Protein expression, purification and mouse antiserum preparation of monkeypox virus A23R / 细胞与分子免疫学杂志

Objective To express the monkeypox virus (MPXV) A23R protein in Escherichia coli and purify by Ni-NTA affinity column, and to prepare mouse antiserum against MPXV A23R. Methods The recombinant plasmid pET-28a-MPXV-A23R was constructed and transformed into Escherichia coli BL21 to induce the expression of A23R protein. After optimizing the conditions of expression, A23R protein was highly expressed. Recombinant A23R protein was purified by Ni-NTA affinity column and identified by Western blot analysis. The purified protein was used to immunize mice for preparing the A23R polyclonal antibody, and the antibody titer was detected by ELISA. Results The expression of A23R recombinant protein reached the peak under the induced conditions of 0.6 mmol/L isopropyl-β-D-thiogalactoside (IPTG), 37 DegreesCelsius and 20 hours. The purity of the protein was about 96.07% and was identified by Western blot analysis. The mice were immunized with recombinant protein, and the titer of antibody reached 1:102 400 at the 6th week after immunization. Conclusion MPXV A23R is expressed highly and purified with a high purity and its antiserum from mouse is obtained with a high titre.

Arterial Reconstruction in Liver Transplantation with Abnormal Donor Hepatic Artery:Clinical Analysis of 42 Cases / 中国医科大学学报

Objective To explore the influence of abnormal donor hepatic artery on hepatic artery and biliary complications after liver transplanta?tion,and summarize the hepatic artery reconstruction procedures during transplantation. Methods The clinical data of 210 cases of liver transplan?tation conducted in our hospital from May 2005 to April 2015 were retrospectively searched for the study,including 42 with abnormal donor hepatic artery. Results Among the 210 liver transplantation,42 cases exhibited abnormal donor hepatic artery,and the aberration rate was 20.0%. Mean volume of blood flow of abnormal group and normal group was 4.7±95.1 mL/min and 190.9±101.6 mL/min,respectively. There was no statistic differ?ence(P=0.519). Twelve cases had arterial complications,the incidence rate was 5.71%,and there was no statistic difference between each group (χ2=0.72,P>0.05). Twenty five cases got biliary complications,the incidence rate was 11.9%,and there was no statistic differences between each group(χ2=0.05,P>0.05). Conclusion There was no statistic difference of mean volume of blood flow after arterial reconstruction between two groups. Liver transplantation with abnormal arterial reconstruction will not increase the incidence rate of arterial and biliary complications.

Impact of warm ischemia injury on mitochondrial morphology and function of rat donor liver after cardiac death / 中国组织工程研究

BACKGROUND:How to control functional activity of donor liver after cardiac death and maintain the optimal function of grafts are the key issues in organ transplantation study. OBJECTIVE:To preliminarily explore the effect of warm ischemia injury on the morphology and function of rat donor liver after cardiac death. METHODS:Cardiac death model was established in Sprague-Dawley rats and the successful models were divided into six groups:control group (warm ischemia for 0 minute), warm ischemia 10 group (warm ischemia for 10 minutes), warm ischemia 20 group (warm ischemia for 20 minutes), warm ischemia 30 group (warm ischemia for 30 minutes), warm ischemia 40 group (warm ischemia for 40 minutes) and warm ischemia 50 group (warm ischemia for 50 minutes). The rat liver specimens in each group were cut into ultrathin sections. The structure of liver cells was observed and photographed by electron microscopy. Flameng score was applied to analyze the degree of mitochondrial damage. Liver mitochondria were extracted and then spectrophotometry was used to assess the viability of cytochrome C oxidase. RESULTS AND CONCLUSION:Under electron microscopy, there were no significant changes in liver cells within 30 minutes of warm ischemia, nuclear membrane was intact, mitochondria mildly swel ed, no mitochondrial crista ruptured, and Flameng score was<2 points. With the extension of warm ischemia time, the cells became swel ing, nuclear chromatin condensated, apoptotic body was clearly visible, mitochondrial matrix coagulated, mitochondria exhibited vacuolation, and Flameng score was 3-4 points. The viability of cytochrome C oxidase showed no significant difference within 30 minutes of warm ischemia, but began to significantly decrease at 40 and 50 minutes. The mitochondrial structure and function after liver injury is not obviously affected by 30 minutes of warm ischemia, and significant changes appear after 40 minutes.

