1.CONSTRUCTION OF ACTIVE RECOMBINANT CASPASE-3 EUKARYOTIC EXPRESSION PLA SMID AND EFFECT OF r-CASPASE-3 ON APOPTOSIS OF PANCREATIC CARCINOMA CELLS
Wei WANG ; Zhiguo LIU ; Zhaoyin QIN
Journal of Pharmaceutical Analysis 2001;13(2):119-122
Objective To construct active recombinant cas pa ses-3 gene(r-caspases-3)eukaryotic expression plasmid and observe the apoptos is inducing activity of r-caspase-3 in pancreatic carcinoma cells. Methods pcDNA3.1(+)/r-caspase-3 was constructed and pan creatic carcinoma cells(PC-Ⅱ)were transfected with the pcDNA3.1(+)/r-caspases -3 by liposomes(LipofectAMINE).The expression of r-Caspase-3 mRNA in pancreat ic carcinoma cells was detected by reverse transcription process of the polymera se chain reaction(RT-PCR), and the signs of apoptosis were examined in pancreat ic carcinoma cells by the methods of the DNA electrophoresis and flow cytometry analysis(FACS).Results The sequence inserted in pBlueSKM/r-Caspase-3 p lasmid was coincident with that of the r-caspases-3. The evaluation result of pcDNA3.1(+)/r-caspases-3 through enzyme cutting was correct. A 894bp strap was observed by RT-PCR after pancreatic carcinoma cells being transfected with the pcDNA3.1(+)/r-caspases-3 by liposomes. No strap was found in control groups. A characteristic DNA ladder was observed in pancreatic carcinoma cells DNA elect r ophoresis, and transparent hypodiploid karyotype peak was found by FACS. Conclusion The plasmid of pcDNA3.1(+)/r-Caspase-3 was c onstructed successfully, the expression of r-Caspase-3 mRMA in pancreatic carc inoma cells was confirmed by RT-PCR, and pcDNA3.1(+)/r-Caspase-3 can induce a poptosis in pancreatic carcinoma cells.
2.The protective effect of estradiol preconditioning on the myocardium of isolated ischemic reperfused rats
Wei WANG ; Chuncheng LI ; Zhaoyin QIN ; Huifang WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 1981;0(02):-
Objective To explore the protective effect of estradiol preconditioning on the myocardium of isolated ischemic reperfused rats. Methods The instrument of Langendorff was used to establish the model of isolated ischemic reperfused heart and male rats were randomly divided into three groups: control, treatment and preconditioning groups. Results Compared with that of control and treatment groups, estradiol preconditioning decreased the release of LDH, CK and intracellular TnI more effectively. Conclusion Estradiol preconditioning produces more protective effect on the myocardium of isolated ischemic reperfused rats.
3.siRNA AGAINST SURVIVIN SUPPRESSES THE PROLIFERATION OF PANCREATIC CANCER CELL PC-2 AND INDUCES ITS APOPTOSIS
Haitao GUAN ; Xinghuan XUE ; Xijing WANG ; Ang LI ; Zhaoyin QIN
Journal of Pharmaceutical Analysis 2006;18(1):78-82
Objective Blocking the expression of Survivin with siRNA, and the effects of suppling the proliferation of PC-2 cell and inducing its apoptosis were investigated. Methods Constructed the siRNA against Survivin plasmid expression vector and transfected it into PC-2 cell with lipofectamineTM 2000, the changes of Survivin expression were detected by semi-quantitive RT-PCR and immunohistochemical method, The effect of suppressing the proliferation of PC-2 cell was detected by the method of MTT; the effect of inducing PC-2 cell apoptosis was detected by flow cytometry. Results The sequence specific siRNA can effectively block the expression of Survivin both at themRNA and protein levels, the expression inhibition ratio was 81.25% at mRNA level and 74. 24% at protein level;blocking the expression of Survivin can suppress the proliferation of PC-2 cell significantly, 24, 48 hours after the cell was reseeded, the proliferation inhibition ratio was 28. 00% and 33. 38% respectively; 24, 48 hours after the transfection, 8.46 % and 7.53 % cells were induced to apoptosis respectively. Conclusion The siRNA against Survivin plasmid expression vector constructed in the study can blocking the expression of Survivin in PC-2 cell effectively and specifically; blocking the expression of Survivin can signiilcantly suppress the proliferation of PC-2 cell and induce a certain degree cells apoptosis; RNAi against Survivin is of a certain value in the gene therapy of pancreatic cancer.
