Objective To insert VEGF-R3(FIT-4)gene fragment into cloning vector and identify it.Methods By T-A cloning,the VEGF-R3 gene fragment amplified by RT-PCR was cloned on the PGEM-T Easy plasmid vector and transfected into E.coli JM109.the recombinant clones were screened by"white-blue plaque selection" and tested by the methods of single restrictional enzyme.Results The VEGF-R3 gene fragment amplified by RT-PCR showed a smear after the electrophoresis on agarose Gel and obtained white clones by"white-blue plaque selection".The electrophoresis result of the recombinant clones tested by the single restrictional enzyme was accordant with expectant result.Conclusion The recombinant clones through the methods T-A may be used in the further gene expression and gene therapy studies.

The morphological and pathophysiological manifestations of theheart after abdominal gun-shot wound created by a model-53 smooth bore gun of 6.35 mm in caliber with 1.03 g steel ball were observed in 15 pigs,which were randomized into 3 groups.Five animals of the first group were killed immediately after wounding and all the animals showed small spotty flat hemorrhages on the endocardium of the left ventricle under gross inspection.Focal or small flat hemorrhages on the myocardium and under the endocardium were found under optical microscopy.Under electron microscopy,myofibrilla gaps were widened and filled with fluid,and mitochondrial swelling or vacuolar degeneration in the myocardium could be seen.7 animals of the second group were infused and monitored until they were killed 72 hours after wounding.It was found that there were varying degrees of functional disorder of the left ventricle in every animal.The serum level of creatine kinase isozyme MB increased significantly.The 3 pigs of the third group were killed with large andrapid blood-shedding without gun-shot wound and showed no similar pathological changes.

Objective To systematically summarize and compare results and related advantages of different methods in the management of adult mid-shaft clavicle fractures. Methods The data of adult midshaft clavicle fracture from 1989 to 2009 was retrieved. The database were included Cochrane Database,med & Ovid Medline, OTA, AAOS, High Wire Press, Springer Link, CBM, CNKI, WANFANG DATA and with hand-search journals in related fields. Internalizing randomized controlled trials and retrospective cohort studies which compared the operative and nonoperative treatment for adult mid-shaft clavicle fracture. Evaluating the quality of included studies and using the Rvaman 4.2 for Meta-Analysis. Results 5053 literatures had been reviewed. After screening, there were 8 studies brought into the review that included 519 patients.There were 5 studies which took union rate as observed indicator. The results had statistically significant (P＜0.00001). A total of 4 studies used malunion rate as indicator, the results had statistically significant (P ＜0.0001). A total of 4 researches selected satisfaction of patient as indicator, the results had statistically significant(P＜0.00001). A total of 5 studies selected symptoms of nerve stimulation as indicator, the results had statistically significant (P＜0.0001). A total of 4 studies selected residual pain as indicator, the results had no significant statistically difference (P=0.56). A total of 2 studies selected barriers of joint activities as indicator, the results had no significant statistically difference (P=0.25). A total of 2 researches used refracture as indicator, the results had no significant statistically difference (P＞O.05). Conclusion This study evaluation results shows that operation can be used as the first choice of treatment of adult midshaft clavicle fractures.

Objective To improve the diagnosis and treatment of Q fever endocarditis.Methods From 2008 to 2013,four cases of Q fever endocarditis were diagnosed in Peking Union Medical College Hospital.Clinical features,laboratory test,management and prognosis were analyzed with literature review.Result All four cases had long period of fever and heart murmur.Two patients represented with respiratory symptom and one with non-specific rash.General laboratory tests including complete blood cell count,ESR,C-reactive protein(CRP),liver function and radiology of lung did not show specific abnormalities.Signs of endocarditis were shown by ultrasound and important for diagnosis.Repeated blood culture was negative.All of the diagnoses were confirmed by serum antibody detection and the patients recovered well with treatment based on doxycycline or minocycline.Conclusions Endocarditis is the most common form of chronic Q fever,which is easily misdiagnosed because its blood culture is negative and may accompanied with varied manifestation such as pneumonia and liver injury.For the patients with chronic fever and blood culture negative endocarditis,chronic Q fever should be considered as differential diagnosis.The confirmatory method for diagnosis is serum antibody detection.Early and sufficient treatment may improve the prognosis.

