1.Study on the expression of ?_2-adrenergic receptor gene and enhanced green fluorescent protein gene in cardiomyocytes transferred by adeno associated virus vector
Wenqian GAO ; Xiaoying LI ; Xiaobing WU
Chinese Journal of Geriatrics 2000;0(04):-
Objective To investigate the way of gene therapy on congestive heart failure and other heart diseases. Methods Recombinant adeno associated viruses (AAV) encoding the human ? 2 adrenergic receptor(AR) and enhanced green fluorescent protein (EGFP) were constructed and the ? 2 AR and EGFP cDNA were transfected in vitro to the neonate rat ventricular myocytes. The expression of ? 2 AR and EGFP on cultured neonate rat ventricular myocytes and the ability of ? 2 AR to potentiate the beta adrenergic signaling were assessed. Results The presence of human ? 2 AR mRNA was demonstrated by Northern blot, the expression of the ? 2 AR gene was demonstrated by Westernimmuno blot. The density of ? AR in the myocytes was greater than that in the control 〔(204.0 ? 6.4) fmol/mg protein vs (76.0 ? 2.8) fmol/mg protein〕 according to a ligand binding assay. Human ? 2 AR gene significantly increased isoproterenol stimulated cAMP as compared with the control〔(116.2?5.8) pmol/10 6 cell vs (58.4 ? 4.6) pmol/10 6 cell〕. EGFP was also expressed in the transfected cardiomyocytes. Conclusions Human ? 2 AR gene could be transfected into cardiomyocytes by AAV vector and could potentiate the beta adrenergic signaling.
2.Observation of the Additional Efficiency of Aldosterone Blockers During Angiotensin Ⅱ Receptor Blockers Treatment on Hypertensive Left Ventricle Hypertrophy
Wenqian GAO ; Ping ZHU ; Qiang WU ; Yutao GUO
Journal of Medical Research 2006;0(01):-
Objective To evaluate the additional efficiency of aldosterone blockers (AB) during angiotensin Ⅱ receptor blockers (ARB) treatment on hypertensive left ventricle hypertrophy in the elderly. Methods Forty-five cases of hypertensive left ventricular hypertrophy detected by Doppler ultrasonic cardiogram examination were enrolled,and were randomized to receive treatment with ARB plus AB (Group A) or ARB plus diuretics (Group B) for 48 weeks. The indicators of left ventricle hypertrophy were checked before and after the treatment. Results The both regimens could significantly decrease the blood pressure,and there was significant difference in the effects on lowering blood pressure between the regimens. Both regimens could alleviate left ventricle hypertrophy,but the regimen of ARB plus AB was superior to the regimen in Group B. Conclusion The addition of AB treatment during the ARB treatment was a good antihypertensive regimen for reducing left ventricle hypertrophy.
3.Circulating microRNA-21 in hepatocellular carcinoma patients
Lan GAO ; Huifang LIU ; Mingzhi LING ; Wenqian TIAN ; Mingjing ZHAO ; Wenxin ZHAO ; Poshi XU
Chinese Journal of Laboratory Medicine 2015;(7):484-486
Objective To evaluate the plasma level of miR-21 in hepatocellular carcinoma (HCC) patients and its clinical diagnostic significance .Methods Case-control study was used .The plasma level of miR-21 was measured by real-time quantitative PCR.The relative expressions of miR-21 were calculated. This study included 60 cases of patients with HCC , 71 patients with liver cirrhosis ( LC ) , 52 healthy volunteers ( HV) from January to June in 2014 in Henan Province People′s Hospital.The receiver operating characteristic ( ROC) curves were analyzed to determine the sensitivity and specificity of miR-21 expression levels in HCC diagnosis. Differences between groups were assessed by the t-test.Results Plasma microRNA-21 level in the 60 patients with hepatocellular carcinoma ( 2.6 ±1.1 ) was significantly higher than in patients with chronic hepatitis (1.6 ±0.9) and healthy volunteers (1.0 ±0.6) (t=5.322,P=0.004;t =8.349, P =0.000 3, respectively ) .Plasma microRNA-21 level in the HCC patients were positively correlated with tumor size and differentiation (tdif=3.366,P=0.019;tsize =3.490,P=0.012). ROC analysis of plasma microRNA-21 yielded an AUC of 0.796 with 70.0% sensitivity and 65.3%specificity when differentiating hepatocellular carcinoma from chronic hepatitis , and an AUC of 0.934 with 89.5% sensitivity and 81.8% specificity when differentiating hepatocellular carcinoma from healthy volunteers.Conclusion The plasma level of miR-21 in HCC patients has high specificity , and maybe help to diagnose of HCC.
