Brain Neoplasms ; metabolism ; Cell Line, Tumor ; Glioma ; metabolism ; Humans ; Neoplastic Stem Cells ; drug effects ; metabolism ; Reactive Oxygen Species ; metabolism ; Sodium Selenite ; pharmacology

Objective To construct the eukaryotic expression plasmid containing glyceraldehydes-3-phosphate dehydrogenase (GAPDH) and cysteine protease inhibitor ( CPI ) gene from periodic Brugia malayi (Bm) and to lay foundation for studying multivalent vaccines. Methods Total RNA was extracted from periodic Bin. The BmGAPDH and BmCPI genes were amplified by RT-PCR. The PCR product was cloned and then subeloned into eukaryotic recombinant plasmid vector pcDNA3.1 (+). pcDNA3.1 (+)/BmGAPDH/BmCPI was constructed. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification, and were transformed into HeLa cell subsequently. The transient expression of BmGAPDH and BmCPI were examined by RT-PCR. The expressed protein was identified by sodium dodeeylsulphate-polyacrylamide gel electrophoresis(SDS-PAGE). Results Two specific bands of around 877 bp of BmGAPDH and 621 bp of BmCPI were amplified, consistent with the expected value. The same bands were obtained by double restriction enzyme digestion of recombinant plasmids or PCR using recombinant plasmid as template. BmGAPDH and BmCPI mRNA were highly expressed in transfeeted HeLa cell. The relative molecular mass (Mr) of the recombinant protein was about 54 × 103. Conclusion The recombinant eukaryotic expression plasmid pcDNA3.1 (+)/BmGAPDH/BmCPI has been constructed successfully and the protein is expressed correctly in mammalian cell.

OBJECTIVE: To investigate the application of pulsed-wave Doppler tissue imaging ( PW-DTI) in evaluating left ventricular systolic and diastolic function in patients with chronic heart failure (CHF). METHODS: Mitral annular velocities (MAV) were measured by PW-DTI in 35 patients with CHF and 25 healthy subjects. Traditional indices for evaluating the global left ventricular function by conventional echocardiography were also studied as a comparison. RESULTS: Peak systolic, peak early diastolic, peak late diastolic mitral annular velocities ( Sa, Ea, Aa), and Ea/Aa ratio progressively decreased in CHF patients compared with the healthy subjects (P <0.01 ). Sa of the mitral annulus correlated linearly with the left ventricle ejection fraction (LVEF) (r =0.890, P < 0.01). Compared with the healthy subjects, Ea in all 3 subgroups of diastolic dysfunction in the CHF group significantly decreased (P <0.01). Aa in 2 subgroups (pseudonormal filling and restrictive filling) decreased (P < 0.01 ) and the decreased Ea/Aa was found in the delayed relaxation and pseudonormal filling subgroups compared with the healthy subjects (P<0.001). CONCLUSION MAV measured by PW-DTI can be used for assessing the left ventricular systolic and diastolic function in CHF patients.
Diastole ; Echocardiography, Doppler, Pulsed ; Female ; Heart Failure ; diagnostic imaging ; physiopathology ; Humans ; Male ; Systole ; Ventricular Function, Left ; physiology

OBJECTIVETo simulate the chemical microenvironment of injured brain tissue, and to explore the effect of this chemical microenvironment on temperature sensitive umbilical cord mesenchymal stem cells (tsUC).

METHODSRat models of traumatic brain injury (TBI) were made by fluid percussion injury, and then the brain tissue extracts of the injured regions were acquired. Human umbilical cord mesenchymal stem cells (UC) were isolated and cultured, and the tsUC were obtained through the infection of temperature-sensitive Simian 40 Large T- antigen (ts-SV40LT) retrovirus. After that, both the two kinds of cells were cultured on the polyacrylamide gels which mimicking the elastic modulus of brain. Four groups were included: UC cultured under normal temperature (UC group), UC cultured added brain tissue extract under normal temperature (UC plus extract group), tsUC cultured under mild hypothermia (tsUC group), and tsUC added brain tissue extract under mild hypothermia for 3 days, then normal temperature for 4 days (tsUC plus extract group). After 24 hours, the apoptosis level was checked. Cell growth and morphological changes in each group were given dynamic observation. Seven days later, cell immunofluorescences were implemented for examining neural differentiation level.

RESULTSCompared with UC plus extract group, the apoptosis and proliferation in UC plus extract group were significantly reduced (P < 0.01) and increased (P < 0.01) respectively. Cell immunofluorescence showed that the both GFAP and Neuron positive cells were significantly enhanced in UC plus extract group than those in tsUC plus extract group.

