Objective To explore the killing effect of dielectric barrier discharge (DBD) plasma on tumor cells and to analyze the DBD-induced apoptosis mechanism.Methods Thiazolyl blue tetrazolium bromide (MTT) assay method was used to detect the killing effect of low temperature plasma on the cytotoxicity of normal spleen leukocytes and acute promyelocytic leukemia cells (LT-12) at different doses.The changes of reactive oxygen species (ROS) level were measured after plasma treatment.The cell apoptosis rate was detected by Annexin V/PI double staining at different doses.The expression of apoptosis-related genes and proteins was detected by qRT-PCR and Western Blot.Results MTT results showed that the killing effect of plasma treatment was dose-dependent and time-dependent.The cell survival rate after 8 hours of treatment decreased from 98％ to 63％ with the dose increasing from 30 s to 240 s.The survival rate decreased from 78％ (2 h) to 39％ (24 h) after the treatment with a same dose (e.g.240 s).Annexin V/PI double staining results demonstrated that the plasma effect can induce apoptosis,and the apoptosis rate was not only positively correlated with the plasma dose,but also with the post-plasma time.The longer the post-plasma time,the higher was the apoptosis rate.The apoptotic rate of the 60 s dose treatment after 12 h was 48％ that increased to 55.3％ with the dose of 120 s.The production of reactive oxygen species (ROS) detected by flow cytometry also showed a time correlation of the plasma treatment.After the plasma treatment,the ROS level immediately increased to 1.24 times,and sharply increased to 5.39 times after 20 h post-plasma.The experimental results of qRT-PCR and Western blot showed that the expression of the genes and proteins of Caspase family and Bcl-2 family was very active at 8 to 12 h post-plasma treatment.Conclusions Low-temperature plasma can effectively kill tumor cells,and apoptosis is the main mechanism of death.The molecular mechanism of apoptosis of tumor cells induced by low temperature plasma was preliminary confirmed.

Objective To construct the third-generation self-inactivating lentiviral vectors including BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a,then integrate them into the genome of mouse embryonic osteoblast cell,MC3T3-E1,and to explore the capability of osteogenic differentiation of individual bone morphogenetic control factor and the effective approaches to further improve the capability of osteogenic differentiation.Methods The plasmid vectors of gene expression including BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a were constructed,which were identified through enzyme cutting and further confirmed through sequencing.After packing pELNS-BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a,mouse embryonic osteoblast cell,MC3T3-E 1 was transfected,and the transfection efficiency was confirmed by GFP fluorescence imaging.The expression level of Runx2 mRNA and the transfection efficiency of individual bone morphogenetic control factor was detected by Real time PCR.Eight groups of MC3T3-E1 were dual-gene co-transfected,and the transfection efficiency was verified by GFP fluorescence imaging.ELISA was adopted to detect the expression level of BGP and ALP in MC3T3-E1culture supernatants;Real time PCR was adopted to detect the expression level of Runx2 mRNA;Western blot was adopted to detect the expression level of protein of BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a.Thus,the effectiveness of osteogenic differentiation of dual-gene co-transfection were evaluated.Results The recombination of lentiviruses,pELNS-BMP-2,pELNS-BMP-4,pELNS-BMP-6,pELNS-BMP-7,pELNS-BMP-9 and pELNS-Wnt3a were successfully constructed.MC3T3-E1 was successfully transfected.The expression levels of Runx2 mRNA were:BMP-2 ＞ BMP-4 ＞ BMP-9 ＞ BMP-7 ＞ Wnt3a ＞ BMP-6.Successful transfection of the dual-gene co-transfection of eight groups of MC3T3-E1 were verified by GFP fluorescence imaging.The expression level of Runx2 mRNA,the expression of BGP and ALP showed BMP-2 and BMP-7 co-transfection group was the most efficient in osteogenesis transfection.Western blot revealed thatthe protein expression of BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a increased after cotransfection of MC3T3-E1 by BMP-2 and BMP-7.Conclusion The third-generation lentiviral vector,pELNS can lead BMP-2,BMP-4,BMP-6,BMP-7,BMP-9 and Wnt3a into MC3T3-E1,mouse embryonic osteoblast cell,and stabilize its expression.Individual bone morphogenetic control factors can promote MC3T3-E1's differentiation to osteoblasts.The dual-gene co-transfection of BMP-2 and BMP-7 can effectively promote osteoblast conversion,which provides significant theoretical basis and technical support for remodeling of tissue engineering bone.

