1.Effects of RNA interference targeting HIF-1α on location and metastasis in HeLa cells
Weiguang WU ; Yaqiong CHEN ; Xiuqin CAO
Basic & Clinical Medicine 2010;30(3):284-288
Objective To construct an eukaryotic vector expressing short hairpin RNA(shRNA) of HIF-1α,and to observe its effects on location and metastasis of HeLa cells under hypoxic condition.Methods shRNA templates was developed based on HIF-1α gene sequence and then cloned into pSilencer2.1-U6-neo vector.The resultant plasmid was transfected into HeLa cells with Lipofectamine 2000.The cells were incubated in hypoxic condition.The HIF-1α protein and mRNA were detected by Western blot and RT-PCR.The colony formation assay and Transwell cabin assay were performed to measure the colony formation and metastasis.Results The plasmid pSilencer2.1-U6-neo-HIF-1α was successfully constructed and transfected into HeLa cells.The expression of HIF-1α in HeLa cells decreased,and the number of colony formation in soft agar and cells penetrating matrigel also decreased under hypoxic condition.Conclusion The shRNA expressing plasmid targeting at HIF-1α may suppress the location and metastasis of cervical carcinoma cells under hypoxic condition.
2.Efficacy and safety of tirofiban treatment combined with percutaneous coronary intervention in the elderly with acute ST segment elevation myocardial infarction
Weiguang HUANG ; Jingyun LUO ; Jin CUI ; Qiang ZHAO ; Tongguo WU
Journal of Chinese Physician 2011;13(7):883-886
Objective To analyze the efficacy and safety of tirofiban treatment combined with percutaneous coronary intervention (PCI) in the elderly with acute ST segment elevation myocardial infarction prospectively. Methods From May 2008 to May 2010, 106 patients who presented with acute STEMI within 12 hours from onset and received successful primary PCI were enrolled into the study. All patients had angiographic evidence of initial total occlusion of infarct-related artery and finally restored toTIMI3 flow after PCI. All patients were divided into tirofiban group (n = 54) and control group (n = 52) according to whether tirofiban was used or not. Assessment of myocardial perfusion included Myocardial Blush Grades (MBG), and the resolution of the sum of ST-segment elevation (sumSTR) at 90 minutes after the procedure. Left ventricular ejection fraction (EF) was measured one week later. Major adverse cardiac events in hospital and bleeding complications were also assessed. Results Baseline clinical and angiographic characteristics of the two groups were similar. Significant higher rates of MBG 3 were observed in the tirofiban group (88. 9% vs57. 7%, P < 0.05). Patients received tirofiban were more likely to achieve higher sumSTR (70. 3% vs 42. 3%, P <0. 05). Ejection fraction was also markedly increased in tirofiban group than control group (56. 2 ± 7.6 vs 46. 7 ± 8. 5, P < 0. 05). In-hospital major adverse cardiac events, it was not different between the two groups(P >0. 05). There were slightly more minor bleeding complications in tirofiban group compared with control(11.1% vs 6. 0%, P >0. 05). No patient had major bleeding or thrombocytopenia.Conclusions Tirofiban can further ameliorate microvascular perfusion and it is safe and feasible for patients with STEMI undergoing primary PCI.
