Objective Using M2-Gi1α fusion protein expressed by baculovirus-Sf9 cell system to find the specific ligand for M2 receptor and detect the interaction of the two parts of the fusion protein.Methods The fused M2-Gi1α cDNAs were generated in a two-step PCR and then expressed in Sf9 cells.[3H]QNB and[35S]GTPγS binding experiments were employed to study the function of M2-Gi1α fusion protein.Results The expression level of M2-Gi1α fusion protein was 8.44±0.39 nmol·g-1 protein.The affinity of GDP to the Gi1α part changed under the affection of different ligands.The IC50 value in the appearance of acetylcholine,oxotremorine,arecoline,atropine,fangchinoline,levitimide were 21.35 μmol·L-1,23.86 μmol·L-1,11.91 μmol·L-1,0.13 μmol·L-1,1.05 μmol·L-1,1.75 μmol·L-1,and 2.5 μmol·L-1 when there was no ligand.Conclusion The M2-Gi1α fusion protein expressed in baculovirus-Sf9 cell system has pharmacological specificity for M2 receptor and the efficient coupling function between the two parts.The M2-Gi1αfusion protein is a helpful tool for detecting the new specific ligands of the M2 receptor.

ObjectiveTo investigate the influence of timing of definitive operation on prognosis of patients with iatrogenic bile duct injury (IBDI). MethodsThe clinical data of 38 patients with IBDI were retrospectively analyzed. Of the 38 patients, the injury happened in 7 patients in our hospital while 31 patients were transferred in from other hospitals. We analyzed the previous operative records, the clinical manifestations of the patients, the radiological images and our operative findings in the definitive operations. According to the timing of the definitive operation, the patients were divided into the intraoperative repair group (n=26), the early repair group (n=15) and the delayed repair group (n=17). ResultsPatients in the intraoperative repair group had the best short-and long-term results, the shortest length of hospital stay and the lowest total cost. The early repair group, though having worse Results than the intraoperative repair group was still better than the delayed repair group. However, early detection of injuries and the ability to carry out timely corrective procedures depended on the availability of surgical expertise in the hospital. ConclusionsThe timing of definitive operation to provide corrective surgery to IBDI was a vitally important factor which affected prognosis.It is best to repair the injury within the same operation. The repair should be carried out by a doctor with ample experience in bile duct surgery.

Aim To generate m3AChR-G11 fusion protein in baculovirus-Sf9 cells and test the couping function,the interation and the influence factors be-tween m3AChR and G11 protein,as well as screen the specific ligands for m3AChR. Methods m3AChR-G11 fused DNA was generated through a two-step PCR and then expressed in Sf9 cells to produce fusion protein. The total concentration for membrane protein was de-tected by BCA method,[3H]QNB and [35 S]GTP?S binding experiment as perfomed to study the function of m3AChR-G11 fusion protein. Results The expression level of m3AChR-G11 was 7. 76 ? 10 -9 mol?g -1. The affinity of GDP to G11 partner changed in the presence of different muscarinic ligands. IC50 values of GDP in the presence of ACh,Pilo,CCh,MCN-A-343,Atro,4-Damp and Dafi were 82. 2,93. 70,12. 10,14. 30, 1. 93,1. 37,0. 72 ? 10 -6 mol ? L -1 respectively,and that in the absence of muscarinic was 1. 99 ? 10 -6 mol ?L-1.Concluslons The m3AChR-G11 fusion protein has the pharmacological specificity of m3 receptor and the efficient coupling interaction of the two partners. Affinity of GDP to ligand-bound fusion protein represents the species of muscarinic ligands. This is helpful in screening and detecting the new specific ligands to muscarinic receptors.

Autoimmune pancreatitis (AIP) is a rare form of chronic pancreatitis which is closely related with abnormal autoimmune.To some extent,it is too difficult to identify the diagnosis between AIP and pancreatic cancer.The treatment includes hormone therapy and surgery.In this article,based on accumulating the experience in the diagnosis and treatment of AIP cases for many years and reviewing the related literatures,we evaluate its type,the etiology and the clinical presentations,as well as summarize the typical characteristics of pathology,radiology and serology.