OBJECTIVETo explore the pattern of lymphatic metastasis and risk factors of esophageal carcinoma that invades less than adventitia.

METHODSClinical data of 484 patients receiving esophagectomy from January 2011 to August 2014 were reviewed, whose carcinoma invaded less than adventitia. The lymph node metastasis pattern of the primary tumor and corresponding influence factor were analyzed.

RESULTSTotal lymph node metastatic rate was 32.0% (155/484). Sixteen of 61 upper thoracic esophageal carcinoma patients (26.2%) had lymphatic metastasis. Fifty-five of 201 middle thoracic esophageal carcinoma patients (27.4%) had lymphatic metastasis. Eighty-four of 222 lower thoracic esophageal carcinoma patients(37.8%) had lymphatic metastasis. The deeper tumor invaded, the easier lymph node metastasis occurred, as well as the lower of the tumor differentiation and the larger of the tumor diameter. Multivariate analysis revealed lesion diameter (P=0.005), differentiation degree (P=0.007) and invasion depth (P=0.001) were independent risk factors of lymphatic metastasis in esophageal cancer that invaded less than adventitia.

CONCLUSIONDepth of tumor invasion, diameter of tumor and tumor differentiation are risk factors of lymph node metastasis of esophageal carcinoma that invades less than adventitia.

Adventitia ; pathology ; Esophageal Neoplasms ; pathology ; Esophagectomy ; Humans ; Lymph Node Excision ; Lymph Nodes ; Lymphatic Metastasis ; Multivariate Analysis ; Retrospective Studies ; Risk Factors

Objective To explore the pattern of lymphatic metastasis and risk factors of esophageal carcinoma that invades less than adventitia. Methods Clinical data of 484 patients receiving esophagectomy from January 2011 to August 2014 were reviewed , whose carcinoma invaded less than adventitia. The lymph node metastasis pattern of the primary tumor and corresponding influence factor were analyzed. Results Total lymph node metastatic rate was 32.0%(155/484). Sixteen of 61 upper thoracic esophageal carcinoma patients (26.2%) had lymphatic metastasis. Fifty-five of 201 middle thoracic esophageal carcinoma patients (27.4%) had lymphatic metastasis. Eighty-four of 222 lower thoracic esophageal carcinoma patients (37.8%) had lymphatic metastasis. The deeper tumor invaded, the easier lymph node metastasis occurred, as well as the lower of the tumor differentiation and the larger of the tumor diameter. Multivariate analysis revealed lesion diameter (P=0.005), differentiation degree (P=0.007) and invasion depth (P=0.001) were independent risk factors of lymphatic metastasis in esophageal cancer that invaded less than adventitia. Conclusion Depth of tumor invasion, diameter of tumor and tumor differentiation are risk factors of lymph node metastasis of esophageal carcinoma that invades less than adventitia.

Objective To explore the pattern of lymphatic metastasis and risk factors of esophageal carcinoma that invades less than adventitia. Methods Clinical data of 484 patients receiving esophagectomy from January 2011 to August 2014 were reviewed , whose carcinoma invaded less than adventitia. The lymph node metastasis pattern of the primary tumor and corresponding influence factor were analyzed. Results Total lymph node metastatic rate was 32.0%(155/484). Sixteen of 61 upper thoracic esophageal carcinoma patients (26.2%) had lymphatic metastasis. Fifty-five of 201 middle thoracic esophageal carcinoma patients (27.4%) had lymphatic metastasis. Eighty-four of 222 lower thoracic esophageal carcinoma patients (37.8%) had lymphatic metastasis. The deeper tumor invaded, the easier lymph node metastasis occurred, as well as the lower of the tumor differentiation and the larger of the tumor diameter. Multivariate analysis revealed lesion diameter (P=0.005), differentiation degree (P=0.007) and invasion depth (P=0.001) were independent risk factors of lymphatic metastasis in esophageal cancer that invaded less than adventitia. Conclusion Depth of tumor invasion, diameter of tumor and tumor differentiation are risk factors of lymph node metastasis of esophageal carcinoma that invades less than adventitia.

