1.Construction and identification of bicistronic retrovirus expression vector for Delta1 and EGFP gene coexpression
Fei HE ; Yinghui TAN ; Zhengron YANG
Journal of Third Military Medical University 2002;0(12):-
Objective To construct a bicistronic retrovirus expression vector to coexpress Delta1 and EGFP genes. Methods Delta1-IRES-EGFP bicistronic retrovirus vector was constructed by linking full-length Delta1 cDNA with IRES-EGFP retrovirus vector. Transfected with the retrovirus vector by electroporation, the ecotropic- and amphotropic-packaging cell lines GP+E86 and PA317 were selected by addition of puromycin. Positive cell clones were obtained and tested for the efficiency of the retroviral supernatants. Two positive cell clones with relatively highest titers were picked up and used for ping-pong amplification assay to improve the retroviral supernatants titer. At day 2 after transfection, GFP expression in NIH3T3 was detected by FACS, and Delta1 expression was determined by RT-PCR assay. Transfected or wild type NIH3T3 were co-cultured with myoblasts cell line C2C12 to test whether the exogenous Delta1 protein was functional or not. Results The bicistronic retrovirus expression vector coexpressing Delta1 and EGFP genes was successfully constructed and verified by PCR and DNA sequencing. A high-level expression of functional Delta protein was detected in transfected NIH3T3 cells. Conclusion Retrovirus is a safe, highly efficient gene transfer system.
2.Therapeutic effect and safety of finasteride for aged patients with isolated systolic hypertension compli-cated benign prostatic hyperplasia
Zhiwei BAO ; Feng YANG ; Yinghui WANG
Chinese Journal of cardiovascular Rehabilitation Medicine 2014;23(3):306-308
Objective:To analyze the therapeutic effect and safety of finasteride on aged patients with isolated systolic hypertension (ISH)complicated benign prostatic hyperplasia (BPH).Methods:A total of 84 aged ISH + BPH pa-tients admitted from Jun 2011 to May 2013 were selected.According to medication plan,they were divided into fin-asteride group (n=44)and routine treatment group (n=40,only received routine treatment because they cannot use finasteride).Course of treatment was all six months.International prostate symptom score (IPSS),quality of life (QOL) questionnaire score,prostate-specific antigen (PSA),incidence rate of acute urinary retention,maximum u-rinary flow rate (Qmax),postvoid residual urine (PRV),prostate volume (PV)and incidence rate of adverse reac-tions were compared between two groups.Results:Compared with routine treatment group,there were significant reductions in IPSS [(13.2±2.4)scores vs.(9.4±1.1)scores],PRV [(21.4±4.6)ml vs.(9.0±2.3)ml],PV [(36.8±3.8)ml vs.(29.5±3.0)ml],PSA [(2.45±0.74)μg/L vs.(1.26±0.48)μg/L]and incidence rate of acute urinary retention (17.50% vs.9.09%),P<0.05 or <0.01,and significant increase in Qmax [(10.2±2.4) ml vs.(13.7±3.8)ml]and QOL score [(3.1±1.0)scores vs.(4.7±1.5)scores]in finasteride group (P<0.05). There were no significant difference in incidence rates of adverse reactions between two groups,P>0.05 all.Con-clusion:Therapeutic effect of finasteride is significant and adverse reactions don’ t increase significantly in aged pa-tients with isolated systolic hypertension complicated benign prostatic hyperplasia.
3.Advances in the application of gene therapy for Parkinson's disease with adeno-associated virus.
Yang CHEN ; Yinghui Lü ; Zhaofa LI
Acta Pharmaceutica Sinica 2014;49(5):576-81
Vectors used to carry foreign genes play an important role in gene therapy, among which, the adeno-associated virus (AAV) has many advantages, such as nonpathogenicity, low immunogenicity, stable and long-term expression and multiple-tissue-type infection, etc. These advantages have made AAV one of the most potential vectors in gene therapy, and widely used in many clinical researches, for example, Parkinson's disease. This paper introduces the biological characteristics of AAV and the latest research progress of AAV carrying neurotrophic factor, dopamine synthesis related enzymes and glutamic acid decarboxylase gene in the gene therapy of Parkinson's disease.