Clinical islet transplantation for type 2 diabetes mellitus: 3 cases report / 中华器官移植杂志

Objective To evaluate the effect of islet transplantation for patients with type 2diabetes mellitus (DM). Methods Since December 2007, 4 cases of islet transplantations were performed on 3 patients with type 2 DM and end-stage renal disease (ESRD). Two patients received simultaneous islet-kidney transplant from single-donor (SIK), and one received 2 consecutive islet transplants 5 months following kidney transplantion (IAK). All recipients given insulin with a dose of percutaneous transhepatic portal catheterization. Anti-CD25 monoclonal antibody was used as induction. For SIK, low-doses of Tacrolimus and sirolimus were used as maintenance immunosuppression protocol. For IAK, the maintenance protocol included cyclosporine and MMF.Insulin dose, the level of blood glucose, C-peptide and the value of HbA1 were observed. Results The first patient of SIKhad normal glucose level 3 days after surgery and became insulin independent within the first month, but insulin was administered gradually and the dose reduced to 1/3. The second patient of SIK died of bleeding and secondary infection of liver puncture site 5 days following operation, the blood glucose level recovered to normal 24 h after operation. The insulin dose of the patient of IAK was reduced to 1/2 after the first transplant. The patient became insulin free after the second operation. The level of fasting and postprandial C-peptide of the surviving recipients increased by 600 pmol/L. The value of HbA1 of the SIK was 6.7 %～7.3 %, while that of the IAK was 5. 5 %～ 5. 9 %. Conclusion Islet transplantation is an effective treatment for patients with type 2 DM.

Improved islet isolation by three-cannula method for collagenase infusion / 中华器官移植杂志

Objective Pancreas perfusion is an essential step in human islet isolation.To develop the new methods for introductal canulation,collagenase infusion and to observe their effects on islets isolation.Methods A total of 17 pancreases were digested from March 2005 to April 2010.The pancreases were distended by three methods:the standard method (n =3),the one-cannula method (n =11) and three-cannula method (n =3).In the standard group,the pancreases were completely cut into half at the mid-body.Two catheters were inserted into the main duct:one directed toward the tail and the other to the head.In the one-cannula method group,a long tube was inserted into the duct at the head,advancing to the tail In the three-cannula method group,pancreatic parenchyma was then minimally cut at the mid-body and three catheters were inserted into the main pancreatic duct:one at the head (the first catheter) and two at the mid-body,one toward the tail (the second catheter) and the other toward the head (the third catheter).The pancreases were digested by improved Ricordi technique.Ficoll continuous density/grads centrifuge method was performed to purify the islets.DTZ staining was adopted to identify islets and count islet equivalent (IEQ). AO/EB fluorescence examination was used to count active islet percentage.Static glucose stimulating test (SGS) in vitro was designed to estimate islet function and calculate SI.Results The distension volume of the threecannula method group was 1.24 rnl/g pancreas,and higher than the other groups (for the standard group:0.71 ml/g pancreas; for one-cannula method group:0.96 ml/g pancreas,P＜0.05).The yield of islet in the three-cannula method group and the one-cannula method group was 2514 and 2270 IEQ/g,which was significantly more than that in the standard group (1914 IEQ/g pancreas,P＜0.05).The purity and viability of the islets were 74 ％/79.3 ％,75.6 ％/79.4 ％ and 78.3 ％/84.0 ％ respectively in the three groups with the difference being not significant among the groups.SI in the one cannula method group (4.74) and the three-cannula method group (5.27) was significantly higher than that in the standard group (3.46).ConclusionThe three-cannula method improved collagenase infusion and the islet yields.

