1.Effect of HIF-1α expression inhibition by RNA interference on radiosensitivity and autophagy of hypoxic human lung adenocarcinoma cell line A549
Yanmei ZOU ; Hua XIONG ; Zhiping XIAO ; Shiying YU ; Xianglin YUAN
China Oncology 2013;(6):413-419
Background and purpose:Hypoxia induced the decreased radiosensitivity of tumor cells, which was the cause of tumor radioresistance and relapse and metastasis. During the course, HIF-1a played the most important role in the regulation of hypoxia. However, it’s still unknown about the effect of HIF-1a on the radiosensitivity of hypoxia tumor cells and the relationship with autophagy. This study was to inhibit HIF-1αexpression in hypoxic lung adenocarcinoma cell line A549 with RNA interference (RNAi), and explore its effect on hypoxic cell radiosensitivity and autophagy. Methods: Plasmids pHIF-1α-shRNA and Neg-shRNA (negative control) were constructed and transfected into hypoxic A549 cells, this positive clone was named A549/HIF-1α-shRNA. Clone formation array was applied to calculate the value of D0, SF2, SER. The expression of HIF-1α, LC3, c-parp was detected by Western blot. Results:The SF2 of hypoxic A549 cell was 0.62, which was higher than that of normoxic A549 cell, SER was 1.45. The level of LC3Ⅱincreased significantly and the level of c-parp decreased after the radiation of hypoxic A549 cell. The level of HIF-1a increased in hypoxic A549 cells. The expression of HIF-1αin hypoxic A549 cells was suppressed markedly after transfection of HIF-1α-shRNA;this clone was named A549/HIF-1α-shRNA. The SF2 and SER were significantly lower in A549/HIF-1α-shRNA cells, 0.45 and 0.72 respectively. Under the hypoxic condition and after the inhibition of HIF-1α, the expression of LC3Ⅱ decreased significantly and the expression of c-parp increased. Conclusion:We successfully established a cell model that HIF-1αexpression was suppressed almost completely by RNAi. The inhibition of HIF-1αby shRNA may raise the radiosensitivity and decrease the autophagy of hypoxic A549 cells in vitro.
2.Meta-analysis of antibiotic prophylaxis use in transrectal prostatic biopsy
Minggen YANG ; Xiaokun ZHAO ; Zhiping WU ; Ning XIAO ; Chen LV
Journal of Central South University(Medical Sciences) 2009;34(2):115-123
ObjectiveTo determine whether antibiotic prophylaxis can reduce the risk of postoperative infective complications in men undergoing transrectal prostatic biopsy (TPB) who had sterile preoperative urine.MethodsMEDLINE, EMBASE, Cochrane Collaboration Reviews, Chinese Medical Current Contents (CMCC), and National Knowledge Infrastructure (CNKI) were searched for rando-mized controlled trials that compared the effect of antibiotic prophylaxis with placebo or active controls for men undergoing TPB with preoperative sterile urine. Two reviewers independently extracted the data of patient characteristics and outcomes based on a prospectively developed protocol.ResultsA total of 12 trials (3 placebo controlled, 3 non-treatment controlled, and 6 activly controlled) involving 1 987 patients, met the inclusion criteria. Prophylactic antibiotic use in patients at low risk undergoing TPB significantly decreased bacteriuria and middle degree fever incidence, but could not decrease the incidence of bacteremia. The relative risk for post-TPB bacteriuria, middle degree fever, and bacteremia were 0.32 (95% CI 0.23 to 0.46), 0.37 (95% CI 0.17 to 0.77), and 0.96 (95% CI 0.61 to 1.50), respectively. Effective antibiotic classes included quinolone, co-quinolone and nitroimidazole, and co-trimethoprim and sulfamethoxazole. Treatment protocols of any duration were effective.ConclusionAntibiotic prophylaxis obviously decreases the incidence of bacteriuria and middle degree fever but not bacteremia in men with preoperative sterile urine undergoing TPB. A significant decrease in bacteriuria incidence can be achieved with a range of antibiotic agents, including quinolones and co-quinolone and nitroimidazole. Treatment protocols of any duration are effective with no heterogeneity.