3.SAHA inhibition of the human ovarian cancer cell line SKOV3 in vitro
Zheng WANG ; Weiguang WU ; Hongliang YAN ; Zhaoling SUN ; Yajuan TANG
Chinese Journal of Clinical Oncology 2013;(12):698-701
10.3969/j.issn.1000-8179.2013.12.004
5.Inhibition of STAT3 expression on chemosensitivity of ovarian cancer cell line SKOV3
Yongmei WANG ; Weiguang WU ; Hongyu GE ; Jianqiu HAN
China Oncology 1998;0(04):-
Background and purpose:The signal transducers factor and activator in transcription 3(STAT3) is a recently studied gene from a variety of solid tumors such as breast, stomach, and ovarian cancer, in which the increase of abnormal expression and activity are accompanied.The aim of this study was to investigate the effects of eukaryotic vectors that express short hairpin RNA(shRNA) in signal transducers and activators of STAT3 on chemosensitivity of human ovarian cancer SKOV3 cells.Methods:Vectors containing shRNA targeting STAT3(pSTAT3-siRNA) were constructed and transfected into SKOV3 cells.These vectors were then divided into 3 groups:SKOV3, SKOV3NS, and SKOV3siRNA.The mRNA and protein expressions of STAT3 were determined by RT-PCR and Western blot, respectively.The growth inhibition and apoptosis rates of the different group cells under chemotherapeutic agents such as cisplatin(20 ?mol/L) were measured by MTT assay and flow cytometry(FCM).Results:MTT assay growth inhibition rates in the tumor cells of the SKOV3 group, SKOV3NS group and SKOV3siRNA group had inhibition rates of 0.46?0.13, 0.44?0.11 and 0.71?0.12.Compared with the SKOV3, SKOV3NS and SKOV3siRNA group, there was a marked increase of SKOV3siRNA group in inhibition rate of cells.The differences were also statistically significant(P0.05).The SKOV3 group, SKOV3NS group and SKOV3siRNA group's cell apoptosis rates were 18?4, 18?3 and 35?4, respectively.However, the SKOV3 group, SKOV3NS group, and SKOV3siRNA group cell apoptosis were significantly increased and the differences were statistically significant(P0.05).The results for the SKOV3 group, SKOV3NS group and SKOV3siRNA in cell STAT3 mRNA were 0.50?0.08, 0.48?0.07 and 0.31?0.09.With the SKOV3 group, SKOV3NS group, SKOV3siRNA group of cells in STAT3 mRNA, its expression in the lungs were significantly lower and the differences were statistically significant(P0.05).SKOV3 group, SKOV3NS group and SKOV3siRNA group of cells checked the results of STAT3 protein were 0.54?0.09, 0.56?0.08 and 0.32?0.09, respectively.The SKOV3 group, SKOV3NS group, and SKOV3siRNA group in STAT3 protein expression was significantly lower and the differences were statistically significant(P0.05).Conclusion:The STAT3 specific shRNA expression vector could effectively suppress the expression level of STAT3 gene in SKOV3 cells as well as enhance their sensitivity to cisplatin.
6.Evaluation of the efficacy and safety of human coagulation factor Ⅷ in the treatment of hemophilia A patients
Ruyi CHEN ; Yan WU ; Yiyun LIU ; Mingxia HOU ; Qingshuang SONG ; Xuanlin ZHONG ; Xueyun WANG ; Wenjie XIE ; Caiping GUO ; Zhan ZHANG ; Yunjia ZHANG
Chinese Journal of Blood Transfusion 2022;35(12):1220-1225
【Objective】 To evaluate the efficacy and safety of human coagulation factor Ⅷ developed by Shenzhen Weiguang Biological products Co, Ltd in the treatment of patients with hemophilia A. 【Methods】 A prospective, multi-center, open, single-group clinical study was conducted. A total of 65 subjects with hemophilia A were enrolled, and human coagulation factor Ⅷ(FⅧ) was injected according to the patients’ bleeding severity. The improvement score of bleeding symptoms and signs after the first infusion of the first bleeding event and the transfusion efficiency of FⅧ activity at 10 min and 1 hour after infusion were taken as the main efficacy indexes. The improvement scores of bleeding symptoms and signs after the first infusion and the increase of FⅧ activity at 10 min and 1 hour after infusion were the secondary efficacy indexes. 【Results】 The 65 subjects were enrolled in safety analysis set (SS) and full analysis set (FAS), and 58 of them were enrolled in protocol analysis set (PPS). Ten minutes and one hour after the first infusion, the level of factor Ⅷ activity in the subjects increased significantly, and the FⅧ activity increased by 100% or more in more than 79% of the subjects. The average infusion efficiency of FⅧ activity in all subjects was more than 100%. In 70% of the subjects, the pain was relieved rapidly and /or the bleeding symptoms were significantly improved 8 hours after each bleeding infusion, and the improvement rate of bleeding symptoms and signs reached 100% 72 hours after infusion. 【Conclusion】 After infusion of human coagulation factor Ⅷ, the activity level of factor Ⅷ in patients with hemophilia A significantly increased. The infusion efficiency can reach a optimal level, and the bleeding symptoms can be significantly improved.
