Application value of three-dimensional printing technology assisted laparoscopic anatomic liver resection of segment 8 / 中华消化外科杂志

Objective:To investigate the application value of three-dimensional (3D) printing technology assisted laparoscopic anatomic liver resection of segment 8 (Lap-S8).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 8 liver cancer patients including 7 cases with hepatocellular carcinoma and 1 case with intrahepatic cholangio-carcinoma who underwent 3D printing technology assisted Lap-S8 in the Hunan Provincial People′s Hospital from January 2019 to December 2020 were collected. There were 7 males and 1 female, aged from 49.0 to 80.0 years, with a median age of 56.5 years. Of the 8 patients, 6 cases underwent laparoscopic anatomic liver resection of the entire segment 8, 1 case underwent laparoscopic anatomic liver resection of ventral subsegmental of the segment 8 and 1 case underwent laparoscopic anatomic liver resection of dorsal subsegmental of the segment 8. 3D printing technology was used to assist preoperative evaluation and intraoperative navigation for all 8 patients. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination, internet or telephone interview to detect survival and tumor recurrence of patients after operation up to March 2021. Measurement data with normal distribution were represented as Mean±SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Surgical situations: all the 8 patients underwent 3D printing technology assisted Lap-S8 successfully, without conversion to open surgery. The operation time, hepatic portal occlusion time and volume of intraoperative blood loss of the 8 patients were (216±41)minutes, (56±11)minutes and 75 mL(range, 50 to 300 mL), respectively. There was no intraoperative blood transfusion in 8 patients, and the surgical margin of the 8 patients was negative. (2) Postoperative situations: the duration of postoperative hospital stay of the 8 patients were (9±3)days. There was no complication such as postoperative hemorrhage, biliary fistula, liver abscess or abdominal infection occurred. (3) Follow-up: all the 8 patients were followed up for 3.0?24.0 months, with a median follow-up time of 12.5 months. During the follow-up, 1 of 8 patients with preoperative diagnosis of recurrent hepatocellular carcinoma developed tumor recurrence at 5 months after operation. The patient underwent laparoscopic surgery followed with the transcatheter arterial chemoembolization and target therapy, and survived with tumor. There was no tumor recurrence in the other 7 patients.Conclusion:3D printing technology assisted Lap-S8 is safe and feasible.

Clinical exploration of laparoscopic liver resection for intrahepatic cholangiocarcinoma of 58 cases / 国际外科学杂志

