Objective To investigate the therapeutic effect of microencapsulated xenogenic hepatocytes transplantation on rat fulminant hepatic failure(FHF).Methods Isolated guinea pig hepatocytes were microencapsulated with alginate-chitosan-polyethyleneglycol(ACP).The microencapsulated hepatocytes(MCHC) were implanted into the peritoneal cavity of Wister rats 24 hrs before the FHF model,which was surgically induced by 90% hepatectomy.After 24hrs of the FHF,the serum ALT,total bitirubin(TB),blood suger(BG) and PT were determined,and the microscopic appearance,function and nember of the implanted microencapsulated hepatocytes were observed,as well as the survival time and life quality of the FHF rats. Results Compared with the control group,the rats in the MCHC implantation group showed milder symptoms of central nervous system and less serious ascites.The ALT,BS, TB,and PT in the MCHC implantation group were all improved significantly (P

This paper is to study the applicability of the air disinfection equipment in general hospital. With the method of plate exposure adopted, five or three sample points are selected. The sample, culture and classification of bacteria are performed 30mins, 60mins, 120mins and 180mins before and after the equipment is turned on, and then the values of cfu/m3 are worked out respectively. The result shows that different departments should adopt different disinfection equipments according their characteristics and functions to meet the requirements of ″Disinfection Technology Specification″. Through this research and corresponding regulation, the management of air disinfection in our hospital is improved.

Objective To evaluate the therapeutic effect of abdominal multiple visceral organ transplantation for hepatic cirrhosis combined with diabetes by using donation after cardiac death (DCD).Methods Two patients suffering from hepatitis B-related liver cirrhosis,hepatocellular carcinoma combined with insulin dependent type 2 diabetes mellitus were given multiple visceral organ transplantation from May to June 2012.The transplanted organs including the liver,pancreas and duodenum were obtained from two donors after cardiac death which accorded with C-Ⅲ criteria.The donor internal and external iliac arteries were anastomosed to celiac axis and superior mesenteric artery and then the donor arteria iliaca communis was anastomosed to recipient abdominal aorta.The portal vein reconstruction was performed by end-to-side anastomosis between the donor and recipient portal vein.The pancreatic juice drainage was achieved by side to side anastomosis between donor duodenum and recipient jejunum.The pancreases of recipients were retained.Results The alanine aminotransferase,aspartate aminotransferase and total bilirubin of two patients were recovered to normal level at 2nd week after operation.The blood glucose and serum amylase returned to normal levels at 7th d and 4th d respectively.The fasting serum C-peptide and insulin were also at normal level at 2nd week.One patient with local intestinal anastomotic fistula was given percutaneous puncture drainage for four weeks and recovered.One patient recovered smoothly one month after transplantation without surgical complications.Conclusion Abdominal multiple visceral organ transplantation is an effective treatment for hepatic cirrhosis combined with diabetes by using DCD donor.

BACKGROUND: Effects of embryonic stem cell-derived hepatocyte-like cell transplantation on oncogenicity of differentiated hepatocyte-like cells and biochemical metabolism of liver should be further studied.OBJECTIVE: To evaluate the therapeutic efficacy of embryonic stem cell-derived hepatocyte-like cell transplantation on the acute liver failure.DESIGN: Randomized controlled study.SETTING: the First Affiliated Hospital of Sun Yat-sen University.MATERIALS: This study was performed at the Central Laboratory, the First Affiliated Hospital of Sun Yat-sen University from January 2005 to February 2006. D3-ES cells extracted from the mice which underwent transfection of green fluorescent protein were graciously presented by professor Huang, Ophthalmology Center of Sun Yat-sen University. Forty 6-week-old D3-129 mice of clean grade and irrespective of gender were provided by Experimental Animal Center of Sun Yat-sen University [certification: SCXK (yue) 2004-0011]. The experimentzl animals were disposed according to ethical criteria.Transforming growth factor, basic fibroblast growth factor, and hepatocyte growth factor were provided by Gibco BRL Company, USA.METHODS: Transforming growth factor, basic fibroblast growth factor, and hepatocyte growth factor were combined to differentiate D3-ES cells into hepatic cells. Cell suspension was poured into liver capsule of 20 mice with 2.0×10(6)cells per mouse. Another 20 mice that determined as the controls were injected with saline. Twenty-four hours later, intraperitoneal injection of 5 μL/20 g carbon tetrachloride was used to induce acute liver failure and to observe quality of life and mean survival time. Twenty-four hours after acute liver failure, vena cava posterior blood was drawn to detect total bilirubin,glutamate-pyruvate transaminase, albumin, blood glucose, pro-time prothrombin time, and other hepatic functional parameters. By scarification, hepatic samples were obtained to evaluate oncogenesis condition, and then HE staining and immunohistochemistry were adopted to detect growth of transplanted cells and albumin expression.MAIN OUTCOME MEASURES: Quality of life, average survival time, hepatic functional parameters, growth of transplanted cells, and oncogenesis condition.RESULTS: Quality of life and average survival time: After the onset of acute liver failure, mice in the control group had incoordination and other symptoms of central nervous system. In addition, 14 mice in the control group and 8 in the transplantation group had abdominal dropsy. Average survival time in the control group was significantly shorter than that in the transplantation group (23, 62 hours, P<0.05). Hepatic functional parameters: Levels of total bilirubin and glutamate-pyruvate transaminase in the control and transplantation groups were higher than those before modeling; levels of albumin and blood glucose were lower than those before modeling; pro-time prothrombin time was significantly longer than that before modeling(P<0.01). Furthermore, levels of total bilirubin and glutamate-pyruvate transaminase in the transplantation group were lower than those in the control group; blood glucose in the transplantation group was higher than that in the control group, and pro-time prothrombin time in the transplantation group was significantly shorter than that in the control group (P<0.05). Growth of transplanted cells and oncogenesis condition: Pathological section demonstrated that structure of liver tissue was not changed remarkably, and tumor was not formed. Moreover, transplanted cells and hepatocyte-like cell were well arranged and combined to express albumin.CONCLUSION: Embryonic stem cell-derived hepatocyte-like cell transplantation can improve quality of life, prolong survival time of model mice with acute liver failure; additionally, transplanted cells may well support biochemical metabolism of liver tissue.

