1.Effects of X-ray irradiation combined with RNAi against STAT3 on radiosensitivity of human esophageal carcinoma cells
Huanyu ZHAO ; Weiming ZHANG ; Jinfei CHEN
Chinese Journal of Radiological Medicine and Protection 2011;31(2):180-184
Objective To explore the effects of X-ray irradiation combined with RNAi against signal transducer and activator of transcription 3(STAT3)on the radiosensitivity of human esophageal carcinoma cells.Methods Human esophageal carcinoma cells of the line Eca-109 were euhured.Three pairs of DNA template aiming at the base sequences of the coding regions 2037-2055,1243-1261,and 455-473 of the STAT3 mRNA were synthesized(siRNAI,siRNA2,and siRNA3),and a negative sequence was synthesized to be used as control.STAT3-siRNA positive recombinant plasmids(pRNAT-U6.1-siRNAI,pRNAT-U6.1-siRNA2, and pRNAT-U6.1-siRNA3), and a STAT3-siRNA negative recombinant plasmid (pRNAT-U6.1-negative)were thus constructed and then transfected into the cultured Eca-109 cells,which were divided into transfection reagent control group,pRNAT-U6.1-siRNAl-3 transfection groups,and pRNAT-U6.1-negative centrel group.The positive eell clones were screened.RT-PCR and Westem blotting were used to detect the STAT3 mRNA and protein expression.The transfected Eca-109 cells were exposed to 0,2,4,6,and 8 Gy of X-rays,respectively,and the survival fraction of the cells was analyzed by clone formation assay.Flow cytometry was applied to analyze the cycle arrest and cell apoptosis 4 Gy post-irradiation.Results Agarose gel electrophoresis confirmed the successful construction of the plasmid pRNAT-U6.1-siRNA.RT-PCR and Western blotting demonstrated that the mRNA and protein expression levels of STAT3 transfected with sTAT3-siRNA3 were both significanfly lower than those of the control groups.At 2-8 Gy, the survival fractions of the siRNA3 group were aU significantly lowered than those of the control group(t=-0.228--0.051,P<0.05).Flow cytometry showed that the percentage of the cell cycle G0/G1 phase and the apoptosis rate of the siRNA3 group were both significantly higher than those of the control groups at 4 Gy post-irradiation(t=-13.137-16.350,P<0.01).Conclusions X-ray irradiation combined with RNAi against sTAT3 could inhibit the proliferation of the human esophageal carcinoma cells,induce cell cycle arrest and apoptosis,improve the radiosensitivity in Eta-109 cells.
2.The Influence of STAT3 Silencing by RNA Interference on the Biological Characteristics of Eca-109 Cell Line
Huanyu ZHAO ; Weiming ZHANG ; Jinfei CHEN
Chinese Journal of Clinical Oncology 2009;36(24):1420-1423
Objective: To study the cell proliferation, cell cycle and apoptosis of esophageal carcinoma Eca-109 cells treated with RNA interference technique to silence signal transducers and activators of transcription 3 (STAT3) gene. Methods: Three pairs of DNA template coding siRNA specific for human STAT3 gene mRNA were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pRNAT-U6.1/neo plasmid to construct STAT3-siRNA expression vector which was then transfected into Eca-109 cells. The expression of STAT3 mRNA and protein in cancer cells was detected by RT-PCR and Western blot, respectively. The cell proliferation, cell cycle distribution and apoptosis were examined by MTT and flow cytometry. Results: STAT3-siRNA expression vector was successfully constructed and identified by sequencing. The results of RT-PCR and Western blot demonstrated that STAT3 expression in Eca-109 cells transfected with STAT3-siRNA expression vector was significantly higher than that in the control group (P< 0.01). MTT showed that after transfection of the siRNA vector into Eca-109 cells, cell proliferation was obviously reduced and the cell growth inhibition ratio in the siRNA3 group was 35.68%, significantly higher than that in the control group (P<0.01). Flow cytometry results suggested that cell cycle arrest and more apoptosis were observed in the siRNA3 group. Cell cycle was arrested at G_0/G_1 phase, and the rate of apoptosis was 13.26%, much higher than that in the control group (P<0.01). Conclusion: Silencing STAT3 gene by RNA interference technique can effectively inhibit STAT3 expression, suppress the proliferation of Eca-109 cells, induce cell cycle arrest at G_0/G_1 phase, and promote apoptosis.