Kidney transplantation from donation after cardiac death donor / 中华器官移植杂志

Objective To summarize the clinical experience of harvesting and using the kidneys from donation after cardiac death (DCD) donors.Methods Fourteen kidney transplantations were successfully performed on 14 patients with end-stage renal diseases.The kidneys were harvested from 7 volunteer donors (age 30～53 years) diagnosed with cardiac death,who were scored 19～23according to the University of Wisconsin donation after cardiac death evaluation.Primary diseases of the donors were cerebral hemorrhage,brain injury,ischemic cerebral vascular disease and brain tumor.Warm ischemia time ranged from 5 to 45 min,and cold ischemia time was 4.5 ～ 12.5 h.Results After transplantation,three patients had delayed graft function (DGF),one had primary non-function (PNF),and two patients developed acute rejection.In the patient with PNF,the transplanted kidney was removed one day after operation and the patient went back to hemodialysis.One patient with DGF was still in recovery with serum creatine 149 μmnol/L (within 3 months after operation).The above two cases both utilized the kidneys with 45 min of warm ischemia time.The rest 12 patients were discharged with normal renal function.Conclusion Under the condition of our country,kidneys strictly harvested from DCD donors can be used as one of the main sources of kidney grafts for kidney transplantation.

Liver transplantation from donation after cardiac death (report of 3 cases) / 中华器官移植杂志

Objective To summarize our experience in the liver transplantation from the donation after cardiac death (DCD).Methods The livers from three DCD donors (2 cases of brain trauma and 1 case of cerebral hemorrhage) were harvested according to the Guidelines for Donation after Cardiac Death in China.These grafts were orthotopically transplanted into three recipients including 2 cases of decompensative hepatic cirrhosis and 1 case of primary liver cancer.The warm ischemic time ranged from 7.5 to 10 min and the cold ischemic time was 4.5,8.2 and 6.5 h respectively.Postoperative immunosuppressive regimens included prednisone,FK506 and mycophenolate mofetil (MMF).Antibiotics and anticoagulatants were used accordingly.Results All of the 3 recipients obtained normal liver function within 3 weeks since the grafts were implanted without PNF,thrombosis and rejection.No postoperative complications occurred in 3 recipients during the follow-up period of 2 to 9 months with normal liver function.Conclusion The liver transplant from DCD donor showed good results in our center.Chinese group Ⅲ of DCD donor,UW score above the middle level and the short warm ischemic time are three keys ensuring the success of the liver transplant from DCD donors.

Analysis of risk factors of acute renal failure after surgery for simple abdominal trauma / 中华创伤杂志

Objective To investigate the risk factors associated with acute renal failure (ARF)after surgery in patients with simple abdominal trauma. Methods A retrospective case-control study was carried out in patients with simple abdominal trauma after surgery from January 2003 through May 2008 in our hospital. Twenty patients with ARF were set as study group and 68 patients without ARF as control group. Clinical data including age, gender, injury type, number of injured organs, blood transfusion volume, intra-abdominal pressure, abdominal infection, shock and related factors were analyzed by using univariate and Logistic regression to identify the independent risk factors of ARF after surgery in patients with simple abdominal trauma. Results Univariate analysis revealed statistical difference in aspects of blood transfusion volume ( ＞ 1 600 ml), operation timing ( interval between trauma and operation ＞12 h), preoperative shock duration ( ＞6 h), postoperative abdominal infection and intra-abdominal hypertension ( ＞ 12 mm Hg) between two groups ( P ＜ 0. 05 ). Logistic regression analysis showed that these five indices were the independent risk factors of ARF after surgery in patients with simple abdominal trauma ( P ＜ 0. 05 ). Conclusions Massive blood transfusion, delayed operation, long preoperative duration of shock, postoperative abdominal infection or intra-abdominal hypertension are the risk factors of ARF after surgery in patients with simple abdominal trauma. We should pay attention to these factors and take effective measures to prevent occurrence of ARF.

