Xenotransplantation can probably overcome the critical shortage of human organ donors for clinical transplantation.Recently,research progresses in the biology of xenograft rejection and zoonotic infections,and the generation of α1,3-galactosyltransferase-deficient pigs have moved this approach closer to clinical application,but immunological rejection is also the primary barrier in the study of xenotransplantation.This review sumarries the research progress of immunological rejection of xenotransplantation,providing some advices for the future of clinical xenotransplantation.

Objective To construct a sequence tag library of HepG2 cells by serial analysis of gene expression (SAGE). Methods Total RNA of HepG2 cells was extracted using a Trizol agent. The double strand cDNA were reverse-translated from mRNA which was isolated from total RNA. According to the protocol of SAGE, a sequence tag library of HepG2 cells was constructed. The transcript, which was represented by each tag, was analyzed by consulting the GenBank and UniGene, and the abundance of the tags was analyzed by SAGE analysis software. Results A total of 14 181 SAGE tags out of 15 586 tags were steadily and effectively amplified and the lengths of the tags were between 200-3000 bp. After sequence analysis of the tags, 2023 specific genes were revealed. Conclusions The use of improved reverse transcription agent kit may guarantee obtaining the full length cDNA, which provides materials for the construction of SAGE library. SAGE is a powerful method to investigate the gene expression profile, and it establishes a foundation for gene researches.

Objective　To investigate whether or not the antisense oligodeoxynucleotide (ODN) of vascular endothelial growth factor (VEGF) can supresses the protein expression of VEGF.Methods　SMMC 7721 is a cell line that expresses VEGF.Therefore the medium conditioned by SMMC 7721 stimulates proliferation of bovine aortic endothelial cell (BAEC).Synthesized antisense ODN complementary to one region of VEGF mRNA was added into the medium of SMMC 7721.The expression of VEGF and the stimulation effect of SMMC 7721 was then determined.Result　It was found that by means of inhibition of VEGF expression,the stimulation effect of SMMC 7721 was inhibited by the VEGF antisense ODN in a dose-dependent manner.At a dosage of 1,2.5,5,10μmol/L,the inhibition rate was 63.2%,82.4%,210% and 287.5%,respectively.Conclusion　It is promising that VEGF antisense ODN works as a new gene therapeutic modaliey for antiangiogenesis of HCC.

Acupuncture Therapy ; Adipose Tissue ; injuries ; Adolescent ; Adult ; Combined Modality Therapy ; Female ; Humans ; Male ; Massage ; Young Adult

Objective To study the proliferation and differentiation patterns of hepatic stem cells from mice cultured in vitro isolated identify their characteristics.Methods Mice were divided into 5 groups according to the pregnant (embryo days,ED13,ED15 and ED18) or born age (day0 and 3).Hepatic stem cells were isolated and cultured in vitro.The amount of the stem cells as well as their growing situation and differentiation pattern were observed and compared among different groups and markers of stem cells (CD117,CD90.1,CD49f,c-Met),hepatic cells(AFP) were used to identify the cultures.Resulls The cells with best situation of growing as colonies were obtained from ED15 group.Their expression of specific markers suggested that they were hepatic stem cells.The stem cells isolated from ED15 mice in subculturing proliferated in line pattern and differentiated in reverse line pattern.The expression of AFP varied as normal distribution as cell differentiation development.Conclusion Most cells have characteristics of hepatic stem cells isolated from fetal liver and the number of these cells decreases gradually as embryo duration prolongs.The hepatic stem cells proliferate in line pattern and differentiate into hepatic cells after in vitro culture.

Objective To determine the value of detection of micrometastasis in peripheral blood to hepatocellular carcinoma (HCC) metastasis or recurrence. Methods Reviewed the related literatures, the methods and significances of the detection of HCC micrometastasis in peripheral blood were analyzed. Results Currently, there are mainly two methods, hematogenous dissemination cell detection and HCC specific mRNA biomarker detection, for detection of HCC micrometastasis in peripheral blood. Theoretically, although they are considered as early detections of HCC metastasis or recurrence, researches still not have a abroad agreeable conclusion from different studies. After adjusting and improving the methods and detection time, different studies also have not gotten a quite consistent conclusion. Conclusion There is a great significance in detection of HCC micrometastasis in peripheral blood to understanding the mechanisms of HCC metatasis and recurrence, and also to improving the clinical therapy. Theoretically and practically, the method should be improved for facilitating the mechanism research of HCC metastasis and recurrence, and the application of detection.