3.Therapeutic effects and pathways of human umbilical cord mesenchymal stem cells transplantation for rat acute liver failure model
Sheng ZHENG ; Juan YANG ; Fang YIN ; Qiongyi XIAO ; Zhiping GUO
Chinese Journal of Organ Transplantation 2014;35(12):747-752
Objective To explore the therapeutic effectiveness and pathways of human umbilical cord mesenchymal stem cells (hUCMSCs) transplantation for acute hepatic failure in rats.Method hUCMSCs were isolated from umbilical cord with attachment culture method,and the surface antigens were tested by flow cytometry.Forty-eight male Sprague Dawley rats were randomly divided into four groups.The animal model of acute liver failure was induced by injecting intraperitoneally with 50% olive oil solution of carbon tetrachloride (2.5 ml/kg).The treatment groups were injected with hUCMSCs suspension separately through the tail vein or injected into the liver 24 h post-modeling.Blood serum and liver tissues were collected at several time points to analyze the improvement of liver function and histological repair.Real-time PCR was used to detect the expression of human CK8,CK18 and AFP mRNA in liver tissues.Immunohistochemistry was used to detect the expression of human CK18 in liver tissues.Result There were statistically significant differences among liver functions after transplantation (P<0.05).hUCMSCs improved histological status through enhancing hepatocellular regeneration and reducing inflammatory cells.Real-time PCR results showed that the expression of CK8,CK18 and AFP mRNA was obviously increased in the tail vein transplantation group and hepatic lobe injection transplantation group as compared with the model group (P<0.05).Immunochemistry results revealed that transplanted hUCMSCs in animal liver could differentiate into functional hepatocyte-like cells that expressed human CK18 as hepatocyte-specific marker in the tail vein transplantation group and hepatic lobe injection transplantation group.No significant differences in histological repair and grade of differentiation were examined between the tail vein transplantation group and hepatic lobe injection transplantation group (P>0.05).Conclusion hUCMSCs can prompt the repair of acute liver failure and enhance pathological repair.Transplanted cells in animal liver can differentiate into functional hepatocyte-like cells that expressing hepatocyte-specific markers.Transplantation of hUCMSCs via the tail vein or direct injection into the liver had the similar therapeutic effects.
4.Expression of silence information regulator 1 (Sirt1) in B cells in peripheral venous blood of subacute cutaneous lupus erythematosus
Nan HU ; Qing LI ; Zhiping XIAO ; Qin QIN ; Cuie KUANG
Journal of Chinese Physician 2015;17(12):1836-1838
Objective To investigate alterations in epigenetic regulation in B cells in subacute cutaneous lupus erythematosus (SCLE).Methods Peripheral blood mononuclear cells (PBMC) were isolated from the peripheral venous blood of SCLE patients and healthy donors with density gradient centrifugation.B cells were isolated with CD19 micro beads and protocols provided by the manufacturer.Total RNAs were isolated with RNA kits.The expressions of silence information regulator 1 (Sirt1) mRNA were investigated with quantitative real time polymerase chain reaction (PCR).Results The levels of Sirt1 mRNA were significantly increased in SCLE B cells relative to healthy controls (P < 0.01).Conclusions Histone modifications appear abnormal in B cells in SCLE.
5.Inhibitory Effect of Ethanol Extract of Oldenlandia Diffusa on Colorectal Cancer Angiogenesis in BALB/c Mice
Yun XIAO ; Zhiping WU ; Congguo JIN ; Jia LI ; Xiaoqun CHEN ; Qian YAO ; Qishun ZHU
Journal of Kunming Medical University 2013;(10):53-57
Objective To explore the inhibitory effect of Oldenlandia diffusa extract on colorectal cancer angiogenesis in BALB/c mice. Methods Thirty-two BALB/c mice with subcutaneous CT26 colon cancer animal model were randomly equally divided into four groups,including the control group (groupⅠ,saline 0.1 mL/(10. d), O. diffusa ethanol extract of 90 mg/(kg.d) (groupⅡ), O. diffusa ethanol extract of 180 mg/(kg.d) (groupⅢ) and O. diffusa ethanol extract of 360 mg/(kg.d) (group Ⅳ) . Each group of mice were treated with intragastric administration of law administration 12 days after vaccination, then stopped and continue fed to 32 days, and the mice were killed. Micro-vascular dense ( MVD) was observed and countered under the microscopy by immunohistory chemistry. Results The murine colon tumor volumes of GroupⅡ,ⅢandⅣwere significantly less than that of groupⅠ,with significant difference ( <0.05) . The tumor microvessel density values of four groups was (7.83±2.87), (5.32±1.27), (1.77±0.70) and (1.87±0.68),respectively. The number of tumor blood vessels in GroupⅡ,Ⅲ and Ⅳ were significantly less than that of Ⅰ group, with significant difference ( <0.05) . Conclusion Within a certain dose range, the ethanol extract of O. diffusa can significantly inhibit the mouse colon cancer and the mechanism may be realated to inhibiting tumor angiogenesis.
6.Mental Health Literacy in Three Cities of China:A Survey Study
Fei LI ; Shuiyuan XIAO ; Zhiping HUANG ; Jianguo SHI ; Zaohuo CHENG ; Wenfeng LUO ; Fangru YANG ; Liang ZHOU
Chinese Mental Health Journal 2009;23(12):883-887
Objective:To assess mental health literacy of urban porpulation and its related factors.Method:Subjects were sampled from 3 cities in the east(Wuxi),central(Changsha)and west(Xi'an)part of China.Subjects were asked to answer a questionnaire which included 5 vignettes of mental disorders and related questions.Results:54.1% of 7309 participants correctly recognized the cases of mania,but only 11.2% correctly recognized the cases of schizophrenia with negative symptoms.The average rate of correct recognition for all 5 vignettes was 41.7%.Participants with higher education level recognized the vignettes more correctly.Ill-character,stress and the pressure from work were reported to be the 3 main reasons of mental disorders.There were strong negative attitudes toward psychiatric patients,especially those with schizophrenia and mania.Conclusion:While the rate of correct recognition of mental disorders is acceptable when compared with similar studies in other parts of the world,negative attitude toward patients with mental disorder is still prevalent in China.
