Objective To evaluate the effect of alginate polysaccharide JM on rats impaired by brain ischemia,and to elucidate its mechanisms.Methods By using the middle cerebral artery occlusion(MCAO) induced by nylon surgical thread inserted through the internal carotid artery into the anterior cerebral artery in rats,the effects of JM on neurological dysfunction and infarct volumn in rats brain were investigated.Neuroblastoma SH-SY5Y cells were employed to study the inhibitory effects of JM on cytotoxicity induced by hypoxia-hypoglycemia.Results JM at the doses of 12.5 and 25 mg?kg-1,iv,30 min before ischemia impressed neurological dysfunction,characterized by the decreased behavioral obstacle scores of MCAO rats.The infarct volumn declined 24h after ischemia as well.Further investigation by flow cytometry revealed that JM significantly reduces the overloading of intracellular free calcium on and suppresses apoptosis induced by hypoxia-hypoglycemia in human neuroblastoma SH-SY5Y cells.Conclusion JM showed protective effects on brain ischemia,probably related to its inhibitory effect on the overloading of intracellular free calcium ion(\i) and cell apoptosis.

Objective To prepare an active anti-tumor component, compound K (C-K), from saponins in leaves of Panax notoginseng (SLPN) using immobilized β-glucanase. Methods Two entrapments, alginate gel-1 (Alg 1) and alginate gel-2 (Alg 2), were evaluated for their ability to immobilize β-glucanase. The amount and purity of C-K obtained from the transformation process were analyzed by HPLC, and the immobilizing parameters were optimized. Results β-Glucanase can be immobilized and reused with either of the entrapment. However, using AIg 1 resulted in higher enzyme activity than Alg 2. The optimal concentration of the immobilized enzyme was 10%; The optimal crosslinking time was 4-6 h; and the optimal concentration of the crosslinking agent was 6%-7%. Conclusion Immobilized β-glucanase shows sustained enzyme activity, good ethanol tolerance, and was reusable for the preparation of C-K from SLPN.

Objective: To compere OQL of recruits of armed police in south area fore-and-aft the basic military training.Methods: The WHOQOL-BREF Scale was used to investigate 539 recruits before the training and at the end of the 3rd month of the basic military training.Results: The score of physiological,psychological,social,and environmental domain of the OQL of recruits before the basic military training were68.90?13.36,65.65?14.28,67.16?16.71,61.06?14.23,The score of physiological,psychological,social,and environmental domain of the OQL of recruits before the basic military training were 71.52?12.16,70.34?14.78,74.10?22.00,66.07?14.36,the differences were salient.Conclution:The Basic Military Training can improve the OQL of recruits of armed police in south area.The improvement are bear on the education of adapting and improved training.

This paper introduces the designing concept of the ECG treadmill system and discusses the methods of its realization.
Amplifiers, Electronic ; Computer Simulation ; Computer Systems ; Coronary Disease ; diagnosis ; Electrocardiography ; instrumentation ; methods ; Equipment Design ; Exercise Test ; instrumentation ; methods ; Humans ; Microcomputers ; Signal Processing, Computer-Assisted ; Software

OBJECTIVE(1) To investigate the promoter methylation status of gene p16(INK4a) and gene RB in breast carcinoma and the adjacent non-neoplastic hyperplastic epithelial tissue. (2) To study the correlation of p16(INK4a) gene expression at protein level with the abnormal gene methylation, the clinical manifestation and the pathological parameters.

METHODSMethylation status of promoters of p16(INK4a) gene and RB gene was detected by using methylation specific PCR in 46 cases of breast cancer, 22 cases of the adjacent non-neoplastic hyperplastic epithelium tissue and 7 cases of normal breast tissue. In addition, the p16(INK4a) gene protein expression level was also detected using immunohistochemical technique(SP method) in 46 cases of breast cancer and 22 cases of the adjacent hyperplastic epithelial tissue.

RESULTSThe methylation rate of p16(INK4a) gene was 23.9% (11/46) in breast cancer, 18.2% (4/22) in the adjacent non-neoplastic hyperplastic epithelial tissue and 1/7 in normal breast tissue, respectively. The methylation rate of RB gene was relatively low, which was 10.8% (5/46), 9.1% (2/22) and 0(0/7) in the above 3 groups, respectively. Methylation rate of p16(INK4a) gene and RB gene was not significantly different among the breast cancer, the adjacent non-neoplastic hyperplastic tissue and the normal tissues (P > 0.05). However, the methylation status of p16(INK4a) gene was closely correlated with its protein expression level and the negative ER expression result of the breast cancer (P < 0.05), but not correlated with the size of the cancer, differentiation status, lymph node metastasis, and age. The methylation status of RB gene was correlated with lymph node metastasis, but not with the size, the differentiation status, ER expression of the breast cancer and the age of the patients.

CONCLUSIONSThe abnormal methylation of p16(INK4a) gene may not play a significant role in the early stage of breast cancinogenesis, but may play a role of in the progression of the cancer. RB gene methylation may also be a indicator in choice to identify the progression and prognosis of breast cancer.