CONCLUSIONtsUC combining with mild hypothermia could significantly reverse injury induced cell apoptosis, improve cell proliferation and neural differentiation under chemical microenvironment after brain injury, which confirmed the adaptation and resistance of tsUC under mild hypothermia after TBI.

Animals ; Apoptosis ; Brain ; cytology ; pathology ; Brain Injuries ; pathology ; Cell Proliferation ; Humans ; Mesenchymal Stromal Cells ; chemistry ; Neurons ; cytology ; Rats ; Temperature ; Umbilical Cord ; cytology

Objective To clone and sequence the cysteine protease inhibitor gene of periodic Brugia malayi(BmCPI) and predict B-cell epitopes in amino acide sequence of BmCPI in order to provide basis for further study the expression of BmCPI and its function. Methods Total RNA was extracted from periodic Brugia malayi.A couple of specific primers were designed on the basis of known sequences of cysteine protease inhibitor gene from BmCPI. The desired gene was amplified by PCR technique from cDNA. The PCR products were purified and cloned into plasmid pGEM-T by T-A cloning method, transformed into Escherichia coli(E, coli) strain DH5α. The recombinant plasmids were screened and identified by digestion with restriction enzyme and PCR amplification. Five parameters and methods were used to predict B-cell epitopes in amino acide sequence of BmCPI. Results For RT-PCR, a specific band of around 621 bp was amplified. The same band was obtained by double restriction of recombinant plasmids or PCR using recombinant plasmid as template. The result of DNA sequencing showed that BmCPI shares 99% nucleotide sequence identity with that of published sequence. It showed that B-cell epitopes were probably at or adjacent to 23 - 32, 50 - 79 and 117 - 126 in its amino acide sequence. Conclusions pGEM-BmCPI is successfully constructed and sequenced, anticipated objective is reached and conditions is provided for further study of BmCPI expression and its function.

Objective To observe the effects of triptolide on drebrin and cofilin expression in the hippocampus of rats with Alzheim-er's disease (AD). Methods Sixty male Sprague-Dawley rats were equally divided into control group, model group and triptolide-treated group with 20 cases in each group. The AD model was established with unilateral injection of beta amyloid 1-40 (Aβ1- 40) into hippocampus in rats. The control group was established with unilateral injection of normal saline with the same volume into hippocampus in rats. The triptolide-treated group was administered triptolide intraperi-toneally, 0.4 mg/kg, once a day, for 15 days after modeling. Spine density of hippocampal neurons was assayed by Golgi staining. Drebrin and cofilin expression of hippocampal neurons was assayed by immunohistochemical staining and reverse transcription polymerase chain reaction (RT-PCR). Results The spine density of hippocampal neurons was higher in the triptolide-treated group than in the model group (P<0.05). The average optical density of drebrin was higher in the triptolide-treated group than in the the model group (P<0.01), while the cell number and average optical density of cofilin were lower (P<0.05). The drebrin mRNA expression was higher in the triptolide-treated group than in the model group (P<0.05), and the cofilin mRNA expression was lower (P<0.01). Conclusion Triptolide may delay the degeneration of dendritic spines in hippocampal neurons of AD rats by regulating the expression of drebrin and cofilin.

UNLABELLEDTo reduce the huge labor-cost in the screening in traditional monoclonal antibody generation, We established a new system for monoclonal antibody generation integrating with protein array. BALB/c mice were immunized by eight recombinant proteins respectively, and the positive hybridoma cells were obtained by cell fusion and ELISA screening. All the eight kinds of positive hybridoma cells were mixed, cloned, screened by protein array, and definite dilution cloned.

RESULTS175 single cell clones were obtained by complex cloning, and 119 of those were positive clones. Then 8 positive cell lines were generated by the following 2 rounds definite dilution cloning. By comparing with the traditional method, we got 8 monoclonal antibodies using the combined protein array screening and multiplex cloning method in 1 cycle, and fewer amounts of antigens were used. As a result, the combined protein array and multiplex cloning method could be used as an economical, rapid and simple tool applying in high throughput monoclonal antibody generation.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; isolation & purification ; Female ; Humans ; Hybridomas ; metabolism ; Mice ; Mice, Inbred BALB C ; Protein Array Analysis

OBJECTIVETo clarify the change of vegetation types and its relationship between the density of alive-snails in the areas of "breaking dikes or opening sluice for water store" in Jicheng.