7.A multicenter phase Ⅲ clinical study of human prothrombin complex concentrate in treatment of hemophilia B
Wei ZHANG ; Yirun LIU ; Yan WU ; Xuanlin ZHONG ; Qingshuang SONG ; Shitao CHEN ; Xueyun WANG ; Caiping GUO ; Zhan ZHANG ; Yunjia ZHANG
Chinese Journal of Blood Transfusion 2022;35(9):915-919
【Objective】 To evaluate the clinical efficacy and safety of one kind of human prothrombin complex concentrate in treatment of patients with hemophilia B. 【Methods】 The clinical data of 36 patients with hemophilia B treated with human prothrombin complex concentrate produced by Shenzhen Weiguang Biological Products Co. Ltd. from May 2018 to April 2019 were retrospectively analyzed, and its clinical efficacy and safety were analyzed. 【Results】 A total of 35 subjects entered the full analysis set (FAS)and safety set (SS), 33 subjects entered the per protocol Set (PPS). Thirty minutes after the first infusion of FAS subjects, the activity of coagulation factor Ⅸ increased from (3.93±0.975) IU/dL to (25.61±9.337) IU/dL, and the infusion efficiency was (96.43±22.007)%. The increased value of coagulation factor Ⅱ activity was (73.25±14.874) IU/dL. The activity of coagulation factor Ⅶ was (42.79±16.847) IU/dL. The increased value of coagulation factor Ⅹ activity was (65.29±17.042) IU/dL. The increased value of coagulation factor Ⅸ activity was (21.68±9.434%) IU/dL. Twenty-four hours after the first infusion of FAS subjects, the improvement of bleeding symptoms and signs was excellent in 21 cases (60%), improved in 14 cases (40.0%), and the effective rate was 100%. The incidence of adverse reactions was 2.9%(1/35), and there was no antibody to human coagulation factor Ⅸ and new virus infection. 【Conclusion】 Infusion of human prothrombin complex concentrate produced by Shenzhen Weiguang Biological Products Co. Ltd. in the treatment of hemophilia B has significant clinical efficacy and good safety.
8.Methodological evaluation of nephelometric assay for the determination of IgA residues in human intravenous immunoglobulin
Mingxia HOU ; Yan WU ; Meiling DING ; Xi′e ZHEN ; Qinghui FU ; Huan ZENG ; Wenjie XIE ; Zhan ZHANG ; Yunjia ZHANG
Chinese Journal of Blood Transfusion 2021;34(10):1090-1093
【Objective】 To establish and evaluate a nephelometric assay for the determination of immunoglobulin A (IgA) residues in human intravenous immunoglobulin(IVIG). 【Methods】 BN ProSpec© automatic protein analyzer and its supporting immunoglobulin A determination kit (nephelometry) produced by German Siemens and the national standard of human IgA were used to establish the nephelometric assay to determine IgA residue in test products and verify the methodology. The test products include IVIG (pH4) prepared by low-temperature ethanol protein separation process and a novel IVIG prepared by chromatography. 【Results】 The average deviation of three calibration curves for IgA residues determination by the nephelometric assay were 1.08%, 0.95% and 1.54%,, and the three deviations of the quality control were 4.00%, -2.30% and -0.20%, respectively, which indicated good calibration and quality control. In the specificity test, the average recovery rates of IgA for reference substance 1 containing 100g/L maltose and reference substance 2 containing 20g/L glycine were 102.7% and 105.8%, respectively. The relative standard deviation (RSD) values of the repeatability tests of the two test products were 3.9% and 1.9%, and the RSD values of the intermediate precision test were 3.6% and 2.3%, respectively.The difference values at each time point in the durability test of test products′ storage time were all less than 10%, and the RSD values of the two test products in the durability test of kits of different batches were 2.8% and 2.2%, respectively. In the accuracy test, the average recovery rates of IVIG (pH4) added to the standard were 94.2%, 101.7% and 96.2%, respectively, and the average recovery rates of the novel IVIG added to the standard were 102.8%, 106.3% and 99.7%, respectively. The average recovery rate of the limit quantification test was 101.0%, and the RSD was 4.0%. 【Conclusion】 Nephelometric assay has the advantages of strong specificity, high precision and accuracy, good repeatability, simple and rapid operation, and automation, and can be used for the determination of IgA residue in IVIG (pH4) and novel IVIG products.
