Animals ; Brain ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Morphinans ; pharmacology ; Morphine Dependence ; metabolism ; Nitric Oxide Synthase Type I ; metabolism

Objective To investigate the potential effects of rosuvastatin on white matter lesion and spatial memo?ry function in chronic hypertensive rats. Methods Fourty-nine male Sprague-Dawley rats were randomly divided into sham-operation group, vehicle-treated group, and rosuvastatin-treated group (10 mg/kg).A model of stroke-prone reno?vascular hypertensive rat (RHRSP) was induced by using the two-kidney two clip method in the vehicle-treated group and the rosuvastatin-treated group. Blood pressure was monitored regularly. Morris water maze experiment was conduct?ed to assess spatial memory function. Luxol fast blue stanning was used to examine the degree of leukoaraiosis. Immuno?fluorescence and electron microscopy was used to detectβ-amyloid deposits. TUNEL staining was used to assess apopto?sis. Results The blood pressure of RHRSP increased progressively after operation.Blood pressure was significantly high?er in RHRSP than in sham-operation group (P<0.01). The escape latencies of the rosuvastatin-treated group were mark?edly shorter in RHRSP than in sham-operation group (P<0.01). The numbers of crossing hidden platform in the 3 groups of rats were 4.55±1.23, 1.00±0.80 and 3.79±0.95 times. There were significantly differences in numbers of crossing hid? den platform among three groups (P<0.01). Luxol fast blue stanning showed that the grading scores for WML were lower in the rosuvastatin-treated than in the vehicle-treated group (P<0.01). Rosuvastatin significantly decreased the burden of Aβdeposits(17.47±3.59 vs. 4.42±1.57,P<0.01)and the TUNEL+cells(37.84±4.73 vs. 14.42±2.43,P<0.01)in the fron?tal cortex when compared with the vehicle-treated group. Conclusions Rosuvastatin may ameliorate spatial memory func?tion through attenuation of white matter lesion, Aβdeposits and apoptosis .

Objective To analyze the results of HBsAg weakly positive and both HBsAg and HBsAb simultaneously positive in 5 items of hepatitis B detected by ELISA .Methods 115 cases of HBsAg weakly positive and 95 cases of both HBsAg and HBsAb simultaneously positive were screened out from 35 280 cases of 5 items detection results of hepatitis B .210 screened samples were performed the electrochemiluminescence immunoassay (ECLIA) quantitation .Results 115 cases of HBsAg weakly positive were re‐detected by using ECLIA ,90 cases had the consistent results with the coincidence rate of 78 .3% .After ECLIA re‐detection in 95 cases of HBsAg and HBsAb double positive results ,11 cases had the consistent results with the coincidence rate of 11 .6% .Conclu‐sion The results of HBsAg weakly positive and both HBsAg and HBsAb double positive in 5 items of hepatitis B detected by ELISA must be cautious .In the detection results of HBsAg weakly positive ,the majority are the samples of HBsAg ,HBeAb and HBcAb positive and HBsAg and HBcAb positive .The results of HBsAg and HBsAb simultaneously positive have poor reliability , which should be careful to issue the detection reports .

Child ; China ; Humans ; Paramyxoviridae Infections ; physiopathology ; Respiratory Tract Infections ; physiopathology ; virology

Inthis study, the DNA fragment encoding the regions Ⅰ to Ⅱ of CSP gene from Plasmodium falciparm isolate FCC1/HN was cloned into an. expression vector pcDNA3 contained cytomegalovius (CMV) and transformed into human Hela cell line. The expressed protein PfCSP (condidate vaccine) was used to immunize BALB/c mice by subcutaneous、 intravenous or intraperitoneal administration respectively. The splenocyte of BALB/c mice immumized with the condidate vaccine released significantly IL-2 and IFN-γ following stimulation with this vaccine. It is associated with increase of the splenic T lymphocyte proliferation stimulated by this vaccine and enhancement of NK cell killing activity in the former studies. These results suggested that the vaccine could stimulate T cell response and enhance the cell-mediated immunity.

Objective Using the technology of siRNA to inhibit gene expression of T cells'nonreceptor tyrosine protein kinase Lck in asthmatic mice,and to study the effect of siRNA inhibited Lck to the function of T cells in asthmatic mice.Methods The 21 - 23 bp RNA fragments of mouse T cell Lck were made by chemosynthesis.INTERFERinTMsiRNA Transfection Reagent was used as transfection reagent to transfect the siRNA into the spleen T cells of asthmatic mice for 48 hours.Then T cells were mixed with bone marrow dendritic cells (DC) of asthmatic mice for another 48 hours.Cell culture suspension was collected and the level of IL-4,IL-13,IL-2,INF-γ were detected with respondent ELISA kits; Western Blot was used to identify if the expression of Lck was blocked.Results The expression of Lck in T cells almost could not be detected in siRNA interference group.The levels of IL-4 and IL-13 in siRNA interference group( 10.19 ± 1.66,12.34 ±0.79) were lower than no-siRNA interference(28.06 ±2.88,27.87 ± 1.61 )and control group ( 22.07 ± 2.5 1,20.47 ± 2.37 ),and the difference was statistical significant ( P ＜0.01 ).Conclusions Special siRNA could block the expression of special gene,and Lck specific siRNA could block the activation and differentiation of T cells and reduce the secretion of inflammatory cytokines in asthmatic mice.