It has been shown, however, that in terms of immune function, a transitional stage of dendritic cell maturation exists between immature myeloid dendritic cells (imDCs) and mature dendritic cells (DCs), which were named semimature dendritic cells (smDCs). Therefore, the theory that DCs can be classified as imDCs and mDCs has been challenged because of finding smDCs. SmDCs and imDCs are regarded as tolerogenic dendritc cells while what concrete mechanism taken by smDCs and imDCs in transplantation immunity has yet to know. The authors analyzed the important roles and the advanced molecular mechanism of smDCs and imDCs in transplantation immunity in order to update and widen understand underlying tolerogenic dendritic cells.

BACKGROUND: During the embryonic stage, E-cadherin expression plays a critical role in the formation of hepatic tissue.OBJECTIVE: E-cadherin gene was transfected into mouse embryonic stem cells (ESCs) to observe its effects on adhesive capacity of differentiated cells.DESIGN, TIME AND SETTING: A cytological in vitro observation was performed at the Laboratory of Surgery, First Hospital Affiliated to Sun Yat-sen University between December 2007 and December 2008.MATERIALS: BALB/c mice at gestational 13 days, of clean grade, were provided by Laboratory Animal Center, Sun Yat-sen University. BALB/c mouse ESCs were preserved by professor Huang Bing from the Department of Ophthalmology, Sun Yat-sen University. CMV promoter-containing eukaryotic expression plasmid pEGFP-N1 was gifted by doctor Lu Zhi-yue from Medical College, Sun Yat-sen University.METHODS: Total RNA was extracted from BALB/c mouse fresh hepatic tissue and synthesized into cDNA by reverse transcription (RT). The synthesized cDNA was used as a template to perform a polymerase chain reaction (PCR) that amplifies a targeted fragment. Following double enzyme digestion, pEGFP-E-cadherin plasmids were reconstructed and transfected into mouse ESCs. In vitro differentiation of transfected mouse ESCs was performed.MAIN OUTCOME MEASURES: Detection of E-cadherin expression in the differentiation system using RT-PCR and immunocytochemistry and observation of adhesive capacity of differentiated cells.RESULTS: E-cadherin gene-transfected ESCs could stably express E-cadherin during differentiational 1-17 days, while non-transfected ESCs expressed a decreasing amount of E-cadherin. The adhesive capacity of differentiated cells that stably expressed E-cadherin was markedly enhanced. Compact cell connection and multi-layer growth state remained at 19 days. While non-transfected ESCs gradually changed from embryoid bodies into noncohesive cell populations.CONCLUSION: Differentiating E-cadherin ESCs exhibit markedly enhanced adhesive capacity and maintain multi-layer growth state.