4.Ultrastructure changes of cochlea and cochlear nerve after maxillofacial blast wound
Hua YANG ; Yinghui TAN ; Zumeng YA ; Zhongyu LI
Chinese Journal of Tissue Engineering Research 2005;9(10):228-229
BACKGROUND: There have been reports about the cochlea injury after high-velocity projectile wounding. The effect on ultrastructure of cochlea after blast was still unclear.OBJECTIVE: To investigate ultrastructural changes of cochlea and cochlear nerve after maxillofacial blast wound.DESIGN:A randomized controlled observational trail with dogs as subjects.SETTING: Otolaryngeal Department and Maxillofacial Surgery Laboratory of the Third Military Medical University of Chinese PLA.MATERIALS: The study was conducted from August 1995 through July 1997. The animal model in maxillofacial blast wound was established in Animal Center of Xinqiao Hospital of the Third Military Medical University of Chinese PLA. Specimens were treated in Maxillofacial Laboratory and observed in Electron Microscope Laboratory. Totally 15 dogs of either sex (weighting 9. 5 - 13.5 kg, mean 11.3 kg ) were randomly divided into three groups with 5 in each group. Two groups served as trauma groups 1 and 2 and the other group as control.METHODS: The maxillary and mandibular regions of 10 dogs in trauma groups were wounded by model 8 cardboard-shelled detonators to establish animal model of maxillofacial blast wound. At the 1st and 6th hour after trauma the wounds were examined and specimens of cochlea and cochlear nerve were dissected out for electron microscopic observation to study the ultrastructural changes. The specimens in the control group were treated the same way as those in the trauma groups except for blast injury.RESULTS: After wounded, the cochlea and cochlear nerve in the early period manifested cilia disorder, edema of the nerve and mitochondrial degeneration. At the 6th hour after trauma there were extensive degeneration in cochlea and cochlear nerve, cilia fallen off hair cells and dissolution of the structures in nerve sheath.CONCLUSION: The ultrastructural changes of cochlea and cochlear nerve are severed as a result of maxillofacial blast wound, but in early period the injury is reversible. So it is very important that early cure should be emphasized in treatment.
5.The clinical features of neurological disorders after pulmonary thromboendarterectomy for chronic thromboembolic pulmonary hypertension
Yonggang HAO ; Song GU ; Yinghui ZHA ; Wenli HV ; Yuanhua YANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2011;27(7):416-419
Objective Investigate the clinical features of the neurological disorders in patients after pulmonary thromboendarterectomy (PTE) for chronic pulmonary thromboembolism , analyze the factorial. Methods A retrospective study was made of 26 patients who underwent PTE between 2002 and 2010 in Beijing Chao-Yang hospital. The symptom of neurological system disorders occurred in the survivors were been investigate. The preoperative conditions and the perioperative conditions of all survivors were investigated. Compared the differences between the groups with neurological system disorders and the groups without. Results There were 22 patients of 26 survived after the surgery. Cardiac function of the survivors improved, and the quality of life improved significantly. 5 cases of the 22 survivors had symptoms of neurological system disorders. 3 patients showed lethargy, delirium, memory disorders, brain CT found no abnormal, symptoms recovered within 2 weeks. The fourth patient showed ataxia, unsteady gait, dance-like movements symmetry, in addition to the above symptoms, brain CT no abnormal showed,brain MRI showed bilateral abnormal signal in the midbrain, basal ganglia, symptoms improved and discharged after 8 weeks, the symptoms completely disappeared 6 months, and the abnormal signals in brain in MRI disappeared. The fifth patient with similar symptoms to the fourth, brain CT no abnormal found, be discharged 10 weeks after the operation, recovery is poor, living part of themselves. The postoperative neurological disorders occurred in the patients of Jamieson surgery type Ⅲ type, Ⅳ type of larger proportion, (P = 0.024), longer circulatory arrest surgery (P = 0.034). Conclusion The neurological disorders postoperative PTE often showed diffuse symmetric cerebral cortex and basal ganglia nerve dysfunction, brain MRI showed abnormal signal corresponding region, the majority of symptoms disappeared in 2 - 8 weeks, abnormal signal in brain MRI could disappeared after 6 months. Neurological dysfunction occurs in patients with more difficult surgical procedure, longer circulatory arrest, suggesting that with the surgery cerebral ischemia and hypoxia related.
