Objective: Construction mouse GM-CSF gene effective eukatyotic expressive vector, selection high GM-CSF expressing mouse leukemia cell line RMA after transfected with the constructed vector, and study the method of treatment leukemia with tumor cells transfected with GM-CSF gene.Methods:770 bp of GM-CSF 3' end cDNA was amplified by PCR and inserted into pcDNAS vector.The constructed vector was transfected into RMA cells by electroporation. After screening by G418 and cloning by limiting dilution, a relative high GM-CSF expressing cell clone was selected by RT-PCR, hematopoietic progenitor cell proliferative assay and hematopoietic progenitor cell colony formation assay. The cells of this clone were inactivated by mitomycin-C and vaccinated mice to induce antitumor immune reaction. Results: The orientation and sequence of the insert was found to be correct, and a GM-CSF high expressing cell line RMA-GM was selected , which can induce mice obtain anti-tumor protective immune ability after inactivated by mitomycin-C.Conclusion:Tumor cells transfected with GM-CSF gene may be used an effective anti-T lymphycoma tumor vaccine.

objective To evaluate endoscopic surgery for high risk patients over 70 years old with severe acute cholangitis. Methods From October 1991 to October 2003, 84 high risk patients over 70 years old suffering from severe acute cholangitis were included and divided into laparotomy surgery (ST) and endoscopic surgery (ET) group. Results 80 patients were cured. The care rate was 81.2% in ST group and 91.6% in ET group. The mortality rate was lower in ET compared with ST (9.1% vs 18.8%). The rate of complication was reduced from 37.5% in ST group to 13.9% in ET group. The average duration of drainage was reduced from 49.4 days in ST group to 18.6 days in ET group. The total treatment days were shortened from 46.2 days to 22.8 days comparing ST with ET. Conclusions Endoscopic surgery should be the first choice for the senile and high risk patients with severe acute cholangitis. Endoscopic surgery was convenient, with mininal injury, safe and effective, and period of treatment was shortened, especially for elderly or critically ill patients, as well as those with MODS or previous billiary operations.

Objective To replace the imported vacuum pump with a Chinese-made instrument using a micro-ELISA negative pressure pump,to further extend the life of the machine.Methods Electric aspirator(YB.Dx23D) was used as source of negative pressure to connect with barrels of waste and the vacuum-phase of imported enzyme immunoassay instrument.The circuit was transformed,and the host matched the load,so the host and negative pressure pumps worked simultaneously.Results Compared with other aspirators,the modified electric aspirator YB.Dx23D has low-noise ratio,high negative pressure,vast flow,small volume,and mobile function.Conclusion The modified aspirator YB.Dx23D electric instrument is well used in washing ELISA plates and the effective use of waste water.Meanwhile,it can be applied to other necessary suction negative pressure(vacuum) equipment.

Objective To evaluate the endoscopic managements of acute cholangitis of severe type (ACST) accompanied with multiple organ dysfunction syndrome (MODS). Methods A total of 122 ACST patients accompanied with MODS from January 2000 to October 2008 underwent endoscopic treatment in two time periods. In critical phase, emergent endoscopic retrograde cholangiopancreatography (ERCP) plus en-doscopic naso-biliary drainage (ENBD) were performed to correct critical situation of the patients. After sta-bilization, endoscopic sphincterotomy (EST) plus stone removal, EST plus stent placement, or laparoscopy was performed according to the causes of ACST. Results Emergent endoscopic managements succeeded in all patients of critical phase. At third day post-operation, a reduction in white blood cell count, serum total bilirubin, body temperature, and rate of patients with shock, mental symptoms and purulent bile juice was a-chieved. Recovery rate of dysfunction organs was 60.2% at one week after emergent procedure, and 82. 6% at 2 weeks post-operation. Selective EST plus stone removal was performed in 36 patients with a success rate n one session at 91.7%. Laparoscopic cholecystectomy was performed in 85 patients with a success rate of 95.3%. Stent was placed in 16 patients with an effective rate of 81.3% at 3 months post the procedure. No severe complication or death occurred during the whole therapeutic course. The 6-month survival rate of 10 cancer cases was 70%. Conclusion Therapeutic ERCP plus ENBD is the first choice for acute severe cholangitis accompanied with MODS, while EST plus biliary lithotomy, or EST plus stent placement, or com-bined laparoscopy are ideal methods for subsequent treatment.

