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.

Objective To investigate the efficiency and safety of laparoscopic hepatectomy (LH) and open hepatectomy (OH) in patients with hepatocellular carcinoma (HCC). Methods A total of 45 HCC cases undergoing hepatectomy from January 2008 to December 2010 in our hospital were divided into LH group ( n =21 ) and OH group ( n =24 ).Their clinical efficiency and safety were compared and analyzed. Results The amount of intraoperative blood loss and blood transfusion in LH group were significantly higher than that of OH ( respectively x2 =3.973,x2 =4.862,all P ＜ 0.05 ).LH is associated with less postoperative complications and duration of hospital stay,compared with OH ( respectively x2 =4.746,t =-2.717,all P ＜0.05).Cumulative survival rates in both groups were not statistically significant ( x2 =0.172,P ＞ 0.05 ).However tumor-free survival rate in LH was significantly higher,superior to that of OH (x2 =4.543,P ＜ 0.05). Conclusions LH and OH are all safe and effective hepatectomy for HCC.LH has the advantage of less complication and shorter hospital stay.

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.

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 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.

Objective To evaluate negative lymph node count (NLNC) in prediction of prognosis of T3 gastric cancer after radical resection.Method 214 T3 patients of radical gastrectomy with complete clinical and follow-up data between Jan 2003 to Dec 2007 were enrolled.Survival was determined by the Kaplan-Metier method and univariate analysis was done by Log-rank test,Multivariate analysis was performed using the COX proportional hazard regression model.-2loglikelihood value and the hazard ratio (HR) value were used to compared the value of number of lymph node-negative (NLNC) staging and pN staging and lymph node metastasis rate (MLR) in gastric cancer prognosis evaluation.Results Univariate analysis showed that,pN stage (x2 =31.664),MLR stage (x2 =34.123),tumor size (x2 =5.025),type of differentiation (x2 =5.993),Borrmann classification (x2 =5.401),NLNC stage (x2 =37.256) were related to survival (P ＜ 0.05).COX multivariate analysis showed that-2loglikelihood of pN staging is 1 336.761,HR value is 1.464,-2loglikelihood value of MRL staging is 1 335.821,HR value is 1.441.-2loglikelihood value of NLNC staging is 1 326.902,HR value is 1.725.The N0 and N1 staging prognosis in different NLNC staging was significant (P =0.008,P =0.014).Conclusions Sufficient number of negative lymph node prolongs survival and reduces the risk of early recurrence in advanced gastric cancer.

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 evaluate a new TNM staging system inclusive of intraperitoneal free cancer cells in terms of postoperative survival of patients with gastric cancer. Methods Free cancer cells (FCC) in the peritoneal washes of gastric cancer patients were estimated by measuring CEA mRNA levels using real-time RT-PCR. After 5-year follow-up, we get the cut-off value of CEA mRNA level by using MedCalc software to analyze the ROC curve. When CEA levels are more than the cut-off value, it may considered as FCC(+), and then using FCC(+) as distant metastasis (MI) to make a new TNM staging and analyze patients life-span. Results (1) Under the ROC curve analysis, when the cut-off value of CEA mHNA level was at 31.21 copies/ml, the Youden's index is the highest. (2) When FCC (+) considered as M1 to make a new TNM staging, the 5-year survival rate showed as below: Ⅰ-Ⅱ, P=0. 134; stage Ⅱ-Ⅲ P=0.004 and Ⅲ-Ⅳ P=0.022,repecetively. Conclusion (1) The best cut-off value of CEA mRNA levels for FCC in peritoneal washes is 31.2 copies/ml. (2) Our study demonstrated that application of FCC(+) in the TNM staging may have a better estimation of prognosis of patients suffering from advanced gastric cancer.

OBJECTIVE: To summarize the classification, property of skull repair material and the processing of related complications in the application of repairing and remodeling the skull.METHODS: The PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) and CNKI Database (www.cnki.net/index.htm) was searched by the first author using key words of "codubix, synmesh, and bone cement" both in English and Chinese. Articles whose content is related to the types, property, biocompatibility and its application effect of skull repairing materials were selected. In the same field, the articles published by authoritative journals or different races were preferred. The repetitive or obsolete literatures were excluded. After that 26 documents were included in this paper.RESULTS: The bone cement exhibited good histocompatibility, however, it is difficult to be absorbed, thus, it only be used for repairing part of skull defects. The study found that titanium had good biocompatibility and could combine with the skull. Its application had a promising prospect, but there are many inadequacies. With the continuous deepening of bio-engineering, bone tissue engineering, and cartilage tissue engineering, it will provide a broader perspective for the study of skull repair materials.CONCLUSION: There are many kinds of materials for skull repairing, and this paper only introduces some of the widely used ones. Actually, the selection of repairing materials should consider the pathogenetic condition, economic condition, local equipment and technical levels. If possible, titanium mesh and titanium screw are preferred, which is characterized by easy operation, few complication and beautiful appearance: When using autogenous bone or bone cements, if related complications can be reduced and handled effectively, the results will be satisfactory.