Objective To investigate the stability and comparability of recombinant human muscle creatine kinase isozyme(CK-MM)in different matrix,and to find out a suitable matrix as a reference material for application of CK measurement system.Methods The stability of the recombinant CK-MM was observed by measuring its catalytic activity in self-made matrix,anti-freeze additive matrix,the certified reference material(CRM)matrix and human serum matrix respectively.For understanding the commutability among different measurement system total CK catalytic activity in various concentration of human serum and the catalytic activity of recombinant CK-MM in different matrix specimen were measured with IFCC recommended reference method and routine reagent produced by Roche and Zhongsheng Diagnostic Company respectively.Results The recombinant CK-MM was stable for 25 days in the self-made matrix,23 days in CRM matrix and at least 240 days in antifreeze additive matrix stored at-20 ℃.The recombinant enzyme protein in self-made matrix and CRM matrix was commutable.Low concentrations of recombinant CK-MM in human serum and in anti-freeze additive matrix were also commutable.Conclusion The recombinant CK-MM had good stability and commutability in either self-made matrix or CRM matrix.It might be used as a candidate reference material for the determination of total creatine kinase in serum.

Objective To summarize the experiences of the treatment of extrahepatic bile duct injury caused by cholecystectomy.Methods A retrospective analysis was made on the treatment of 11 cases of extrahepatic bile duct injuries in our hospital from 1989 to 2001.The position of injuries,types,the time from onset to diagnosis, operation methods and therapeutic effects were analyzed.Results Three of the 11 cases received an emergency operation,while 8 underwent selective operation.Seven cases (62.7%) of extrahepatic bile duct injury were diagnosed during the cholecystectomy,and the remaining 4 cases (37.3%) were diagnosed after operation.Seven cases of extrahepatic bile duct injury diagnosed during operation were treated immediately and 4 cases treated 3-4 days after the first operation.All the patients were cured and discharged.Conclusion The correct principles and methods of handling the cases of extrahepatic bile duct injury caused by cholecystectomy should be mastered;Roux-en-y cholangiojunostomy is the best option for the treatment of extrahepatic bile duct injury.

Objective To study the inhibitory effect of human antisense high mobility group box 1 (HMGB1) gene on the growth of human pancreatic adenocarcinoma cell line (PANC-1).Methods HMGB1 was cloned and reversely inserted into eukaryotic expression vector pcDNA3.1 to get pcDNA3.1/anti-HMGB1, which was then transfected by liposome method into PANC-1 cells.After G418 selection, HMGB1 expression of PANC-1 cells before and after transfection was detected by RT-PCR and Western blot, the cell proliferating activity in vitro was examined by MTT analysis, and cell cycle and apoptosis were assessed by flow cytometry.Results pcDNA3.1/anti-HMGB1 vector was obtained.After transfection with pcDNA3.1/anti-HMGB1, HMGB1 gene expression of PANC-1 cells was inhibited in both mRNA and protein level, and the proliferation of cultured pancreatic adenocarcinoma cells was suppressed with a suppression rate higher than 40 % on the sixth day.The cells of G1 phase increased obviously while the cell number of S phase decreased, the number of apoptosic cell increased.Conclusion Antisense HMGB1 gene can inhibit the proliferation of human ancreatic adenocarcinoma cells and increase the cell apoptosis rate.

Objective To construct a prokaryotic expression system for human creatine kinase(CK) MM isozyme,purify the recombinant protein of CK expressed in Eschericheia coli(E.coli) and examine the stability of the recombinant protein for its application in CK measurement system as the quality control.Methods Total RNA of extracted from fetal cardiac muscle was reversetranscripted and cDNA encoding human CK was amplified which was inserted into pET-15b plasmid vector.The recombinant plasmid was transfered into E.coli BL21(DE3) and induced by isopropyl-?-D-thiogalactopyranoside(IPTG).Recombinant CK-MM was separated from bacterial proteins by affinity chromatography on a Ni2+-Sepharose column.The activity of the recombinant enzyme was observed in different matrixes.Results The enzymatic activity of the crude extracts of CK-MM was up to 280,000U/L.After one step affinity chromatography,the fusion protein showed a single band in SDS-PAGE gel.The purified protein was stable and the enzymatic activity remained unchanged for a month in the matrix containing bovine serum albumin,EDTA and 1,4-Dithiothreitol(DTT).Conclusion The recombinant CK-MM showed key properties of the native creatine kinase isozyme and may be used as control and calibrator for determination of serum CK-MM.

Objective To compare the success rate, time of tumor formation and number of tumors in three methods of tumor transplantation, in order to seek an ideal animal model for molecular imaging study. Methods Forty-eight BALB/C-nu/nu nude mice were randomly divided into three groups (each n=16). Tumor tissue mass of 2 mm3 was injected into subcutaneous of nude mice in experiment group. Tumor tissue mass of 1 mm3 was applied in control group 1. Tumor cells suspension liquid was injected into subcutaneous of nude mice in control group 2. The tumor formation rate, the time of tumor formation and the number of tumors were observed. MRI scanning were performed 3-6 weeks after implantation. Results The rate of tumor formation of three groups was 93.75%, 75.00% and 43.75%, respectively. The time of tumor formation was (21.7±2.4), (29.8±2.9) and (34.6±3.9) days, respectively. The rate of solitary nodule implanted tumor was 93.33% in experiment group, higher than that in control group 1 (75.00%) and control group 2 (14.29%). The tumors were hypointense on T1WI and hyperintense on T2WI. Conclusion Transplantation 2 mm3 tumor tissue mass is effective to set up the subcutaneous implanted tumor models with a high success rate of tumor formation, a short time of tumor formation and high rate of solitary tumors, being suitable for the study of molecular imaging. The models can undertake conventional T1WI, T2WI and T2-mapping imaging, and the imaging qualities are good.