Intraoperative vessel Doppler evaluation of vessel coniplications of adult orthotopic liver trauspiantation / 中华器官移植杂志

Objective To analyze the role of the introperative vessel Doppler sonographic evaluation of the hepatic artery and portal vein. Methods Intraoperative vessel Doppler sonograms of 116 patients were analyzed for peak systolic velocity of hepatic artery and blood flow of the portal vein.In patients having abnormal findings on sonography (peak systolic velocity of hepatic artery less than 30 cm/s, blood flow of the portal vein less than 800 ml/s), the vascular anastomoses were checked.Results Fourteen of 116 cases revealed less hepatic arterial peak systolic velocity than 30 cm/s. In 9 of the 14 cases, the hepatic arterial peak systolic velocity was normal after injection of 0. 5 % lidocaine into celic trunk root, and papaverine and 654-2 into artery, 3 of the 9 cases endured artery thrombosis. In the other 5 of the 14 cases, by-pass anastomoses were done, and the hepatic arterial peak systolic velocity was normal, and no hepatic arterial complication occurred. Five of 116 cases revealed less hepatic portal vein blood flow than 800 ml/rnin. 4 of the 5 cases revealed shunt between portal vein and vena cava. The blood flow was normal after ligation of the shunt, and thrombosis occurred in 1 case of the 4. The another 1 of the 5 cases was presented with portal vein thrombosis of grade m, and the blood flow remained lower than normal when side-to-side anastomosis was done after resection of thrombosis. Then vein by-pass of the superior mesenteric vein to portal vein with donor iliac vein was done, the blood flow became normal, and no complication occurred. Conclusions The vessel Doppler sonography during liver transplantation was of pivotal values in preventing and diagnosing vessel complications. For the patients with abnormal findings though intraoperative vessel Doppler sonography, the close monitoring should be done in order to find out vessel complication as

Three cases report of malignant fibrous histiocytoma of thyroid with reference review / 中华内分泌外科杂志

Objective To investigate the clinical characteristics, diagnosis, treatment and prognosis of malignant fibrous histiocytoma (MFH) of thyroid. Methods The clinical case and follow-up data (n=3) of MFH of thyroid from our hospital between 2000 and 2007 was analyzed, with the correlating references reviewed. Results MFH of thyroid accounted for 0.574% (3/523) of malignant tumor of thyroid. MFH of thyroid progressed so fast as to invase recurrent laryngeal nerve (RLN) and /or cervical lymph nodes. Conclusions The diagnosis of MFH of thyroid is difficult, for it is easy to be misdiagnosed as thyroid carcinoma. Immunohistochemistry pathology can provide a good differential diagnostic value, with CK negative, Tg negative, and Vimentin positive to support the diagnosis of MFH. Extensive and thorough excision should be the main choice to treat MFH of thyroid. Patients of MFH of thyroid often died of recurrence or metastasis in 2 years after operation. MFH of thyroid is a kind of rare thyroid primary tumor, with malignancy of high grade and poor prognosis. The disgnosis of MFH of thyroid depends on immunohistochemistry staining, while excision should be the main choice for treatment.

Hepatic artery reconstruction in orthotopic liver transplantation / 中华普通外科杂志

Objective To summarize experience for hepatic artery reconstruction in orthotopic liver transplantation(OLT).Method A retrospective analysis was made for 183 cases of orthotopic liver transplantation performed in our institute from May 1995 to december 2006.All the arterial reconstructions were performed with 6-0 polypropylene sutures in an interrupted fashion under a 3.5 magnification surgical loupe.Donor hepatic arteries were anastomosed at the origin of the celiac artery with a Carrel's patch or at the level of splenic artery confluence.Extra-anatomic arterial reconstruction was based on recipient aorta using donor iliac artery graft.OLT with routine anatomic arterial construction served as control.Heparin or low-molecule-weight heparin as a prophylactic anticoagulation therapy was maintained during and after operation if prothrombin time is less than eighteen seconds.Follow-up Doppler ultrasonography was used daily in the early postoperative period.Results Overall incidence of hepatic artery complications was 3.28%.Hepatic artery thrombosis(HAT)was observed in 5 cases.Hepatic artery stenosis(HAS)occurred in 1 patient.Routine anatomic arterial construction was performed in 89.07%(163)of cases,and HAT developed in 3(1.84%)cases.Extra-anatomic arterial reconstruction was carried out in 10.93%(20)of the patients;the presence of HAT was identified in 3(15.0%)cases(X2=9.73,P＜0.01).Thrombolysis,balloon angioplasty,and vascular stenting via hepatic artery were performed.One patient suffering from identified hepatic artery thrombosis died of liver failure 19 days post-op.The other 5 patients were cured successfully with patent blood flow by interventional therapy.Mortality related to hepatic artery complication was 16.7%.Conclusion HAT and HAS may be minimized by using gastroduodenal branch-patch anastomosis and postoperative anticoagulation.Close follow-up by Doppler ultrasonography helps to make a prompt diagnosis and reduce HAT-and HAS related graft loss.