ObjectiveTo observe and analyze the synapses developing process of newly generated connections of autologus activated Schwann cells (AASCs) in combination with fetal spinal cord cell suspension(FSCS) in the surrounding area of the spinal cord injury site.MethodsA total of 42 Wistar rats underwent unilateral ligation of the saphenous nerve.The portion of nerve tissues distal to the ligation site were harvested 1 week after operation.AASCs were isolated,cultured and purified.Spinal cord injury model produced in 42 Wistar rats on T7 by modified Allen impact method.Three days after injury,20 μl FSCS with a density of 1×105/μl prepared from pregnant rats (El4) in combination with AASCs were injected into the epicenter of the traumatized cavity.Animals were sacrificed at 2,4,6,8,10,12 weeks post transplantation.Light and electronmicroscopic studies as well as immunohistochemical assay were carried out to evaluate the graft survival,its differentation and integration with the host.ResultsIn the transplantation area,AASCs showed good growth and differentiation,and glial scarring surrounding the lesions was less.The neuroblast stretched out the terminal endings 4 weeks after implantation,followed by the presenting of the pre- and post-synaptic membrane.Eight weeks post transplantation,the dense or developed projections were observed in the pre- and post-synaptic membrane,the high electron dense substance full filled the synaptic cleft.All the spherical cleat vesicles,granular vesicles,elliptical vesicles and flattened-f type vesicles were discovered under the electron microscope.Ten weeks after injury,the axosomatic,dendrosomatic,dendro-dendritic,axoaxonic,and dendro-axonic synapses coexisted.Light microscopy showed that the graft cell grew gradually.Immunohistochemical assay showed that NF,5-HT,CGRP and GFAP positive fibers were in the graft.Synapses,glia fibers and blood brain barrier integrated each other.Conclusion1) The transplanted FSCS combined with AASCs can develop mature synapses with miscellaneous synaptic vesicles in the acute injured spinal cord.2) Co-existing indicate the possibility of synaptic connection between FSCS and host.

Objective To explore the clinical characteristics and risk factors of re-fracture in patients suffering from osteoporosis-related fractures as well as effective interventions.Methods From January 2006 to January 2008,a total of 273 patients with osteoporosis-related fracture were entered in the study,including out-patients and in-patients who were over 50 years old.The patients were divided into fracture group(n=225)and re-fracture group(n=48).The re-fracture rate was followed up for 2 years,during which 11 patients developed re-fracture.General data including age and sex,fracture types,femoral neck bone mineral density(BMD)T-scores tested by dual-energy X-rays absorptiometry(DEXA),Charlson index,timeinterval between two fractures as well as mobility skill assessment were collected and analyzed.Results The average age at the first fracture was 67.7±8.5 years vs.72.7±9.5 years for the re-fracture cases.Female accounted for 70.2% of the fracture group and 77.1% of the re-fracture group.The most common re-fracture type was vertebral fracture for the first time and femoral neck fracture for the second time during the followup.Risk factors for a second fracture in osteoporotic fractures patients include age(＞75 years,HR=1.23; ＞85years,HR=1.68),female sex(HR=1.36),prior vertebral fractures(HR=1.62),prior hip fractures(HR=1.27),BMD T-score＜-3.5(HR=1.38)and weakened motor skills(HR=1.27).The refracture rate in osteoporosis-related fractures was 4.9% followed up for 2 years.The second fracture happened 3.7 years after the first one on average.Conclusion The risks of second fracture among patients with initial brittle fracture are substantial.Mobility skill assessment is an important risk factor for osteoporosis fractures recurrence.There is adequate time between fracture and re-fracture for effective interventions to prevent or reduce the risks of refracture,especially for the old women with a vertebral or hip fracture.Medication,motor function rehabilitation and fall-down prevention training would be helpful.