This paper presents the knowledge on fixing the heartbeat machine to the patients in such aspects as fixing,testing,using,notice in fixing,etc.

Objective: To clone human T cell receptor (TCR) ? chain gene and express its encoding protein by baculoviral expression system in insect cells. Methods: TCR ? chain cDNA was cloned from normal human peripheral blood mononuclear cells (PBMC) by RT-PCR and inserted into baculoviral transfer vector. This vector was co-transfected with baculovirus into insect sf 9 cells. The recombinant protein expressed was identified by SDS-PAGE and flow cytometry with mouse anti-human ? chain monoclonal antibody. Results: Human TCR ? chain cDNA cloned and inserted into baculoviral vector specifically expressed protein in the insect sf 9 cells, accounting for about 11% of the total protein yield in the supernatant of cell lysate. The molecular weight of the recombinant protein determined by SDS-PAGE was identical to what we anticipated. The insect cells transfected with recombinant baculovirus were demonstrated by intracellular labeling flow cytometry to express ? chain protein. Conclusion: Human T cell receptor ? chain gene was successfully cloned from human PBMC, and its encoding protein was highly expressed in insect cells. The TCR ? chain protein obtained by bioengineering technique is useful for in depth biological function study.

BACKGROUND:Autologous bone marrow mesenchymal stem cel s can treat decompensated liver cirrhosis, however, little evidence has addressed the control ed clinical research in hepatitis B patients with decompensated live cirrhosis.
OBJECTIVE:To evaluate the clinical efficacy and safety of autologous bone marrow mesenchymal stem cel s in the treatment of hepatitis B with decompensated live cirrhosis.
METHODS:A total of 67 hepatitis B patients with decompensated live cirrhosis were divided into two groups according to their wishes to receive stem cel transplantation. The control group (34 patients) only received oral administration of nucleoside analog antivirus and supportive treatment. The treatment group (33 patients) received autologous bone marrow mesenchymal stem cel s transplantation via hepatic artery plus antivirus and supportive treatment. The liver functional index, clinical signs and symptoms, adverse reactions were observed and compared at 4, 12, 24 weeks after treatment.
RESULTS AND CONCLUSION:After treatment, al patients’ symptoms were improved to varying degrees. After 4 weeks of treatment, the liver functional indexes were al significantly improved compared with before treatment, the levels of alanme aminotransferase, cholinesterase and prothrombin activity in treatment group were significantly ameliorated compared with control group (P<0.05). At 12 and 24 weeks of treatment, the alanme aminotransferase, albumin, total bilirubin, cholinesterase and prothrombin activity in control group and treatment group showed statistical y significant differences compared with before treatment (P<0.05). At the same time point, al the indicators in the treatment group were significantly ameliorated compared with control group (P<0.05). The Child-pugh score and model for end-stage liver disease score declined at 4, 12, 24 weeks after treatment, showing significant differences compared with before treatment. The difference was also significant at the same time point between two groups. The treatment of nucleoside analogue antivirus combined with autologous bone marrow mesenchymal stem cel s transplantation on hepatitis B patients with decompensated liver cirrhosis is an effective method to improve liver function and blood coagulation function, with symptom improvement, safety and low risk.

AIM: To explore the mechanism underlying inducible nitric oxide (NO) caused injury of endothelial cells during inflammation. METHODS: The activity of iso-enzymes of NO synthase (NOS), NO level and iNOS expression were examined using NADPH method, Griess reaction and RT-PCR, respectively. Furthermore, the lactate dehydrogenase (LDH) release rate, malondialdehyde (MDA) content were also measured. RESULTS:Co-administration of cytokines (TNF-? 5?10 5 U/L, IL-1? 2?10 5 U/L, INF-? 2?10 5 U/L) and LPS (10 mg/L) caused an obvious increase in NOS activity, NO levels (about two-fold) and a significant injury of the cells. At the same time, a significant increase in iNOS mRNA was also detected. Wheareas, treatment of the cells separately with cytokines or LPS for 24 h had no significant effect on NOS activity and NO level in cell lysates, however, it caused a significant increase in LDH release and MDA content. Also, the effect of cytokines and LPS on cell viability was concentration-and time-dependent. L-NMMA, a inhibitor of NOS, can suppress inducible NO production and protect cells against NO induced injury. CONCLUSION:Co-administration of cytokines (TNF-?, IL-1? and INF-?) and LPS significant activated iNOS and NO production which, in turn, induced oxidative reaction in endothelial cells.