3.Effects of X-ray irradiation combined with RNAi against STAT3 on proliferation and apoptosis of human esophageal carcinoma cells
Huanyu ZHAO ; Weiming ZHANG ; Jinfei CHEN
China Oncology 2006;0(11):-
Background and purpose:Several reports demonstrated that the expression of STAT3 has been found to be an oncogene in solid tumors such as head and neck squamous cell carcinoma,esophageal carcinoma and prostate carcinoma.This study was done to explore the effects of X-ray irradiation combined with RNAi against STAT3 on proliferation and apoptosis of human esophageal carcinoma cell line Eca-109.Methods:Three pairs of DNA template coding siRNA were synthesized against STAT3 to reconstruct pRNAT-U6.1-siRNA-STAT3,which was transfected into Eca-109 cells,the positive cell clones were screened with G418.Inhibitory effect of STAT3 mRNA and protein in Eca-109 cells was detected by RT-PCR and Western blot,respectively.The transfected cells were exposed to 0,2,4,6 and 8 Gy X-ray respectively;the survival fraction of Eca-109 cells was analyzed by clone formation assay,and flow cytometry was applied to analyze cell apoptosis at the dose of 4 Gy.Results:pRNAT-U6.1-siRNA-STAT3 was reconstructed and identifi ed as correct by sequencing.RT-PCR and Western blot analyses demonstrated that STAT3-siRNA could obviously reduce the expression of STAT3 mRNA and protein in Eca-109 cells.Clone formation assay and flow cytometry results showed that irradiation at different doses combined with STAT3-siRNA could inhibit the proliferation of Eca-109 cells,irradiation with 4 Gy X-ray could induce apoptosis.Conclusion:X-ray irradiation combined with RNAi against STAT3 could inhibit the proliferation of Eca-109 cells and induce apoptosis.
4.Features of coronary angiography in type 2 diabetic patients combining various risk factors
Huanyu ZHAO ; Lian CHI ; Lihua ZHONG ; Yan CUI
Chinese Journal of Tissue Engineering Research 2005;9(3):234-235
BACKGROUND:With the increasing morbidity of type 2 diabetes, people should pay more attention to the early intervention of cardiovascular complications in order to reduce the incidence of its complications. OBJECTIVE:To observe the risk factors of type 2 diabetes combining coronary heart disease(CHD) and the images of coronary angiography. DESIGN:A cross-sectional observational study. SETTING:Internal medicine department of a municipal hospital. PARTICIPANTS:The study was completed in the Department of Endocrine Harbin First Hospital from May 1998 to May 2003.Inclusive criteria:Type 2 diabetic patients aged above 30 years old and combined with CHD with either sex, who met the diagnostic standard of diabetes set by ADA in 1997 and Naming and Diagnostic standard of ischemic heart disease set by WHO in 1979. Exclusive criteria:Patients suffering from severe heart,liver,kidney disease and serious infection,rheumatoid heart disease,pulmonary heart disease and so on.There were 98 inpatients, who met the above mentioned inclusive criteria, were set as the diabetic group with 56 males and 42 females,aged from 35 to 70 years old;and 85 CHD patients without combining diabetes admitted to hospital at the same time were chosen as the control group with 53 males and 32 females, aged from 40 to 75 years old. INTERVENTIONS:The blood glucose was observed with the glucose oxidase method,fasting insulin and C peptide with radioimmunoassay (RIA),glycosylated hemoglobin with affinity chromatography, triglyceride(TG),total cholesterol(TC),high density lipoprotein-cholesterol(HDL-C) and low density lipoprotein-cholesterol (LDL-C) with automatic biochemical analysis,and selective right coronary angiography was applied by JUDKINS method. Of coronary arteries. RESULTS:The comparison of the coronary angiographic images between the two groups showed that there was no significant difference in the lesion of left main trunk and circumflex artery between the diabetic group and control group(P >0.05);While there were more cases with lesion of anterior descending artery and right coronary artery in the diabetic group than in the control group(P< 0.05). The pathological changes in the diabetic group mainly manifested with lesion of three branches and single branch in which represented diffusive changes.Lesion of three branches was the main pathological change of coronary artery in type 2 diabetic patients with diffusive changes. CONCLUSION:With the extension of course of type 2 diabetes in patients,the incidence of CHD will gradually increase with extensive pathological changes and much severe conditions.
