China ; epidemiology ; Humans ; Occupational Diseases ; epidemiology ; prevention & control ; Surveys and Questionnaires ; Trimethyltin Compounds ; analysis ; poisoning

Chromatography, Gas ; Humans

Objective To screen and identify the dengue virus-specific antigens,then establish the ELISA detection method for dengue virus antibody.Methods Using bioinformatic software DNAStar and ANTHEPROT to analyze the hydrophilicity,flexibility,surface probability and antigenicity of dengue virus type 1-4,Japanese encephalitis virus and Yellow fever virus M,E and NSI protein amino acid sequence and also consider the influence of secondary structure.Then in accordance with epitopes localion and amino acid sequence similarity,forecast the share and specific epitopes.Reference the sequence information of different dengue virus strains in GenBank to analyze the epitopes conservative.Based on the results of bioinformatic analysis,5 specific epitopes were amplified and inserted into prokaryotic expression vector pMAL-C2x or pET32a.Then the vectors was transferred into E.coli Rosetta( DE3 ).lsopropyl-β-D-thiogalactoside(IPTG) was used to induce the expression of gene segments.SDS-PAGE were used to identify the expression proteins,and the antigenicity were tested using Western blot.Using the antigen selected by Western blot,ELISA method for dengue virus antibody detection was established.Results Eighty shared epitopes and 25 specific epitopes were forecasted,and 5 antigenic fragments encloude analyzed epitopes from dengue virus type 2 and 3 were expressed in E.coli successfully.One dengue virus type 1-4 shared antigens (Den-Ag5),one dengue virus type 2 and 4 shared antigens( DenAg3),one dengue virus type 1-3 shared antigens(Den-Ag2) and two dengue virus type 1,2 and 4 shared antigens( Den-Ag1,Den-Ag4)were conformed using Western blot.Using antigens Den-Ag5,Den-Ag1 and DenAg2,the ELISA method for dengue virus antibody detection were established.Conclusion Based on the bioinformatic analysis and Western blot verification,5 dengue virus specific antigen were conformed,and the ELISA detection method for dengue virus antibody were established.

Objective To observe the changes of the skeletal development related cells after dihydroorotate dehydrogenase (DHODH) deficiency.Methods The DHODH expression in MC3T3-E1 cells derived from mouse calvaria osteoblast precursor cells was inhibited by specific small interfering RNAs (siRNAs),and cell proliferation,ATP production and expression levels of bone-related genes were investigated in these cells.Results After reducing the DHODH expression by using specific siRNAs,cell proliferation was inhibited and cell cycle was arrested at G1/S stage.In addition,the ATP production was reduced in whole cells,especially in mitochondria.Furthermore,the expression levels of Runt-related transcription factor 2 (Runx2) and osteocalcin (Ocn) mRNAs in the DHODH inhibition group were decreased compared with the control group.Conclusion Inhibiting DHODH protein affects the differentiation and maturation of osteoblasts.The mitochondrial dysfunction in osteoblasts may be one of causes leading to the abnormal bone formation in Miller syndrome.

AIM: To explore the relationship between dynamic changes of population spike (PS) and morphologic alterations in hippocampal CA1 region and morphology after transient ischemia/reperfusion and the improving effects of Chinese herbs 9602. METHODS: Changes of evoking population spike ware investigated by electrical stimulating Schaffer collateral in CA1 region of hippocampal slice after ischemia/reperfusion in vivo . Apoptosis and morphologic alterations at different time points after cerebral ischemia/reperfusion were detected by using TUNEL and Nissl staining. RESULTS: The threshold voltage of CA1 region in evoking population spike increased markedly as compared with sham control. The enhancement of wave amplitude was reduced significantly after tetanic stimulation. The duration of enhancement in amplitude decreased with the passage of reperfusion. Above all were observed from 8 h after ischemia/reperfusion. They became remarkable and got to its top at 7 day after ischemia/reperfusion treatment. TUNEL positive cells were observed in hippocampal CA1 region at 8 h, got to the top at 24 h and then gradually reduced after ischemia/reperfusion. A lot of abnormal cells in CA1 region was found, and the number of pyramidal cell reduced progressively by Nissl staining after ischemia/reperfusion. Chinese herbs 9602 reduced the threshold voltage of CA1 region in evoking population spike remarkably, enhanced the wave amplitude and prolonged the duration of PS enhancement; decreased the number of TUNEL positive cell, prevented the reduction of pyramidal cell in CA1 region. CONCLUSIONS: The excitability and reactivity were decreased and there was a gradual functional disturbance of synaptic transmission in CA1 pyramidal cell and most notable changes happened at 7 d ischemia/reperfusion, suggesting that was partly due to delayed neuronal death induced by ischemia/reperfusion. Apoptosis plays an important role in the functional deficiency of CA1 region of hippocampus induced by cerebral ischemia/reperfusion. The effects of 9602 on ameliorating the excitability and reactivity of CA1 pyramidal cells relate to inhibiting apoptosis, attanuating delayed neuron death induced by ischemia/reperfusion.

