The 6.5kb specific fragment containing the T7 promoter and the transcription vector was cut down from the full-length cDNA clone of Newcastle disease virus strain ZJI of goose origin,and thereafter it was self-ligated to form the high guality plasmid for mutagenesis.Site-directed mutagenesis technique was used for inserting three additional G nucleotides(nts) into the region between the T7 promter and the leader sequence of the NDV.The RT-PCR was employed to amplify the F/HN genes fragements,and then they were ligated by the shairing restriction enzyme BsmBI and finally the corresponding fragment in the mutatant full-length cDNA was substituted by the new one.The sequencing results showed that the three additional G nts were successfully inserted and the mutant nts in the full-length cDNA were corrected and all these studies lay a foundation for the research on the reverse genetics of NDV strain ZJI.

[Objective] To explore the characteristics and achievements of YU Chang'sYuyi Grassin treating typhoid disease. [Method] Based on studying, thinking and analysing theYuyi Grass, combined with related literature research, it summed up the YU Chang characteristics and achievements in treating typhoid disease. [Results] YU Chang treating typhoid fever syndrome had accuracy differentiation, forcefully rectifying children with typhoid fever being mistaken for the fallacies of present concern of charlotte, fine solution of typhoid rehabilitation treatment; slow treatment for typhoid critical patients, illustrating the appropriate use of ginseng theory for typhoid fever, having the original opinion in such use. [Conclusion] The Yuyi Grass treating typhoid disease has rich experience and unique achievements. These insights and achievement in learning by using the theory of Treatise on Febrile Disease for treating typhoid disease have important guiding significance in Chinese medicine clinic.

Objective To prepare monoclonal antibody specifically against carcinoembryonic antigen glypican-3 (GPC3) and its preliminary application. Methods GPC3 was cloned with PCR to pET16b vector and expressed in E. co-liBL21. Spleen cells were obtained from Balb/c mice embedding immunized with purified antigen intraperitoneally, and fused with Sp2/0 cells. Hybridoma cells were screened by indirect ELISA, and identified by Western blot assay using puri-fied protein after the cell fusion. The indirect immunofluorescence method was used to detect the GPC3 expression in HepG2 cell line. Results The prokaryotic expression vector of GPC3 was successfully constructed, and GPC3 was stably expressed in E. coliBL21. A mouse hybridoma cell line secreting monoclonal antibody to GPC3 was obtained. Western blot analysis showed that monoclonal antibody specifically recognized recombinant protein. Monoclonal antibody could be used to detect GPC3 protein expression in HepG2 cell line by indirect immunofluorescence. Conclusion The monoclonal antibody against GPC3 was successfully obtained.

AIM:To determine the right ventricular angiotensin converting enzyme2 (ACE2) activity in adult SD rats under normoxia and hypoxia environment,and to detect the relationship between ACE2 and high altitude heart disease initially. METHODS:Adult male Sprague Dawley (SD) rats were raised under hypoxia environment simulated to high altitude (5 000 m,23 h/d),then divided into hypoxia 1 d,15 d and 30 d groups randomly. The control group was set up under normoxia environment. The right ventricular function,ponderal index,pulmonary artery pressure and right ventricular ACE2 activity were determined. The effect of captopril or nitrendpine on cardiac ACE2 activity in hypoxia 30 d group was also observed. RESULTS:The dramatic up-regulation of cardiac ACE2 expression of mRNA and protein and its activity in hypoxia 30 d group was observed,together with obvious increase in right ventricular function,ponderal index and pulmonary artery pressure. Although captopril or nitrendipine depressed pulmonary artery pressure and cardiac function dramatically,no significant alteration of right ventricular ACE2 activity was detected. CONCLUSION:Chronic hypoxic exposure promotes the ACE2 expression and activity in right ventricular,indicating that ACE2 may be correlated to the changes of cardiac architecture and function induced by hypoxia. Up-regulation of ACE2 expression maybe contribute to the increase in right ventricular ACE2 activity,and pulmonary artery hypertension may not be the main reason for the changes of ACE2 activity under hypoxia environment.

Objective To explore the effect of ω-3 fish oil on nutrition improvement and inflammatory reaction of patients with gastrointestinal tumor after operation.Methods 60 cases with gastrointestinal tumor were divided into control group(30 cases) and study group (30 cases),both groups were provided with parenteral nutrition treatment[ 104.6 kJ · kg-1 · d-1 ].Fish-oil fatty emulsion was given to the study group.The postoperative for the first day received half of the total energy and the total energy The remaining four days.Blood samples were gained on the morning of day 1,on the morning of day 3 and day 6 after operation respectively to measure albumin ( ALB),prealbumin (PA),total protein(TP),transferrin(TRF),the neutrophilic granulocyte count,lymphocyte count (TLC),serum C-reactive protein(CRP).Results Both groups of patients was comparable(all P ＜ 0.05 ).Both groups of patients was treated after five days of postoperative and ALB,TP,TRF were not significantly different.PA in both groups on day 6 was significantly increased,and the study group was higher than control group.there were statistical differences between them ( all P ＜ 0.05 ).The neutrophilic granulocyte count and CRP of both groups were significant reduced,and the study group was lower than control group.There were statistical differences between them( all P ＜0.05 ).Conclusion ω-3 fish oil on nutrition could improve nutritional quality and modulate inflammatory reaction of patients with gastrointestinal tumor after operation.

