1.Evaluation of the viability and energy metabolism of canine pancreas graft subjected to significant warm ischemia damage
Chunhui YUAN ; Yongfeng LIU ; Guichen LI
Chinese Journal of Organ Transplantation 2005;0(07):-
Objective To investigate the energy metabolism state and the posttransplantive function under different warm ischemia time (WIT). Methods The left lobe of pancreas was resected, and according to WIT (30, 60, 90, 120 min), the canines were randomized into 4 groups. The graft was perfused and preserved by the UW solution (0-4 ℃) cold storage method for 24 h. And then the heterotopic segmental autotransplantation of pancreas was performed, while the normal control group and no warm ischemia(NWI)group were also set up. Blood sugar (BG) and intravenous glucose tolerance test (IVGTT) K value and the plasma insulin level were examined in the first week after pancreas transplantation. The amylase level of pancreatic juice 24 h and one week after transplantation were detected either. The tissue concentrations of adenosine triphosphate (ATP) and total adenine nucleotides (TAN) were determined using high performance liquid chromatography (HPLC). The pancreas cortex was taken for morphological analysis and ultrastructure study 1 h after transplantation. Results Plasma glucose in groups of 30 min and 60 min was returned to normal 3-5 days after transplantation, while groups of 90 min and 120 min could not one week after transplantation. IVGTT K values in groups of NWI, 30 min and 60 min were more than 1, while those in groups of 90 min and 120 min were less than 1 one week after transplantation. The plasma insulin levels in groups of 90 min and 120 min were much lower than those in groups of NWI, 30 min and 60 min (P
2.Protective effects of protease inhibitor on liver function after ischemia reperfusion and orthotopic liver transplantation in rats
Guichen LI ; Yongfeng LIU ; Jialin ZHANG
Chinese Journal of Hepatobiliary Surgery 1998;0(06):-
Objective To investigate the role of neutrophils and protease inhibitor in the pathogenesis of hepatic ischemia reperfusion injury. Methods The models of liver ischemia reperfusion (LIR) and orthotopic liver transplantation (OLT) were established in rats. The contents of serum ALT and LDH and liver myeloperoxidase(MPO) were determined. Meanwhile, the hepatic pathohistological changes were observed and the number of infiltrating neutrophils counted. As for the LIR model, the animals were administered with aprotinin (AP) before ischemia. Then reperfusion was achieved 45 min after ischemia. The samples were collected after reperfusion for 4 h. In OLT model, the animals were administered with AP 1 h before the transplantation and the samples were collected 4 h postoperatively. Results The contents of serum ALT and LDH and liver MPO were significantly higher in the control group than in the experimental one (P
3.Studies on Mineral Elements for Cultivated Halenia Ellipitica D. Don in Qinghai Province
Tiancai LI ; Guichen CHEN ; Guoying ZHOU
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(02):-
Objective To compare the change of mineral element in Tibet herb Halenia ellipitica D. Don before and after the seeds matured. Method We separately collected the cultivated Halenia ellipitica D. Don of several different districts in Qinghai province before and after the seeds matured and analyzed the contents of mineral elements as potassium, sodium, calcium, magnesium by atom absorb spectrum instrument. Results and Conclusion The calcium, magnesium and potassium, sodium elements nutriment has an antagonistic characteristic. The sodium and phosphorus elements nutriment has a coordination characteristic of the cultivated Halenia ellipitica D. Don in Qinghai province during the growth period.