4.In vitro antifungal activity of four antifungal agents alone or in combination against Exophiala dermatitidis biofilms
Wenqian ZHENG ; Yi SUN ; Lujuan GAO ; Qingzhi WU ; Ming LI ; Tongxiang ZENG
Chinese Journal of Dermatology 2018;51(1):51-53
Objective To evaluate the in vitro antifungal activity of 4 antifungal agents alone or in combination against Exophiala dermatitidis (E.dermatitidis) biofilms.Methods E.dermatitidis biofilms were prepared by using a modified 96-well plate-based method.The in vitro antifungal activity of amphotericin B,voriconazole,itraconazole and caspofungin alone or in combination against E.dermatitidis biofilms were investigated via the broth microdilution checkerboard technique.Results The sessile minimum inhibitory concentration ranges resulting in 50% (SMICS0) and 80% inhibition (SMIC80) of E.dermatitidis biofilms were all > 32 mg/L for itraconazole,voriconazole and caspofungin,and the SMIC50 and SMIC80 ranges of amphotericin B were 1-2 mg/L and 4-8 mg/L respectively.The combination of amphotericin B with voriconazole showed synergistic inhibitory effects against E.dermatitidis biofilms,while the combination of amphotericin B with itraconazole or caspofungin,as well as the combination of voriconazole with caspofungin,revealed no synergistic effects.No antagonistic effect was observed in any of the combinations.Conclusion Amphotericin B appears more active against E.dermatitidis biofilms,and the combination with voriconazole can enhance the anti-biofilm effects against E.dermatitidis biofilms.
5.GRK 2 level in peripheral blood lymphocytes of elderly patients with acute myocardial infarction
Wenqian GAO ; Chunguang HAN ; Xiaochun LU ; Yongxue LIU ; Haipeng HUI ; Hao WANG
Journal of Geriatric Cardiology 2013;(3):281-285
Objective To investigate the G protein-coupled receptor kinase 2 (GRK 2) level in peripheral blood lymphocytes with cardiac func-tion in elderly patients with acute myocardial infarction. Methods This study enrolled 40 patients with acute ST-segment elevation myo-cardial infarction (STEMI) and 40 patients with unstable angina. All patients were 65 years or older. Cardiac function was evaluated by echocardiography, and the GRK 2 level in peripheral blood lymphocytes was measured. Patients with STEMI were followed up for 2 years. Results The GRK 2 level in peripheral blood lymphocytes was significantly higher in patients with STEMI than in patients with unstable angina, and was negatively correlated with left ventricular ejection fraction, cardiac output, stroke volume, and left ventricular fractional shortening. The GRK 2 level was significantly elevated in some patients with acute STEMI and poor cardiac function. Conclusions In-creased GRK 2 level in patients with acute STEMI may contribute to poor myocardial systolic function and myocardial remodeling. Meas-urement of the GRK 2 level in peripheral blood lymphocytes may assist in the evaluation of cardiac function and myocardial remodeling in elderly patients with acute STEMI.
