1.Establishment and identification of a mitochondrial tracking system
Lin LYU ; Sihan WANG ; Quan ZENG ; Han DUAN ; Zhuang MAO ; Changyao WANG ; Xuetao PEI ; Hua WANG ; Yanhua LI
Chinese Journal of Pharmacology and Toxicology 2023;37(12):928-935
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.
2.Progress of concurrent chemoradiotherapy in esophageal cancer
Yunxiao WANG ; Zhongchao HUO ; Xiu JIN ; Xuetao HAN ; Yun DONG
Cancer Research and Clinic 2020;32(6):436-441
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.
3.Effect of allogeneic platelet transfusion on migration and invasion of human lung cancer A549 cells and its mechanism of action
HAN Lina ; ZHAO Xuetao ; MA Ming ; WU Bo ; ZHAO Liang ; ZHANG Cong ; SHAN Baoen
Chinese Journal of Cancer Biotherapy 2020;27(9):1018-1023
[Abstract] Objective: To observe the effect of allogeneic platelets transfusion on the invasion and metastasis of human lung cancer A549 cells, and to preliminarily explore its mechanism of action. Methods: Eighty-nine patients with advanced lung cancer, who had received platelet transfusion in the Chemotherapy Department of Fourth Hospital of Hebei Medical University between January 2017
and December 2018, were enrolled in this study. The study cells were randomized into Ctrl group (A549 cells co-incubated with culture medium), Before group, and After group (A549 cells co-incubated with plasma Before and After platelet transfusion, respectively). The migration and invasion of A549 cells co-cultured with plasma before and after platelet transfection were detected by Scratch and
Transwell experiments. The expression of MMPs, TIMPs and epithelial-mesenchymal transition (EMT) related proteins E-cadherin, N-cadherin and Vimentin, as well as vascular endothelial growth factor (VEGF) and its receptor 2 (VEGFR2) were detected by Western blotting (WB) method. Results: The scratch healing ability of A549 cells in After group was significantly higher than that of Ctrl group
and Before group [(73.67±2.60)% vs (58.33±2.33)%, (35.33±2.03) %; P<0.01, vs Ctrl group; P<0.05, vs Before group], and there was also a significant difference between Before group and Ctrl group (P<0.05). The results of cell migration experiment showed that the number of transmembrane cells in After group was significantly higher than that in Ctrl group and Before group [(69.67±7.84) vs (18±2.08) and (39.33±2.03), all P<0.01]. The cell invasion experiment showed that the number of transmembrane cells in After group was significantly higher than that in Ctrl group and Before group [(59.34±3.46) vs (18.34±1.56) and (37.58±2.79), all P<0.01]. When A549 cells were co-incubated with plasma before and after platelet transfusion for 48 h, it was found that the expressions of MMP9 and MMP2 were increased (P<0.05), while their inhibitors TIMP1 and TIMP2 were decreased (P<0.01); the expressions of EMT-related proteins N-cadherin and Vimentin were increased (P<0.05), but E-cadherin was decreased (P<0.01); the expressions of angiogenesis related proteins VEGF and VEGFR2 were increased (P<0.05). Conclusion: Alloplatelets transfusion can promote the invasion and metastasis of lung cancer A549 cells, which may be realized by regulation of the expressions of EMT, metallomatrix protease and vascular growth factor-related proteins.
4.Roles and mechanisms of Wnt/β -catenin signaling related lncRNA in tumor progression
LI Zemeng ; HAN Dan ; CAO Xuetao
Chinese Journal of Cancer Biotherapy 2020;27(3):315-320
长链非编码RNA(lncRNA)是一类长度大于200 nt、且不编码的RNA。lncRNA已被证明与人类疾病紧密相关,尤其 是肿瘤发生发展。研究表明,肿瘤中一些异常表达的lncRNA可以通过不同的信号通路, 如Wnt/β-catenin信号通路,促进肿瘤进 展过程。在不同肿瘤组织中具有特异性表达特征的lncRNA与Wnt/β-catenin信号通路之间的相互作用显示出其作为新的生物标 志物和治疗靶点的潜能。本文就Wnt/β-catenin信号通路相关lncRNA通过调控Wnt/β-catenin信号转导,影响不同肿瘤类型发生 发展的作用进行综述。本文结果或可为临床肿瘤诊断和治疗提供新的思路。
5.The clinical significance of MGMT promoter methylation in the treatment and progno-sis of glioma patients
Xuetao HAN ; Huandi ZHOU ; Xiaoying XUE
Chinese Journal of Clinical Oncology 2019;46(12):645-648
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.
6.Research progress on the relationship between the canonical Wnt signaling pathway and tumor radiation resistance
Yu YANG ; Huandi ZHOU ; Xiaoying XUE ; Ge ZHANG ; Xuetao HAN
Chinese Journal of Radiological Medicine and Protection 2017;37(4):316-320
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.
7.One-stage anterior-posterior decompression and internal fixation for severe fracture-dislocation of lower cervical spine in "beach chair position"
Jiqiang TANG ; Xuetao SUN ; Minglin SUN ; Fengmin TANG ; Binbin YUAN ; Yue HAN ; Jidong ZHANG ; Qun XIA
Chinese Journal of Trauma 2017;33(9):779-784
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.
8.Polylactic acid-polyglycolic acid lumbar interbody fusion cage full of broken bones versus autologous bone: an influence on the spinal stability?
Hongge SONG ; Xuetao LI ; Guanghui HAO ; Qinan ZHANG ; Bing HAN ; Li CHEN ; Yujie HAI ; Huafeng LIU ; Yanchao CHEN ; Jiashuang WANG
Chinese Journal of Tissue Engineering Research 2017;21(22):3445-3451
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.
9.Polytetrafluoroethylene segment implantation for arteriovenous internal fistula with outflow tract obstruction
Yu ZHANG ; Zhiqiang ZHU ; Xin ZHENG ; Xuetao MA ; Jianwei LIU ; Zhiyou HAN
Chinese Journal of Tissue Engineering Research 2014;(30):4911-4916
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.
10.Wnt3a is Important in The Differentiation From Neural Stem Cell Into Dopaminergic Neuron In vitro
Shu HAN ; Wei SHI ; Yanhua LI ; Hailei YAO ; Xiaoyan XIE ; Lin CHEN ; Shuangshuang SHI ; Cixian BAI ; Xue NAN ; Fang YAN ; Yunfang WANG ; Xuetao PEI
Progress in Biochemistry and Biophysics 2006;0(10):-
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

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