Objective To observe the levels of cAMP and adenylyl cyclase(AC) in hippocampus of mice with vascular dementia(VD) and the effect of Dihydroergocriptine(DHE),and to explore the molecular pathogenesis of VD.Methods The mice were subjected for ischemia-reperfusion three times on bilateral common carotid arteries by knots to establish models of VD and the changes of learning and memory were tested on d29/d30 after operation.DHE was administrated to another group of mice,which was taken as treatment group.The cAMP level was evaluated by the radioimmunoassay;AC mRNA positive neurons of hippocampus CA1 area were examined through in-situ hybridization.Results Compared with shamed-operation group,the learning and memory of model group was worse(P

Objective To investigate the calcium signal transduction pathway in hippocampal neurons of mice with vascular dementia (VaD) and the intervention effect of dihydroergocriptine (DHE). Methods The mice were subjected for ischemia-reperfusion repeatedly on bilateral common carotid arteries by knots to establish the VaD models. Animals with sham-operation were taken as control group. The treating group was administered with DHE after the establishment of VaD model. The behavior changes were observed through the step-down avoidance test and water maze test on the 29th and 30th days after operation. The resting 〔Ca 2+〕 i level of hippocampal neurons was evaluated by laser scanning confocal microscopy. RT-PCR technique was used to measure mRNA expression of CaM an CaMPKⅡ in hippocampal neurons. Results The resting 〔Ca 2+〕 i level in model group(43.50?3.00) was significantly higher than those in the sham-operation group (25.50?3.50) (P

OBJECTIVETo observe the effect of acupuncture along affected meridian on the mem- brane metallo-endopeptidase (MME) gene expression of migraine patients without aura (MO) of Gan-yang hyperactivity syndrome (GYHS).

METHODSTotally 20 MO patients of GYHS were randomly assigned to the acupoint group (acupuncture along affected meridian) and the non-acupoint group, 10 cases in each group. Needling was performed once per day for 10 consecutive days. Gene chip technology was used to obtain two sets of gene expression profiles and analyzed using Gene Ontology (GO).

RESULTSIn the acupoint group, MME gene expression decreased after needling (P = 0.0023).That gene was rich in the beta-amyloid metabolic process (P = 3.16E-05) and the peptide metabolic process (P = 0.009612). Its expression was not seen in the non-acupoint group.

CONCLUSIONThe effect of point selection along affected meridian could be achieved possibly by regulating the MME gene expression.

Acupuncture ; Acupuncture Points ; Acupuncture Therapy ; Endrin ; analogs & derivatives ; metabolism ; Humans ; Meridians ; Migraine Disorders ; therapy ; Syndrome

Objective To investigate the feasibility of ultrasmall superparamagnetic iron oxide- enhanced(USPIO)-enhanced MR imaging for monitoring synovitis of antigen-induced arthritis in rabbit model and explore the optimal MR imaging sequences.Methods Nine female white rabbits with antigen(0.5 ml mBSA,2 mg/ml)induced arthritis of the right knees were used in the study.The left knees of these rabbits and both knees of another 3 rabbits served as the control.Nine to 28 days(mean 21.3 d)after successful model induction,all knees were imaged before and 24 h after intravenously injection of USPIO (0.3 ml/kg),among which 2 rabbits were also imaged at 48 and 72 h after administration of USPIO respectively.The MR protocol included spin-echo(SE) T_1WI,fast spin-echo(FSE)T_2WI,gradient echo (GRE)T_2~* WI and short tau inversion recovery(STIR).Images were analyzed quantitatively and qualitatively based on signal characteristics and patterns of the synovium.Paired t-test was used for the analysis of the signal intensity of inflammatory synovial membrane before and 24 h after injection of USPIO. MR findings were correlated with histopathology.Results Arthritis was successfully induced in all 9 right knees with intraarticular injection of mBSA.Pathological examination revealed hyperplasia of synovium with infiltration of USPIO-loaded-macrophages.MR depicted synovial thickening(thickness 2.07?0.97 mm) and joint effusion.Synovium and joint fluid appeared as slightly hypo- or iso-intense on T_1 WI and hyper- intense on T_2 WI or T_2~* WI.Twenty four hours after USPIO injection,significant T_1 enhancement(ASNR 41.91%?27.94%),negative T_2 and T_2~* enhancement(△SNR -34.92%?11.77% and -57.24%? 16.05%)were demonstrated in the region of synovial inflammation respectively.The signal at 48 h and 72 h changed less than that at hour 24.No signs of arthritis occurred in all left knees and in all knees of the artificial model group.Conclusion Iron oxide phagocytized into macrophages can be a root cause resulted in signal change on USPIO-enhanced MR images.The gradient echo sequence should be the optimal sequence to be used in USPIO-enhanced MR imaging in antigen-induced arthritis.

By means of genetic cloning and recombinant techniques, full genome cDNA sequences of rotavirus strain TB-Chen were isolated from an infantile hospitalized with acute gastroenteritis. Nucleotide sequences analyses showed that the full genome of strain TB-Chen contains 18613 nucleotides, encoding 5791 amino acids. Genotyping results showed that the strain TB-Chen belongs to genotype G2P[4]/NSp4[A]. This is the first report on a full genome of Group A rotavirus in China, and has important significance for deep understanding structure and functions of rotaviruses and developing rotavirus vaccines.