Objective To study the feasibility and safety of coronary sinus (CS) ventricular pacing comparing with traditional right ventricular (RV) pacing in ordinary pacing treatment.Methods Sixty-one patients with an indication of pacemaker implantation were randomized into traditional RV pacing group and CS ventricular pacing group.The success rate,complications,preoperative and postoperative QRS wave duration and pacemaker parameters were compared between the two groups.Results (1) The instant success rate in traditional RV pacing and the CS pacing groups were 100％ and 68.97％ (P＜0.01);the surgical success rate was significantly higher by using left ventricular electrode than the ventricle tined electrode in CS pacing group (85.71％ and 25％,P＜0.05).There was no significant difference in the incidence of complications between the two groups (P＞0.05);(2) The increase of QRS complex width at postoperative 1 month in CS pacing group is significantly less than the RV pacing group (P＜0.05);(3) Although the pacing threshold and electrode impedance during operation,postoperative 1 month and 3 months in CS pacing group were higher than RV pacing group (P＜0.05),the CS pacing was effective.Conclusion The CS ventricular pacing is as safe and effective as traditional RV pacing,and is more consistent with physiological ventricular activation sequence.Using left ventricular electrode can significantly improve the success rate of CS ventricular pacing.

Objective To investigate the results and complications in the treatment of the mechanically unstable fractures of the distal femur when Less Invasive Stabilization System (LISS) is used.Methods From September 2011 to July 2014,81 patients with mechanically unstable fractures of the distal femur were treated with the LISS,according to the inclusion criteria and exclusion criteria,59 patients meet the condition including 31 male and 28 female.The mean age of the patients was 49.8 years (range 18-80 years).The fractures occurred on the left side in 40 cases and on the right side in 19 cases.According to AO classification,27 type 33A2,14 type 33A3,13 type 33C2,5 type 33C3.2 cases combined with ipsilateral fractures of the femoral shaft,according to AO classification,1 type 32A1,1 type 32C1.The mechanism of injury was a fall from the height in 8 cases,a traffic accident in 18 cases,a crush injury in 7 cases,a fall injury in 26 cases.55 fractures were closed,and 4 were open.According to the Gustilo-Anderson classification,there were 3 type Ⅰ,1 type Ⅱ.Results The 7-hole plate were used in 21 patients,9-hole plate in 26 patients and 13-hole plate in 12 cases.The 3.5 mm or 6.5 mm lag screw were placed around the LISS plate to stabilized the articular fracture fragment in 11 cases.The mean operation time was 105.2 min (85-145 min),the mean bleeding volume was 203 ml (130-315 ml).All patients were followed up 11-27 months (average 13.2 months).1 delayed union(13 months),the average time to union was 16 weeks (range 12-21 weeks).The average flexion of the knee was 116 degree,0 degree in extension.The functional outcome:32 had an excellent result,19 had a good result and 8 had a fair result,with 86.4％ excellent and good results.No deep infections occurred.No loss of reduction.3 cases had malalignment,2 failed fixation,4 patients with symptomatic hardware irritation.27 cases underwent implants removal after union,cold-welding occurred in 4 cases (9 screws) which resulted in difficult removal.Conclusion LISS is one of the reliable and effective methods in fixation of mechanically unstable fractures of the distal femur.However,its operation indications and operating instructions should be strictly followed.

Objectives: To study and prepare monoclonal antibody against glycophorin A of human erythrocyte(GPA McAb).This antibody is a key reagent in preparation of bispecific antibodies for rapid whole-blood immunoassay.　Methods: BALB/c mice received GPA antigen injection. Hybridoma was produced by traditional techniques. Hybridoma was determined with ELISA and indirect agglutination(IA) method.　Results: All 3 GPA McAb reacted with GPA, and they did not autoagglutenate with four types of red cells(type A,B,O,AB).Of the 3 McAb-GPA, two antibodies were IgG1,one IgG2 subtypes. Two IgG1 McAb-GPA can be used in making bispecific antibody in preparation of rapid whole-blood immunoassay.　Conclusions: Immunogenity of GPA was enhanced after coupling with the native serum albumin of the immunized animal, and high titer GPA McAb could be easily obtained. This result is important in making bispecific antibodies in preparation of rapid whole-blood immunoassay.

[Abstract ] Objective Cardiac shock wave therapy (CSWT) can promote arteriogenesis in ischemic myocardia , but the mo-lecular mechanism remains unclear .The study aimed to explore the effect of CSWT on arteriogenesis in human cardiac microvascular endothelial cells ( HCMEC ) and the role of focal adhesion kinase (FAK) and Calcium-activated potassium channels (KCa) in the sig-nal conduction pathway of CSWT arteriogenesis . Methods HC-MEC cells cultured in vitro were randomly divided into control group , CSWT group , CSWT +T ( FAK inhibitor PF-573228 ) group and CSWT+F( SCa inhibitor iberiotoxin ) group.Each group received one CSWT(0.09 mJ/mm2, 200Times) 48 h after added stimulant.24 hours'conventional culture later , tests were made on the levels of endothelial nitric oxide synthase ( eNOS ) and vascular endothelial growth factor ( VEGF) mRNA as well as the changes of related protein expression . Results ①QPCR test showed that eNOS , VEGF mRNA expressions increased in CSWT group compared with control group (4.61 ±0.19 vs 3.99 ±0.17, P<0.05), while compared with CSWT group, eNOS, VEGF mRNA expressions in CSWT +T group were decreased (0.62 ±0.10 vs 0.40 ±0.02, P<0.05), eNOS, VEGF mRNA expressions in CSWT +F group were also decreased (0.53 ±0.02 vs 0.64 ±0.02, P<0.05), all the differ-ences were of statistical significance .②Western blot showed that eNOS , VEGF protein expressions increased in CSWT group compared with control group(0.63 ±0.02 vs 0.43 ±0.02, P<0.05), while compared with CSWT group , eNOS, VEGF protein expressions in CSWT+T group were decreased (0.36 ±0.01 vs 0.29 ±0.02, P<0.05), eNOS, VEGF protein expressions in CSWT +F group were also decreased (0.37 ±0.02 vs 0.30 ±0.02, P<0.05), all the differences were of statistical significance . Conclusion CSWT can improve eNOS , VEGF mRNA and protein expressions in HCMEC cells and FAK and KCa may participate in the signal conduction pathway of CSWT arteriogenesis .