6.Role of miR-26a/b in AngⅡ-induced hypertensive vascular remodeling
Yishan ZHANG ; Miao YUAN ; Yuan HUANG ; Qiaozhu WANG ; Wenqian ZHANG ; Dengfeng GAO
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(5):649-654
Objective To detect the expression of miR-26a/b in the aorta and serum of mice so as to explore the role of miR-26a/b in vascular remodeling of hypertension.Methods C576L/BJ male mice were randomly divided into AngⅡ group and control group.Mini-osmotic pump was implanted subcutaneously into the back of mice, and the model of blood vessel remodeling in mice was established by continuous infusion of AngⅡ(2.0mg/kg·d).The mice in control group were injected with saline.Blood pressure was taken before the intervention and at 3, 5, 7, 10 and 14 days after the intervention.After 2 weeks, the mice were killed, the serum and aorta tissues were collected, and the expression of miR-26a/b was determined by RT-PCR.HE staining, Masson staining and immunohistochemistry were performed to observe changes in vascular morphology, fibrosis and protein expression.Results After the intervention, systolic blood pressure and diastolic blood pressure were significantly higher in AngⅡ group than in control group (P<0.05).HE staining showed that the vessel wall of AngⅡ group was thicker than that of control group.Masson staining showed more blue collagen deposition in the middle of aorta in AngⅡ group but no obvious collagen deposition in control group.RT-PCR showed that the expression of miRNA-26a/b in the serum and aorta of AngⅡ group was significantly lower than in control group (P<0.05).Immunohistochemistry indicated that the expressions of CTGF, collagen Ⅰ and collagen Ⅲ all increased after AngⅡ infusion (P<0.05).Conclusion MiR-26a/b, CTGF, collagen Ⅰ and collagen Ⅲ may be involved in AngⅡ-induced vascular remodeling in hypertension.MiR-26a/b may be a new therapeutic target of vascular remodeling in hypertension.
7.Role of angiotensin Ⅱ type 2 receptor in repeated propofol anesthesia-induced neuroapoptosis in basal ganglia of newborn rats
Jinmin LIU ; Hemao ZHANG ; Wenqian LI ; Hong GAO ; Qingfan ZENG
Chinese Journal of Anesthesiology 2018;38(5):548-551
Objective To evaluate the role of the angiotensin Ⅱ type 2 receptor (AT2R) in repeated propofol anesthesia-induced neuroapoptosis in the basal ganglia of newborn rats.Methods Fiftyfour pathogen-free Sprague-Dawley rats,aged 7 days,weighing 10-15 g,were divided into 3 groups (n=18 each) using a random number table:control group (group C),repeated propofol anesthesia group (group P) and AT2R agonist CGP42112A group (group G).In group C,0.9% sodium chloride injection 3 ml/kg was intraperitoneally injected,and half of the initial dose 1.5 ml/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.In group P,propofol 30 mg/kg was intraperitoneally injected,and half of the initial dose 15 mg/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.In group G,CGP42112A 1 mg/kg was intraperitoneally injected,propofol 30 mg/kg was intraperitoneally injected 5 min later,and half of the initial dose of propofol 15 mg/kg was given every 20 min for 5 times in total,lasting for 3 consecutive days.Six rats were sacrificed at 2 h after emergence from anesthesia,and brains were removed for detection of neuroapoptosis in the basal ganglia by TUNEL assay.The apoptosis index was calculated.Another 6 rats were sacrificed,and the basal ganglia were isolated from brains to detect the expression of activated caspase-3,AT2R and peroxisome proliferator-activated receptor gamma (PPARγ) (by Western blot) and the expression of AT2R and PPARγ mRNA (by real-time polymerase chain reaction).The other 6 rats were fed until 28 days old,and the cognitive function was then assessed using Morris water maze test.Results Compared with group C,the escape latency was significantly prolonged,the time of staying at the target quadrant was shortened,the frequency of crossing the platform was decreased,the apoptosis index of the basal ganglia was increased,the expression of activated caspase-3 was up-regulated,and the expression of AT2R and PPARγprotein and mRNA was down-regulated in group P (P<0.05),and no significant change was found in the parameters mentioned above in group G (P>0.05).Compared with group P,the escape latency was significantly shortened,the time of staying at the target quadrant was prolonged,the frequency of crossing the platform was increased,the apoptosis index of the basal ganglia was decreased,the expression of activated caspase-3 was down-regulated,and the expression of AT2R and PPARγ protein and mRNA was up-regulated in group G (P<0.05).Conclusion Inhibited activation of AT2R is involved in repeated propofol anesthesia-induced neuroapoptosis in the basal ganglia of newborn rats.
