2.Study of Effect of CtBP2 Knockout through shRNA on Proliferation of Prostate Cancer Cells
Yan LIU ; Yong XU ; Chao GAO ; Zhihong ZHANG
Tianjin Medical Journal 2014;(10):977-979,980
Objective To study the effects of shRNA-CtBP2 on the growth of prostate cancer PC3 cells. Methods There were three experimental groups in this study,which include blank control group, empty plasmid transfected group and transfected shRNA group. CtBP2 mRNA sequence is targeted by 3 pairs of designed interfering shRNA to built shRNA-Ct-BP2 recombinant plasmid then it is transfected into PC3 cells. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot assays were used to detect the transcription and expression levels of CtBP2 mRNA and protein, respective-ly. PC3l proliferation was measured by MTT assay. Results Builting shRNA-CtBP2 recombinant plasmid and transfect-ing PC3 cells were successful. Transcription and expression levels of CtBP2 mRNA and protein were significantly decreased in shRNA-CtBP2 transfected PC3 cells. After CtBP2 silencing, cell proliferation was blocked in the shRNA-CtBP2 cells compared to that of blank control group (P<0.01). Conclusion shRNA-CtBP2 could significantly inhibit CtBP2 expres-sion, suppress the growth of PC3 cells, which suggests that CtBP2 may be a new target for PCa gene therapy.
3.Evaluation of reversal effect of 2-DG on multidrug resistance by detecting uptake of 99mTc-MIBI in HNE-1/DDP cells
Yong SHEN ; Weili SUN ; Chao YUAN ; Huiqin XU ; Bin LIU
Chinese Pharmacological Bulletin 2015;(10):1433-1438
Aim To evaluate the reversal effect of 2-deoxy-D-glucose ( 2-DG ) on multidrug resistance ( MDR) by observing the uptake change of 99m Tc-MIBI in HNE-1/DDP cells, and to explore its mechanism. Methods The uptake of 99m Tc-MIBI in HNE-1/DDP cells under different concentrations of 2-DG was detec-ted by γ-counter, and the clearance rates of 99m Tc-MI-BI in HNE-1 cells and HNE-1/DDP cells after treated with 2-DG (10 mmol·L-1 ) were compared. The con-tent of ATP in HNE-1/DDP cells was detected after treated with 2-DG. P-glycoprotein ( P-gp ) and multi-drug resistance-associated proteins ( MRP ) expression were measured by Western blot. Apoptotic HNE-1/DDP cells treated with DDP alone or combined with 2-DG (10 mmol·L-1 ) were detected by propidium io-dide ( PI ) staining. Results The clearance rate of 99m Tc-MIBI in HNE-1/DDP cells was 54. 8%, which was significantly higher than that ( - 41. 3%) in HNE-1 cells (P<0. 01). The clearance rate of 99mTc-MIBI in HNE-1/DDP cells was -203. 7% after treat-ment with 2-DG ( 10 mmol · L-1 ) , which could be significantly reduced compared with the control group ( P<0. 01 ) . The level of ATP was 55 . 69% compared with the negative control group and the expression of P-gp and MRP protein decreased dramatically in HNE-1/DDP. With the combination of 2-DG and DDP, the ap-optotic rate of HNE-1/DDP cells reached 49 . 4%which was significantly higher than DDP treated group (22. 5%) . Conclusion Multidrug resistance and the reversal effect of 2-DG on multidrug resistance could be evaluated effectively by detecting the uptake change of 99m Tc-MIBI in HNE-1/DDP cells. The mechanism may be related with the inhibition of ATP level and the re-duced expression of P-gp and MRP protein in cancer cells.
4.Biomechanical effects of osteoporosis vertebral augmentation on the adjacent intervertebral levels using a three-dimensional finite element analysis
Sheng LU ; Yong-Qing XU ; Mei-Chao ZHANG ;
Chinese Journal of Orthopaedic Trauma 2004;0(09):-
Objective To investigate effects of volume,distribution and leakage of bone cement in ver- tebroplasty on the adjacent intervertebral bodies using an experimentally calibrated and anatomically accurate fi- nite-element model of elderly L4-L5 vertebral bodies.Methods Computed tomography(CT)scanning was done,at 1 mm intervals,on L4-L5 vertebral bodies of the lumbar spine of an old man cadaver that had no abnormal findings on rnentgenograms.The L4-L5 motion segment data were obtained from the CT scans to establish in computer a three-dimensional finite element model of L4-L5 functional spinal unit(FSU)with the software Ansys 7.0.Compressive load after virtual vertebroplasty on the damaged model,unipedicular and bipedieolar injections and leakage of cement into the intervertebral space,and the resulting endplate and disc stresses of the adjacent vertebral bodies were analyzed in various spatial distributions of the filling material and different loading conditions. An anatomically accurate finite-element model of elderly L4-L5 vertebral bodies was developed.Results The FSU study suggested that changes in stress and strain at adjacent levels were minimal.Furthermore,endplale and disc stresses of the adjacent vertebral bodies were not influenced by bone cement filling volumes or distributions except under bending,whereas asymmetric distribution and leakage of bone cement to the disk increased the stress of adjacent endplates.Conclusion Since asymmetric distribution and significant leakage of cement into the intervertebral space can increase the stresses of endplates of adjacent vertebral bodies and may lead to a fracture, symmetric placement of cement in operation should be achieved and leakage of cement avoided.