Objective@#To explore the safety and feasibility of laparoscopic liver resection in the treatment of intrahepatic cholangiocarcinoma (ICC).@*Methods@#The retrospective study was adopted. The clinical data of 58 patients with ICC who underwent laparoscopic liver resection in the Department of Hepatobiliary Minimally Invasive Surgery of the First Affiliated Hospital of Hu′nan Normal University were collected From January 2016 to December 2018. Among them, 34 patients were males and 24 were females, aged from 34 to 71 years with a median age of 54 years. Observation indicators: (1) Surgical treatment: surgical methods, operation time, intraoperative blood loss, intraoperative blood transfusion rate, intraoperative hepatic portal blocking time, conversion rate, postoperative complications, postoperative hospital stay. (2) Postoperative pathological conditions. (3) Follow-up.Follow-up visits were conducted using an outpatient clinic and telephone to understand patient survival after surgery. The follow-up period was until June 2019. Measurement data with normal distribution were expressed as (Mean±SD), count data was expressed as frequency and percentage.@*Results@#A total of 58 patients were included in this study, of which 48 patients underwent laparoscopic radical surgical resection of intrahepatic cholangiocarcinoma and 10 patients underwent laparoscopic conversion to laparotomy. (1) Surgical treatment: laparoscopic resection of the left liver (segments Ⅱ, Ⅲ and Ⅳ), laparoscopic resection of the right liver (segments Ⅴ, Ⅵ, Ⅶ and Ⅷ), laparoscopic resection of the right posterior lobe (segments Ⅵ and Ⅶ), laparoscopic extended resection of the right posterior lobe, laparoscopic resection of the middle lobe (Ⅳ, Ⅴ and Ⅷ), laparoscopic resection of the V and Ⅵ, laparoscopic resection of the left liver (segments Ⅱ, Ⅲ and Ⅳ) combined with the caudate lobe (segments I and Ⅸ), laparoscopic extended left hemihepatectomy, laparoscopic resection of the VI, laparoscopic resection of the Ⅶ and Ⅷ, laparoscopic resection of the left lateral lobe (segments Ⅱ and Ⅲ) and laparoscopic resection of the right hepatic mass; operation time: (320.38±107.68) min; intraoperative blood loss: (262.34±76.06); intraoperative blood loss: 0 (0/58); Intraoperative hepatic portal occlusion time: (48±15) min, the conversion rate was 17.2% (10/58); the incidence of postoperative biliary fistula was 6.8% (4/58), and the patient was discharged after conservative treatment and unobstructed drainage (T-tube vacuum suction); the postoperative gastrointestinal recovery time was (1.84±0.57) d; no other serious complications occurred.Postoperative hospital stay: (9.34±3.39) d; there were no deaths and unplanned surgeries during the perioperative period. (2) Pathological conditions: 32 cases received lymph node dissection during the operation, and 26 cases showed cholangiocarcinoma without lymph node dissection; pathological examination showed that the pathological reports of all tumor margins were negative, and 4 cases showed lymph node dissection and positive lymph node metastasis. (3) Follow-up results: of the 58 patients with ICC, 49 were followed up for 6 to 36 months. The tumor survival time was (4 to 36) months. 28 patients survived without tumor. 17 patients had intrahepatic metastasis with multiple lymph node metastasis. 4 patients were treated with microwave ablation after intrahepatic metastasis was found. 9 patients were lost to follow-up.@*Conclusion@#Laparoscopic treatment of intrahepatic cholangiocarcinoma is safe and feasible in experienced centers.

Clinical exploration of laparoscopic liver resection for intrahepatic cholangiocarcinoma of 58 cases / 国际外科学杂志

Objective To explore the safety and feasibility of laparoscopic liver resection in the treatment of intrahepatic cholangiocarcinoma (ICC).Methods The retrospective study was adopted.The clinical data of 58 patients with ICC who underwent laparoscopic liver resection in the Department of Hepatobiliary Minimally Invasive Surgery of the First Affiliated Hospital of Hu'nan Normal University were collected From January 2016 to December 2018.Among them,34 patients were males and 24 were females,aged from 34 to 71 years with a median age of 54 years.Observation indicators:(1) Surgical treatment:surgical methods,operation time,intraoperative blood loss,intraoperative blood transfusion rate,intraoperative hepatic portal blocking time,conversion rate,postoperative complications,postoperative hospital stay.(2) Postoperative pathological conditions.(3) Followup.Follow-up visits were conducted using an outpatient clinic and telephone to understand patient survival after surgery.The follow-up period was until June 2019.Measurement data with normal distribution were expressed as (Mean ± SD),count data was expressed as frequency and percentage.Results A total of 58 patients were included in this study,of which 48 patients underwent laparoscopic radical surgical resection of intrahepatic cholangiocarcinoma and 10 patients underwent laparoscopic conversion to laparotomy.(1) Surgical treatment:laparoscopic resection of the left liver (segments Ⅱ,Ⅲ and Ⅳ),laparoscopic resection of the right liver (segments Ⅴ,Ⅵ,Ⅶ and Ⅷ),laparoscopic resection of the right posterior lobe (segments WⅥ and Ⅶ),laparoscopic extended resection of the right posterior lobe,laparoscopic resection of the middle lobe (Ⅳ,Ⅴ and Ⅷ),laparoscopic resection of the Ⅴ and Ⅵ,laparoscopic resection of the left liver (segments Ⅱ,Ⅲ and Ⅳ)combined with the caudate lobe (segments Ⅰ and Ⅸ),laparoscopic extended left hemihepatectomy,laparoscopic resection of the Ⅵ,laparoscopic resection of the Ⅶ and Ⅷ,laparoscopic resection of the left lateral lobe (segments Ⅱ and Ⅲ) and laparoscopic resection of the right hepatic mass;operation time:(320.38 ± 107.68) min;intraoperative blood loss:(262.34 ± 76.06);intraoperative blood loss:0 (0/58);Intraoperative hepatic portal occlusion time:(48 ± 15) min,the conversion rate was 17.2％ (10/58);the incidence of postoperative biliary fistula was 6.8％ (4/58),and the patient was discharged after conservative treatment and unobstructed drainage (T-tube vacuum suction);the postoperative gastrointestinal recovery time was (1.84 ± 0.57) d;no other serious complications occurred.Postoperative hospital stay:(9.34 ± 3.39) d;there were no deaths and unplanned surgeries during the perioperative period.(2) Pathological conditions:32 cases received lymph node dissection during the operation,and 26 cases showed cholangiocarcinoma without lymph node dissection;pathological examination showed that the pathological reports of all tumor margins were negative,and 4 cases showed lymph node dissection and positive lymph node metastasis.(3) Follow-up results:of the 58 patients with ICC,49 were followed up for 6 to 36 months.The tumor survival time was (4 to 36) months.28 patients survived without tumor.17 patients had intrahepatic metastasis with multiple lymph node metastasis.4 patients were treated with microwave ablation after intrahepatic metastasis was found.9 patients were lost to follow-up.Conclusion Laparoscopic treatment of intrahepatic cholangiocarcinoma is safe and feasible in experienced centers.

