Objective:To eastablish the efficient presentation of antigen from apoptotic cells by human DC from peripheral blood. Methods: using recombinant human granulocyte/macrophage colonystimulating factor(GM- CSF) and interleukin 4 (IL- 4 ) we have established dendritic cells (DC)from peripheral blood monocyte that maintain the antigen capturing and processing capacity characteristic of immature dendritic cells in vivo. GM - CSF 50ng/ml , IL- 41 000ng/ml once two days(total four). on the 3 rd day of culture, immature DC and apoptotic hepatochlangioma cells were in coculture lasting 7 days. Results:these cells had typical dendritic morphology, express high levels of CD1a ,B7 and acquired antigen from apoptotic cells and induced an increased T cell stimulatory capacity in MLR. Conclusions:we have established DC from blood mononuclear tells using GM- CSF and IL- 4 and DC can be efficiently drived from apoptotic cells and can induce the increase of T cells obviously. It probably becomes an effective approach of antigen transduced with DC.

BACKGROUND: Dendritic cells play an important role in antigen present in vivo, and the mechanism of tumor cells in escaping the antigen presentation of dendritic cells existed in the patients with tumor.OBJECTIVE: To sensitize dendritic cells from human peripheral blood with apoptotic hepatoma cells induced by mitomycin.DESIGN: A randomized control trial by taking apoptotic hepatoma cell sensitized dendritic cells as the observed subjects.SETTINGS: Institute of Field Surgery, Daping Hospital, the Third Military Medical University of Chinese PLA; Institute of Radiation Medicine,Chinese PLA Academy of Military Medical Sciences.MATERIALS: The experiments were carried out in the Institute of Radiation Medicine, Chinese PLA Academy of Military Medical Sciences from April 1998 to May 1999. The cell strain was the QBC939 bile duct cancer cell strain, and mitomycin was used as the antitumor drug.METHODS: Mononuclear cells were isolated from the peripheral blood of normal people, 50μg/L granulocyte-macrophage colony stimulating factor(GM-CSF) and 1 000 U/mL interleukin-4 (IL-4) were added, once every other day for 4 times. On the 3rd day of culture, the apoptotic bile duct cancer cells induced by mitomycin was added, and then cultured in vitro for 4 days, finally the dendritic cells were collected.MAIN OUTCOME MEASURES: ① The identification of the cultured dendritic cells was observed; ② The dendritic cells were co-cultured with necrotic and normally cultured bile duct cancer cells respectively, and the phagocytized apoptotic body loaded antigens were observed; ③ The immunostimulatory activity of dendritic cells (1×103, 5×103 and 1×104/well)and that after loaded by antigen were detected, and the mononuclear cells were taken as controls.RESULTS: ① The cultured and amplified dendritic cells expressed high levels of costimulatory molecules of CD1a and B7, and there were typical irregular processes on the surface. ② The tumor cells formed apoptotic bodies when they were induced by mitomycin, which were arrested and phagocytized by dendritic cells. ③ The ability of the antigen loaded dendritic cells in stimulating the proliferation of allogenic lymphocyte T was further enhanced.CONCLUSION: The apoptotic tumor cells induced by mitomycin can induce the mononuclear cells from human peripheral blood differentiating into the dendritic cells with the concomitance of GM-CSF and recombinant IL-4 and amplify dendritic cells. Meanwhile, the dendritic cells can effectively present the antigens of apoptotic bile duct cancer cells, and it probably becomes a new effective approach for tumor antigen to sensitize dendritic cells.

Radiotherapy is one of major cancer treatment methods.However,radiation resistance is an important reason to restrict the efficacy of radiotherapy and lead to treatment failure.In recent years,the relationship between the canonical Wnt signaling pathway and tumor radiation resistance has more and more attention of the scholars.This review summarized recent ten years findings concerning the canonical Wnt signaling pathway and tumor radiation resistance and tried to find some valuable rules or some internal relationships among different pathways by systemically analyzing.

BACKGROUND:Arteriovenous internal fistula is the first choice for hemodialysis. In the process of hemodialysis, many patients suffer from venous outflow stenosis. The methods including thrombolysis, intervention, surgical repair and fistula reconstruction al have their disadvantages. OBJECTIVE:To compare the midterm effects of polytetrafluoroethylene segment implantation and exclusively surgical repair in arteriovenous internal fistula with outflow tract obstruction. METHODS:Eighty patients with venous outflow stenosis, aged 22-80 years, were divided into test group (n=50;polytetrafluoroethylene segment implantation) and control group (n=30;simple surgical repair). The post-operative infection rate, postoperative time til recurrence of fistula dysfunction, and accumulate survival rate were compared between the two groups. RESULTS AND CONCLUSION:During the fol ow-up period of 10-28 months in the test group, there were nine patients with vascular access dysfunction, and the accumulate survival rate was 100%for 6 postoperative months, 92%for 12 months, and 82%for 18 months. In the control group, there were seven cases of vascular access dysfunction at 8-28 months of fol ow-up, and the accumulate survival rate was 93%for 6 postoperative months, 87%for 12 months, and 77%for 18 months. No statistical y significant difference in the postoperative infection rate was observed between the two groups. The Kaplan-Meier survival curves showed that the accumulate survival rate was slightly higher in the test group than the control group, but there was no significant difference based on log-rank test (P=0.44). These findings indicate that polytetrafluoroethylene segment implantation for arteriovenous internal fistula with outflow tract obstruction has the similar effects as the surgical repair if it does not alter the autologous behavior of the initial access and maximal y reserve the vessels for puncture.