BACKGROUND: Clinical studies demonstrate that the durable stability of the prosthesis depends on a close geometric fit between the prosthesis and femoral medullary cavity.OBJECTIVE: To study the law of the parameters of medullary canal section shape in proximal femur DESIGN: Repeated measurement observation.SETTING: Department of Orthopaedics, Ninth People's Hospital, School of Medicine, Shanghai Jiaotong University MATERIALS: Ten femoral samples without any damage was obtained from corpse in the Staff Room of Anatomy of the Shanghai Second Medical University METHODS: The experiment was carried out in the Department of Or thopaedics, Ninth People's Hospital between January 2000 and March 2000. Section morphology of medullary cavity of proximal femur was dealt with image-processing, and conus curve fitted parameter mathematical method was proposed; and at the same time, section of medullary cavity of proximal femur of 10 patients was measured manually.MAIN OUTCOME MEASURES: Computer-aided method and manual method were applied to measure the coordination at the end of extrude and the coordinate of link joint RESULTS: 10 femoral samples entered the stage of result analysis. Com puter was used to measure the coordination of the end of extrude of the section medullary cavity of proximal femur (X,Y) and the coordination of the connection point (X,Y).There was no significant difference of the measuring results between computer-aided method and manual CONCLUSION: Computer-aided imaging-processing method not only reduces the error but also can be completed by computer automatically in measuring section morphology of medually cavity in proximal femur. It is suitable for a variety of morphology measurement.

In a hip replacement case when a standard hip stem is chosen, it is rough to select hip stems before operation, thus some stand-up standard hip stems should be prepared. The operation time is usually prolonged. In order to solve this problem, this paper presents a method to automatically select optimal standard hip stems by computers. Femoral anatomical data of a patient are acquired from X-ray films. Based on these anatomical data and a database of average cross-sections in proximal femurs, proximal femur of the patient can be reconstructed. This proximal femur model makes it possible to optimally select a standard hip stem. Theory analyses indicate that the method, presented in this paper, is practicable.
Arthroplasty, Replacement, Hip ; instrumentation ; Computer-Aided Design ; Hip Joint ; diagnostic imaging ; surgery ; Hip Prosthesis ; standards ; Humans ; Radiography

BACKGROUND: Properties of cancellous bone is significant for metabolic diseases, fracture and joint retrograde diseases.OBJECTIVE: To study the biomechanical properties of cancellous bone.DESIGN: Experimental observation based on femur specimens as the single sample.SETTING: Laboratory of Orthopaedic Biomechanics Ninth Affiliated Hospital Shanghai Second Medical University.PARTICIPANTS: The study was completed in the Laboratory of Orthopaedic Biomechanics, Shanghai Ninth People's Hospital from May to July in 2001,totally 19 male cadaveric femoral heads were selected.METHODS: Totally 19 cadeveric cancellouce bone specimens(about 30years old) from the bearing area of fresh femur head were tested by compression test.MAIN OUTCOME MEASURES: Yield load limiting stress, limiting strain, energy value and elastic modulus of specimens.RESULTS: The compressive data of cancellous bone were measured, the average value of yield load was (410.64 ± 190.29) N , limiting stress was (8.69 ± 3.75) MPa, limiting strain was (10. 84 ±6.58)%, energy value was (2.54 ± 1.89)J, modulus of elasticity was (40.77 ± 32. 12)MPa, At the early time of loading, the cancellous bone showed elastic deformation, then CONCLUSION: The trial indicates that biomechanical properties lighten the pressure of external compression.