4. Expression and mechanism of Twist2 in glioma
Lizhen WANG ; Weijia WANG ; Yifeng XIONG ; Shan XU ; Shanshan WANG ; Yi TU ; Zhaoyin WANG ; Xinling YAN ; Jinhong MEI ; Chunliang WANG
Chinese Journal of Pathology 2017;46(12):832-836
Objective:
To investigate the significance of Twist2 in glioma and whether it is involved in the malignant transformation of glioma by epithelial-mesenchymal transition (EMT).
Methods:
Using immunohistochemical method detected the expression level of Twist2 in 60 cases of gliomas (including WHO grades Ⅱ, Ⅲ and Ⅳ, each for 20 cases) and 20 cases of non-tumor brain tissues. Real-time fluorescence quantitative PCR and Western blot were used to detect the expression level of Twist2 mRNA and protein in 61 cases of fresh glioma tissue (WHO grade Ⅱ 16 cases, Ⅲ 21 cases, Ⅳ 24 cases) and 12 cases of adjacent tissues, and the expression levels of E-cadherin, N-cadherin and vimentin were also investigated in fresh glioma tissue.
Results:
Immunohistochemistry results showed that the percentages of Twist2 expression in glioma was 90%(54/60) compared with 30%(6/20) in non-tumor brain tissues(
5.Occupational Status of Physiotherapists in Four Regions in Asia
Ying WANG ; Zhaoyin ZHU ; Yi ZHU
Chinese Journal of Rehabilitation Theory and Practice 2018;24(9):1103-1109
After retrieving the databases, as well as the official websites of institutes and administrations of physiotherapists, the qualifications, working conditions and professional development in Chinese Taiwan, Chinese Hong Kong, Singapore and Japan were investigated and analyzed.
6.Application of bilateral lateral pre-decompression of distal and proximal digits in replant of degloving injury
Zhaoyin JIN ; Jie SUI ; Chaoqian ZHANG ; Qiao ZHU ; Guoping ZHANG ; Congbin TIAN ; Hui WANG ; Jinbo LIU
Chinese Journal of Microsurgery 2022;45(4):361-365
Objective:To explore the clinical application of distal and proximal bilateral lateral pre-decompression in replant of digit degloving injury.Methods:From March 2012 to May 2021, 14 patients with 29 digits had replantation surgery of degloved soft tissue and severed digits in Section II, Department of Orthopaedics, Changzhou Medical District of 904th Hospital of PLA Joint Logistic Support Force. There were 2 types of injuries: degloving injury of soft tissue but with intact digital tip, and digital tip degloving injury with intact distal phalanx and nail bed. With the technique of distal and proximal bilateral lateral pre-decompression, pre-decompression incisions were made to the subdermal on both sides of the degloved skin over the proper palmar digital arteries. From where, the distal stump of the proper palmar digital arteries and nerves for anastomosis were found and had them anastomosed with the proximal proper palmar digital arteries and proper palmar digital nerves, then anastomosed digital pulp and digital dorsal veins (11 cases were direct anastomosis and 3 cases were bridged anastomosis). Follow-up was carried out by outpatient consultation, telephone and WeChat APP. The appearance of the digit body, the shape of digital pulp, the nails grow and the motions of the digits were observed, and the Evaluation Standard of Replantation of Severed Fingers by the Society of Hand surgery of Chinese Medical Association was used to evaluate the recovery of function.Results:All 29 segments of degloving digit survived. Small necrotic areas was found in 4 digits and healed after the change of dressing. The followed-up time was lasted for 3-36 months. There was no obvious atrophy found in all the digits. Appearances of digit pulp and nails were satisfactory with good sensational recovery for TPD at 6-10 mm, 7 mm in average. Motions of all the repaired digits were good. According to the Evaluation Standard of Replantation of Severed Fingers by the Society of Hand surgery of Chinese Medical Association, 23 digits were in excellent, 4 in good and 2 in poor, with a satisfactory rate at 93.1%.Conclusion:For a distal digit injury with intact distal soft tissue and relatively mild injury of blood vessels and nerves, using the method of distal and proximal bilateral lateral pre-decompression can help to achieve good appearance in survived digital pulp, good sensational recovery and good nail growth. An individualised treatment intra-and-after the surgery could offer an ideal therapeutic effect.