BACKGROUND:Along with the widespread application of biodegradable materials in the field of medicine and the in-depth research of biomechanics,the drawbacks of traditional medical metal materials are increasingly appearing.In recent years,researchers at home and abroad focus on biodegradable materials that are represented by high molecular polymer to seek new breakthroughs in the field of spinal instability.OBJECTIVE:To investigate biomechanical changes of polylactic acid-polyglycolic acid (PLGA) lumbar interbody fusion cage in the body and discusses its feasibility for treating segmental instability of the spine.METHODS:Forty-two healthy pigs (9 months old) were randomly divided into two groups (n=21),and L4/5 intervertebral disc nucleus pulposus was removed in all animals.In experimental group,PLGA lumbar interbody fusion cage filled with broken bone was implanted;and in control group,autologous bone was implanted.X-ray was performed to observe the fusion of operation segments at 4,12 and 72 weeks postoperatively.Feasibility of fibrous fusion was measured by biomechanical test.Histologically,bone graft fusion at the surgical site and material degradation were detected.RESULTS AND CONCLUSION:(1) Imaging examination:Bone graft fusion in two groups was not visible at 4 weeks after operation.Evidence of increasing fusion was found in the experimental group at 12 weeks after operation;a visible part of the bone bridge was found in the control group,in which there was one case of fusion.Degradation of the fusion cage with one case of fusion in experimental group was found after 72 weeks after operation,and two cases of fusion in the control group.(2) Biomechanical test:There was no difference in the spinal range of motion between the two groups in different states at 4 weeks after operation (P ＞ 0.05).The spinal range values of motion at most of the states at 72 weeks after operation were significantly lower than those at 4 weeks after operation.(3) Cell histology observation:With the passage of time,the materials in the experimental group degraded gradually;new bone grew slowly and then fast,with bone fusion step by step.Fusion results were similar in the two groups.Our experimental findings indicate that the PLGA lumbar fusion cage has good biocompatibility.In addition to the individual state (left flexion),the mechanical properties of the fusion cage are similar to that of autogenous bone,and the fusion cage enables the segmental reconstruction of the pig spine to the maximum extent.

Wnt signaling is implicated in the control of cell growth and differentiation during neural stem cell(CNS) development.Wnt3a, one of wnt gene family members, has effect on regeneration neurospheres and differentiation into neurons.Wnt3a inhibits regeneration of neurospheres, and promotes its differentiation. In vitro neurosphere was cultured in a serum-free defined medium DMEM/F12 supplemented with bFGF and EGF. Dissociated cells were plated onto poly-d-lysine-coated coverslips and propagated in medium containing recombined Wnt3a-adenovirus. Plenty of Nurr1 were detected by RT-PCR after 3 days. Wnt3a combined AA would improve NSC differentiation into dopaminergic (DA) neuron. The quantity of DA neuron is obviously more than the AA alone group's. Moreover, the expression of TH mRNA is 1.86 fold in Wnt3a combined AA group. Induced cells were immunostained for TH and DAT. The proportion of TH-positive was (37.42 ? 2.54) % (P

Esophageal cancer is the most common malignant tumors in the digestive system. The treatments include surgery, radiotherapy, chemotherapy, targeted therapy and immunotherapy. In order to prolong the survival time and reduce the recurrence rate, most of the patients with locally advanced esophageal cancer were treated with combined radiotherapy and chemotherapy. There are many ways of concurrent chemoradiotherapy, together with the application of molecular targeting and immune drugs in esophageal cancer, there are different modes of combined therapy, which are of great significance to improve clinical efficacy and treatment compliance. This article reviews the progress of concurrent chemoradiotherapy for esophageal cancer in recent years.

Adjuvant temozolomide-based chemotherapy has become the standard of care for most postoperative glioma patients. However, a large proportion of these patients do not respond to temozolomide. DNA repair enzyme O6-methylguanine-DNA methyl-transferase (MGMT) promoter methylation has emerged as an important molecular marker in patients with gliomas. It is associated with prognosis and resistance to alkylated drugs such as temozolomide. MGMT promoter methylation is the key mechanism of MGMT gene silencing, thereby inhibiting DNA repair and increasing the sensitivity of chemotherapy. We reviewed current data on the prog-nostic and predictive relevance of MGMT testing and clinical trials, summarized the clinical application of MGMT promoter methyla-tion, in order to provide reference for the individualized treatment of glioma patients.