ObjectiveUsing the technology of siRNA to inhibit the gene expression of no-receptor　tyrosine protein kinase Lck in T cells of asthmatic mice,and to study the therapeutic effect of Lck specific　siRNA in asthmatic mice.MethodsReceptor tyrosine protein kinase Lck specific siRNA fragments were　taken from chemosynthesis.In vivo-jetPEITM was used to transfect the siRNA into mice body through tail　vein injection.The mice were killed 48 hours later,and the levels of IL-4,IL-17 in bronchoalveolar lavage　fluid (BALF) were detected with respondent ELISA kits.The change of inflammatory histopathology in lung　was observed with H.E.staining.The expression of Lck in lung was detected with immunohistochemistry　(IHC),and the level of Lck in lung tissue homogenate was detected with Western Blot.Results Compared with asthmatic group[ (234.68 ± 11.15 ) pg/ml,( 96.76 ± 8.28 ) pg/ml],the levels of IL-4,IL-17　[ (234.68 ± 11.15)pg/ml,(96.76 ±8.28) pg/ml] in the BALF of siRNA interference group decreased,　and the inflammation in the lung relieved.IHC indicated that the expression of Lck in lung decreased and　the level of Lck in lung tissue homogenate decreased ( P ＜ 0.05 ).Conclusions Lck specific siRNA　could reduce the level of IL-4,IL-17 in the lung tissues of asthmatic mice,and relieve the inflammatory reaction in lung.

Objective:To screen for the pathogenesis-related genes of osteosarcoma and to assess their roles for the de- velopment of osteosareoma.Methods:Total RNA was extracted from 3 ATCC osteosarcoma cell lines and an osteoblastic cell line and was used to synthesize biotinylated cRNAs;the latter were hybridized to Affymetrix~(?)GeneChip~(?)U133A ar- rays and a gene with more than 2 folds of change was selected.Ten of the differentially expressed genes were chosen and the primers were designed and the synthesized.Then SYBR~(?)Green real-time PCR(RT-PCR)method was used to detect the expression of the 10 genes in 9 fresh osteosarcoma specimens.ABI Prism 7 000 system was used to analyze the differ- ent expression between osteosarcoma cell line and osteoblastic cell line.Results:We identified 58 up-regulated and 142 down-regulated genes in the 3 osteosareoma cell lines.Many of the genes were firstly reported to be related to the patho- genesis of osteosarcoma.These differentially expressed genes were mainly involved in energy and material metabolism,on- cogene,signal transduction gene,transcription- related genes,cell cycle-related genes,cell apoptosis-related gene,im- mune response gene,tumor suppressor genes,etc.The array results of 10 randomly selected genes were further verified by the RT-PCR in 9 fresh osteosarcoma specimens.Conclusion:Many genes are involved in the pathogenesis of osteosarcoma. Gene microarray can help to discover the genes related to the pathogenesis of osteosarcoma,which may lay a foundation for studying the molecular mechanism of osteosarcom.

Objective: To obtain polycystin-1 intracellular region. Methods: cDNA of polycystin-1 intracellular region was generated by PCR and then cloned into pProEX Hta, which was prokaryotic expression vector. After verified by sequencing, the recombinant was transformed into E.coli host to express and purify the fusion protein by affinity chromatography. Results: 660 bp cDNA of polycystin-1 intracellular region and 2.6×104 fusion protein were obtained. Conclusion: The fusion protein containing polycystin-1 intracellular region is obtained and is helpful for preparing anti-polycystin-1 monoclonal antibody.

Objective: To obtain polycystin-1 intracellular region. Methods: cDNA of polycystin-1 intracellular region was generated by PCR and then cloned into pProEX Hta, which was prokaryotic expression vector. After verified by sequencing, the recombinant was transformed into E.coli host to express and purify the fusion protein by affinity chromatography. Results: 660 bp cDNA of polycystin-1 intracellular region and 2.6×104 fusion protein were obtained. Conclusion: The fusion protein containing polycystin-1 intracellular region is obtained and is helpful for preparing anti-polycystin-1 monoclonal antibody.