5.Analysis of 244 ADR Cases in Beijing Tongren Hospital in 2005
Xiaokun JIN ; Chiying XIU ; Huanyu ZHAO ; Li WEN
China Pharmacy 1991;0(05):-
OBJECTIVE:To investigate the characteristics of adverse drug reactions(ADR) in our hospital so as to promote rational clinical drug use.METHODS:244 ADR cases collected in 2005 in our hospital were analyzed retrospectively.RESUL TS:Among the 244 cases,the female exceeded the male;79.83% of the patients were more than 40 years old.The cardiovascular system medicines took the lead which amounted to 45.69% of the total,then came the antimicrobial medicines.The intravenous administration caused most of ADRs.Alprostadil Injection accounted for 9.02% of the total, the highest.Skin and its appendages impairment cases took the biggest percentage at 28.14%.CONCLUSION:we should pay more attention to the ADR monitoring and the spread of ADR knowledge and enhance the ADR awareness of medical staff.sen or avoid the occurrence of ADR,monitoring of which and publicizing of ADR knowledge should be emphasized.
6.Clinical analysis of pregnancy complicated with cervical epithelial neoplasia-18 cases
Tao Lü ; Huanyu ZHENG ; Jian ZHAO ; Zhaohui LIU ; Qinping LIAO
Chinese Journal of Perinatal Medicine 2009;12(4):277-280
Objective To discuss the diagnosis,treatment and following-up of cervical intraepithelial neoplasia (CIN) during pregnancy.Methods Eighteen pregnant women with CIN,presented to the hospital from Jan.2004 to May.2008,were retrospectively reviewed.Pap smear,HPV,copolscopy and cervical biopsy were performed for diagnosis and all were followed up by Pap smear and copolscopy during pregnancy every 12 weeks,while 15 were followed up until 6-8 weeks postpartum.Results The incidence of CIN during pregnancy was 0.16%.All of the 18 women had abnormal Pap smear,followed by colopscopy and biopsy.Conservative management during the pregnancies every 3 months showed no progress in CIN.Among the 18 cases,17 delivered at term and only 1 preterm (36+4 weeks).Two out of the 18 women had normal delivery (11.1%),2 by forceps(11.1 %) and 14 by cesarean sections (77.8%).No neonatal apnea or neonatal respiratory distress syndrome had been observed.Pathological diagnosis was confirmed in all cases prenatally,including 4 CIN1,4 CIN2 and 10 CIN3.Four out of the 10 CIN3 remained after delivery,while 1 changed to CIN2.These 5 cases underwent leep electrical excision procedure and Pap smear results turned to normal at 6 to 11 months after delivery.One HSIL case were lost.The rest 4 CIN3 and all of the 8 CIN1 and CIN2 cases turned to normal or lesser degree.Fourteen of the 18 women received HPV test during pregnancy,and 12 (85.7%) were positive,among which 7 women were followed up at postpartum and 6 of them were HPV positive.Conclusions The three steps method for diagnosis of CIN during pregnancy is safe and effective.Conservative management of CIN during pregnancy is recommended.
7.The effect of ziprasidone and risperidone on schizophrenia patients and the change of leptin ,adiponectin levels
Ao ZHAO ; Qiping LI ; Huiteng LI ; Chunyang LI ; Zaiping HUANG ; Huanyu XU ; Jieping GUAN
International Journal of Laboratory Medicine 2016;37(4):506-507,510
Objective To observe the effects of ziprasidone and risperidone on schizophrenia patients and the change of serum leptin and adiponectin levels .Methods Totally 80 cases of schizophrenia patients were randomly divided into ziprasidone group and risperidone group ,which were treated for 8 weeks .Measure the positive and negative symptoms scale (PANSS) score and body weight of that number ,leptin and adiponectin at baseline ,treatment 4 weeks and 8 weeks respectively for patients ,at the end of the experiment ,the results for statistical analysis .Results Two groups of 4 ,8 weeks after treatment scores compared with baseline scores dropped significantly ,the difference was statistically significant(P<0 .05) .Risperidone group after treatment ,leptin levels significantly increased body mass index ,and adiponectin levels significantly decreased ,compared with the baseline before treatment was statistically significant difference(P<0 .05) .Conclusion Ziprasidone and risperidone in treatment of schizophrenia have similar efficacy .Ziprasidone has no significant effect on body weight ,leptin and adiponectin levels in treatment of schizophrenia patients . However ,risperidone has a significant effect ,long-term use should pay attention to the side effects .