Objective Big endothelin-1(big ET-1) is associated with the prognosis of various cardiovascular diseases. This study is to explore the predictive value of plasma big ET-1 level for long-term outcome after mitral valvuloplasty in patients with moderate to severe chronic ischemic mitral insufficiency .Methods A total of 142 consecutive patients with moderate to severe chronic ischemic mitral regurgitation who underwent coronary artery bypass grafting and mitral valvuloplasty were enrolled at our center from January 2009 to December 2015.The clinical baseline data were collected and recorded.All patients were followed up.The mortalities and major adverse cardiovascular and cerebrovascular events during the follow-up period were re-corded.Results A total of 142 patients were enrolled, with mean age of(59.4 ±8.3) years.During(51.9 ±22.6) months follow-up, 19 cases(13.4％) died and 35 cases(24.6％) had major adverse cardiovascular and cerebrovascular events.Uni-variate and multivariate Cox regression analysis showed that big ET-1 was an independent risk factor for all-cause mortality (HR=3.970, 95％CI:1.535-10.268) and major adverse cardiovascular and cerebrovascular events(HR=2.754, 95％CI:1.238-6.129).In addition, left ventricular ejection fraction, pulmonary hypertension and stroke history were independent risk factors for all-cause mortality , while left ventricular ejection fraction , stroke history were independent risk factors for major adverse cardiovascular and cerebrovascular events.Conclusion Long-term outcomes of simultaneous coronary artery bypass grafting and mitral valvuloplasty for moderate to severe ischemic mitral insufficiency are satisfactory and big ET-1 is an inde-pendent risk factor for long-term outcomes of these patients.

Objective To assess the clinical value of cross raft screws technique in preventing postoperative collapse of tibial plateau fracture.Methods From September 2014 to November 2017,data of 14 patients with tibial plateau fracture who were treated by cross raft screws technique were retrospectively analyzed.There were 9 males and 5 females aged from 30-65 years old (average,44.4± 1.7 years).There were 8 patients that the thickness of subchondral cancellous bone measured preoperatively by CT data was less than 4 mm,and 6 patients that the thickness of subchondral cancellous bone measured intraoperatively was less than 4 mm.1/4 tubular plate was placed along the anterior rim of lateral tibial plateau,and the 3.5 mm cortical bone screws were fixed as bamboo raft from anterior to posterior through the plate.The collapse of tibial plateau after surgery were measured by CT scan after union of the fracture.The function of knee was evaluated by Rasmussen Anatomical and Functional Grading.Results The time of tubular plate procedure was 18 to 35 min (average,24.1±5.4 min).All 14 patients were successfully followed-up for 13.8±5.1 months.The height of collapse preoperatively by CT scan was 5-21 mm (average,8.00± 1.40 mm).3 days after the operation,the height between articular line and lateral articular surface was 0-2 mm (average,0.80±0.06 mm).Compared with CT data preoperatively,the collapse was corrected postoperatively that was proved by CT scan (P＜ 0.05).After the fracture was healed,according to CT data,the height between articular line and lateral articular surface was 0-2 mm (average,0.70±0.08 mm).Compared with CT data postoperatively,there was no postoperative collapse happened (P=0.466).The position and length of nails were placed appropriately.The average healing time of fracture was 3.6 months.There were no infection,nonunion and pain of tendon happened.The plate could be touched subcutaneously in 2 patients,who had no discomfort feelings.The patient's postoperative Rasmussen Anatomical Grading were 13-18 (average,16.7),including 8 cases excellent and 6 cases good.The postoperative Rasmussen Functional Grading was 18-28 (average,25.7),including 11 cases excellent and 3 cases good.Conclusion The cross raft screws technique is a good way to prevent the postoperative collapse of the lateral articular surface of tibial plateau.