Exploration on laparoscopic hepatectomy on central liver tumor: a report of 40 cases / 中华外科杂志

Objective@#To assess the safety and feasibility of the application of the laparoscopic modality in the perioperative treatment of central liver tumors.@*Methods@#Collecting all the clinical information of a total of 40 patients with central liver tumors who received laparoscopic resection treatment carried out at Department of Hepatological Surgery of People′s Hospital of Hunan Provincial from January 2016 to December 2018 to take a retrospective review. There were 19 males and 21 females.The age was (59.5±14.5) years (range: 15 to 71 years) . There were 26 cases of primary hepatic carcinoma (24 cases of hepatocellular carcinoma, 2 cases of cholangiocellular carcinoma) , 8 cases of hepatic cavernous hemangioma, 1 case of metastatic hepatic carcinoma, 5 cases of hepatocellular adenoma. The maximum diameter of tumors were (6.2±2.9) cm (range: 2 to 13 cm) . The patient′s information about hepatectomy methods, blocking mode and time of blood flow, operation time, intraoperative blood loss, intraoperative blood transfusion rate, post-operative hospitalization time, perioperative reoperation and postoperative complications were collected.@*Results@#A total of 40 patients all were treated with laparoscopic surgery. The surgical procedure was as follows: 2 patients received the right hepatic lobectomy (Ⅴ, Ⅵ, Ⅶ and Ⅷ segments) , 2 patients received the left hepatic lobectomy (Ⅱ, III and Ⅳ segments) , 13 patients received mesohepatectomy (Ⅳ, Ⅰ and Ⅷ segments) , 2 patients received left hepatic trisegmentectomy (Ⅱ, Ⅲ, Ⅳ and Ⅷ segments) , 2 patients received right hepatic trisegmentectomy (Ⅳ, Ⅴ, Ⅵ, Ⅶ and Ⅷ segments) , 7 patients received Ⅷ segmentectomy, 1 patient received Ⅳ segmentectomy, 3 patients received Ⅴ and Ⅷ segmentectomy, 5 patients received hepatic caudate lobe resection (Ⅰ, Ⅸ segments) , and 3 patients received local tumors resection.Pathological results: there were 26 cases of primary hepatic carcinoma (24 cases of hepatocellular carcinoma, 2 cases of cholangiocellular carcinoma) , 8 cases of hepatic cavernous hemangioma, 1 case of metastatic hepatic carcinoma, 5 cases of hepatocellular adenoma; the pathological reports of all malignant tumor cases all showed negative incisal edge. The operative time was (333±30) minutes (range: 280 to 380 minutes) ; the intraoperative hepatic portal occlusion period was (58±13) minutes (range: 30 to 90 minutes) ; the intraoperative hemorrhage was (173±129) ml (range: 20 to 600 ml) ; the intraoperative blood transfusion rate was 2.5% (1/40) ; the postoperative incidence of bile leakage was 2.5% (1/40) , the hospital discharge of 1 patient with bile leakage was approved after conservative treatments like T pipe decompression and adequate drainage; there was 1 case of abdominal infection and 1 case of pulmonary infection, both of which were discharged from the hospital with conservative treatments; there were no other serious postoperative complications. The postoperative hospital stay was (10.7±2.7) days (range: 6 to 16 days) ; there were no perioperative mortality and reoperation cases.@*Conclusion@#In the centers with abundant laparoscopic hepatectomy experiences, the laparoscopic resection is proved to be safe and feasible in the perioperative treatments of central liver tumors by the highly selective cases, the adequate preoperative assessment and reasonable surgical techniques and approach.