METHODSSynthesized false color images of Jicheng before and after 1998 (1994 and 2003) were classified without supervision and results were compared. Vegetation types were identified on the spot.

RESULTSNormalized difference vegetation index (NDVI) of snail habitats before 1998 were between 126 and 183 in Jicheng, whose vegetation types were mainly paddy, cotton and cabbage. NDVI of snail habitats in Jicheng after 1998 were between 152 and 193 whose vegetation types were mainly poplar forest, bulrush and grass. Areas of snail habitats increased from 64.64% to 66.47%. Snail habitats were mostly composed of mixed vegetation types and mono-typed vegetation was hardly found. According to the density of alive-snails orders from high to low were poplar forest and bulrush, poplar forest and grass, bulrush.

CONCLUSIONVegetation types would not be identified by unsupervised classification only. Poplar forest, bulrush and grass were closedly related to the density of alive-snails.

Animals ; Breeding ; China ; epidemiology ; Disease Vectors ; Ecology ; Environmental Monitoring ; Epidemiological Monitoring ; Fresh Water ; Plants ; Satellite Communications ; instrumentation ; Schistosoma japonicum ; isolation & purification ; Schistosomiasis japonica ; epidemiology ; transmission ; Snails ; growth & development ; parasitology ; physiology

OBJECTIVETo investigate the hypocholesterolemic activity of red yeast rice (RYR) and its underlying mechanism.

METHODSThree groups of hamsters were fed either the control diet or one of the two experimental diets containing by weight 0.1% RYR (0.1RYR) or 0.3% RYR (0.3RYR). Blood (0.5 mL) was collected from the retro-orbital sinus into a heparinized capillary tube at the end of week 0, 3, and 6. Plasma lipoproteins were measured using enzymatic kits, while fecal neutral and acidic sterols were quantified using a gas-liquid chromatography.

RESULTSPlasma total cholesterol was reduced by 12% in 0.1RYR group and by 18% in 0.3RYR group compared with the control value. Similarly, plasma triacylglycerol was decreased by 11% in 0.1RYR group and by 24% in 0.3RYR group. Western blotting analysis demonstrated that RYR had no effect on sterol regulatory element binding protein 2, liver X receptor, 3-hydroxy-3-methylglutary-CoA reductase, LDL receptor, and cholesterol-7alpha-hydroxylase. HPLC analysis confirmed that RYR contained 0.88% monacolin K. It was recently found that RYR supplementation increased excretion of fecal acidic sterols by 3-4 folds compared with the control value.

CONCLUSIONHypocholesterolemic activity of RYR is mediated at least partially by enhancement of acidic sterol excretion.

Animals ; Bile Acids and Salts ; secretion ; Biological Products ; pharmacology ; Blotting, Western ; Body Weight ; drug effects ; Cholesterol ; metabolism ; Cholesterol 7-alpha-Hydroxylase ; metabolism ; Cricetinae ; Dietary Supplements ; Feces ; chemistry ; Feeding Behavior ; drug effects ; Hydroxymethylglutaryl CoA Reductases ; metabolism ; Lipoproteins ; blood ; Liver ; enzymology ; Liver X Receptors ; Naphthalenes ; analysis ; Organ Size ; drug effects ; Orphan Nuclear Receptors ; metabolism ; Receptors, LDL ; metabolism ; Sterol Regulatory Element Binding Protein 2 ; metabolism ; Weight Gain ; drug effects

OBJECTIVETo provide the fittest model for forecasting schistosomiasis prevalence in Haokou village of "breaking dikes or opening sluice for waterstore" in Dongting Lake areas by comparing the results of Moving Average, Exponential Smoothing, Autoregressive Model and Autoregressive integrated moving average model (ARIMA model) from 1990 to 2002.

METHODSError sum of square of four statistical methods was compared and the fittest model was chosen.

RESULTSError sum of square of predicted schistosomiasis prevalence rates in Haokou village from 1994 to 2002 were 39.40, 39.86, 26.63, 22.54 respectively.

CONCLUSIONARIMA model seemed to be the fittest one in the prediction of schistosomiasis prevalence in Haokou village of "breaking dikes or opening sluice for waterstore" in Dongting Lake from 1990 to 2002.

Animals ; China ; epidemiology ; Epidemiologic Methods ; Forecasting ; Humans ; Models, Statistical ; Prevalence ; Risk Assessment ; Schistosomiasis japonica ; epidemiology ; Time Factors