8.Hepatitis B virus (HBV) infection and postpartum haemorrhage:A meta-analysis
Haiying LIANG ; Qitao HUANG ; Lin ZHOU ; Qianqian MA ; Qiumin SHE ; Wenqian CHEN ; Yunfei GAO ; Yanhong YU ; Mei ZHONG
The Journal of Practical Medicine 2016;32(12):2035-2040
Objective To investigate the risk of postpartum haemorrhage in HBV-infected pregnant women. Methods Cohort or case-control studies that discussed the relationship between hepatitis b virus infection and pregnancy outcome were searched in PubMed , EMBASE , Wiley Online Library , Cochrane Library , Google Scholar, CBM, WanFang database and CNKI etc. (till August 2015). The quality of included Cohort or case-control studies was evaluated , and Meta-analysis was performed with Rev Man5.2 software. Results Four observational case-control studies and 17 cohort studies , involving 19 549 women in observation , were identifled. Meta-Analysis results displayed that the incidence of postpartum haemorrhage in HBV-infected women was 9.3%, while 2.8% in women without HBV [RR = 2.97, 95% CI (2.25 ~ 3.92),P < 0.01]. Compared with normal-risk women , the incidence of postpartum haemorrhage of HBV-infected women with normal hepatic function was also higher [RR = 2.56, 95% CI (2.01 ~ 3.25),P < 0.01]. HBV-infected women with hepatic dysfunction had higher incidence of postpartum haemorrhage than those with normal hepatic function [RR = 2.67, 95% CI (2.17~ 3.28),P < 0.01]. Conclusions HBV-infected women are at higher risk of postpartum haemorrhage than normal pregnancy women and further hepatic dysfunction would lead to a continuing increase of the risk.
9.Effects of photodynamic therapy alone or in combination with antifungal agents on the apoptosis of planktonic and biofilm cells of Exophiala dermatitidis
Yuting XU ; Wenqian ZHENG ; Lujuan GAO ; Yi SUN ; Linyun LI ; Ming LI ; Tongxiang ZENG
Chinese Journal of Dermatology 2018;51(7):515-518
Objective To evaluate the in vitro effects of photodynamic therapy alone or in combination with antifungal agents on the apoptosis of planktonic and biofilm cells of Exophiala dermatitidis (E.dermatitidis).Methods The planktonic suspensions of E.dermatitidis were prepared,and the biofilms of E.dermatitidis were prepared via a modified 96-well plate-based methods.Planktonic and biofilm cells of E.dermatitidis were separately divided into several groups:antifungal agent groups treated with antifungal agents alone,photodynamic therapy group receiving photodynamic therapy alone,combination groups receiving photodynamic therapy followed by the treatment with antifungal agents,and blank control group receiving no treatment.These antifungal agents included amphotericin B,posaconazole,voriconazole and itraconazole.The concentrations of these antifungal agents were all 1 mg/L,and the treatment with antifungal agents lasted 2 hours.Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to detect the apoptosis of planktonic and biofilm cells of E.dermatitidis in all the groups.Results The antifungal agents and photodynamic therapy both affected the apoptosis of planktonic (both P < 0.001) and biofilm cells (beth P < 0.05) of E.dermatitidis.The apoptosis rates of E.dermatitidis planktonic cells in the control group,amphotericin B group,posaconazole group,voriconazole group and itraconazole group were 11.67% ± 0.21%,13.30% ± 1.78%,14.30% ± 3.61%,14.51% ± 1.91%and 36.17% ± 4.00% respectively.The apoptosis rate of E.dermatitidis planktonic cells was significantly higher in the itraconazole group than in the control group (P < 0.05),but no significant differences were observed between the other 3 antifungal agent groups and control group (all P > 0.05).The photodynamic therapy group also showed a significantly higher apoptosis rate of E.dermatitidis