Animals ; Gene Expression Regulation, Neoplastic ; Gene Silencing ; Gene Targeting ; Genes, Tumor Suppressor ; Genetic Therapy ; Humans ; Neoplasms ; therapy ; Neovascularization, Pathologic ; genetics ; Proto-Oncogenes ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; therapeutic use

BACKGROUND:Bone tumors around the Ⅱ section of pelvis are difficult to treat due to complicated anatomic structures.Using computer-aided technique,the excision range and prosthesis preparation can be individualized,which may obtain notable therapeutic efficacy in treating pelvic fractures in the clinic.OBJECTIVE:To discuss the application and the clinic effect of computer-aided technique in bone tumors therapy around the Ⅱsection of pelvis.METHODS:The pelvis model was generated with its CT data by rapid prototyping.Simulated bone resection and reconstruction were performed on the models.Then we designed surgical extension and made hemi-pelvic.Eight cases received resection of pelvic tumor and reconstruction based on computer-aided technique.RESULTS AND CONCLUSION:The resection of tumor and implantation of prosthesis were easily accessible.Two cases relapsed and 1 case loosened at 2 years after operation.According to Harris scoring criteria after total hip replacement,the scores of cases were well.The simulated resection and reconstruction of bone tumors around the Ⅱ section of pelvis based on computer-aided technique makes the operation easy and reconstruction precise,which produces good clinic results and offers a good promise for the application.

Objective To synthesize and fusion express the predicted T-cell epitopes of Schistosoma japonicum, and analyze their immunogenicities. Methods The plus and minus oligo-nucleic acid strands of epitopes P7, P17, P18 were synthesized following their DNA sequence, respectively. The Nco I restriction enzyme sites were added to the 5′ end of epitope gene and the Xho I restriction enzyme sites were added to the 3′ end of epitope gene. The plus and minus strand of each epitope gene was annealed to form double strand DNA fragments. Then the double strand DNA fragments encoding epitope peptide were cloned into the site between Nco I and Xho I of plasmid pET32c(+) to construct recombinant plasmid which was transformed into E.coli DH5?. The recombinant plasmid containing P7, P17, P18 genes respectively was identified by PCR, restriction digestion and DNA sequencing, and then transformed into E.coli BL21 (DE3) for expressing the fusion protein. The fusion protein of peptide-thioredoxin(Trx) was expressed by inducing with IPTG and analyzed with SDS-PAGE. The fusion proteins were purified with Ni2+ column affinity chromatography. Meanwhile, the peptides P7, P17, P18 were synthesized artificially following their amino acid se-quence. By using the purified epitope peptide fusion proteins and synthesized epitope peptides, the splenic cells of C57BL/6J mice immunized with ultraviolet-attenuated cercaria of Schistosoma japonicum were stimulated respectively. The stimulation activity of fusion proteins and synthesized peptides were assayed by detecting the incorporation rate of 3 H-thymidine. Results The double strand DNA fragments of epitopes P7, P17, P18 were successfully cloned to form recombinant plasmids, all of which could express a 20 kDa fusion protein. Both the fusion protein and synthesized epitope peptides of P7 and P17 were able to stimulate the lymphocyte cells to proliferation effectively. Conclusion The peptide P7 and peptide P17 are testified as T-cell epitopes of Schistosoma japonicum.

Objective To explore the protective effect of recombinant superoxide dismutase(SOD)fusion protein against the infection of Schistosoma japonicum Chinese strain.Methods The recombinant SOD fusion protein was expressed and purified with Glutathione sepharose 4B.C57BL/6J mice were immunized with the recombinant SOD fusion protein mixed with Freund adjuvant.Four weeks after the final immunization,the mice of the experiment and control groups were challenged with(45?2)S.japonicum cercariae.All the mice were sacrificed on the forty-fifth day after the challenge to calculate the worm reduction rate and egg reduction rate,and to observe the pathologic changes of liver tissue of the mice.Results The worm reduction rate was 35.63% and the egg reduction rate was 31.17% in the experiment group.The number of granuloma in the live tissue of the experiment group was less than that of the control group,and the mean diameter of single granuloma in the experiment group reduced by 22.32% compared with that of the control group.The IgG subclass levels of IgG1,IgG2a,IgG2b were higher than those of the control group.Conclusion The recombinant SOD fusion protein has a protective effect against Schistosoma japonicum infection.

Objective To express the cytosolic superoxide dismutase(SOD)of Schistosoma japonicum and analyze its antigenicity.MethodsThe DNA sequence of Schistosoma japonicum gene of cytosolic SOD was amplified by reverse transcription polymerase chain reaction(RT-PCR)and subcloned into the expression vector pGEX-4T-3 to construct a recombinant plasmid Sj SOD-pGEX-4T-3.This recombinant plasmid was transformed into component cell of E.coli BL21(DE3).The fusion protein of GST-SOD was expressed by inducing with IPTG and purification by affinity chromatography.The specific antiserum was prepared by immunizing the BALB/c mouse with GST-SOD fusion protein,and the immnuogenicity of GST-SOD was investigated by Western blot analysis.ResultsThe gene of cytosolic SOD was amplified successfully and subcloned into expression vector pGEX-4T-3 forming the recombinant expression plasmid Sj SOD-pGEX-4T-3.The fusion protein GST-SOD was expressed after the recombinant containing Sj SOD-pGEX-4T-3 being induced by IPTG.Immunizing the BALB/c mouse with the fusion protein GST-SOD produced high titer specific antiserum of 1∶12800 and the fusion protein GST-SOD was recognized by sera of severe infection rabbits.ConclusionsThe cytosolic SOD has been expressed successfully and has a preferable immunogenicity.