6.Diagnosis and microsurgical treatment of intramedullary hemangioblastoma in cervical spinal cord
Xu-Hui HUI ; Chao YOU ; Rui-Chun LI ; Si-Qing HUANG ; Kai-Yong YANG ;
Chinese Journal of Microsurgery 2006;0(06):-
Objective To investigate the diagnosis and the microsurgical treatment of intramedullary hemangioblastoma in cervical spinal cord.Methods The signs of MRI,and the results of operations were analysed in 26 patients with the tumors.Rusults The tumors can be classified into two types:Solid type (14 cases)and cystic type(12 eases).All the tumors underwent total removal and were all hemangioblastoma confirmed by histopathologic examinations.Postoperatively,neurological status were improved in 17 patients, remained in 7 cases and worse in 2 cases.Conclusion For intrameduUary hemangioblastoma of cervical spinal cord MRI is of significant importance in the diagnosis of localization and the nature of the tumors which is conductive to selecting appropriate operative methods.There is high risk in operating at cervical section,but microsurgical total resection is the optimal method to stop the development of the clinical presentation.Opera- tive methods varied with the different typer of the tumor.It is the most important principal that dissection is performed along the correct interface and the tumor should be removed en bloc after it is devascularized.
7.The application of anchor nail combined with Krackow suture technique in the treatment of flesh achilles tendon rupture
Jian NA ; Yong XU ; Chao MA ; Hui WANG ; Sitong QIN ; Keyong MA
Chinese Journal of Postgraduates of Medicine 2011;34(35):29-31
Objective To explore and evaluate the advantages of anchor nail in the treatment of achilles tendon rupture.Methods The clinical data of 27 patients with fresh achilles tendon rupture who received the treatment of thread anchor nail from January 2007 to August 2010 was retrospectively analyzed.Among them,14 patients received Krackow method,7 patients received Lindholm method to repair achilles tendon,6 patients received anchor nail combined with Krackow suture technique repair of achilles tendon.Arner-Lindholm clinical evaluation criterion was used for objective evaluation.Results Twenty-seven patients got 1-36 ( 17.35 ± 5.70) months follow-up.According to the corresponding standard in the 14 cases using Krackow method,8 cases were excellent,2 cases were good,4 cases were bad,the excellent and good rate was 71.4%( 10/14);in the 7 cases using Lindholm method,5 cases were excellent,1 case was good,1 case was bad,the excellent and good rate was 85.7 (6/7) ; all of the 6 cases using anchor nail combined with Krackow suture technique repair of achilles tendon were excellent,the excellent and good rate was 100.0%(6/6).Postoperative incisions were primarily healing,joint range of motion were all right,without rigidity,gait were almost normal,without the secondary fracture of ligament.Conclusion Anchor nail combined with Krackow suture technique in the treatment of achilles tendon rupture has significant effect,it is worth to be popularized because of its preonunced effect.