9.Association of OPG gene single nucleotide polymorphisms with susceptibil-ity to rheumatoid arthritis in Chinese Han population
Yueming CAI ; Xia LONG ; Qingwen WANG ; Jing WANG ; Zhicheng WU ; Weiguang WANG ; Huiping ZENG ; Lu ZHANG
Chinese Journal of Pathophysiology 2014;(7):1204-1208
AIM: To investigate the association of osteoprotegerin ( OPG) gene single nucleotide polymor-phisms (SNPs), 163A/G (rs3102735) and 245T/G (rs3134069), with susceptibility to rheumatoid arthritis (RA) in Chinese Han population .METHODS:A total of 205 patients with RA and 171 healthy control subjects were enrolled into this study.Genotyping was performed by polymerase chain reaction-based restriction fragment length polymorphism and subsequently confirmed by DNA sequencing .Odds ratio ( OR) and 95%confidence intervals ( CI) were calculated for the risk genotypes and alleles .RESULTS: OPG gene polymorphisms 163A/G and 245T/G were conformed to the Hardy-Weinberg equilibrium .The statistical differences in the genotypes of AA , AG and GG at 163A/G locus were found in RA and controls.The G allele was associated with an increased risk of RA , with OR of 1.219 (95%CI:1.066~2.339).No significant difference was observed between RA group and control group with respect to genotypic and allelic frequencies of OPG gene 245T/G (P>0.05).CONCLUSION:The OPG gene 163A/G SNP may be associated with RA susceptibility , and G allele may be the risk factor for developing RA .
10.Bone Marrow Mesenchymal Stem Cell Transplantation in the Repair of Rat Spinal Cord Hemisection Injury
Jinsheng WU ; Aiping DONG ; Xiaocui WANG ; Zhixin WEI ; Weiguang LIU ; Zhimin LUAN ; Zengjun ZHU
Acta Laboratorium Animalis Scientia Sinica 2010;18(1):-
objective To investigate the differentiation of bone marrow-derived mesenchymal stem cells into neurons and transplantation of the stem ceils to repair rat hemisection spinal cord injury.Methods Adherent culture was used to isolate and culture rat bone marrow mesenchymal stem cells(MSCs).The rat spinal cord homogenate supernatant was used to induce neural differentiation of the 3rd generation ceils.The nature of ceil differentiation was identified by immunohistochemistry.The rat model of hemisection spinal cord injury was prepared and BrdU was locally injected to label the induced neurons.The distribution of living cells in the injuried spinal cord was observed at 5 weeks after cell transplantation.Results MSCs were spindle and polygonal,with 1-2 nucleoli seen under the inverted microscope.After induction with spinal cord homogenate supernatant there were a number of slender cytoplasmic projections forming interwined network and showing nestin expression,therefore,indicating the neuronal nature.MSCs at 5 weeks after transplantation into the spinal cord injury were surviving and their expression of MAP-2,NF,GFAP was significantly higher than that in the control rats(P<0.05).The rat motor function was improved than before transplantation.Conclusion MSCs induced by spinal cord homogenate supernatant can be transplanted into hemisection spinal cord injury and improve the motor function of the injuries lesions.