Role of oncogene and tumor suppressor gene in tumor metabolic reprogramming / 中国生化药物杂志

With the understanding of tumor metabolism, the process and mechanism of tumor metabolic reprogramming gradually attracted much attention in recent years.Oncogenes and tumor suppressor genes are constantly changing the pathway and flux of tumor metabolism in tumorigenesis to meet the needs of tumor growth and proliferation.The role of c-MYC, TP53, HIF-1αas well as the related signal pathways in tumor metabolic reprogramming would be discussed.

Research on the experimental design of medical molecular biology in the era of precision medicine / 中华医学教育探索杂志

Objective Through the design of comprehensive experiments, the precise medical philo-sophy was put into the medical molecular biology experimental teaching, and to explore its effect. Methods Eight-year medical students of Grade 2012 and Grade 2013 in the Fourth Military Medical University were chosen as the teaching subjects. Experimental group consisted of 36 students of Grade 2013, while control group consisted of 30 students of Grade 2012. Precision medicine-based learning was applied in experimental group while traditional learning method was adopted by the control group. At the end of the course, students of two groups were implemented theoretical and experimental skills assessment; through questionnaire students were required to evaluate the effect of teaching methods and the number of two groups of students who asked questions after class greater than or equal to 1 times was counted to evaluate the students' learning enthusiasm. SPSS 15.0 software was used to make t test and Chi-square analysis for the data of the students. Results The assessment results showed that the experimental group was better than control group, especially in the section of comprehensive experimental design [(16.7 ± 2.04) vs. (13.9 ± 2.87), P=0.000]. The results from questionnaire showed that the satisfaction degree of experimental group was also higher than that of control group in many respects, including learning interests, innovation capability, knowledge mastery, cognition of precision medicine and clinical research, satisfaction with the teaching method (P<0.05). And students' learning enthusiasm and the proportion of the number of students asking questions in the experimental group were higher than the control group (P=0.000). Conclusions Precision medicine-based learning not only changes the situation of slavish imitation and passive acceptance in traditional learning, but also arouses students' interest in study and helps students to cultivate clinical thinking, which is com-mensurate to the characteristics of precision-medicine era.

Multi-angle Explorative Analysis of NSFC Peer Review Evaluative Factors / 中华医学科研管理杂志

Objective To analyze the factors that affecting peer review evaluation for National Science Foundation of China (NSFC) award application.Methods Base on the feedback of NSFC peer review experts,total 10 evaluation factors were extracted for analysis.The percentage of common problem and in application,and percentage of key criteriaconcerned by reviewers,and their effect on funding situation were analyzed.Results 10 evaluatiion factors were divided into 4 groups:core factors including research content and innovation;factors including hypothesis,budget and writing,basic factors including the significance of research and previous study,general factors including experience and capacity of applicants,research team and laboratory condition.Conclusions Analysis and evaluate of peer review factors from different angle may help applicants and administrators to improve the quality of NSFC application and evaluation.