In the past,it is always believed that routine drainage after pancreaticoduodenectomy (PD) is one of the most crucial methods to prevent and detect postoperative complications and to reduce mortality.However,in more recent years,with the development of fast track surgery,scholars in pancreatic surgery have investigated the necessity of drainage after PD together with its merits and faults.Therefore,this paper gives a brief review on this topic.

Objective To investigate the diagnosis and treatment of intraductal papillary mucinous neoplasm (IPMN) of the pancreas.Methods The clinical data of patients with pathologically verified IPMN who underwent surgical treatment between January 2006 to April 2014 in the First Affiliated Hospital of Harbin Medical University were studied retrospectively.There were 27 males and 21 females.The average age was (57.8 ± 8.8) years old.The average caliber of the main pancreatic duct was (1.1 ± 0.6) cm.The average size of the branch duct IPMN was (4.6 ± 1.5) cm.Results 35 patients underwent pancreaticoduodenectomy.Eight patients underwent distal pancreatectomy.Two patients underwent duodenum-preserving pancreatic head resection.Two patients underwent splenic-preserving distal pancreatectomy and one patient underwent total pancreatectomy.No patient died in perioperative period,and the median length of hospital stay after surgery was 14.3 days.Postoperative pathological examination revealed 5 (10.4％) adenoma,12 (25.0％) moderate-grade dysplasia,14 (29.2％) high-grade dysplasia and 17 (35.4％) invasive carcinoma.The postoperative complication rate was 22.9％.The mean follow up period for the noninvasive tumors was 48.9 months,with no recurrence or deaths.The mean follow up period of the invasive tumors was 43.2 months,with 1 death and no recurrence.Conclusions The indications for resection of IPMN should be based on treatment guidelines and on the patient' s general condition.It is suggested that the diagnosis and treatment of IPMN should be conducted in specialized pancreatic surgery centers.

Objective To summarize the experience in surgical management of hilar cholangiocarcinoma.Methods The clinical data of 88 patients with hilar cholangiocarcinoma who received surgical treatment at the First Affiliated Hospital of Harbin Medical University from January 2007 to December 2011 were retrospectively analyzed.All the patients were diagnosed by imaging examination.According to the severity of jaundice and predictive remnant liver volume,19 patients received percutaneous transhepatic cholangial drainage (PTCD) and 4 received portal vein embolization.The fundamental operation consisted of hilar cholangiocarcinoma resection,skeletonization of hepatoduodenum ligament and Roux-en-Y cholangiojejunostomy,and the transanastomotic stent was placed for 6 months.The count data were analyzed using the chi-square test; the survival rate was analysed using the Kaplan-Meier method; the survival was analyzed using the Log-rank test.Results Of the 88 patients,58 patients (including 11 patients who received PTCD) received hilar cholangiocarcinoma resection.Of the 58 patients,43 (including 4 patients who received portal vein embolization preoperatively) received R0 resection,and 15 received palliative resection.Thirty patients received internal and (or) external drainage.Commitant partial hepatectomy was performed on 22 patients (including 9 received left hemihepatectomy,2 received extended left hemihepatectomy,7 received left hemihepatectomy + caudate lobectomy,4 received right hemihepatectomy).Commitant pancreatico-duodenectomy was performed on 7 patients,commitant hepatic artery resection on 3 patients,and commitant portal vein resection on 2 patients.According to the modified Bismuth-Corlette classification,there were 17 patients with type Ⅰ,19 with type Ⅱ,21 with type Ⅲa,20 with type Ⅲb,and 11 with type Ⅳ.Of the 58 patients who received hilar cholangiocarcinoma resection,19 had postoperative complications,and 2 patients died within 30 days after operation.Seventy-three patients were followed up,and the overall 1-,3-,5-year survival rates were 68.5％,28.8％,11.0％,respectively.The 1-,3-,5-year survival rates of patients who received R0 resection were 94.6％,43.2％,18.9％,respectively,which were significantly higher than 78.6％,35.7％ and 7.1％ of patients who received palliative resection (x2=4.77,P ＜0.05).The 1-,3-,5-year survival rates of patients who received palliative resection were significantly higher than 18.2％,0,0 of patients who received biliary drainage (x2 =13.26,P ＜ 0.05).Conclusions R0 resection is the best choice for patients with hilar cholangiocarcinoma,and biliary drainage with no resection is the last choice.Sufficient preoperative treatment,optimized choice of surgical procedure and exquisite surgical techniques are important for the improvement of the prognosis.