To investigate the expression and significance of the ? glucuronidase(? G) gene in the patients with primary hepatocellular carcinoma(HCL), ? G gene was detected in the carcinoma tissue of 54 cases of HCL, 51 cases of cirrhosis tissue of HCL and 10 normal liver tissue by immuno fluorescence, immunohistochemistry and in situ hybridization respectively. The results showed that the expression level of ? G gene in HCL was much higher than that in the normal tissue and the expression level of ? G gene in peri carcinoma tissue was much higher than that in the carcinoma tissue. The data also showed that the expression level of ? G gene was highly correlated with metastasis and differentiation, but not related to tumor size, pathologic type, AFP and serum HBsAg level. It suggested that ? G gene could be used as a predictor for diagnosis and metastasis of hepatocellular carcinoma.

Objective To study the effects of various media, salt intensity, and organic components on the growth of Salvia miltiorrhiza adventitious roots and the synthesis of tanshinone ⅡA and protoca-techuic aldehyde. MethodsThe adventitious roots were obtained through tissue culture by manipulation of various media, salt intensity, and organic components and the contents of tanshinone ⅡA and protocatechuic aldehyde were determined by HPLC. ResultsThe effect of media MS, LS, B5, White, and SH on adventitious roots of S. miltiorrhiza was observed. Adventitious roots grew better under high salt intensity while secondary metabolite biosynthesis was accelerated under low salt intensity in MS basal medium.The reciprocity of five organic components had significant effect on root growth; glycin favored the synthesis of tanshinone ⅡA; scarcity of one of inosital, glycin, VB1, and VB6 inhibited the synthesis of protocatechuic aldehyde. ConclusionMS Basal medium is used for adventitious root culture. The results show that salt intensity and organic components have significant effects on adventitious root culture of S. miltiorrhiza and secondary melabolite synthesis.

Objective To study the effects of carbon, nitrogen, and phosphate sources on the growth of Salvia miltiorrhiza adventitious roots and the contents of tanshinone ⅡA and protocatechuic aldehyde. Methods The adventitious roots were obtained through tissue culture by manipulation of carbon, nitrogen, and phosphate sources and the contents of tanshinone ⅡA and protocatechuic aldehyde were determined by HPLC. Results Carbon, nitrogen, and phosphate sources were necessary for the culture of S. miltiorrhiza adventitious roots. The highest times of root multiplication were achieved at sucrose level of 30 g/L after 20 d culture, 60 g/L sucrose and low level sucrose were favorable for biosyntheses of tanshinone ⅡA and protocatechuic aldehyde, respectively. The highest root yield and tanshinone ⅡA content on day 25 were obtained by intermittent sugar adding during cultivation, and the production of adventitious roots and tanshinone ⅡA were 2.3-and 2.4-fold compared with those of control, respectively. The maximum root growth rate, contents of tanshinone ⅡA and protocatechuic aldehyde were achieved while NH4+-NO3-was 1∶4, 1∶4, and 1∶1, respectively when concentration of total nitrogen source was kept at 60 mmol/L. To compare with the control group, changing of KH2PO4 concentration could favor for the adventilious root growth, but high KH2PO4 concentration inhibited tanshinone ⅡA biosynthesis. ConclusionThe results show that various carbon, nitrogen, and phosphate sources have the significant effects on adventitious root culture of S. miltiorrhiza. The best carbon source and its concentration, nitrogen and phosphate sources for the growth of S. miltiorrhiza adventitious root and the synthesis of secondary metabolite are confirmed.

Objective To discuss the causes,agents and treatment of pancreatic encephalopathy in acute pancreatitis.Method We reviewed 26 cases of acute pancreatitis combined with pancreatic encephalopathy within recent 10 years.Results Pancreatic encephalopathy occurred always accompanied with such agents as hyperpyrexia, waterelectrolyte disturbance,hypoxemia, azotemia and bloodsugar disturbance etc.Conclusions The occurrence of the pancreatic encephalopathy is based on the harm that pancreatin does to the brain.The causes of pancreatic encephalopathy vary.To inhibit the releasing of pancreatin is the principle to prevent pancreatic encephalopathy, and to maintain normal physiological function,to control infection and nutritional support are the important links to prevent the pancreatic encephalopathy from happening.