Objective To seek a stable,rapid and effective method in order to obtain large number of pure adult Schwann cells (Scs) in vitro. Methods Bilateral sciatic nerves of each adult SD rat were crushed several times to establish the degeneration animal model.One week later.the degenerated nerves were detected and prepared to collagenase/dispase double enzyme digestion method,trypsin/collagenase double enzyme digestion method,single enzyme digestion for half-piece method,respectively.The S-100 immunocytochemistry was used to detect the purity of Scs. Results The confluence of cells was happened within 3-5 days in collagenase/dispase double enzyme digestion method.At time of confluence.about 70%-80%cells showed S-100 positive in this method.The quantities of cells from the degenerative nerve pieces cuhured hy single enzyme digestion were not uniform within 3-5 days.cells were confluent within 10-12 d anti less than 40%cells showed S-100 positive at time of confluence.Most of tissue pieces were tangled and cells were lost in trypsin/collagenase double enzyme digestion method.Only 3-5 cells in every 100 fiehts couht smwive in vitro,confluence is diffieult.Conclusion The collagenase/dispase double enzvme digestion method was the most stable and effective method to obtain large number of pure adult Schwann cells in vitro rapidly among the present three methods.

Objective To investigated the biological procedure of allograft decalcified bone matrix(DBM)and bone cement(BC)combined with bovine bone morphogenetic protein (bBMP)used for the repair of femoral defect caused by microwave－ induced hyperthermia in dogsby 99mTc－ MDP bone scintigraphy.Method The canine femoral defect(length 25mm,width 10mm)was caused by microwave－ induced hyperthermia(50℃ ,20minutes)and the composite material was implanted .Then the canine femurs were examined by 99mTc－ MDP bone scintigraphy respectively at different postoperative time and the results were compared with that of X－ ray photography and histological observation.Bone cement was implanted in the other femur as a contrast.Results It could be observed at the first and the second month that the radioisotope was gathered in the place where the composite material was implanted and the amount of radioisotope gathered in was the most abundant at the third month and it was lasted to the fourth month. That of the sixth month was decreased to that of the second month.The radiation count of the first, the second, the third the fourth and the sixth month were 93.9± 12.7, 110.7± 16.4,222.1± 24.0,201.3± 26.9 and 111.6± 20.7 respectively,and the count of the third month and the fourth month were more than that of the first, the second and the sixth month(P<0.01).Conclusion The composite material could be remodeled easily and the new bone could be formed by the induction of bBMP. So it could be merged with the normal bone.While the 99mTc－ MDP bone scintigraphy is the object and reliable index to determine the biological procedure of the composite material in dogs.

ObjectiveTo observe the clinical effect of interventional embolization on hepatocellular carcinoma(HCC) with hepatic arterioportal shunts(APS) ,in the hope of improving patients' survival quality and time. MethodsTwenty-nine patients with HCC patients and APS after a successful PVA,steel coils embolization, all patients were given routine TACE therapy. The changes of gastrointestinal bleeding,ascites,diarrhea and aminotransferase were analyzed retrospectively. ResultsNineteen cases got successful embolization in the first time[achievement ratio 65.5％(19/29)]; 3 cases got recurrence after embolization [patency ratio 10.3％(3/29)],7 cases appeared new APS[incidence ratio 24.1％(7/29),5 cases were embolized 2 times, 2 cases were embolized 3 times]. The effective rate of gastrointestinal bleeding,diarrhoea,aacites were 91.7％ (11/12), 84.6％ ( 11/13 ), 83.3％ ( 15/18 ) respectively, which had significant difference between preoperative and postoperative condition. The survival rate of 3 months,6 months, 1 year,and 2 years after operation were 96.6％ (28/29),89.7％ (26/29),65.5％ (19/29),34.5％ (10/29). Conclusion Interventional therapy is a safe and effective treatment to HCC with APS.

Objective To investigate the clinical and pathological features of eosinophilic diseases with cardiac involvement Methods We analyzed the clinical and cardiac pathological data of 7 patients with eosinophilic diseases with cardiac involvement under endomyocardial biopsy or autopsy.Results Seven patients (5 male, average age 51 years) were enrolled.Four patients were diagnosed as idiopathic hypereosinophilic syndrome and three were Churg-Strauss syndrome.Peripheral blood eosinophila count increased significantly in all patients.Cardiac involvement included angina pectoris, myocardial infarction, heart failure, presyncope and sudden death.Electrocardiogram showed cardiac ischemia, bundle branch block and third degree atrioventricular block.Echocardiography suggested ventricular and atrial enlargement, decreased ventricular systolic function, pulmonary hypertension, valvular prolapse and insufficiency and endocarditis.Pathology displayed infiltration of eosinophils, formation of granulomata, necrotizing vasculitis, myocardial necrosis and endomyocardial fibrosis in heart.Coronary artery could be also affected and led to myocardial infarction.Conclusions Cardiovascular complications of eocinophilic diseases are a major source of morbidity and mortality in these disorders.The manifestations are multiple and early recognition and treatment with steroid and immunosuppressant can improve prognosis.