ObjectivecircRNAs play an important role in tumor development, but the relationship between circRNAs and hepatocellular carcinoma remains to be further explored. The present study aimed to bioinformatically analyze the target gene of microRNA-1. Another aim was to screen circRNAs that are associated with target genes and differentially expressed in hepatocellular carcinoma cells, as well as provide theoretical basis for clinical screening of molecular markers and targeted therapies related to hepatocellular carcinoma.MethodsThe miRNA related database used for the prediction of microRNA-1 target genes, and the bioinformatic analysis of the target genes of microRNA-1 involved functional enrichment analysis and signal transduction pathway enrichment. Then, the circRNAs, which are related to the downstream target genes of microRNA-1, are screened through the circRNA database.ResultsThe number of microRNA-1 target genes was 230 in miRNA related database. Through GO analysis, it was found that the target genes of microRNA-1 had a strong tendency in regulation, and were mainly enriched in three aspects: biological function, biological process and cell localization.The target genes of microRNA-1 are involved in the function of proteins, regulation of biosynthesis, cofactor binding, enzyme regulation and other biological processes. Predicted target genes of miRNA-1 were significantly enriched in cancer signaling pathways, hepatitis B occurrence, endocytosis and splicing pathways. Further, 21 circRNAs related to the target gene of microRNA-1 were found in three circRNA databases, wherein hsa_circ_0004651 was highly expressed in hepatocellular carcinoma cell line HepG2 and its pavent gene was hnRNPD.ConclusionMicroRNA-1 influence the occurrence of hepatocellular carcinoma development through the regulation of protein and enzyme. Hsa_circ_0004651 may affect the development of hepatocellular carcinoma with microRNA-1 and its parental gene hnRNPD.

Objective To study the prevalence,treatment policy and control of hypertension in patients with maintenance hemodialysis, and to analyze the influencing factors of hypertension control.Methods We studied the current status of 1382 patients with maintenance hemodialysis in 11 dialysis centers in Shanghai, among them 809 were male, and 573 were female.Hypertension was defined as systolic blood pressure(SBP) ≥ 140 and/or diastolic blood pressure (DBP) ≥90 mm Hg ( 1 mm Hg = 0.133 kPa).Those who had a history of hypertension and requiring antihypertensive therapy were also diagnosed as hypertension though their blood pressure was within normal range during the survey.Hypertension control was defined as blood pressure ＜ 140/90 mm Hg before each dialysis session.Results The prevalence of hypertension in the hemodialysis patients was 86.3%.The treatment rate and control rate in those patients were 96.8% and 25.5% respectively.More than half (50.4% ) of patients were treated with only one kind of anti-hypertensive drug, and 34.4% with 2 kinds, 14.2% with 3 kinds, 1.0% with 4 kinds or more.Calcium channel blocker (CCB) was the most frequently prescribed drug (61.0%), followed by angiotensin Ⅱ receptor blockers ( 56.4% ), centrally acting anti-hypertensive agent ( 26.4% ), beta blockers and alpha, beta-blockers( 14.0% ).The control rate of hypertension in those hemodialysis people was aggravated by the existence of coronary artery disease.The patients who need more kinds of antihypertensive agents have a poorer control rate of hypertension.The hypertension control rate elevated significantly with the adequate hemodialysis.Conclusions There is a very high prevalence of hypertension in maintenance hemodialysis patients.Although the treatment rate is high, the control rate is unsatisfactory.So the control of hypertension in hemodialysis patient is still a clinical challenge.Appropriate dialysis adequacy, reasonable use of erythropoietin, treatment of heart disease and judicious use of antihypertensive drugs may be helpful to improve the clinical outcome.