Aim To investigate the up-regulation of miR-22 through Wnt pathway inhibits the proliferation,migration and invasion in human gastric MGC803 cells induced by diallyl disulfide(DADS).Methods The effects of proliferation,migration,and invasion of gastric cancer cells were evaluated by MTT,wound-healing and invasion assays.Online prediction software was applied to search the target gene of miR-22.Luciferase report gene assay was used to assess the target genes Wnt-1 of miR-22.The expressions of Wnt-1,β-catenin and TCF-4 were tested by qRT-PCR and Western blot,respectively.Results MTT showed that DADS and miR-22 notably decreased the proliferation compared with control group(P<0.05).Wound-healing assay showed that DADS and miR-22 could significantly inhibit the migration of MGC803 cells compared with the control group, especially in miR-22+DADS(P<0.05). Invasion assay showed that DADS and miR-22 could markedly inhibit the invasion of MGC803 cells compared with the control group, especially in miR-22+DADS(P<0.05). Online prediction software to search the target gene exhibited that Wnt-1 may be a target gene of miR-22. Luciferase report gene assay disclosed that Wnt-1 was identified as a direct target of miR-22. Qrt-PCR showed that the expression of Wnt-1 Mrna was respectively down-regulated by DADS and miR-22 compared withcontrol group, especially in miR-22+DADS(P<0.05). Western blot exhibited that DADS and miR-22 obviously suppressed the expressions of Wnt-1, β-catenin and TCF-4 proteins, especially in miR-22+DADS(P<0.05).Conclusion Up-regulation of miR-22 through Wnt pathway can remarkably suppress the proliferation, migration and invasion in MGC803 cells by DADS.

OBJECTIVETo investigate the phenotype change of airway smooth muscle cell (SMC) in bronchial asthma and the effect of Zhichuan Capsule (ZCC) on it.

METHODSSixty male SD rats were randomized into 5 groups equally, the normal control group, the model group, and three treated groups treated with ZCC, Western medicine (WM) and ZCC + WM respectively. Excepting rats in the control group, they were all made into bronchial asthma model rats and received respective treatment. Histomorphology of the airway SMC was observed by electron microsope at the 4th week, and the expressions of smalpha-actin and sm-MHC were examined at the 2nd and the 4th week after provocation.

RESULTSAs compared with the control group, moyfilaments in SMC were lesser, secretive organelles were more and levels of smalpha-actin and sm-MHC expressions at the 4th week were lower in the model group (P < 0.05). While histomorphology of SMC in the WM group was as narmal, but some hypoxia changes were seen, such as the fracture of mitochondria cristae and deplation of matrix; histomorphology of SMC in the ZCC treated group and the ZCC + WM treated group were similar to that in the normal control group, and their expressions of smalpha-actin and sm-MHC were enhanced as compared to those in the model group (P < 0.05).

CONCLUSIONAirway SMC changes from contractile phenotype to synthetic phenotype in asthma rats, but the progress of change may be inhibited by ZCC.

Animals ; Anti-Asthmatic Agents ; therapeutic use ; Asthma ; drug therapy ; pathology ; Bronchi ; cytology ; pathology ; Drugs, Chinese Herbal ; therapeutic use ; Male ; Myocytes, Smooth Muscle ; metabolism ; pathology ; Phenotype ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

Objective To investigate the effect of carbon monoxide(CO) on hydrogen sulfide(H_2S)/cystathionine-?-lyase system(CSE) in vascular smooth muscle cells(VSMC) of spontaneous hypertensive rats(SHR).Methods The SHR VSMC were divided into 2 groups:experimental group(hemin was added to the culture media at a concentration of 10 ?mol/L)and control group (dimethyl sulfoxide was added to the culture media at an equal volume).The content of H_2S in the cultrue media was measured by sulfide electrode method,and the CSEmRNA level was assayed by competitive reversed transcription-polymerase chain reaction(RT-PCR).Results Compared with control group,the content of H_2S in the culture media of hemin-treated SMCs was significantly lower[(16.03? 2.14) ?mol/L vs (13.31?1.74)?mol/L],and the CSEmRNA level in hemin-treated SMCs was decreased markedly(0.17?0.04 vs 0.13?0.03).Conclusion CO can down-regulate the H_2S/CSE system in SMC of SHR.

Objective To investigate the effect of nitric oxide on hydrogen sulfide/ cystathionine-?-lyase(CSE) system in the vascular smooth muscle cells of spontaneously hypertensive rats(SHR).Methods The SHR aortic smooth muscle cells(SMCs) were divided into 2 groups: sodium nitroprusside(SNP) group and control group.The content of hydrogen sulfide in the culture media was measured by sulfide electrode method,and the CSE mRNA level was assayed by competitive reversed transcription-polymerase chain reaction((RT-PCR)).Results Compared with control group,the content of hydrogen sulfide in the culture media of SNP group was significantly higher [(16.16?3.71) ?mol/L vs(22.71?4.84) ?mol/L],and the CSE mRNA level in SMCs in SNP group was lower than that of control group.Conclusion Nitric oxide exerted complicated effect on the hydrogen sulfide/CSE system in the SHR smooth muscle cells.J Appl Clin Pediatr,2006,21(3):140-141

Objective To explore the relationship between dynamic changes in ultra-micro-structural of pulmonary arteries and endogenous hydrogen sulfide in rats with left-right shunt.Methods Rats in shunt group were subjected to an abdominal aorta-inferior vena cava shunt to create an animal model of pulmonary artery structural remodeling. After 1 day, 3 days, 1 week, 4 weeks and 8 weeks of experiment, the ultra-micro-morphologic changes of pulmonary arteries of rats were observed under electronic microscope and H_2S concentration in serum was evaluated by modified sulfide electrode method.Results The changes of ultra-micro-structure of pulmonary arteries were progressively exacerbated, endothelial cells became swollen and large in size on 3 days, smooth muscular cells increased in size as well as the change of endothelial cells in 1 week, and they changed from contractile phenotype to synthetic phenotype in 4 weeks.Conclusions Shunt exhibited changes of ultra-micro-structure of pulmonary arteries are accompanied by the changes of endogenous H_2S. It is suggested that endogenous H_2S might play a protective role in changes of ultra-micro-structure of pulmonary artery.