Objective To construct the short hairpin RNA(shRNA) expression vector of survivin and down-regulate the expression of survivin through RNA interference in neuroblastoma cell line SH-SY5Y. Methods Two pairs of oligonucleotide sequences specific for human survivin mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pBSHH1 plasmid. After being identified by restriction enzyme digestion and sequencing, the recombinant plasmids pBSHH1-S1 and pBSHH1-S2 were transfected into SH-SY5Y cells, respectively. Survivin expression in the transfected cells was assayed by both RT-PCR and western blot. Results Enzyme digestion analysis and DNA sequencing showed that the oligonucleotide fragments were correctly inserted into pBSHH1 plasmid, and survivin expression in the transfected cells was knocked down significantly by pBSHH1-S1 or pBSHH1-S2 at both the protein and mRNA level. Conclusion The shRNA expression vectors of survivin were successfully constructed, and could down-regulate survivin expression in SH-SY5Y cells, which lay a foundation for further research on gene therapy for tumors such as neuroblastoma.

Objective To investigate the clinical effect and security of spraying Zhikang Capsules guided by gastroscopy in treatment elderly upper gastrointestinal bleeding.Methods Eighty-four elderly gastric duodenal ulcer bleeding patients were randomly assigned to group A,B and C (n =28 for each group).Patients in group A were used noradrenalin to spray with gastroscopy,in group B were used thrombin and in group C were used Zhikang Capsules.The total effective,the immediate homeostasis rate,the incidence of rebleeding,the emergency operation rate and the incidence of adverse reaction were recorded and analyzed.Results Total effective rate of group C was 96.4％ (27/28),significantly higher than that in group A (71.4％,20/28),B (75.0％,21/28) and the difference was significant(P＜0.05).The incidence of rebleeding in group A,B,C were 28.6％ (8/28),17.9％ (6/28),3.6％ (1/28) respectively,and the difference was significant (P ＜0.05).The same trend was seen regarding of the incidence of adverse reaction among three groups (25.0％,7/28;10.7％,3/28;0,0/28;P＜0.05).Conclusion Spraying Zhikang Capsules guided by gastroscopy is showed with good therapeutic effect in treating elderly upper gastrointestinal bleeding and high safety,and with less adverse reaction.

Objective To investigate the effect of adrenomedullin(ADM) on hypoxic pulmonary vascular structural remodeling and proadrenomedullin N-terminal 20-peptide(PAMP) in rats with hypoxic pulmonary hypertension.Methods Twenty-four male Wistar rats were randomly divided into control group(n=8),hypoxia group(n=8) and hypoxia with ADM group(n=8).ADM was subcutaneously administered into rats of hypoxia with ADM group by mini-osmotic pump(300 ng/h).The concentration of plasma PAMP was measured by radioimmunoassay,and the expression of PAMP in pulmonary artery was detected by immunohistochemical assay.Results Mean pulmonary arterial pressure,the ratio of right ventricular mass to left ventricular plus septal mass,and relative medial thickness and relative medial area of pulmonary arteries significantly increased in hypoxic rats as compared with that of controls(P

We analyze the characteristics of foreign students and the problems in biochemistry teaching process by summarizing the practice of medical biochemistry teaching for undergraduate foreign students in Wenzhou Medical College. After careful thought on these problems,we brought forward some ideas and methods which may be useful to reform the teaching mode and improve the teaching results of medical biochemistry for foreign students.

AIM: To examine the alteration of pathologic structure and gaseous molecules in rats with pulmonary hypertension induced by high pulmonary blood flow.METHODS: Aortocaval shunting was produced for 11 weeks in rats, and pulmonary hemodynamics was evaluated.Pulmonary vascular micro- and ultra- structure was also examined.Meanwhile,the concentration of plasma nitric oxide (NO) and carbon monoxide (CO) was measured by spectrophotometry.The expression of endothelial nitric oxide synthase (eNOS) and heme oxygenase-1 (HO-1) in pulmonary arteries was detected by immunohistochemistry.RESULTS: After 11- week aortocaval shunting,pulmonary artery mean pressure was significantly increased.Muscularization of small pulmonary vessels and relative medial thickness and area of pulmonary arteries were obviously increased in shunting rats compared with controls.Ultrastructure of intrapulmonary arteries changed obviously in shunting rats.Meanwhile,plasma NO concentration was increased and eNOS expression in pulmonary artery endothelial cells was significantly augmented in rats of shunting group.Plasma carbon monoxide level and HO-1 expression in puomonary artery smooth muscle cells,however,were not altered in shunting rats.CONCLUSIONS: Pulmonary vascular structural remodeling is the important pathologic basis of pulmonary hypertension induced by a left-to-right shunt,and NO other than CO might play an important regulating role in the development of high pulmonary blood flow-induced pulmonary hypertension.