8.Expression and significance of vasculogenic mimicry and matrix metalloproteinase-2 in prostate cancer
Kuo YANG ; Chao MENG ; Yong XU ; Yan LIU ; Zhihong ZHANG ; Ranlu LIU ; Baojie MA
Chinese Journal of Urology 2012;33(10):791-794
Objective To find out if an unique blood mode-vasculogenic mimicry (VM) exist in prostate cancer and analyze its relationship with prognosis. Methods The age of the patients was 59 to 72 years,with a mean age of (66.7 ± 11.0) years.All had been confirmed as prostate cancer by biopsy.The level of serum PSA was 15.6 to 76.7 μg/L,with a mean level of (34.6 ± 1.7 ) μg/L.A histochemical and immunohistochemical dual staining method for PAS-CD31 and MMP-2 in 96 prostatic adenocarcinomas were conducted to explore if VM exist in prostate cancer,describe its morphology and distribution.The different expression of MMP-2 in VM positive and VM negative tissue were.The clinical data were collected and analyzed to explore the relationship between disease progression and VM. Results Among 96 cases,24 cases were found VM positive,the others were negative.The boundary membrane of VM was PAS positive,which was continuous or incontinuous. A positive correlation between VM density and high Gleason score were found.The tPSA were higher in the VM positive cases than that in VM negatives.The VM positive patients had poorer prognosis than that of the negatives.The MMP-2 expression was found correlated with VM positive (rs =0.60,P < 0.01 ). Conclusions VM exists in the prostate cancer as an independent structure,and it is more common in the high-grade cancer.The progression-free survival of VM positive patients could be worse than the negative.MMP-2 and VM could be the indicators of poor prognosis.
9.Long segment pedicle screw fixation for the thoracolumbar fracture and dislocation
Yong QI ; Hongtao SUN ; Wangyang XU ; Xiaozhong ZHOU ; Dixin LUO ; Chao LI ; Weijian CHEN ; Xunjie JIN
International Journal of Surgery 2011;38(8):543-545,封3
Objective To discuss the value of the posterior long segments pedicle screws fixation in surgical treatment of thoracolumbar fracture-dislocation,and summarize the experience.Method s We reviewed the clinical information of 27 cases of thoracolumbar fracture-dislocation since July 2007 to March 2010; all the cases were treated with the posterior long segments pedicle screws fixation.In these cases,20were male,7 were female.And the age ranged from 18 to 58 years old (average age was 35 years).The posterior approach was used,and the pedicle screws were inserted into the 2 vertebrae above and below the fractured vertebrae.Posterior decompression and bone grafts for fusion were also accomplished during the operation.Results Follow-up ranged from 7 to 25 months,averaging 14 months.Completely restoration of the dislocation was got in 25 cases and the spine sequence resumed to nearly normal.One case with lateral dislocation failed to complete correction; another 1 case operated 20 days after surgery failed to complete correction of forward dislocation.At the last follow-up,the mean Cobb angle loss was 3 degrees,the height of the vertebral had obvious loss compared with that of post-operation.The loosen of screw nut was found during the follow-up,but no dislocation and secondary deformity were observed after fixation.There was no death,no infection.Conclusions The technique of posterior long segments pedicle screw fixation is of great value in the clinical applications,since it has the following advantages:the operation is simple,trauma is small,the fixation is firm,good restoration of fracture and dislocations can be easily got,and most importantly,it can effectively prevent long-term vertebral height loss and the backward protruding deformity.
10.Osteogenic differentiation of bone mesenchymal stem cells regulated by osteoblasts under EMF exposure in a co-culture system.
Ji-zhe, YU ; Hua, WU ; Yong, YANG ; Chao-xu, LIU ; Yang, LIU ; Ming-yu, SONG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):247-53
This study examined the osteogenic effect of electromagnetic fields (EMF) under the simulated in vivo conditions. Rat bone marrow mesenchymal stem cells (BMSCs) and rat osteoblasts were co-cultured and exposed to 50 Hz, 1.0 mT EMF for different terms. Unexposed single-cultured BMSCs and osteoblasts were set as controls. Cell proliferation features of single-cultured BMSCs and osteoblasts were studied by using a cell counting kit (CCK-8). For the co-culture system, cells in each group were randomly chosen for alkaline phosphatase (ALP) staining on the day 7. When EMF exposure lasted for 14 days, dishes in each group were randomly chosen for total RNA extraction and von Kossa staining. The mRNA expression of osteogenic markers was detected by using real-time PCR. Our study showed that short-term EMF exposure (2 h/day) could obviously promote proliferation of BMSCs and osteoblasts, while long-term EMF (8 h/day) could promote osteogenic differentiation significantly under co-cultured conditions. Under EMF exposure, osteogenesis-related mRNA expression changed obviously in co-cultured and single-cultured cells. It was noteworthy that most osteogenic indices in osteoblasts were increased markedly after co-culture except Bmp2, which was increased gradually when cells were exposed to EMF. Compared to other indices, the expression of Bmp2 in BMSCs was increased sharply in both single-cultured and co-cultured groups when they were exposed to EMF. The mRNA expression of Bmp2 in BMSCs was approximately four times higher in 8-h EMF group than that in the unexposed group. Our results suggest that Bmp2-mediated cellular interaction induced by EMF exposure might play an important role in the osteogenic differentiation of BMSCs.