BACKGROUND:E-cadhedn plays an important role in development of liver tissue in the embryonic stage.Therefore,it is importance for investigating the feasibility of dynamic expression of E-cadherin in embryonic stern cell differentiation to in vitro development of liver tissue.OBJECTIVE:To observe the dynamic expression of E-cadherin in embryonic stem cell differentiation and the effect on cell adhesion.DESIGN,TIME AND SETTING:An in vitro cytological observation was performed at Surgical Laboratory of the First Affiliated Hospital of Sun Yat-sen University from December 2007 to December 2008.MATERIALS:Embryonic stem cells of BALB/c mice were obtained from Professor Huang (Department of Ophthalmology,Sun Yat-sen University).Twenty 13-day-old pregnant BALB/c mice of clean grade were provided by the Experimental Animal Centar of Sun Yat-sen University.METHODS:Following trypsinization,embryonic stem cells were suspend-incubated in DMEM culture medium containing fetal bovine serum,2-mercaptoethanol,HEPES,penicillin,and streptomycin.Embryoid body was formed 5 days after normal development and incubated in the culture plate at day 6.Liver tissue which was obtained from 13-day-old pregnant BALB/c mice was prepared for fetal liver cells which were frozen-sectioned as the controls.MAIN OUTCOME MEASURES:E-cadherin expression was detected using reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry st varying time points of 1,5,9,13,and 17 days in stages of initial differentiation of embryonic stem cells,formation of embryoid body,and formation of differentiated clusters.Additionally,the effect of E-cadherin expression on cell adhesion was also detected.RESULTS:RT-PCR showed that E-cadherin mRNA expression was not observed at day 1 but peaked at day 5;gradually,the expression was decreased until the expression was stopped at day 17.E-cadherin mRNA expression was strong in fetal liver cells in the control group.Immunocytochemistry showed a similar outcome.Morphologically,embryonic stem cells developed from unicells into compact three-embryonic layer embryoid body and into incompact cell population.CONCLUSION:E-cadherin expression correlates with differentiated cell adhesion;additionally,the lost expression in an in vitro environment may be an important cause for unable regularization of differentiated cells.

AIM: To construct prokaryotic expression vector of human angiogenesis inhibitor arresten gene and express recombinant arresten in Escherichia coli. METHODS: Human arresten gene was amplified from recombinant plasmid pGEM-Arr with polymerase chain reaction (PCR), and then cloned into prokaryotic expression vector pRSET by means of recombinant gene technology. The recombinant plasmid pRSET-Arr was transformed into E.coli BL21(DE3), and recombinant arresten was expressed in the bacteria under induction of IPTG. The expressed products were detected by SDS-PAGE analysis. RESULTS: Restriction analysis indicated that the arresten gene was successfully inserted into the expression vector, and DNA sequencing verified that the reading frame of the recombinant vector was correct. Recombinant arresten was successfully expressed in Escherichia coli; its molecular weight was about 26 kD and its amount was approximately 30% of total bacterial proteins.CONCLUSION: The successful construction of prokaryotic expression vector containing human arresten gene and the effective expression of recombinant arresten in Escherichia coli laid the foundation for further study on its biological functions.

Objective Sirolimus is a new, potent immunosuppreasant considered to be nonnephrotoxic. There is limited experience with the use of sirolimus in liver transplant recipients. This study was to investigate the clinical experience of conversion from tacrolimus-based to sirolimus-based immunosuppression in liver transplant recipients. Patients switched to cyclosporine-based immunosuppression during the same period were also enrolled as controls. Methods This retrospective study examined liver transplant recipients who had been switched from tacrelimus-based to sirolimus-based or cyelosporine-based immunosuppressive therapy between January 2004 and January 2008 in the First Affiliated Hospital of Sun Yat-sen University. Patients were divided into 2 groups: those switched to sirolimus-based immunosuppression (group A; n=32); and those switched to cyclosporine-based immunosuppression (group B; n=15). Results The rate of successful conversion was 34.5% in group A (10/32) compared with 45.5% in group B (7/15); this difference was not statistically significant (P>0.05). After conversion, renal function in patients in group A remained normal, while the renal function in patients in group B become abnormal 4 months after conversion (P<0.05). In group A, some simlimus-associated adverse effects occurred but were mild and easy to control. Conclusion Sirolimus can be used safely in place of tacrolimus in liver transplant recipients.

Objective To observe the early effect of organ donation after pancreas-kidney transplantation.Methods Eight cases of diabetic nephropathy received combined pancreas kidney transplantation.There were 8 donors,including 6 males and 2 females,with an average age of (26 10) years old (range from 15 to 42 years).There were 4 cases of donors with China during the transition period of brain heart double death organ donation (C-Ⅲ) standard,3 cases of donors in line with the international standard of brain death organ donation (C-Ⅰ) standard,1 case of international standard of heart death organ donation (C-lⅡ M-Ⅲ) citizen donors.There were 6 men and 2 women for recipients of the same blood type.Results Eight cases were awake 4-6 h postoperation and the ventilator was removed 8-14 h after operation.The rehabilitation therapy began 2 days postoperation from surgery intensive care unit (SICU) to the common wards.Serum C-peptide and insulin levels achieved normal range in 1-2 weeks after transplant.Blood glucose returned to the normal level in 2-3 weeks,and the creatinine level decreased to the normal level in 2 weeks postoperation.Duodenal intramural hematoma occurred in one patient intraoperatively,and the pancreatic graft was removed for safe consideration.Other patients had no serious surgical complications within 2 weeks after transplantation.Conclusion For organ donation after death of pancreas kidney transplantation,early organ function recovered well.Under the strict preoperative evaluation,the young donors can be safely used in combined pancreas and kidney transplantation.