Objective To study the effects of A?25-35 on the expression of gene P21,CDK4,E2F1 and BAX in cultured PC12 cells.MethodsPC12 cells were treated with 25 ?mol/L A?25-35,the relation between cell cycle redistribution and apoptosis was analyzed by flow cytometry(FCM).Protein and mRNA expression of P21,CDK4,E2F1 and BAX were detected by RT-PCR and Western-blot respectively.Results About 90% PC12 cells were found arrest on G0/G1 by FCM being deprived serum.Treated with 25 ?mol/L A?25-35 for 8,16,24 h,the percent of S phase cells was raised remarkably(P

Aim To study the effective mechanism of curcumin on abnormal cell cycle and apoptosis of serum-deprived PC12 cells induced by ?-amyloid peptide 25-35(A?25-35).Methods Synchronized PC12 cells were pretreated with 5 ?mol?L-1 Cur for 1 h,and then 25 ?mol?L-1 A?25-35 for 24 h.Protein and mRNA expression of p21,CDK4,E2F1 and bax were detected by RT-PCR and Western blot respectively.Results After synchronized PC12 cells being pretreated with 5 ?mol?L-1 Cur for 1 h,the mRNA and protein expression of p21 gene were increased gradually,while CDK4,E2F1 and bax gene were decreased.Conclusion Cur maybe effects the redistribution of cell cycle and apoptosis of serum-deprived PC12 cells induced by A?25-35,through increasing the mRNA and protein expression of p21,and decreasing the mRNA and protein expression of CDK4,E2F1 and bax gene.

Objective To evaluate the clinical significance of serum level of vascular endothelial growth factor (VEGF),angiopoietin (Ang)-1 and Ang-2 in patients with rheumatoid arthritis (RA).Methods Serum levels of VEGF,Ang-1 and Ang-2 were measured with enzyme linked immunosorbent assay (ELISA).Twenty-one healthy subjects,24 osteoarthritis patients and 82 rheumatoid arthritis patients were included.We defined active and inactive group according to RA disease active score,while early active RA and late active RA group were defined on the basis of disease course.There were 28 early active patients,32 late active patients and 22 inactive patients with rheumatoid arthritis.At the same time 29 RA patients were examined with ultrasound.Synovial hypertrophy (US joint count SH,US index SH),synovial fluid(US joint count SF,US index SF),resistance index and power Doppler signal (US joint PD,TSS) were scored.The correlation was analyzed.We also detected the serum levels of VEGF,Ang-1,Ang-2,ESR,CRP and DAS28 in 25patients with active RA after 3 month regular treatment.We used one-way ANOVA to compare the differences between groups,and Wilcoxon test to compare the differences between before and after treatment.We analyzed the correlation with linear correlation or Spearman rank test.Results The serum level of VEGF [(1285 ±272) pg/ml],Ang-1 [(0.55±0.25)ng/ml] in patients with rheumatoid arthritis were higher than osteoarthritis patients [(934±80) pg/ml,(0.32±0.16) ng/ml] and normal controls [(565±115) pg/ml,(0.24±0.21) ng/ml],and the serum level of Ang-2 [(1.36±0.40) ng/ml] was higher than normal controls [(0.52±0.32) ng/ml].The serum level of VEGF [(1355±194) pg/ml] in early active patients was higher than late active patients [(1096±477) pg/ml] and inactive patients [(862±91) pg/ml].The serum level of Ang-1 in early active patients,late active patients and inactive patients with rheumatoid arthritis had no statistically significant differences.The serum level of Ang-2 in inactive patients [(2.0±2.0) ng/ml] was significantly higher than late active patients [0.9±0.8) ng/ml].The serum level of VEGF was positively correlated with US joint SH,US index SH,US joint PD,and TSS.The serum level of Ang-1 was positively correlated with US joint SH,US joint PD,and TSS.The serum level of VEGF and Ang-1 were negatively correlated with RI.The serum level of Ang-2 was not correlated with US joint SH,US index SH,US joint SF,US index SF,US joint PD,TSS and RI.In the active RA patients,the serum level of VEGF,Ang-1 and Ang-2 was positively correlated with each other.In the inactive RA patients,the serum level of VEGF,Ang-1 and Ang-2 was not correlated with each other.The serum level of VEGF and Ang-1 before treatment was slightly higher than that of after treatment,but the difference was not statistically significant.The serum level of Ang-2 after treatment was significantly higher than that before treatment.ESR,CRP and DAS28 of after treatment were lower than those before treatment.Conclusion The serum VEGF and Ang-1 level could be used as useful index to reflect RA synovial thickening and angiogenesis The serum level of Ang-2 could be used as one of the efficacy indices.They may influence each other,and they may be the key factors that mediate the onset and development of RA angiogenesis and synovial inflammation.