Adult ; Aged ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; genetics ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic ; Genes, p16 ; Humans ; Lymphatic Metastasis ; Middle Aged ; Receptors, Estrogen ; metabolism ; Retinoblastoma Protein ; genetics ; metabolism

The regulation of foreign gene expression in Insect-Baculovirus Expression System is very complex. In this report, the effect of 5'-UTR in the expression of hGH gene in cultured Sf9 cells was examined. A 18 bp length in the end of 5'-UTR of hGH (human Growth Hormone, hGH) cDNA including a stem-loop structure was deleted by PCR. The truncated hGH cDNA, delta 1hGH was cloned in pFastBac1, named pFast-Bac-delta 1hGH. After transforming into E. coli. DH10Bac, which have a shuttle vetor-Bacmid, the delta 1hGH was integrated into Bacmid by site-specific transposition, and an expression vector, rBacmid-delta 1hGH DNA was acquired. By transfecting the cultured Sf9 cells with the recombinant expression vector DNA, pure recombinant virus, rAcV-Bac-delta 1hGH was obtained, and hGH gene was expressed. Immuno-blot and Chemiluminescent assay revealed that the expressed hGH had normal immunological activity, the amount of hGH expression level in Sf9 cell supernatant infected with rAcV-Bac-delta 1hGH containing the truncated 5'UTR was four to five times higher than that infected with rAcV-Bac-hGH.
5' Untranslated Regions ; genetics ; Animals ; Base Sequence ; Cells, Cultured ; DNA, Complementary ; genetics ; Gene Expression Regulation ; Genetic Vectors ; genetics ; Human Growth Hormone ; genetics ; metabolism ; Humans ; Immunoblotting ; Insecta ; cytology ; genetics ; Recombinant Proteins ; isolation & purification ; metabolism

Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.

OBJECTIVETo observe the polymorphism and gene frequency of interleukin 6 (IL6) gene -572C/G in Chinese Han nationality population, that associating with susceptibility to myocardial infarction(MI) and impacting on the extent of coronary artery lesions; to analyze the function of IL6 gene -572C/G polymorphism.

METHODSWith PCR-RFLP method, IL6 gene -572C/G polymorphism was genotyped to 232 MI patients and 260 healthy adults. The effect of IL6 gene -572C/G polymorphism was observed to the extent of coronary artery lesions and the ability of IL6 production from peripheral blood mononuclear cells (PBMC).

RESULTSThere was IL6 gene -572C/G polymorphism in Chinese Hans. -572CG+GG genotype and G allele were more frequent in patients than in controls (P< 0.01). The relative risk for G allele carrier to suffer from MI was 1.68 times of CC genotype individual (95%CI 1.17-2.41, P< 0.01). However, the distribution of IL6 gene -572C/G polymorphism was no significant difference among patients with single-vessel, two-vessel and three-vessel lesions (P> 0.05). After PBMC cultured for 24 hours, the IL6 concentration in supernatant was significantly higher in subjects with CG genotype than those with CC genotype (P< 0.05).

CONCLUSIONIL6 gene -572G allele may be a genetic susceptibility factor to MI attack of Chinese Hans population, and related to the high expression of IL6.

Aged ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Interleukin-6 ; genetics ; Male ; Middle Aged ; Myocardial Infarction ; genetics ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; genetics

OBJECTIVETo determine whether fatty acid synthase (FAS) is expressed in human multiple myeloma( MM) cells and investigate the proliferation inhibition effect of fatty acid synthase inhibitor cerulenin on multiple myeloma cell line U266 and its mechanism.

METHODSFAS mRNA expression in human MM cell line U266, RPMI8226 cell was assayed by RT-PCR. The proliferation inhibition rate of U266 cells was assayed by MTr analysis. Cell apoptosis and cycle distribution were evaluated by flow cytometry (FCM).

RESULTSFAS mRNA was highly expressed in human multiple myeloma cell lines as compared with healthy donor PBMNCs. After U266 cells were treated with cerulenin (the concentrations from 5 microg/ml to 640 microg/ ml) for 24 h, the cell proliferation was markedly inhibited with a dose related manner, while the inhibition rate of human skin fibroblast cells were all lower than 30%. When U266 cells were treated with 20 pjg/ml cerulenin for 12 h and 24 h, the early apoptosis rate revealed by Annexin V/PI were 56. 9% and 69. 3% respectively, being higher than that of the blank controls (4. 3% and 1.8%, P < 0. 01). Cell cycle analysis showed it was blocked in S phase. Conclusion FAS is highly expressed in human MM. Cerulenin could induce apoptosis and inhibit proliferation of U266 cells. FAS might be a new potential target for multiple myeloma treatment.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cerulenin ; pharmacology ; Dose-Response Relationship, Drug ; Fatty Acid Synthases ; antagonists & inhibitors ; biosynthesis ; Humans ; Multiple Myeloma ; metabolism ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured

BACKGROUNDTo investigate the significance of blood HCV RNA screening in the prevention of post-transfusion hepatitis C.

METHODSTotally 56,400 anti-HCV negative blood samples collected from Jan. 2000 to Dec. 2003 were tested for HCV RNA by RT-PCR, and the patients who received the HCV RNA negative blood were followed up.

RESULTSThe HCV RNA positive rate was 2.5 per thousand (146/56,000) and none of the patients followed up suffered from HCV infection.

CONCLUSIONHCV RNA screening for the anti-HCV negative blood samples is very effective and feasible for prevention of post-transfusion hepatitis C.

Feasibility Studies ; Follow-Up Studies ; Hepacivirus ; genetics ; Hepatitis C ; blood ; etiology ; prevention & control ; Humans ; Mass Screening ; methods ; RNA, Viral ; blood ; genetics ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; Transfusion Reaction