Objective To prepare the 99Tcm-labeled human epidermal growth factor receptor type 2 (HER2) affibody molecule ZHER2:342 and evaluate its receptor binding specificity in vitro.Methods The molecular ZHERa:342 was labeled with 99Tcm using the ligand exchange method.The labeling efficiency and radiochemical purity were measured by HPLC.The major factors,such as the mass of SnC12 and NaOH and reaction time were analyzed,and the optimal method was summarized.Cell binding kinetics and cellular retention of the probe were investigated in HER2-expressing SKOV-3 cells and MDA-MB-231 cells with low HER2 expression respectively.HER2 binding specificity of 99Tcm-ZHER2:342 was analyzed by a pre-injection of excess unlabeled ZHER2:342 to saturate HER2 receptors.One-way analysis of variance and two-sample t test were used.Results The optimal labeling procedure was as follows:5 μg (1 g/L) of ZHER2:342 was mixed with 5 μg of NaOH (1 g/L),then 8.8 μg SnC12(1 g/L,solution) was added,followed by 150 μl (37 MBq) 99TcmO4-solution,and finally the mixture was slightly vortexed and incubated for 1 h at room temperature.99TcmZHER2:342 was stable in vitro with a high labeling efficiency of (98.10± 1.73)％.The radiochemical purity was ＞ 98％,and was more than 85％ after the incubation for 24 h in saline and fresh human serum.The cell binding of 99Tcm-ZHER2:342 with HER2-expressing SKOV-3 cells gradually increased over time with a peak of (9.95± 1.02)％ at 6 h.The binding of 99Tcm-ZHER2:342 in SKOV-3 cells was significantly higher than that in MDA-MB-231 cells at every time point (5.68-9.88 vs 0.56-2.11 ; t:from-34.50 to-13.14,all P＜0.01).The labeled molecular probe retained the capacity to bind specifically to HER2-expressing SKOV-3 cells since the cell binding decreased from (9.95 ± 1.02) ％ to (2.11 ±0.27) ％ after receptor saturation (t =-13.14,P＜0.01).Conclusions 99Tcm-ZHER2:342 has a high labeling efficiency,good stability and optimal binding specificity.These characteristics enable it to be a promising molecular probe for HER2-targeting imaging.

Objective To observe the curative effect of catheter thrombolysis in the treatment of subacute lower limb arterial occlusion.Methods Eighty-five patients with subacute lower limb arterial occlusion (98 limbs) and treated with catheter thrombolysis were enrolled and the removal of thrombus and patency of vessels were evaluated by angiography.Then,balloon dilatation or stent implantation was performed according to the angiographic findings after thrombolysis. Results After catheter thrombolysis, the degree and length of occlusion were significantly improved (P < 0.01). To judge the curative effect: significantly effective in 5 limbs (5.1%), effective in 85 limbs (86.7%), ineffective in 8 limbs(8.2%),total effective rate was 91.8%.Catheter thrombolysis combined with balloon dilatation and stent implantation were performed according the vascular occlusion status and all the occluded segments were open, the postoperative angiography showed no distal vascular occlusion. No bleeding occurred. Conclusions Catheter thrombolysis is effective and safe in the treatment of lower extremity arterial occlusion in subacute stage.Endovascular treatment combined with endovascular treatment can achieve good results.

