The complete length of P gene from rabies virus was amplified by RT-PCR using a pair of specific primers designed according to the relevant sequences from GenBank. The PCR product was cloned into cloning expression vestor pGM-T to obtain the cloning expressed plasmid pGM-T-P. After double-digestion by NotI and EcoRI, the product was transferred into prokaryotic expression vetor pET-32a(+)to obtain the prokaryotically expressed plasmid pET-32a-P. The target gene was then expressed in the E.coli BL21(DE3) cell with IPTG induction. The highest expression of target protein was analysed by SDS-PAGE, and the good immunoreactivity to rabies virus antibodies was proved by Western-blot analysis. By using purified protein, the indirect ELISA assay for the detection of rabies virus antibodies in canine serum was applied after management of the optional working condition.

Objective To explore the effects of co-culture of chondrocytes and bone mesenchymal stem cells (BMSCs) on constructing engineered cartilages, and confirm the most suitable ratio of chondrocytes to BMSCs. Methods Chondrocytes and BMSCs were isolated from articulars cartilages of rabbit (1 month old)cells (40 μl 4×107/ml) were seeded into a poly(lactic-co-glycolic acid) (PLGA) scaffold and cultured statically for 2 days. They were transferred into the cyclic pressures system, and cultured under cyclic pressures for 3 weeks. The engineered cartilages were harvested and examined by gross observation, histological staining, immunohistochemisty of collagen Ⅱ, the content of glyeosaminoglycans, GAGs, DNA and the percentage of collagen Ⅱ dyeing area. Results The engineered cartilages of the co-cultured groups grew bigger than those of the chondrocytes alone group, and their surfaces were smooth and glossy. The distributions of cartilaginous extracellular matrices in the co-cultured groups were more homogenous than those of the chondrocytes alone group.gen Ⅱ dyeing area of the co-cultured groups were higher than those of the chondrocytes alone group. The conConclusion Co-culture of chondrocytes and BMSCs could improve the quality of engineered cartilages. The

Objective: To observe the improvement of symptom and cardiac function after intervention on depressive mood in patients with myocardial infarction (MI) Methods: Among 76 patients with MI, 47 with depressive mood (according to results of SDS and HRSD) were randomly divided into intervention group (n=26) and control group (n=21), the former received health education and supportive therapy, some with major depression received antidepressant for 3 months Three months later, both groups were assessed with SDS and HRSD again, as well as assessments of symptoms of heart disease and cardiac function Results: The scores of SDS and HD in both groups were lower 3 month later (P

AIM: To study the effects of artemisunate on anoikis resistance in human breast cancer cells. METHODS: After breast cancer MCF-7 cells were treated with artemisunate, the growth of anchorage independence of breast cancer MCF-7 cells was examined in soft agar colony formation, and apoptosis of breast cancer were evaluated by terminal deoxynucleatidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and DNA ladder. RESULTS: Artemisunate significantly inhibited the growth of anchorage independence of breast cancer MCF-7 cells in soft agar colon formation in a dose-dependent manner. Artemisunate induced apoptosis in breast cancer cells was conformed by DNA ladder and TUNEL assay, which was in a time-and dose-dependent manner. CONCLUSION: Artemisunate inhibits the growth of anchorage independence of breast cancer MCF-7 cells, which is related to anoikis.

Objective To investigate the operation experience of reducing time of heart clamp in double valve replacement and tricuspidplasty in 66 patient. Methods From May 1999 to March 2003, 66 patients received mitral valve replacement(MVR), aortic valve replacement(AVR) and tricuspidplasty. Mitral valves were replaced by continuous suture. Aortic valves were replaced by continuous suture in 59 cases and interrupted suture in 7 cases. Results 132 artificial valves were grafted. The time of aortic clamp and pump was 34.5?4.3 minutes and 56.1?7.8 minutes respectively. There were no postoperative severe complications and death in all 66 cases in hospital. Conclusions Decreasing the time of aortic clamp and pump is good for myocardium protection and may reduce postoperative complications.