Modulation of MEK/ERK and PI3K/Akt pathways to the expression of DDR2 and MMP-13 in choroidal neovascularization / 中华实验眼科杂志

Background It is estimated that discoidin domain receptor 2 (DDR2) and matrix metalloproteinase-13 (MMP-13) play an important role in the development of tumor angiogenesis.However,their effects on choroidal neovascularizaiton (CNV) have not been clarified yet.Objective This study was to observe the expression pattern of DDR2 and MMP-13 in CNV and to further investigate the regulation role of MEK/ERK and PI3K/Akt pathways to the expression of DDR2 and MMP-13 in CNV.Methods CNV models were established in 78 Brown Norway (BN) rats by retinal photocoagulation with 532 nm laser.Then the animals were randomly divided into the normal control group (n =7),the model control group (n =39),PD98059 (MEK1 inhibitor) group (n =16) and LY294002 (PI3K inhibitor) group (n =16),and 5 mmol/L PD98059 or 5 mmol/L LY294002 3 μl was intravitreally injected 1 day and 7 days after photocoagulation in the PD98059 group or LY294002 group.The expression of DDR2 and MMP-13 mRNA and proteins in the CNV area were detected by using reverse transcription PCR (RT-PCR),and the expression levels of p-ERK/ERK and p-Akt/Akt protein were detected by Western blot assay.CNV thickness was measured by pathological examination 14 days after photocoagulation,and the changes of CNV thickness,the expression levels of DDR2 and MMP-13 in CNV were compared among the model control group,PD98059 group and LY294002 group.Results Three days after photocoagulation,the cells within the lasered lesions proliferated,then CNV formed 7 days after photocoagulation and became stable 14 days after photocoagulation.Immunohistochemistry staining indicated that DDR2 was weakly expressed in the cells of ganglion cell layer,inner nuclear layer and vascular endothelial cells;while MMP-13 was strongly expressed in the cells of the inner limiting membrane layer,photoreceptor layer and sclera layer.Both DDR2 and MMP-13 were strongly expressed in CNV area.Double immunofluorescence staining revealed that MMP-13 and DDR2 co-expression in CNV area.RT-PCR revealed that the relative DDR2 mRNA levels at 1 day,3 days,7 days and 14 days after photocoagulation were 55.22±4.03,47.74±2.23,14.82±4.56 and 5.59±2.47 respectively,while the relative MMP-13 mRNA levels were 25.54±3.83,43.51±4.36,10.90±4.00 and 5.23±3.23 respectively.Compared with the normal control group,the expression of DDR2 and MMP-13 were significantly increased (all at P＜0.05).Immunofluorescence staining showed that the relative fluorescence unit (RFU) values at 1 day,3 days,7 days and 14 days after photocoagulation were 2.73±0.53,5.21±0.31 and 3.22±0.33 for DDR2 and 1.66±0.17,3.57±0.44,2.67±0.21 for MMP-13,respectively.The RFU values in the PD98059 group and LY294002 group 14 days after photocoagulation were 1.14±0.19,1.03±0.14 for DDR2 and 1.37±0.25,1.24±0.20 for MMP-13,respectively.Compared with the model control group,the differences were statistically significant (both at P＜0.05).Western blot results showed that,compared to the normal control group,the expression levels of p-ERK and p-Akt pretein increased at day 7 after photocoagulation (both at P＜0.05),and returned back to the baseline at day 14 after photocoagulation (both at P＞0.05).Both PD98059 and LY294002 treatment were able to attenuate the thickness of CNV to 57.21％ and 50.34％ at day 14 after photocoagulation and further decrease the expression levels of DDR2 and MMP-13 in CNV (all at P＜0.05).Conclusions The expressions of DDR2 and MMP-13 up-regulate in laser-induced CNV.MEK/ERK and PI3K/Akt pathways suppress the development of CNV by regulating the expression of DDR2 and MMP-13.

Practice in experimental teaching reform of molecular biology course for postgraduates / 中华医学教育探索杂志

Molecular biology experiment is an important course for postgraduates in medical university.Through updating and optimizing what to teach(emphasizing study design,setting up comprehensive experiment and updating practical techniques),improving teaching mode(paying attention to experimental preparation,teacher qualification and postgraduates' capability training) and perfecting evaluation approaches(combining the experiment report with experimental summary and regular grades with experimental practice),good results have been obtained.