Objective: To evaluate the application of frontal flap in reconstructing the defect of nose, the clinical data of 12 cases were reported. Methods: Twelve patients with nasal skin cancers undertaking radical dissection of tumor accompanied frontal flap reconstructing one stage were reviewed retrospectively.Results: The overall following\|up rate was 100%, 10 of these patients curled slightly after reconstructing the nose, but influence of the nasal outline was little. Two cases were recurrent locally, one occurred 24 months after operation, the other occurred 38 months. Both of them received radiotherapy. Malformation arised after radiotherapy. The total successful rate of reconstruct nasal outlook was 83.33%, the 3\|year locally recurrent rate was 8.33%,5\|year survival rate ws 100%, 5\|year survival rate without tumor was 83.33%.Conclusion:The blood supply of frontal flap is abundance;healing up soon after reconstruction; the manipulation of surgery is simple and safe. It is worth applying this operation generally in clinical practice.

Objective To analyze the surgical treatment of pancreatic duct stones.Methods The clinical data of 46 patients with pancreatic duct stones treated in our hospital from January 2008 to January 2013 were retrospectively analyzed.Results The most common symptoms were abdominal pain in 42 patients,diarrhea in 4 patients,diabetes in 6 patients,increased level of amylase in 4 patients,high level of CA19-9 in 9 patients and concomitant pancreatic cancer in 5 patients.4 patients had history of acute pancreatitis.All patients were diagnosed with pancreatic duct stones by preoperative imaging.The stones were located in the head of the pancreas in 21 cases,in pancreatic body and tail in 17 cases,and in the whole length of the pancreas in 8 cases.7 cases had single stone,28 cases had 2 to 3 stones,and 11 cases had more than three stones.21 cases had stones with a maximum diameter more than 1.0 cm,and 25 cases with a maximum diameter less than 1.0 cm.Pancreatic lithotomy plus pancreaticojejunostomy was performed in 33 cases,pancreatoduodenectomy in 8 and resection of the body and tail of pancreas plus splenectomy in 5 cases.6(13.0％) patients had postoperative complications,and there was no mortality.3(6.5％) patients had postoperative residual stones.39 cases were followed up with follow-up time ranging from 3 months to 57 months.Pain relief rate was 85.7％,Stone occurred in 2 (4.3％) patients.Conclusions Surgery is an important treatment for pancreatic duct stones,and treatments should be adopted based on the situations of individual patients.

Objective To investigate the effect of postoperative prophylactic transcatheter arterial Chemoembolization(TACE) on preventing recurrence in 54 high-risk patients with hepatocellular carcinoma.Methods These 54 HCC cases were greded as high risk for recurrence and put on close follow up after radical resection.Risk factors included tumor thrombus,cancer satellite or multiple cancer nodules,postoperative high AFP level.Among them 42 cage (target group) received TACE,in which liver function was of Child A,and hepatitis virus replication was controlled in 1000 copies/ml.12 cases(centrol group) didn't receive TACE.The recurrent rate of HCC was compared between the cases with prophylactic TACE and those without through two years of follow-up. Result The recurrence rate of HCC was significantly lower in the cases with prophylactic TACE(19.O%) than those without(50%) within 1 year after the radical operation,and 2 years (52.3% vs 83.3%). Conclusion Postoperative TACE contributes to reducing the short-term HCC recurrence rate.

Chronic kidney disease (CKD) is becoming a global social problem. It is important to slow down the progression of CKD for economic and social concerns. In recent years, it has been found that colon is one of the vital organs which produce uremic toxins. And enterogenous uremic toxins are closely related to the prognosis of CKD. Theory of gut-kidney axis for the slowdown of CKD progression was raised by foreign scholars and became the research hot spot. Colon therapy with traditional Chinese medicine (TCM) has been widely used in clinical practice and is believed to slow down the progression of CKD by numerous clinical reports. However, low re-search quality and ambiguous results limited its further application. Under the guidance of senior TCM Professor Huang Chunlin, who emphasized the method of draining turbidity through bowels in the management of CKD, from the Nephrology Center, Guangdong Provincial Hospital of Chinese Medicine, as well as the modern theory of gut-kidney axis, we had carried out a series of exploratory researches which will provide data and methodology support for further confirmatory studies and improve its effectiveness.

Liver transplantation is the only effective method to treat various end-stage liver diseases. The shortage of standard donor liver is the main factor limiting the development of liver transplantation, whereas the application of marginal donor liver has significantly expanded the source of donor liver. New technologies, such as extracorporeal membrane oxygenation (ECMO), mechanical perfusion and hypothermia, can significantly improve the quality of marginal donor liver, which has good development prospects in organ function maintenance. This article reviews the common types of marginal donor liver and the application of related novel technologies in the maintenance of marginal donor liver function.