Prevention and management of hepatic artery thrombosis in orthotopic liver transplantation / 中国普通外科杂志

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.

Eavluation of different surgical procedures and their therapeutic effects in treatment of pancreatic cyst:a report of 118 cases / 中华肝胆外科杂志

Objective To evaluate the effect of operation method in surgical treatment of pancreatic cyst. Method Retrospectively stastical analysis on 118 cases of pancreatic cyst was carried out.Result Thirty-one cases underwent external drainage,54 cases were subjected to internal drainage,and 33 cases to resection.Con- clusion For a genuine cyst,it is better to have it reacted.External drainage is only indicated for pseudocysts in emergent situations such as infection,hemorrhage or rupture,but is often accompanied with multiple complica- tions.Internal drainage is indicated for unresectable cysts and is superior to the external one,but it has some com- plications too.Resection of the pancreatic body and tail is the first choice for cysts located there,and it shows a good and thorough therapeutic effect with seldom recurrence.

Simultaneous pancreas-kidney transplantation:a report of 19 cases / 中国普通外科杂志

Objective To study the application of intraoperative drug administration in simultaneous pancreas-kidney transplantation(SPK), in order to achieve favorable postoperative results. Methods In the 19 cases of SPK,we primarily administered albumin 1.5~2.0g/kg to increase blood volume. Intraoperative electrocyte fluid dosage was 30~50mL/kg. Isoptin, 5mg/12h and prostaglandin E1(PGE1) 100?g/12h was given to lessen preservation injury and somatostatin(3mg/12h) to inhibit exocrine secretion of the transplanted pancreas. Results After reperfusion,the 19 transplanted pancreas grafts secreted amylase averaging 20 800U/L,and the transplanted kidneys produced urine,through the ureters,in 2~10min.In the 19cases,exogeneous insulin was withdrawn within 1.5h~9d and kidney function returned to normal within 2~4d. Conclusions In the SPK recipients, the rational use of albumin, vasodilators and somatostatin plays an important role in obtaining better function of transplanted pancreas and kidney.

Surgical treatment of hilar cholangiocarcinoma: report of 192 cases / 中国普通外科杂志

Objective To evaluate the diagnostic method for and surgical effect on hilar cholangiocarcinoma. ] Methods The clinical features, diagnosis, surgical therapy and long term result were retrospectively analysed on 192 cases of hilar cholangiocarcinoma admitted into our hospital from Jan.1984 to Jun.1999Results] MRCP delineated 100%(44/44) of the hilar tumors. 153 cases underwent exploration. 88 cases treated by internal or external drainage have survived for 3～15 months. Tumor resection was performed on 53 cases, and orthotopic liver transplantation on 2 cases, with a resection rate of 36%(55/153) including radical resection in 38 cases (median survival period was 31 months). 17 cases undergoing palliative resection have a median survival period of 13 months. Conclusion MRCP was an ideal noninvasive preoperative examination for identifying hilar cholangiocarcinoma. Routine frozen section of the bile duct margin should be adopted. Radical resection definitely prolongs survival period. For Bismuth type Ⅲ、Ⅳ without extrahepatic metastasis, orthotopic liver transplantation was an effective surgical therapy.