Banana bunchy top virus (BBTV),family Nanaviridae,genus Babuvirus,is a single stranded DNA virus (ssDNA) that causes banana bunchy top disease (BBTD) in banana plants.It is the most common and most destructive of all viruses in these plants and is widespread throughout the Asia-Pacific region.In this study we isolated,cloned and sequenced a BBTV sample from Hainan Island,China.The results from sequencing and bioinformatics analysis indicate this isolate represents a satellite DNA component with 12 DNA sequences motifs.We also predicted the physical and chemical properties,structure,signal peptide,phosphorylation,secondary structure,tertiary structure and functional domains of its encoding protein,and compare them with the corresponding quantities in the replication initiation protein of BBTV DNA1.

BACKGROUND:Currently, human umbilical cord derived-mesenchymal stem cel s are mainly for local transplantation, which has some shortcomings, such as large trauma, bleeding, complications, that limit its widespread application in clinical practice. OBJECTIVE:To investigate the feasibility of intravenous transplantation of human umbilical cord derived-mesenchymal stem cel s for repair of spinal cord injury. METHODS:Eighty Wistar rats with spinal cord hitting were divided into five groups:blank control group with no transplantation (n=10), DMEM local transplantation group (n=15), DMEM intravenous transplantation group (n=15), cel local transplantation group (n=20), cel intravenous transplantation group (n=20). The functional recovery of spinal cord injury was observed with Basso, Beattie and Bresnahan scores at regular time as wel as hematoxylin-eosin staining and immunohistochemistry staining. RESULTS AND CONCLUSION:During 1 day to 2 weeks after transplantation, there was no significant difference in the Basso, Beattie and Bresnahan scores between the five groups;within 4-12 weeks after transplantation, the Basso, Beattie and Bresnahan scores were significantly higher in the two cel transplantation groups than the other three groups, but there was no difference between these two cel transplantation groups (P>0.05). Histological observation showed that the number of voids and glial scars was less in the cel local transplantation group and cel intravenous transplantation group compared with the other three groups, and there was also no difference between the two cel transplantation groups. These results indicate that the intravenous transplantation of human umbilical cord derived-mesenchymal stem cel s is similar to the local transplantation in the repair of acute spinal cord injury, which is simple and avoids secondary injuries and various complications. It is recommended that this method provide a new approach for cel transplantation.

Telomerase, as a highly specific ribonucleoprotein, is composed by the RNA template and protein subunits. It can delay or prevent the process of disc degeneration by maintaining telomere length homeostasis as well as affecting the p53-p21-pRb pathway, p16Ink41-pRb pathway and Wnt/β-catenin signal pathway. Telomerase can regulate the senescence and apoptosis of intervertebral disc cell, expected to prevention and repair reconstruction of the structure by telomerase gene therapy of intervertebral disc degeneration.

ObjectiveTo investigate the effect and mechanisms of rostral ventrolateral medulla (RVL) on the pressor response of lateral hypothalamus-perifornical region (LH/PF) in rats.Methods30 healthy Wistar rats were randomly divided into four groups: the phentolamine group; propranolol group; atropine group and glutamate diethyl ester group, saline was as the control in every group. After microinjection of Glu into LH/PF, the blood pressure and heart rate were observed. Then phentolamine, propranolol, atropine and glutamate diethyl ester were microinjected into RVL and the blood pressure and heart rate changes induced by microinjection of Glu were observed again.ResultsMicroinjection of Glu into LH/PF can cause the blood pressure elevating and heart rate accelerating. The pressor response of Glu to excited LH/PF could be attenuated after response of phentolamine, propranolol, atropine and glutamate diethyl ester microinjected into RVL. The blood pressures of phentolamine group; propranolol group; atropine group and glutamate diethyl ester group reduced significantly different from those in the saline control group (P<0.01).ConclusionThe α-,β-,M- and Glu-receptors in the RVL induce the pressor response of LH/PF region.

Objective To explore the effect of 1,25-dihydroxyvitamin D3 [1,25-(OH)2VitD3] on insulin resistance and inflammatory response in mice with severe burns under stress. Methods One hundred and thirty mice were randomly divided into healthy group (n=10), experimental groupⅠ(n=40), experimental groupⅡ(n=40) and control group (n=40). The model mice were scalded to 30%of total body surface area (TBSA) burnt (Ⅲ°) in experimental groupⅠ, the experi-mental groupⅡand control group. Mice in experimental groupⅠwere given by gavaging 1,25-(OH)2VitD3 (1μg·kg-1) with 0.6 mL peanut oil at 8 am for every other day. At the same time and by the same way, mice in experimental groupⅡreceived 1,25-(OH)2VitD3 (4μg·kg-1) with 0.6 mL peanut oil, and mice in control group received only 0.6 mL peanut oil. The serum levels of fasting blood glucose (FBG), fasting insulin (FIns), TNF-αconcentration, and the NF-κB positive rate in wound tis-sues were determined respectively on day1, day3, day7 and day14 after burn. Results (1) The values of insulin resistance index (HOMA-IR), serum TNF-αand the NF-κB positive rate in wound tissues were higher at different time points in exper-imental groupⅠ, experimental groupⅡand control group than those in healthy group. (2) The levels of HOMA-IR at the same time points were significantly lower in experimental groupⅠand experimental groupⅡthan those in control group, and the value was significantly lower in experimental groupⅡthan that of experimental groupⅠ(P<0.05). The level of HOMA-IR was the highest at day 3 and then gradually decreased at the different time points in the same group (P<0.05). (3) The se-rum levels of TNF-αand NF-κB positive rate of wound tissues at the same time points were significantly lower in experimen-tal group Ⅰand experimental group Ⅱ than those in control group, and the value was significantly lower in experimental groupⅡthan that of experimental groupⅠ(P<0.05). The serum level of TNF-αand NF-κB positive rate of wound tissues were gradually decreased at the different time points in same group (P<0.05). Conclusion 1,25-(OH)2VitD3 can reduce the insulin resistance and inflammatory response in mice with severe burns under stress.