6.Measurement of CD4~+CD 30~-Th/CD4~+CD 30 ~+Th subgroup in thrombocytopenic purpura patients by flow cytometer
Guisheng ZHOU ; Qiuxia YANG ; Yinghui SONG ; Al ET
Chinese Journal of Immunology 2000;0(11):-
Objective:Investigation about immunological pathogenesis of thrombocytopenic purpura was made.Methods:Th1 and Th2 cell number between thrombocytopenic purpura patients and normal people was detected by flow cemetry.Results:Th2 cell number in thrombocytopenic purpura patients was significantly higher than that in control group,whereas Th2 cell number in thrombocytopenic purpura patients was significantly lower than that in control group.Conclusion:The imbalance between Th1 and Th2 could partly explained the mechanism of thrombocytopenic purprua.
7.3D UTE-T2* analysis of achilles tendons disease and correlation with clinical score
Yang QIAO ; Hongyue TAO ; Yiwen HU ; Yinghui HUA ; Shuang CHEN
Chinese Journal of Medical Imaging Technology 2017;33(3):445-448
Objective To compare T2* value in healthy and diseased achilles tendons (AT) using 3D-ultra-short echo time (UTE),and to analyze the correlation between T2* value and clinical scores.Methods Fifteen patients (patients group) and ten volunteers (control group) were included and underwent an MR scan with UTE sequence.The T2* value of four ROI in AT were calculated.Independent sample t-test was used to compare the differences of T2* value between two groups.The correlation of T2* values with the American Orthopaedic Foot and ankle society ankle hindfoot scale (AOFAS) and achilles tendon total rupture score (ATRS) were analyzed.Results The T2* value of four ROI were statistically significantly higher in patients group than those of control group.There were negative correlation between T2* and AOFAS or ATRS scores (r=-0.92,-0.90,both P<0.01).Conclusion The T2* value of pathologic AT increases.The quantitative MRI UTE-T2* can evaluate AT from the biochemical structure and correlates with clinical outcomes,which is helpful to diagnose AT in the early stage.
8.Induced Cardiomyogenic Differentiation of Bone Marrow Mesenchymal Stem Cells in Vitro
Yan HUANG ; Yinghui LI ; Fen YANG ; Zhongquan DAI
Space Medicine & Medical Engineering 2006;0(06):-
Objective To investigate the effect of 5-azacytidine on cardiomyogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Method BMSCs were isolated from the marrow of adult SD rat’s femoral /tibial bones.Different concentration of 5-azacytidine were added to primary BMSCs on 3 d and cultured for different times.Cardiomyogenic differentiation of BMSCs was observed by immunohistochemistry and RT-PCR. Result After treated with 5-azacytidine, BMSCs proliferated slowly, became spindle-shaped after 10 d and aligned in a striated pattern after 20 d.TnT positive cells were showed by Immunohistochemistry and they expressed two cardiac-marked genes GATA-4 and ?-MHC.Thus 5-azacytidine induced cardiomyogenic differentiation of BMSCs in a time and concentration-dependent manner. Conclusion Our study suggests that bone marrow mesenchymal stem cells can differentiate into cardiomyocytes in vitro. They are ideal donor cells in cellular cardiomyoplasty for treatment of myocardial infarction.