4.Effects of L-arginine on the hypothermic perfusion and on preservation of isolated pancreas in canines
Chunhui YUAN ; Guichen LI ; Yongfeng LIU ; Jiang LIANG ; Sanguang HE
Chinese Journal of General Surgery 1997;0(06):-
Objective To study the effects of L-arginine on the reperfusion injury of hypothermic perfusion and on preservation of isolated pancreas(ISP) in canines . Methods After segmental pancreatic transplantation(SPTS) model were set up,the animals were randomly divided into 3 groups:(1)ISP irregulated and preserved with Euro Colln Solution(ECS)plus L-arginine(L-arg group);(2)IPS irregulated and preserved with ECS plus N-nitro-L-arginine methy ester(L-NAME group);and(3)IPS irregulated and preserved with ECS plus normal saline(Control group).The IPS were irregulated and preserved at 0-4℃ for 24h.Serum concentration of amylase and lipase were detected. NO,NOS and myeloperoxidase (MPO) and expression of iNOS mRNA in the pancreas graft were measured. Histological examination was performed. Results The level of NO in the L-arg group was significantly higher than that in the control group(P
5.Effect of minocycline on ischemia-reperfusion injury in liver transplantation after circulatory death in rats
Anyin WANG ; Ying CHENG ; Qi PAN ; Jianan YU ; Guichen LI
Chinese Journal of Organ Transplantation 2016;37(12):709-715
Objective To study the protective effect of minocycline against ischemia-reperfusion injury after liver transplantation and its molecular mechanism after circulatory death in rats.Methods The rat donation after circulatory death (DCD) liver transplantation model was established by using magnetic-ring method.The donor and recipient were male SD rats.The rats were divided into sham operation group (SHAM group),liver transplantation group,minocycline group (MIN group),atractyloside + minocycline group (ATR + MIN group),24 rats in each group.In the MIN group,10 mg/kg minocycline was injected through the dorsum vein of the penis after reperfusion.The ATR + MIN group was injected with 2 mg/kg atractyloside.The open status of mitochondrial permeability transition pore (mPTP) was detected at 2,6,and 24 h after operation.Western blotting and immunohistochemistry were used to detect the expression of caspase3 and cyt c.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were determined.Liver tissues were stained with hematoxylin-eosin (HE) and pathological changes were evaluated by Suzuki's standard.The survival of each group was calculated.Results As compared with liver transplantation group and ATR+ MIN group,the mPTP opening of MIN group decreased (P<0.05),the expression of caspase3 and cyt c and the serum ALT and AST levels decreased significantly (P<0.05),liver tissues injury was alleviated (P<0.05),and the survival rate increased significantly after 30 days (P <0.05).There was no significant difference between liver transplantation group and ATR + MIN group (P > 0.05).Conclusion Minocycline reduces ischemia-reperfusion injury in DCD liver transplantation in rats probably by inhibiting the mPTP opening,and preventing cyt c release and caspase3 activation to reduce hepatocyte apoptosis.This effect can be blocked by mPTP opener.
6.Effects of cholecystokinin on canine Oddi sphincter function after pancreas transplantation with bladder drainage
Guichen LI ; Chunhui YUAN ; Ying CHENG ; Yongfeng LIU ;
Chinese Journal of General Surgery 1997;0(06):-
Objective To study the effects of cholecystokinin (CCK) on canine Oddi sphincter(SO)function after pancreas transplantation with bladder drainage. Methods Normal canine and transplant canine SO manometry before and after CCK administration was carried out. Results SO basal pressure in control group was(18 5?2 8) mm Hg, frequency was(9 7?1 5)per min, amplitude was (47?6) mm Hg, motility index was(236?56). After CCK administration, basal pressure, frequency, amplitude and motility index decreased significantly to(10 2?2 2) mm Hg ,(5 0?1 2)per minute,(19?5) mm Hg and(50?17), all P
7.Culture of pancreatic duct-derived stem cells of rats in vitro in the dynamic three-dimension cell culture system
Xuchun CHEN ; Ying CHENG ; Guichen LI ; Rui SHI ; Xiaohang LI ; Da LI ; Yongfeng LIU
Chinese Journal of Organ Transplantation 2012;33(6):371-375
Objective To isolate,purify and identify pancreatic duct derived stem cells (PDSCs) from the pancreatic duct of rats,and culture in the three-dimension cell culture system.Methods Adult male Wistar rats underwent perfusion with collagenase V via the pancreatic duct,then the pancreas was surgically procured,digested,followed by discontinued density gradient centrifuge to isolate ductal tissue from islets.The acinar and ductal tissue was cultivated in serumcontaining medium in the three-dimension cell culture to obtain adherent cells,as PDSCs,which were expanded by consecutive passages.The morphology and characterization of PDSCs on phenotype were examined.Results PDSCs could be obtained through in situ collagenase V digestion,discontinued density gradient centrifuge,and culture in the three dimension cell culture system.Morphologically,PDSCs had remarkable size,most with one nucleus.PDSCs grew in many layers in three-dimension cell culture system.PDSCs was revealed to express CD29,CD73,CD90,CD105,but not CD14,CD19,CD34,CD45 by FACS,in agreement with MSCs.Conclusion PDSCs of rats could be obtained through in situ collagenase V digestion,discontinued density gradient centrifuge,and culture in the three-dimension cell culture system.PDSCs lines were successfully established.