8.Risk factors for acute kidney injury in overweight patients with acute Stanford type A aortic dissection following Sun's pocedure
Xudong PAN ; Honglei ZHAO ; Tao BAI ; Jinrong XUE ; Ningning LIU ; Huanyu QIAO ; Yongmin LIU ; Lizhong SUN
Chinese Journal of Thoracic and Cardiovascular Surgery 2017;33(5):295-299
Objective To identify the risk factors for acute kidney injury (AKI) in overweight patients who underwent surgery for the treatment of acute type A aortic dissection(TAAD).Methods A retrospective study including 108 consecutive overweight patients(body mass index(BMI) ≥24) between January 2010 and May 2013 in Beijing Anzhen Hospital was performed with Sun's procedure of total aortic arch replacement and frozen elephant trunk implantation.AKI was as defined according to Acute Kidney Injury Network (AKIN) criteria based on serum creatinine(sCr) or urine output.Results The mean age of the patients was(43.69 ± 9.66) years.During the postoperative period seventy-two patients(66.7%) developed AKI.The overall postoperative mortality rate was 7.4%,8.3% in AKI group and 5.6% in non-AKI group.There is no statistically significant difference between the two groups(P =0.32).A logistic regression analysis was performed to find out the independent risk factors for AKI:elevated preoperative sCr level and postoperative drainage volume.Renal replacement therapy(RRT) was performed in fifteen patients (13.9 %).Conclusion A higher incidence of AKI (66.7 %) in overweight patients following acute TAAD was identified.The logistic regression model found out elevated preoperative sCr level and 72 hour drainage volume as the two independent risk factors for AKI in overweight patients.More attention should be paid to prevent AKI in overweight patients following TAAD.
9.The study of targeted blocking SDF-1/CXCR4 signaling pathway in vivo with T140 on SDF-1 and MMPs levels
Kun WANG ; Fengkai ZHAO ; Yanlin LI ; Yang YU ; Huanyu GAO ; Yu XIAO ; Longteng LI ; Xiangjia YAN ; Di JIA
The Journal of Practical Medicine 2015;(19):3133-3136
Objective To explore the effect of T140 on SDF-1 and MMPs levels through targeted blocking SDF-1/CXCR4 signaling pathway , and to investigate the function of T140 to prevent from cartilage degeneration. Methods Thirty-six 9-month-old male healthy Hartley guinea pigs were divided into three groups: experimental group(group A,n = 12),experimental control group(group B,n = 12) and blank control group (group C,n = 12). In the 2nd,4th,6th,8th,10th,12th week, the levels of SDF-1 in serum were quantified with ELISA. In the 12th week, mRNA levels of MMP-3,MMP-9 and MMP-13 in articular cartilages were measured with RT-PCR. Results The serum levels of SDF-1 of the group A decreased gradually but increased in group B and C. Group A had a statistical significance compared with group B and C at the same time point (P< 0.05).The mRNA levels of MMPs in group A were lower than group B and C (P < 0.05). Conclusion T140 could block the SDF-1/CRCR4 signaling pathway and decrease the secretion of SDF-1 and mRNA expression levels of MMPs and reduce the cartilage degeneration.
10.Comparative study of curative effect and blood glucose and lipid in schizophrenic patients who treated with ziprasidone and risperidone
Qiping LI ; Ao ZHAO ; Huiteng LI ; Chunyang LI ; Zaiping HUANG ; Huanyu XU ; Jieping GUAN ; Tanling XIAO ; Jinhua ZHANG ; Zhanhong FU
International Journal of Laboratory Medicine 2015;(17):2471-2472,2475
Objective To observe the effects of ziprasidone and risperidone on patients with schizophrenia and their influence on bloodglucoseandlipids.Methods 96patientswithschizophrenicenrolledinthestudywererandomlydividedintotwogroups,zi‐prasidone and risperidone group ,and both were treated for 8 weeks .Their blood glucose ,blood lipid of base line and at the end of the 4th ,8th week were determined respectively .Results The positive and negative symptoms scores of the two groups by using Positive and Negative Syndrome Scale(PANSS) before and after treatment were not statistically different(P> 0 .05) .Compared with the baseline scores ,scores at the end of 4th and 8th week in both ziprasidone and risperidone groups significantly decreased(P<0 .05) ,but there was no statistically significant difference between the two groups(P>0 .05) .After 8 weeks′ treatment ,the ef‐fective rate was 91 .7% in ziprasidone groups and 89 .6% in risperidone group .There were no significant differences between the two groups(P>0 .05) .The blood lipids and glucose levels were less increased after ziprasidone treatment ,but was not statistically significant(P>0 .05) .The blood lipids and glucose levels significantly increased after risperidone treatment(P<0 .05) .Conclusion Ziprasidone and risperidone had the same effect on schizophrenia .Ziprasidone had no effect on blood glucose and lipids in schizo‐phrenic patients ,while risperidone could increase blood glucose and lipids level ,we should pay attention to the side effects of long‐term use .