9.Effects of endophytic fungi on growth and two kinds of terpenoids for Euphorbia pekinensis
Yinghui YONG ; Chuanchao DAI ; Fukang GAO ; Qiyin YANG ; Mo ZHAO
Chinese Traditional and Herbal Drugs 1994;0(07):-
Objective To study the producing method for Euphorbia pekinensis promoted by endophytic fungi.Methods The tissue culture plantlets were innoculated with endophytic fungi,then it was transplanted to the pods and cultured for one year.The biomass and content of terpenoids(isoeuphpekinensin and euphol)were determined.Results The growth of the E.pekinensis was promoted by endophytic fungi E4(Fusarium sp.Ⅰ)and E5(Fusarium sp.Ⅱ).The fresh weight of the root increased 49.45% and 59.74% for E4 and E5 treated,respectively.The plants dry weight coefficient increased 2.99% and 6.48% for E4 and E5 treated,respectively.The contents of isoeuphpekinensin and euphol in the plants were increased for tissue cultured in both bottles and pods.For the plants cultured in pods for one year,the isoeuphpekinensin and euphol increased 92.79% and 40%,respectively treated by E4.The isoeuphpekinensin and euphol increassed 105.32% and 241.38%,respectively treated by E5.Conclusion The endophytic fungi E4 and E5 could promote the growth of E.pekinensis and accumulated terpenoids.
10.The combination of dexmedetomidine and propofol for sedation in pediatric patients undergoing regional anesthesia
Yinghui WANG ; Baozhong YANG ; Zhixiang DUAN ; Yongkang CUI
Chinese Journal of Primary Medicine and Pharmacy 2015;(8):1166-1169,1170
Objective To evaluate the effectiveness and safety of the combination of dexmedetomidine and propofol for sedation in pediatrics undergoing regional anesthesia.Methods Forty-eight children aged 2 -6 years old,scheduled for elective surgery under regional anesthesia,were randomly divided into the two groups.The dexme-detomidine plus propofol group(group DP,n=24)received an infusion over 10 minutes of dexmedetomidine 0.7μg/kg and propofol 1.0 to 2.5 mg/kg bolus for induction,then an infusion of dexmedetomidine 0.5μg·kg-1·h-1 and propofol 100-300μg·kg-1·min-1 for maintenance.The propofol group(group P,n=24)received a propofol 1.0 to 2.5mg/kg bolus for induction,then an infusion of propofol 150 -300μg·kg-1 ·min-1 for maintenance.If body movement was detected during surgery,a bolus of propofol 1 .0mg/kg was administered in both groups,and if there was another body movement within three minutes of the propofol bolus,a bolus of ketamine 1 .0mg/kg was adminis-tered.Hemodynamic data,respiratory rate,and oxygen saturation were recorded by researchers blinded to the study drugs.Recovery time and the primary outcome were evaluated by a modified Aldrete score.The occurrence of adverse events and the amount of supplemental ketamine used were also recorded.Results The consumption of propofol in the group DP was (149.00 ±24.93)μg·kg-1·min-1,and in the group P was (217.00 ±17.90)μg·kg-1· min-1.The propofol consumption was significant lower that in group DP(t=117.94,P=0.00).The needs for sup-plemental ketamine bolus in the group DP was 0.50(1.00)/case,and it was 2.00(1.00)/case in group P.The need for supplemental ketamine was less in group DP(Z=-4.48,P=0.00).Hypopnea occurred in 5 cases in group P, and 0 in group DP.The incidence of hypopnea was more in group P(χ2 =5.58,P=0.03).In group DP,the mean arterial pressure and heart rate at discharge were respectively (74.96 ±5.69)mmHg and (101.17 ±9.65)bpm,and their baseline values were (81.13 ±7.23)mmHg and (112.00 ±13.65)bpm.Both mean arterial pressure and heart rate in the group DP remained decreased at discharge(t=10.78,P=0.00;t=10.08,P=0.00;respectively).The discharge time of the group DP was (40.91 ±10.90)min and it was(39.67 ±14.95)min in group P.There was no significant difference in the discharge time(t=0.11,P=0.74).Conclusion The combination of dexmedetomidine and propofol can provide appropriate depth of sedation in pediatric patients undergoing regional anesthesia.