Objective To retrospectively analyze clinical features,treatment methods and efficacy of giant cell tumor of bone in proximal tibia,and to investigate risk factors affecting tumor recurrence and functional outcomes.Methods A total of 250 patients with giant cell tumor of bone in proximal tibia confirmed by pathology,who had undergone surgical treatment from March 2000 to July 2014,were enrolled in this study.There were 132 males and 118 females,with an average age of (34.59±12.86) years.A total of 140 patients who were followed up for more than 3 years were included in this study,and there were 72 males and 68 females,with an average age of (34.46± 11.96) years.There were 11 cases of Campanacci grade Ⅰ,58 cases of grade Ⅱ,71 cases of grade Ⅲ and pathological fracture of 47 cases.According to surgical methods,they were divided into bone grafting group (49 cases),bone cement filling group (34 cases),prosthesis group (46 cases) and others group (11 cases).The epidemiology,clinical and radiographic features and risk factors affecting tumor recurrence and functional outcomes were analyzed.Results A total of 140 patients were followed up,the follow-up period was 36-324 months,with an average of 95.4 months,and the median follow-up time was 88 months.Recurrence was found in 26 cases,and recurrence rate was 18.57％,with an average recurrence interval of 25.85 months.Recurrence was found in 17 cases in the first 2 years.The 5-year free survival rate was 77.60％.The recurrence rates were 18.37％ in bone grafting group,20.59％ in bone cement filling group,15.22％ in prosthesis group and 27.27％ in the others group,no statistically difference was found on recurrence rate and free survival rate (P=0.805,P=0.558).Recurrence was not related to all kinds of factors.A variety of related factors affecting postoperative recurrence were analyzed,sex,the first diagnosis of the original recurrence,left and right side,whether the eccentricity,fracture,cortical bone destruction,soft tissue mass,surgical methods,high-speed grinding,auxiliary application,and there was no significant correlation between recurrence and these factors.The MSTS 93 score was 25.26±4.31.Function of the primary patients was better than that of recurrence (P=0.044).Function of the patients treated with curettage with or without internal fixation was better than that with segmental resection (P=0.011).Function of the patients treated with grafting or bone cement filling was better than that with prosthesis or allograft-prosthesis reconstruction (P=0.004).There were no significant correlation between MSTS function score and gender,left and right side,whether the eccentricity,whether fractures,cortical bone destruction (Campanacci grade),whether there is soft tissue mass,whether the use of assisted inactivation,whether the use of grinding or internal fixation.Conclusion Various surgical methods had no significant effect on the recurrence of proximal tibial GCT,as for GCT in proximal tibia,there is no relation between recucrrence and related factors.Whether primary tumor and surgical methods are two important factors affecting limb function.

Objective To investigate the clinical and epidemiological characteristics of osteosarcoma in China and analyze the incidence and clinical treatment of osteosarcoma.Methods The clinical data of 1 593 patients with osteosarcoma in 7 bone cancer treatment centers from January 2000 to February 2017 were analyzed.We retrieved large samples of documents in the database,extracted the relevant data and compared the data with this study.SPSS 13 software was used for statistical analysis and each factor was tested by x2.Results Of 1 593 patients,984 were males and 609 were females.The ratio of male to female was 1.62∶ 1.The average age was 23.2 years (range from 3-80 years).The peak age of onset was 11-20 years (52.4％).There were 217 elderly patients (＞ 40 years old).The tumors occurred in 1 524 limbs and 69 in axial bone.The most common sites of disease were distal femur 706 cases,proximal tibia 375 cases,distal humerus 117 cases and others 395 cases.There were 1 154 cases (71％) around the knee joint.The axial bone included 18 cases of spine,49 cases of pelvis and 2 cases of rib.Preoperative biopsy was performed in 1 111 cases and incisional biopsy in 280 cases.Preoperative diagnosis could be made in 1 345 cases (84.4％ of all patients,accounting for 96.7％ of biopsy patients).There were 79 cases with metastasis at first visit,accounting for 5％.Preoperative chemotherapy was performed in 1 185 cases (74.4％).With the DIA preoperative chemotherapy 271 cases,DIA+MTX 251 cases,AP 149 cases.220 cases of tumor cell necrosis rate was evaluated after operationaccording to the Huvos classification.There were78 cases of grade Ⅰ,105 cases of Ⅱ grade,35 cases of Ⅲ and 2 cases of grade Ⅳ.There were 1 299 cases undergoingpostoperative chemotherapy (81.5％),1 306 patients undergoinglimb salvage surgery (82％).Thespecific operation with prosthetic replacement is the most common (911 cases,69.8％).The postoperative chemotherapy included DIA+MTX regimen 471 cases,DIA regimen 266 cases and AP regimen 98 cases.Before and after the operation,379 cases were treated with the same chemotherapy regimen and 666 cases were changed.A total of 18 large sample documents were retrieved in Chinese data base (5 684 cases).The sex ratio,age range,peak incidence and location of the disease were similar to those of this study.The average age was 1-2 years old younger.The percentage of lung metastases associated with initial visits washigher.The rate of preoperative and postoperative chemotherapy and limb salvage waslowand the rate of tumor necrosis wasbetter after chemotherapy.A total of 22 large sample literatures were searched in foreign database (12 850 cases).The ratio of men and women is 1.30∶1 and the proportion of women is higher than the domestic data.The average age was 1-2 years older.The ratio of the knee joint was lower.The percentage of lung metastases associated with initial visits washigher.The rate of preoperative and postoperative chemotherapy and limb salvage were similar to that of this study and the rate of tumor necrosis was better after chemotherapy.Conclusion The general situation of the incidence of osteosarcoma (sex,age,location of the disease) is not significantly different from the previous reports both at home and abroad.In the treatment,preoperative and postoperative chemotherapy rate and limb salvage surgery rate have improved significantly compared with the past.The chemotherapy program from the coexistence of various programs,gradually to the DIA+MTX and DIA program as the mainstream program.