Objective To explore the surgical feasibility and clinical efficacy of one-stage anterior-posterior approaches in treatment of severe fracture and dislocation of lower cervical spine in "beach chair position".Methods Sixteen male cases of severe fracture and dislocation of lower cervical spine and with a mean age of 49.8 years (range,36-78 years) treated surgically from May 2012 to May 2016 were analyzed retrospectively by using case series study.The segment of injury was C4-5 in 4 cases,C5-6 in 7 and C6-7 in 5.The degree of spinal cord injury according to the American Spine injury Association (ASIA) score was Grade A in 4 cases,Grade B in 7 and Grade C in 5.Sub-axial injury classification (SLIC) score was 8 points in 9 cases and 9 points in 7.After a general anesthesia,a ring with a hole was hanged on patient's head before the operation.Then,under the protection of hole traction,the upper of operating bed was swung up slowly,so that the patient was restricted in vertical "beach chair position" with traction on the halo in order to immobilize the head and partially reduce the kyphotic deformity.Routine cervical anterior-posterior approach was done with the exposure of damaged section of the front and rear structure.Pedicle screw system or lateral mass screw displacement was conducted.Anterior intervertebral discectomy or fracture vertebral was performed,using collaborative reset prying method before and after the road.In the front of intervertebral cage or titanium net support bone graft,rear pedicle screws or lateral mass screws fixation and bone graft fusion were implemented.The operation time and blood loss were recorded.The healing of the wound was observed.The recovery of neurological function was evaluated according to the ASIA grade.Postoperative review X-ray,CT and MRI were done to evaluate the reset and bone graft in position and fusion.Results All the surgeries were done well without aeroembolism and other related complications.The mean operative time was 153 minutes (range,150-180 minutes),and the mean amount of blood loss was 543 ml (range,400-800 ml).Sixteen cases were followed-up from 6 to 24 months (mean 13.7 months).All the incision were healed at Ⅰ stage.Spinal cord function did not aggravate.The ASIA grade was improved with an average of one to two Grades 6 months after surgery.Postoperative X-ray and CT confirmed that graft object position was favorable and cervical sequence was recovered well.The Cobb angle decreased from (23.6 ± 5.3) ° preoperatively to (4.0 ± 0.4)°postoperatively,and the translational displacement of vertebral body was restored into (2.7 ±0.4) mm (P ＜ 0.01) from (10.9 ± 1.6) mm before operation.The cervical spinal canal was not obstructed and the cervical spinal cord was relieved,showed by MRI.Conclusions One-stage anterior-posterior approaches for severe fracture and dislocation of lower cervical spine circumferential reconstruction in "beach chair position" is a beneficial and effective method,without the need of changing positions in a collaborative reduction and fixation.The method can reduce the interference of spinal cord,shorten the operation time and save anterior extra fixation.

OBJECTIVE To observe whether mitochondria can be transferred from mesenchymal stem cells(MSCs)to irradiated cells by establishing a mitochondrial fluorescent reporting system.METHODS The lentiviral vector pSIN-EF1α-COX8A-DsRed2(named COX8A-DsRed2)that might guide the expres-sion of red fluorescence protein in the membrane of mitochondria was constructed.A lentivirus(named Lv-COX8A-DsRed2)was prepared in 293T cell line.Dental pulp stem cells(DPSCs)(named DPSC-COX8A-DsRed2)was infected with Lv-COX8A-DsRed2.The intracellular expression of the red fluores-cence protein in DPSC was observed under fluorescence microcopy.The mitochondrial localization of the expressed red fluorescent probe in DPSC-COX8A-DsRed2 was confirmed according to TOMM20 immunostaining and MitoTracker Green staining results,which could specifically label mitochondria.The IEC-6 cells that received 10 Gy X-ray radiation were used as an injured cell model.The co-culture system was established by supplementing DPSC-COX8A-DsRed2 into the culture plate with the irradi-ated IEC-6 labelled by CFSE for 24 h.RESULTS The imaging results of fluorescent microcopy obser-vation showed that DPSC-COX8A-DsRed2 expressed the mitochondrial fluorescent reporting system,which was co-located with TOMM20 protein and Mito Tracker Green.The imaging results of confocal fluorescence microcopy showed that the mitochondria with red fluorescent protein were transferred from DPSC-COX8A-DsRed2 to the irradiated IEC-6 cells,suggesting that the established mitochondrial fluorescent reporting system could indicate mitochondrial transfer from donor cells to injured ones.CONCLUSION DPSC-COX8A-DsRed2 stably expressing the mitochondrial fluorescent reporting system is established,which can be used to track mitochondrial transfer.