In vivo dynamical monitoring of rat bone marrow-derived cells participating in choroidal neovascularization under hyperglycemia / 中华实验眼科杂志

Background Our previous study demonstrated that hyperglycemia aggravate the choroidal neovascularization (CNV) by promoting the chemotaxis process of bone marrow-derived cells (BMCs).Bioluminescence imaging (BLI) can dynamically monitor CNV in vivo.However,how diabetes mellitus (DM)participate in CNV is still in research.Objective This study was to dynamically observe the influence of BMCs to CNV under hyperglycaemia by using BLI combined with histopathology.Methods BMCs from luciferase-green fluorescent protein (Fluc-GFP) double transgenic mice were injected to adult wild type C57BL/6J mice (nine mice per group) via caudal vein to create the chimera models with a chimerism degree higher than 85％,and the chimeric mice were randomized into the control group and DM group based on randomized number table.Streptozotocin [60 mg/(kg · d)] was intraperitoneally injected daily for 5 days to establish the DM models in the chimeric mice of the DM group.CNV was induced in the chimeric mice of both control group and DM group with 532 nm laser photocoagulation.BLI signal of BMCsFluc+GFP+ was in vivo examined by IVIS Kinetics system 1,3,5,7,14,21 and 28 days after CNV modeling.At the seventh day after laser,part of mice were sacrificed,and choroidal and retinal sections were prepared for histopathological examination.The length and thickness of CNV were compared between the control group and DM group.The use and care of experimental followed Statement of ARVO.Results The chimerism degree of the chimeric mice was (88.85 ± 2.46) ％ 28 days after BMCs transplantation,and the blood glucose concentration in the DM group was (17.88±0.86)mmol/L.Histopathological examination revealed that CNV broke through the Bruch membrane toward subretinas.The length of the CNV was (338.67±33.17) μm in the DM group and (180.33±24.68)μm in the control group,showing a significant difference between the two groups (t =8.943,P＜0.05).However,no significant difference was seen in the CNV thickness between the two groups (t =1.790,P＞0.05).Light signals appeared 1 day and reach strongest 7 days after CNV modeling in both groups.The Light signals were stronger in the DM group than those in the control group on 5,7,14 and 21 days after CNV modeling (t =3.411,5.594,5.067,2.663,all at P＜0.05).Conclusions Hyperglycemia can promote more BMCs to participate in the pathogenesis and aggravation of CNV.The behavior of BMCs in CNV can be evaluated using BLI in vivo.

To study and structure a preliminary indicators system for evaluating the projects of Hospital in finished phase / 中华医学科研管理杂志

Objective To structure a preliminary indicators system for evaluating the projects of Hospital in finished phase.Methods With Delphi method,collect the experts' opinion of the importance of every item.With AHP (Analytic hierarchy process) method,calculate the weight coefficient of every item.Results The positivity of the experts was fine; The authority coefficient of two rounds were 0.859 and 0.833,the consultation results are reliable; The coordination coefficient of two rounds were 0.254 and 0.553,according to the significance test,p values were less than 0.05,indicating that the results were desirable.According to the score of every item,we got the weight index of every items based on the AHP method.Finally we structure the indicators system including 3 primary indicators and 10 secondary indicators.Conclusion The preliminary results of this study provide a reference for the performance evaluation of projects in finished phase of China-Japan Friendship Hospital.

Evaluation of the written project approval indexes system of Capital Medical Development Research Foundation / 中华医学科研管理杂志