Objective: To examine the alteration of pathologic structure and endogenous hydrogen sulfide pathway in rats with pulmonary hypertension induced by high pulmonary blood flow. Methods: Sixteen SD rats were randomly divided into shunting group and control group. An 11 week aortocaval shunting was produced in rats of shunting group, and pulmonary artery mean pressure (mPAP) was evaluated using right cardiac catheterization. The ratios of right ventricular mass to body weight (RV/BW) and right ventricular mass to left ventricular plus septal mass[RV/(LV+S)] were also detected. Pulmonary vascular micro and ultra structures were examined. Meanwhile the concentration of plasma hydrogen sulfide (H 2S) was measured by spectrophotography. The gene expression of cystathionine ? lyase (CSE)was detected by in situ hybridization, and the activity of CSE in lung tissues was measured by H 2S production according to chemical analysis. Results: After 11 weeks of aortocaval shunting, pulmonary artery mean pressure was significantly increased. Muscularization of small pulmonary vessels and relative medial thickness of pulmonary arteries were obviously increased in shunting rats compared with controls. Ultrastructure of intrapulmonary arteries changed obviously in shunting rats. Meanwhile, plasma H 2S concentration was decreased and the activity of CSE (according to H 2S production) in lung tissues decreased in shunting rats. CSEmRNA expression by pulmonary arteries was significantly suppressed. Conclusion: Pulmonary vascular structural remodeling is the important pathologic basis for pulmonary hypertension induced by high pulmonary blood flow. The down regula tion of endogenous H 2S pathway might play an im portant role in the development of high pulmonary blood flow induced pulmonary hypertension.

Based on the practice of bilingual education in medical biochemistry,this paper discussed the advantages,problems and solutions of bilingual education in medical biochemistry of teaching-centred medical college or university.This study will be helpful for developing bilingual teaching of biochemistry in other teaching-centred medical colleges or universities.

OBJECTIVETo introduce a safe and specific approach of (13)C magnetic resonance spectrum ((13)C MRS) spectroscopy and investigate the alterations in hepatic anabolism.

METHODSRelative anaplerotic, pyruvate recycling and gluconeogenic fluxes were measured by (13)C MRS isotopomer analysis of blood glucose from rats with 40% body surface area burn injury, and from rats exposed to sham injury. A short chain fatty acid, [U (13)C] propionate which was avidly extracted by the liver, was infused intravenously to deliver (13)C into the citric acid cycle. Proton-decoupled (13)C MRS of deproteinized plasma or extracts of the freeze-clamped liver were used to determine the distribution of (13)C in blood or hepatic glucose.

RESULTSThere was no difference in the multiplets detected in the glucose carbon-2 anomer from blood or liver after 45 or 60 minutes of the infusion of the propionate, indicating that steady-state isotopic conditions were achieved. Gluconeogenesis relative to citric acid cycle flux was not altered by burn injury; in both sham and burn groups the rate of glucose production was about equal to flux through citrate synthase. In the sham group of animals, the rate of entry of carbon skeletons into the citric acid cycle was about 4 times than that in the burn group. Similarly, flux through pyruvate kinase (again relative to citrate synthase) was significantly increased after the burn injury.

CONCLUSIONSSince results from analysis of the blood glucose are the same as that of the hepatic glucose, (13)C distribution in the glucose and hepatic metabolism can be assessed based on the (13)C MRS analysis of the blood glucose.

Animals ; Blood Glucose ; analysis ; Burns ; complications ; Carbon Isotopes ; Citric Acid Cycle ; physiology ; Disease Models, Animal ; Gluconeogenesis ; physiology ; Liver Diseases ; etiology ; pathology ; Liver Function Tests ; Magnetic Resonance Spectroscopy ; methods ; Male ; Probability ; Radiographic Image Enhancement ; Rats ; Rats, Sprague-Dawley ; Reference Values ; Sensitivity and Specificity

OBJECTIVETo investigate the mutation characteristics of ATM gene in Chinese patients with ataxia-telangiectasia (AT).

METHODSMutation of ATM gene was screened by polymerase chain reaction, reverse transcription-polymerase chain reaction, polyacrylamide gel electrophoresis combined with DNA direct sequencing in two Chinese AT patients.

RESULTSA missense mutation of 1346(G>C) in exon 11, which was a homozygotic mutation, was identified in one patient; a nonsense mutation of 610 (G>T) in exon 6 combined with a missense mutation of 6679 (C>T) in exon 47, which was a compound heterozygotic mutation, were identified in the other patient. They were co-segregated with the disease and were localized within the functional domain of ATM gene.

CONCLUSIONTotally three novel ATM gene mutations were identified in two Chinese AT patients.

Ataxia Telangiectasia Mutated Proteins ; Base Sequence ; Cell Cycle Proteins ; genetics ; Child ; China ; Codon, Nonsense ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; Female ; Gene Frequency ; Humans ; Male ; Mutation ; Mutation, Missense ; Protein-Serine-Threonine Kinases ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Spinocerebellar Ataxias ; genetics ; Tumor Suppressor Proteins ; genetics

OBJECTIVETo investigate the role of autophagy on the pathogenesis of spinocerebellar ataxia 3/Machado-Joseph disease (SCA3/MJD).

METHODSHEK293 cells expressing polyglutamine-expanded ataxin-3 were used as cell model for SCA3/MJD. The level of polyglutamine-expanded ataxin-3 was detected after cells were treated with different inhibitors or inducer of autophagy.

RESULTSInhibition of autophagy increased aggregate formation and cell death in HEK293 cells expressing mutated ataxin-3, and vice versa.

CONCLUSIONThe data suggested that autophagy is involved in the degradation of mutant ataxin-3, resulting in a decrease in the proportions of aggregate-containing cells and cell death in HEK293 cells expressing polyglutamine-expanded ataxin-3. It is possible that autophagy may be applied as a potential therapeutic approach for SCA3/MJD.

Ataxin-3 ; Autophagy ; Cell Line ; Humans ; Machado-Joseph Disease ; genetics ; metabolism ; physiopathology ; Mutation ; Nerve Tissue Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Peptides ; metabolism ; Repressor Proteins ; genetics ; metabolism