8.Experimental study on the protective effects of adenosine on reperfusion injury following pancreaticoduodenal transplantation in rats
Chunhui YUAN ; Yongfeng LIU ; Ning ZHAO ; Guichen LI ; Ying CHENG ; Sanguang HE
Chinese Journal of General Surgery 1994;0(05):-
8-PT group[(2.05?0.66)?mol/g, ( 3.98 ?0.41)?mol/g] 24h after transplantation(P
9.Prevention and management of hepatic artery thrombosis in orthotopic liver transplantation
Gang WU ; Yongfeng LIU ; Jian LIANG ; Shurong LIU ; Guichen LI ; Donghua CHENG ; Sanguang HE
Chinese Journal of General Surgery 2001;0(07):-
ObjectiveTo investigate the methods of hepatic arterial reconstruction in orthotopic liver (transplantation)(OLT), and the prevention and management of associated complications, so as to improve the therapeatic results and donor survival rate of OLT.MethodsThe clinical data of 85 cases of orthotopic liver transplantation performed in our institute from May 1995 to May 2004 were retrospectively analysed. Hepatic artery reconstruction was made by use of Carrels patch of donor celiac artery or bifurcation of donor common hepatic artery and splenic artery anastoniosed to left and right hepatic artery of recipient in 16 cases((18.82)%), to bifurcation of gastroduodenal and proper hepatic arteries of recipient in 61 cases((71.76)%), or use of donor iliac artery interposition graft to abdominal aorta in 8 cases(9.42%). Regular heparin or low-molecular-weight heparin as a prophylactic anticoagulation therapy was maintained during and after (operation) as indicated by prothrombin time. Intra-and postoperative Doppler ultrasonography was used to (monitor) hepatic arterial blood supply.ResultsHepatic artery thrombosis(HAT) was observed in 1 case (intraoperatively)(overall incidence 1.2%).Thrombectomy and reconstruction of the arterial anastomosis were performed immediately.This patient currently has a survival time of 13 months without HAT. HAT was not observed in any of the other 84 cases during the follow up of 2 to 52 months.Conclusions Hepatic artery thrombosis may be minimized by proper selection of anastomotic site of hepatic artery reconstruction and (effective) use of postoperative anticoagulation.Close follow-up by Doppler ultrasonography may make a prompt diagnosis of HAT, salvage grafts and avoid re-transplantation.
10.Scanning and analysis of the KATP channel mutations in 12 cases of infancy onset type 1 diabetes mellitus
Li REN ; Wenli YANG ; Jie YAN ; Yuyun WU ; Yanmei SANG ; Cheng ZHU ; Guichen NI
Chinese Journal of Applied Clinical Pediatrics 2016;31(8):579-583
Objective To screen the mutation of KATP channel mutations in Chinese pedigrees with infantile onset type 1 diabetes mellitus (T1DM) and neonatal diabetes mellitus.Methods A cohort of 12 children of infant onset T1DM and neonatal diabetes mellitus admitted into Beijing Children's Hospital between March 2004 and June 2013 were selected.PCR amplification and direct sequencing were used to analyze the 39 exons of ABCC8 gene and one exon of KCNJ11.And the mutational sites of the parents of the probands was sequenced in order to identify the inheritance.Results Analysis revealed ABCC8 mutation in 25% (3/12 cases) of the patients,a case of transient neonatal diabetes (TNDM),a case of permanent neonatal diabetes mellitus (PNDM) and a case of infant onset T1DM.All positive patients showed a known heterozygosis mutation in the ABCC8 gene(R1182Q,c.3545G > A,D209E,c.627C > G,E208K c.622G > A).The residue R1182Q,which was located at a position involved in joining transmembrane domain 2 to nucleotide binding domain 2,the mutations E208K and D209E were located in the intracellular region that links the transmembrane domain with the gatekeeper module.All the three mutations were located throughout the cytoplasm part of SUR1 protein.The TNDM successfully transferred from insulin to oral sulfonylureas therapy.Conclusions There is a complex genetic pathogenesis in neonatal and infant-onset diabetes.The KATP channel activating mutations is one of the main causes of neonatal diabetes mellitus and may cause T1DM in infants in China.Oral Glibenclamide therapy seems highly effective for some patients with the KATP channel activating mutations.