Objective The purpose of this study was to evaluate the written project approval Indexes System of Capital Medical Ddevelopment Research Foundation.We analyzed the approval results of application projects and proposed measures to improve the index system.Methods We evaluated its reliability with Cronbach's alpha coefficient,split-half reliability; validity was evaluated with contract validity and content validity,discrimination was studies according to discretion grouping law,and feasibility was studied with the percentage-of-completion method.Results ① Reliability:Cronbach's alpha coefficient of factor and total scores'and split-half reliability were all over 0.80.②Contract validity:the standard factor loading coefficients of the index system were over 0.70 (range:0.57～0.86),indicating path performed well.All factor loading coefficients were statistical significant (T-values were 30.49～54.25,all were over 1.96).Fit Indexes (RMSEA =0.065、NNFI =0.99、IFI =0.99、CFI =0.99、GFI =0.95、AGFI =0.93、X2 =1028.21、X2/DF =11.82) indicated the system's goodness of fit was good.③ Content validity:Scale-level CVI,S-CVI was 0.97 and Interrater agreement was 0.93,over 0.90,which indicated that indexes system's whole Content validity performed well.Except for the item Academic thought item-level CVI being 0.57 all items' item-level CVI were 1.00.④Discrimination:Discrimination index of every primary index,secondary index and total score ranged from 0.51 to 0.76 (all were over 0.40),meaning that discrimination performed extremely well.Conclusion The indexes system's reliability,contract validity,content validity and discrimination conformed to the actual application projects appraisal results.

Application of structural equation model on validity assessment of the evaluation indicator system / 中华医学科研管理杂志

ObjectiveTo explore the application of the Structure Equation Model on validity assessment of the evaluation indicator system,we studied Medical Capital Development Fund projects in 2007 to evaluate their construct validity.MethodsPart of the evaluation results (1200 items) were divided into two parts randomly.PART1 was used for Exploratory Factor Analysis (EFA) to explore the Indicator System' structure,and formed model F1.Confirmatory Factor Analysis (CFA) was used to assess the construct validity of model F1.With considerations of Modification Index(MI) and reality,F1 was modified into F3.At last,we take PART2 as the sample to confirm F3.ResultThe results of model F3 fit indices showed a better fit.All the path coefficients,estimated load factors and other parameters were statistically significant.ConclusionStructural Equation Model is professional,reliable and satisfactory for the evaluation of indicator system's contract validity.

Analysis of 206 ethic reviews in a hospital within 3 years / 中华医学科研管理杂志

Objective To reveal the average pass rate of a general hospital's ethic review and common problems.Methods The records generated by the ethics committee of a hospital in 3 consecutive years were reviewed and analysed with ridit of CMH and Bootstrap statistical methods.Results The pass rate for drug clinical trial is 36.09%,medical instruments 18.42%,and subject evaluation 14.29%.Among the problems,those concerning informed consent take up 63.59%,of protocol 14.56%,of CRF 7.77% and of the others 22.820%.The domestic applicants had more problems than those abroad.Besides,the pass rate of drug trials decreased with year from 45.83% in 2006 to 19.44% in 2008,which was statistically significant (P=0.029).In terms of drug clinical trial,the pass rate of revised projects in foreign countries was significantly higher than the domestic ones (P=0.028).ConclusionEthics Committee in a hospital plays a very important role in clinical researches.And informed consent and research protocol are very important in the ethic review.

Construction and significance of directional expression cDNA library from human NB4 cells / 华中科技大学学报（医学）（英德文版）

Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.

Construction and significance of directional expression cDNA library from human NB4 cells / 华中科技大学学报（医学）（英德文版）

Human acute premyeloid leukemia cell cDNA expression library was constructed to screen acute premyeloid leukemia tumor antigen. Total RNA and purified mRNA were extracted from human premyeloid cell line NB4. First and second strands of cDNA were synthesized by reverse transcription. After blunting, the cDNA fragments were ligated with EcoR I adapters. Then the cDNAs were digested with Xho I, and less than 400 bp cDNA fragment was removed by Sephacryl-S400 spin column, the remaining were ligated with lambdaZAP vector. The recombinants were packaged in vitro, and a small portion of packaged phage was used to infect E. coli XL1-Blue-MRF' for titration. The recombinants were examined by color selection. In order to evaluate the size of cDNA inserts and the diversity of library, the pBK-CMV phagemid was excised from the ZAP express vector by using ExAssist helper phage with XLOLR strain, and then the pBK-CMV phagemid was digested by Xho I and EcoR I. The results showed that the NB4 cell line cDNA library consisting of 1.65 x 10(6) recombinant bacteriophages was constructed with the recombinant ratio of 99.6%. The average length of the recombinant exogenous inserts was about 1.7 kb. It was concluded that the constructed cDNA library are deserved to screen target clones.