Colorectal cancer is currently the world's fourth incidence of malignant tumors,the early clinical manifestations are not obvious,so the early diagnosis is difficult to find,most of them are in progress.Treatment of advanced stage with chemotherapy,interventional therapy,radiotherapy and other methods,in which chemotherapy in the killing of tumor cells at the same time,the normal cells of the body also has a killing effect.In recent years,all kinds of new nano targeting delivery system has been developed,which can be targeted to the tumor tissue,so it is more and more important in the treatment of intestinal tumor,especially with metastasis.The author has made an overview of the types of nano drug carrier materials,the research status and its application in the research of colorectal cancer.

BACKGROUND:Acute toxicity in vivo experiments in previous studies has been confirmed that the median lethal dose of chitosan microcapsules is higher than 2 000 mg/kg, but the specific pathogenic mechanism is unclear.
OBJECTIVE:To explore the influence of nano chitosan on MC3T3-E1 cellgrowth as a bone substitute material, as wel as the physiological function of rats.
METHODS:MC3T3-E1 cells were respectively cultured in Dulbecco’s modified Eagle’s medium with different concentrations of chitosan nanoparticles (0, 10 mg/L, 100 mg/L, 1 g/L, 10 g/L). The absorbance values were determined. Changes in MC3T3-E1 cellmorphology were observed by scanning electron microscope after 24 hours culture. 10 g/L nano chitosan suspension was prepared using PBS. Two different doses of nano chitosan suspension (166.67 and 16.67 mg/kg body weight) with PBS were injected intraperitoneal y into Sprague-Dawley rats, three times a week, for 4 weeks. The control group was injected with equal volume of physiologic saline. Serum biochemical markers were detected to analyze the functions of liver and kidney of rats. Moreover, histopathology slices were observed to evaluate the morphological changes of tissue and inflammatory infiltration.
RESULTS AND CONCLUSION:10 mg/L, 100 mg/L, 1 g/L, 10 g/L nano chitosan suspensions were found growth inhibition in MC3T3-E1 cells as compared with the control group (P<0.05). The reunion of chitosan was observed in the cytoplasm of MC3T3-E1 cells by transmission electron microscopy. On the cellsurface, pseudopodia formed, wavy undulating membrane, nucleus degeneration, fragmentation and condensation were found. Compared with the control group, blood urea nitrogen, Na+levels were significantly increased in rats injected with nano chitosan suspension at two dosages, but the K+level in the high concentration group was decreased significantly (P<0.05). cellapoptosis was found in the liver and renal tissue in a dose-dependent manner. It suggests that apoptosis may be the possible mechanism of nano chitosan toxicity, and normal physiological function may be impacted over a certain dose.

Adult ; Female ; Heart Neoplasms ; Humans ; Pericardium ; Pheochromocytoma

Objective To investigate HIV high risk population size estimation method,analyze the epidemic situation in China and offer scientific evidence for policy making. Methods Appropriate methods to estimate different type high risk populations were ascertained by experimental study. Results Size estimation methods and procedures for different type high risk population were formed.Epidemic estimation and parameters fit for China were ascertained.HIV epidemic situation and characteristics were stated. Conclusion Methods formed in the study are the key points for the understanding of number of PLWHA,for the exact judgment of HIV epidemic,for the improvement of inspection system and examination work,for the formation of "Four Free and One Care".

Objective To establish the model of abdominal heterotopic heart transplantation and grope steady operation methods.Methods The heart transplantation was performed in the control group(group A) and experiment group(group B) with 30 rats in each group.Another 30 animals not performed the operation were as the blank group(group C).During the procurement of the donor heart saline solution containing heparitin was injected into the inferior vena cava of liver.At the same time abdominal aorta was transected.Then heart was perfused until it stopped.The activity of superoxide dismutase(SOD) and content of malondialdehyde(MDA) in blood was measured after transplantation.Results The total perfusion time of the graft was shortened within 2 min and hot ischemia time was shortened within 25 s in the group B.The activity of SOD and content of MDA between two groups was significantly different(P<0.05).Conclusion The perfusing method improved with new technique is quicker and more effective than the traditional one and results in the successful rate improvement.