies.

METHODSThe drug-serum of TTR was prepared by feeding pure-breed New Zealand rabbits at various doses (18 mL/kg, 13 mL/kg, and 8 mL/kg) of TTR, and used to intervene the VSMCs get from the iliac artery of diabetic model rats cultured by tissue block plantation. The impact of drug-serum on VSMCs proliferation was estimated through measuring tritiated thymine deoxy-ribonucleotide (3H-TDR) incorporation using liquid-scintillation detector, and the cell cycle was detected using flow cytometry.

RESULTSThe 3H-TDR incorporation in cells after intervention with various doses of TTR drug-serum were reduced significantly, in a concentration-dependent manner, to the level lower than that in the normal serum control group (P < 0.01). And the percentage of S and G2/M phase cells in the drug-serum treated groups also were markedly lower than that in the normal control group (P < 0.01).

CONCLUSIONDrug-serum of TTR could inhibit the proliferation of VSMC of diabetic rat in vitro in a concentration-dependent manner.

Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Diabetes Mellitus, Experimental ; pathology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Iliac Artery ; cytology ; Male ; Muscle, Smooth, Vascular ; pathology ; Rabbits ; Random Allocation ; Rats ; Rats, Wistar ; Serum

Objective To investigate the clinical manifestation of trachoma,and to select the appropriate laboratory test for clini- cal diagnosis.Design Retrospective case series.Participants Retrospective analysis of medical records from 61 patients with trachoma from Jan 2003 to Aug 2006 in Bejing Tongren Eye Center.Methods Grades of trachoma diagnosis were according to the criteria de- signed by Chinese Ophthalmological Society (1979).The general state of health,case history,and the laboratory investigations of pa- tients were recorded.Laboratory tests included the conjunctiva scraping for inclusions,C.trachomatis immune antigen test and PCR test.Main Outcome Measures Manifestation of corneal and conjunctiva,the results of laboratory tests of C.trachomatis.Results Out of sixty-one patients including 28 males and 33 females,the average old was (29.05?19.99) years.88.5% cases were inⅠstage of tra- choma,8.2% were inⅡstage,and 3.3% were inⅢstage.The C.trachomatis inclusions were found in 7 (11.5%) scraping smears.42 (68.9%) cases were positive in C.trachomatis antigen test.46 (75.4%) cases were positive in PCR tests.The positive rates of antigen and PCR test were significantly higher than that of scraping (P=0.00).Conclusions A majority of clinical patients were inⅠstage of trachoma.The degree of their distress was minimal.It is necessary to apply C.trachomatis,antigen test or PCR test to improve the clin- ical diagnosis.

OBJECTIVETo evaluate the influence of different tranexamic acid administration methods during and after cardiac surgery with cardiopulmonary bypass(CPB) on coagulation function and postoperative bleeding.

METHODSPatients undergoing elective cardiac surgery with use of CPB (n=60) were randomized in a double-blind fashion to one of two treatment groups:group A(n=30) , administered with tranexamic acid 10 mg/kg (intravenous injection slowly before skin incision) , followed by infusion of normal saline until postoperative 12 hours;and group B(n=30) , administered with tranexamic acid 10 mg/kg(intravenous injection slowly before skin incision) , followed by infusion of tranexamic acid 1 mg/(kg·h) until postoperative 12 hours. Hemoglobin, platelet count, and coagulation function were assessed before anesthesia induction, after surgery, 8am next day and 24 hours after surgery. Bleeding, allogeneic blood transfusion, and fluid infusion during the postoperative 24 hours were recorded.

RESULTNo differences were found between groups in terms of coagulant function, postoperative bleeding, allogeneic blood transfusion, and fluid infusion(P>0.05) .

CONCLUSIONCompared with intraoperative administration alone, prolonged treatment with tranexamic acid after cardiac surgery shows no advantage because it can not further improve coagulant function, reduce bleeding, or reduce allogeneic blood transfusion.

Adolescent ; Adult ; Aged ; Antifibrinolytic Agents ; administration & dosage ; therapeutic use ; Blood Coagulation ; drug effects ; Cardiac Surgical Procedures ; Cardiopulmonary Bypass ; Double-Blind Method ; Female ; Humans ; Male ; Middle Aged ; Perioperative Period ; Postoperative Hemorrhage ; prevention & control ; Postoperative Period ; Tranexamic Acid ; administration & dosage ; therapeutic use ; Young Adult

OBJECTIVEThe present study is to observe in vitro the proliferation ability of the muscle cells from permanent myopathy (PM) patients of nomokalaemic periodic paralysis (normKPP), which is caused by mutations of Met1592Val in the skeletal muscle voltage gated sodium channel (SCN4A) gene on chromosome 17q23.1. We also evaluate the possible effect of the foreign basic fibroblast growth factor (bFGF) in preventing and curing PM.

METHODSThe gastrocnemius muscle cells were taken from two male patients with PM of the same Chinese family with Met1592Val mutation of SCN4A, determined by gene screening. Four male patients suffering from the skeletal injury without PM were taken as control. All preparations were protogenerationally cultured in vitro. Proliferation of the cultured preparations was measured by MTT. Activities of the lactic dehydrogenase (LDH), creatine kinase (CK), and protein content in these cells were also detected. The effects of bFGF with different doses (10 ng/mL, 20 ng/mL, 40 ng/mL, 80 ng/mL, 120 ng/mL and 160 ng/mL) on the above mentioned parameters were also evaluated.

RESULTSCells from both PM and control subjects were successfully cultured in vitro. The cultivation of the muscle cells from PM patients in vitro was not yet seen. Results indicated the obvious stimulation of bFGF on cell proliferation, activities of LDH and CK, protein synthesis, in a dose dependent manner. The optimal dose of bFGF was 120 ng/mL (P<0.05), beyond which greater dose caused a less effect. The effect of bFGF on 160 ng /mL was stronger than that on 80 ng/mL, but there was no significant difference (P>0.05).

CONCLUSIONMyoblastic cells from patients with PM had a weaker ability of developing into the myotubules, thus they were unable to perform effective regeneration, which resulted in a progressive necrosis. The exogenous bFGF could promote the division and proliferation of the muscle cells in vitro. These results shield a light on bFGFos potential role in preventing and treating PM.

Adult ; Cell Proliferation ; drug effects ; Cells, Cultured ; Creatine Kinase ; metabolism ; Dose-Response Relationship, Drug ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Male ; Methionine ; genetics ; Middle Aged ; Muscle Development ; genetics ; physiology ; Muscular Diseases ; genetics ; pathology ; Mutation ; genetics ; Myoblasts ; drug effects ; NAV1.4 Voltage-Gated Sodium Channel ; Sodium Channels ; genetics ; Valine ; genetics