Effects of glucose,insulin and uric acid on the expression of hemoglobin oxygenase in pig renal tubule cell line / 西安交通大学学报(医学版)

Objective To investigate the effects of metabolically correlated factors on pig renal tubule in cellular level by observing effects of glucose,insulin and uric acid on activity of haemachrome oxygenase in renal tubular epithelial cells.Methods Pig near renal tubular cell line(LLC-PK1) was selected as experimental object.After stimulating cells respectively with glucose(5,10,22,33mmol/L),uric acid(0,0.1,0.2,0.4mmol/L);and insulin(0,10~(-9),0~(-8),10~(-7)mol/L) in different concentrations for 48 hours,the activity of HO-1 within the cells was determined.Results In uric acid group the activity of HO-1 was significantly increased in a concentration-dependent manner,while in glucose and insulin groups,the activity of HO-1 did not change.Conclusion After LLC-PK1 was stimulated by uric acid of different concentrations,HO-1 was induced evidently,which indicates that uric acid might affect oxidative stress in renal tubule cell line.

Construction and space conformation analysis of the fusion gene of human IgG3 upper hinge region and p53 tetramerization domain / 细胞与分子免疫学杂志

Aim To construct the fusion gene of human IgG3 upper hinge region and p53 tetramerization domain and to analyze its space conformation. Methods The fusion gene was obtained by recursive polymerase chain reaction (R-PCR),and was cloned into vector pUC19. The positive clone was selected and sequenced with PE310 auto-sequencer. The space conformation of the fusion gene expression product was predicted by using computer program Antheprot. Results Restriction endonuclease digestion confirmed that the fusion gene has been inserted correctly into the vector. The result of sequencing showed that the fusion gene is identical with designation. Analyzing with antheprot program showed that the fusion gene expression product could auto-assembly into a tetramer with four long and flexible linkers. Conclusion Successful construction of the fusion gene mentioned above laid the foundation for further preparation of multivalent gene engineering antibody.

Expression and characterization of fusion pro-tein hTNFα -hPgnK5 containing thrombin cleav-age site in E. coli / 细胞与分子免疫学杂志

Aim To express the fusion protein hTNFα -hPgnK5 in E.Coli and to identify its activity. Methods The hTNFα -hPgnK5 gene express vector was constructed and the fusion protein was expressed in E.coli. The activity of fusion protein to inhibit the proliferation of tumor cells or angioendothelial cells in vitro was determined by MTT. Results Fusion protein hTNFα -hPgnK5 possessed immunoactivity in Western blot analysis and could inhibit proliferation of tumor cells or angioendothelial cells in vitro test. Conclusion The hTNFα gene and hPgnk5 gene can be coexpressed in prokaryotic system and expression products can inhibit the proliferation of tumor cells or angioendothelial cells in vitro.

Exploring a New Gene Containing ACP Like Domain in HumanBrain and Expression It in E. coli / 生物化学与生物物理进展

To look for new genes from human brain, get a fragment was obtained using adaptor primer and 3' anchor polymerase chain reaction (PCR) with the human adult whole brain cDNA as template. The fragment was cloned into T easy vector and automatically sequenced with 310 Genetic Analyzer. Later the whole length cDNA of this novel gene was got with the method of 3'rapid amplification of cDNA end (RACE). The whole length of cDNA of this novel gene is 2 024 bp. Chromsome location is at 14q11.2 including 16 extrons and 15 introns. After scanning the sequence against GenBank it is proved that the sequence is a new one. ORF analysis showed that there is a complete coding region in it,it can interprate a protein containing 357 amino acid residules. ProDom analysis result showed that there is an acyl carrier protein (ACP) like domain in it. The gene was banked into GenBank. Then, a pare of primers were designed and were used to amplify the coding region and cloned into pGEX-4T1 expressing vector to express it in E. coli . The Dot blotting and Northern blot showed that this novel gene is highly expressed in the normal adult human brain.