Objective:To explore the method and curative effect evaluation of repairing and rebuilding the long segmental bone defect after resection of long bone malignant tumors in adolescents.Methods:Reviewed our experience in 10 patients. Among them, 4 were male and 6 were female. The age was 10.2 years old (range, 6-16 years old). Including 7 cases of osteosarcoma, 3 cases of Ewing's sarcoma; 2 cases of humerus tumors, 7 cases of femoral tumors, and 1 case of tibial tumors-who presented between April 2015 and October 2019, in which the proportion of bone defect account for 71.4%-89.3% of the total length of diseased bone after tumor resection, with an average value of 80.9%. Considering the epiphyseal growth of young people, on the premise of ensuring enlarged tumor resection, we use traditional fibula autograft and prosthesis combined with steel plate, 3D printing design to retain the super-prosthesis of the epiphysis, and 3D printing bone trabecular was processed for the sake of making it grow successfully into the binding part of prosthesis and bone as we expect, simultaneously, the design of the short-force arm retaining the bilateral epiphysis can assit the steel plate and interlocking screws to enhance the stability.Results:The operation was completed successfully, and there were no complications such as nerve and blood vessel injury during the operation. After follow-up of 4-8 months, with an average follow-up of 22.9 months. One patient's review of X-ray film after 2 years of the operation showed that the transplanted fibula had a sign of bone resorption, and the internal fixation was in it's original position without obvious abnormalities. Two cases' auxiliary steel plates had fractured within one and a half to two years after surgery. With replacing the fractured steel plates, the internal fixation was good until now. According to the evaluation standards of the International Society of Limb Salvation: 2 cases of humerus osteosarcoma were respectively scored 23 points, 26 points, an average of 24.5 points; 4 cases of femoral osteosarcoma and 3 cases of femoral Ewing's sarcoma respectively scored 26 points and 25 points, 19 points, 30 points, 27 points, 24 points, 28 points (average, 25.6 points); 1 case of tibial osteosarcoma scored 26 points; all obtained relative satisfying treatment results.Conclusion:The combination of 3D printing and traditional machining techniques to produce an individualized long segmental prosthesis that preserves the epiphysis which applicated in the limb salvage treatment of bone malignant tumors can achieve excellent therapeutic effect.

Objective:To investigate the prognostic value of molecular subtypes in patients with resected invasive breast cancer.Methods:Between 2015 and 2018 7 869 patients with invasive breast cancer after undergoing surgery were included in this analysis. Breast cancer was classified into four subtypes according to the status of hormone receptor (HR) and HER2: HR+/HER2-, HR+/HER2+, HR-/HER2+, and HR-/HER2-. Kaplan-Meier curves and COX regression were used to compare disease-free survival (DFS) and overall survival (OS) among different subtypes.Results:The 5-year DFS and OS were 86.30% and 94.29%, respectively. Proportions of HR+/HER2-、HR+/HER2+、HR-/HER2+ and HR-/HER2- were 52.9%、17.5%、14.1%和15.5%, respectively. The 5-year DFS of HR+/HER2- subtype (88.12%) was higher than HR+/HER2+ (84.67%, P=0.026), HR-/HER2+ (84.19%, P<0.001) and HR-/HER2- (83.70%, P<0.001). The 5-year OS of HR+/HER2- (95.38%) was not different from HR+/HER2+ (95.17%, P=0.187), while it was higher than that of HR-/HER2+ (92.26%, P<0.001) and HR-/HER2- (91.69%, P<0.001). Subtype was still a significant factor regarding DFS and OS in multivariable analyses adjusting for age, sex, stage, Ki67, types and time of surgery. The DFS ( P=0.257) and OS ( P=0.511) was not different between HR-/HER2+与HR+/HER2- subtypes, while HR-/HER2+ and HR-/HER2- patients had worse DFS ( P<0.05) and OS ( P<0.05) than that with HR+/HER2-. Conclusions:Molecular subtype is a significant independent prognostic factor for DFS and OS in operable invasive breast cancer. HR+ subtypes have better prognosis compared with HR- subtypes. The DFS and OS were not different between HR+/HER2- and HR+/HER2+, or between HR-/HER2+ and HR-/HER2-.