Objective:To retrieve and analyze the relevant literatures on vancomycin added into bone cement to provide the evi-dence for the treatment of osteomyelitis and other orthopedic infections. Methods:Search strategy and criteria of inclusion and exclu-sion for literatures were designed. PubMed, SCI, Embase, CNKI, VIP and the other databases were searched, and the articles from the establishment date to February 2014 were statistically analyzed using bibliometric methods. The final included documents were sta-tistically analyzed in respect of the article type, year, contents, citation frequency and the maln contents of the study. Results:A total of 1 941 articles were searched, and 430 of them were in the final inclusion. The total number of the articles in every year was in an es-calating trend. The paper focused on the research and analysis of clinical studies, and there were 74 clinical studies among the includ-ed literatures, which accounted for 17. 2% of all the included literatures. The highest citation frequency was 97 for one literature. The research included the overall situation, year distribution, publishing country, research type analysis, corresponding author and their in-stitutions, journals, citation frequency, and the maln content of work and clinical studies on vancomycin added into bone cement. The analysis could provide reference for the clinical treatment of orthopedic diseases. Conclusion: The results of the analysis show that vancomycin added into bone cement in the treatment of chronic osteomyelitis is effective with high security, and the technology is ma-ture.

Clinical trials for medical research subjects of protection in China is still the researchers and sub-jects′cognitive biases, informed consent to carry out the difficult and the lack of follow-up tracking examination questions.Based on the experience and the domestic exploration of a hospital, put forward the Suggestions of per-fecting our country′s subjects protection: establish multi-sectoral cooperation of the subjects protection system, improve the legal consciousness and the consciousness of the subjects, the ethics committee should be field trips, build system of compulsory insurance, adverse drug reaction relief system is established.

Objective To study the function of early persistent vacuum suction drainage in patients with high risk of pancreatic fistula after pancreaticoduodenectomy (PD).Methods From Jul.2010 to Jun.2013,the clinical data of 286 patients undergoing PD were retrospectively evaluated.87 patients with high risk of pancreatic fistula were screened and then divided into early persistent vacuum suction drainage group (the observation group) and conventional drainage group (the control group) according to postoperative drainage manners.We statistically analyzed the two groups in terms of general information,blood loss,operative time,medical expenses,hospital stay,mortality and morbidity of complications such as pancreatic fistula.Results There were 40 patients screened into the observation group and 47 patients into the control group.No difference was found between the observation group and the control group in basic clinical data or surgical data.There was no statistical difference between the two groups in delayed gastric emptying,bile leakage,bleeding or the incidence of pancreatic fistula and intra-abdominal infection.The incidence of pancreatic fistula with grade B and C in the observation group was statistically lower than that of the control group(12.5％ vs 34.0％,P＜0.05).The incidence of intraabdominal infection in observation group was statistically lower than those in the control group (20.0％ vs 40.4％,P＜0.05).The incidence of total complications in observation group was statistically lower than that in the control group (60.0％ vs 83.0％,P＜0.05),but no difference was found between the observation group and control group in morality.Early persistent vacuum suction drainage could reduce hospital stay((21.93±7.14) days vs (28.70±12.45) days,P＜0.05) and clinical expense ((64.8±12.0) thousands vs (75.2± 14.6) thousands,P＜0.05) in patients with high risk of pancreatic fistula after PD.Conclusions Early persistent vacuum suction drainage can reduce the rate of grade B and C pancreatic fistula in patients with high risk of pancreatic fistula undergoing PD.The manner can also reduce the incidence of intra-abdominal infection,total complications and cost of hospital stay.This manner can accelerate postoperative recovery of patients undergoing PD and is worth of widely used.