Objective To express UL148 RNA of human cytomegalovirus (HCMV) clinical strains in vitro and to study its functions. Methods Urine of a newborn with HCMV infection was inocula-ted into human embryo lung cells. HCMV clinical strain was isolated and identified by multiplex PCR. UL148 gene was amplified and cloned into pGEM-T-Easy plasmid after double enzyme digestion. A recombi-nant plasmid was constructed and located at the downstream of the T7 promoter. The recombinant plasmid was identified by electrophoresis of the recombinant plasmid,PCR product and double enzyme product. Se-quencing analysis was used for final confirmation. UL148 was transcribed into RNA by 32P labeling. Post-translational modification sites were analyzed by bioinformatics method based on UL148 sequence characteris-tics. Results The clinical strain of HCMV was obtained in vitro. Electrophoresis and sequencing analysis confirmed the successful construction of the recombinant plasmid. UL148 RNA was transcribed in vitro by T7RNA polymerase. Post-translational modification sites showed that UL148 gene contained one cell adhe-sion sequence, one legume lectins beta-chain signature, two N-myristoylation sites, one casein kinase Ⅱphosphorylation site,seven protein kinase C phosphorylation sitse, one cAMP/cGMP-dependent protein ki-nase phosphorylation site, two N-glycosylation sites and one transmembrane region. Conclusion UL148 gene might encode a viral adhesion molecule involving in the signal transduction in host cells.

Objective: To investigate the UL148 gene function of human cytomegalovirus (HCMV) low passage clinic isolate and new strategies for anti-HCMV treatment, the DNA-based external guide sequences (EGSs) were designed to inhibit UL148 RNA expression. Methods: UL148 RNA secondary structure was analyzed by RNA structure technique, an appropriate region was chosen for DNA-based EGS57 synthesis, targeted the UL148 RNA. The M1RNA and UL148 RNA were generated by PCR for transcription in vitro. The UL148 RNA and M1RNA were transcribed in vitro under the function of T7 RNA polymerase. The UL148 was labelled by ³²P. The cleavage reactions were carried out by mixing up EGS, M1RNA with UL148 RNA for 1 h. The products were separated by urea denaturing polyacrylamide gel electrophoresis and detected with Typhoon Phosphor Imager. Results: UL148 RNA ranged from 58 to 72 sites was the binding position, and 57 was a cleavage site. EGS57 was designed and synthesized. EGS57 was combined with UL148 RNA to form the natural substrate of M1RNA. UL148 RNA and M1RNA were synthesized through T7 RNA polymerase catalyzing, and the products were conformed. After cleaving reactions, DNA-based EGS57 was shown to be able to cleave UL148 RNA efficiently in vitro by a complex with M1RNA. Conclusions UL148 RNA was cleaved efficiently by EGS57, and the cleaving site was conformed as expectation. It will provide the gene silent tool effectively for further study the function of UL148 gene.

The effects of cardiomyocyte grafting on left ventricular (LV) remodeling and function in rats with chronic myocardial infarction were evaluated using high-frequency ultrasound. Chronic myocardial infarction was induced in 50 Wister rats by ligating the left anterior descending artery. They were randomized into two groups: a trial group that received neonatal rat cardiomyocyte trans- plantation (n=25) and a control group which were given intramyocardial injection of culture medium (n=25). The left ventricular (LV) geometry and function were evaluated by high-frequency ultrasound before and 4 weeks after the cell transplantation. After the final evaluation, all rats were sacrificed for histological study. The results showed that 4 weeks after the cell transplantation, as compared with the control group, the LV end-systolic dimension, end-diastolic dimension, end-systolic volume and end-diastolic volume were significantly decreased and the LV anterior wall end-diastolic thickness, LV ejection fraction and fractional shortening were significantly increased in the trial group (P<0.01). Histological study showed that transplanted neonatal rat cardiomyocytes were found in all host hearts and identified by Brdu staining. It was suggested that transplantation of neonatal rat cardiomyocytes can reverse cardiac remodeling and improve heart function in chronic myocardial infarction rats. High-frequency ultrasound can be used as a reliable technique for the non-invasive evaluation of the effect of cardiomyocyte transplantation.