planktonic cells (41.37% ±7.80%) compared with the control group (P < 0.05).After the treatment with photodynamic therapy combined with amphotericin B,posaconazole,voriconazole or itraconazole,the apoptosis rates of E.dermatitidis planktonic cells were 29.23% ± 6.71%,37.23% ± 10.86%,43.57% ± 6.42% and 69.87% ± 3.53% respectively.Moreover,the photodynamic therapy + voriconazole group and photodynamic therapy + itraconazole group both showed significantly higher apoptosis rates compared with the voriconazole group and itraconazole group respectively (both P < 0.05).The apoptosis rate of E.dermatitidis biofilm cells was significantly higher in the photodynamic therapy group than in the control group (32.00% ± 0.43% vs.25.30% ± 1.31%,P < 0.05),as well as in the photodynamic therapy + amphotericin B than in the amphotericin B group (P < 0.05).Conclusion Photodynamic therapy combined with antifungal agents can markedly promote the apoptosis of planktonic and biofilm cells of E.dermatitidis.
10.Treatment of patients with orthopaedic trauma during COVID-19 epidemic
Xiao CHANG ; Yang SONG ; Wenqian WANG ; Yu FAN ; Baozhong ZHANG ; Jia ZHANG ; Xiongfei ZOU ; Peng GAO ; Bin YU ; Na GAO ; Jianguo ZHANG
Chinese Journal of Orthopaedic Trauma 2020;22(7):561-566
Objective:To report our experience in treatment of patients with orthopaedic trauma during COVID-19 epidemic.Methods:We retrospectively analyzed the 67 patients with orthopedic trauma who had been treated at Department of Orthopaedics, Peking Union Medical College Hospital from February 1 to March 31, 2020. After screening for COVID-19 infection was performed under strict protection, the patients were diagnosed and assigned to outpatient emergency treatment or hospitalization according to their specific condition. Twenty-six patients were treated at the outpatient emergency department. They were 8 males and 18 females with an average age of 69.5 years. Of them, 6 with vertebral compression fracture were placed on bed brakes, 14 with limb fracture immobilized after close reduction, 2 with skin laceration treated with debridement and suture, and 4 with hip fracture immobilized in bed. In the 41 hospitalized patients, there were 14 males and 27 females with an average age of 68.5 years. In them, hemiarthroplasty was performed for 7 femoral neck fractures, kyphoplasty for 5 vertebral compression fractures, total elbow arthroplasty for one humeral intercondylar fracture, exploration and suture for one case of Achilles tendon rupture, and internal fixation surgery for the remaining 27 cases.Results:Most of the patients had osteoporotic fractures which accounted for 61.5% (16/26) of the outpatients and 68.3% (28/41) of the inpatients, respectively. The duration from injury to surgery averaged 2.3 days and the length of hospitalization 4.5 days for the 41 hospitalized patients, decreased compared with the corresponding data (3.1 days and 11.5 days) for the similar inpatients in the same period last year. In the 41 inpatients, fever was observed upon hospitalization in 4 cases and after operation in 26 cases, and related to their primary injury or surgical trauma in all. Acute pulmonary embolism happened during operation in one patient with femoral shaft fracture.Conclusions:The prevention and control of COVID-19 should be continued. The awareness and behavior of health care providers are expected to be enhanced by strict screening protocol, protection and supervision. The proportion of elderly patients with osteoporotic fracture increased during the COVID-19 pandemic. Shortage of blood was the major problem affecting the treatment. Timely surgical treatment should be indicated for the patients with orthopedic trauma, especially those with lower extremity fracture.