7.Influence of Rapamycin on apoptosis of acute lymphoblastic leukemia EU-4 cells induced by Methotrexate
Zhaoxia ZHANG ; Yixiang ZHANG ; Zhiping LIU ; Lili CHEN ; Zhihui XIAO ; Bo LI ; Xicai TANG
Chinese Journal of Applied Clinical Pediatrics 2017;32(14):1101-1103
Objective To explore the influence of Rapamycin (RAPA) on apoptosis of acute lymphoblastic leukemia EU-4 cells induced by Methotrexate (MTX).Methods EU-4 cells were pretreated 0.5 h with 5 pμg/L,10 μg/L,25 μg/L,50 μg/L and 100 μg/L RAPA.RAPA untreated group was set up as control group.The cells were collected and the expression of LC-3 was assayed by using Western blot.The apoptosis of EU-4 cells was detected by using flow cytometry (FCM).The effects of different concentrations of RAPA pretreatment on autophagy and apoptosis of EU-4 cells were observed,and the pretreatment concentration of RAPA was determined.EU-4 cells were divided into RAPA pretreatment group and untreated group.The cells were treated with 0.05 μmol/L,0.10 pμmol/L and 0.20 μmol/L MTX for 24 h,respectively.The apoptotic rate was detected by using FCM.Western blot was performed to test the expression of LC-3 protein,while the absorbance(A562) was measured by using microplate reader,and the protein concentration in the sample was calculated according to the standard curve.Results The apoptotic rates of EU-4 cells in the control group and the different concentrations of PAPA pretreatment group showed that the control group,5 μg/L,10 μg/L,25 μg/L,50 μg/L and 100 μg/L RAPA were (7.51 ±0.32)%,(7.33 ±0.41)%,(7.71 ± 0.51) %,(7.63 ± 0.38) %,(7.80 ± 0.43) % and (16.66 ± 0.87) %,respectively.The apoptotic rates of EU-4 cells in the 5 μg/L,10 μg/L,25 μg/L and 50 μg/L PAPA pretreatment group had no significant difference from those in the control group(t =0.427,-0.417,-0.297,-3.561,all P > 0.05).The apoptotic rate of EU-4 cells in 100 μg/L PAPA pretreatment group was significantly higher than that in control group,and the difference was significant (t =-28.815,P < 0.01).Combined with the results of Western blot and FCM,50 μg/L was used as the pretreatment of PAPA.The apoptosis rates of EU-4 cells in PAPA pretreatment group were (50.23 ± 2.11) %,(66.88 ± 2.89) % and (73.11 ± 2.67) % after treated with 0.05 μmol/L,0.10 μmol/L and 0.20 μmol/L MTX for 24 h,respectively.The apoptotic rates of EU-4 cells in PAPA unpretreatment group were (22.53 ± 1.67) %,(42.82 ± 2.26) % and (53.22 ± 1.93)% after treated with 0.05 μmol/L,0.10 μmol/L and 0.20 μmol/L MTX for 24 h,respectively.The apoptotic rates of EU-4 cells in RAPA pretreatment group were significantly higher than those in untreated group in the same concentration of MTX treatment after 24 h,and the differences were significant(t =12.693,66.148,14.429;all P < 0.01).Conclusion With RAPA pretreatment,relative low dose MTX can induce a great deal of acute lymphoblastic leukemia EU-4 cells apoptosis.
8.Submental ultrasonongraphy in assessing oropharyngeal swallowing disorders in children with cerebral palsy
Huachun XIONG ; Jiaojiao HUANG ; Ning XIAO ; Dengna ZHU ; Zhiping GAO ; Yunxia ZHAO ; Jun WANG
Chinese Journal of Physical Medicine and Rehabilitation 2017;39(6):433-437
Objective To explore the application of submental ultrasonongraphy (SUS) in the assessment of oropharyngeal swallowing disorders in children with cerebral palsy.Methods Seventeen children with cerebral palsy and oropharyngeal swallowing difficulties (7 on nasal feeding,10 on oral feeding) constituted the treatment group while 20 normal counterparts formed the control group.SUS was applied to measure any changes in the thickness of the tongue muscle and the range of hyoid bone displacement when they swallowed 5 ml of water.The results were compared with those assessed using the functional oral intake scale to decide the best cut-off point for detecting tube-feeding-dependent dysphagia.The intraclass correlation coefficient (ICC) of the 20 children in the control group was calculated to evaluate the intra-rater and inter-rater reliability of SUS.Results The average tongue muscle thickness change and hyoid bone displacement amplitude of the children on nasal feeding were significantly smaller than those of the children without nasal feeding and the normal children.The best cut-off point for the tongue muscle thickness change data was 1.0 cm,and that of the hyoid bone displacement amplitude was 1.5 cm.All of the ICCs were above 0.4,indicating good intra-rater and inter-rater reliability for the SUS examination.Conclusion Submental ultrasonongraphy can help assess the swallowing function of children with oropharyngeal swallowing disorders.
9.The value of serum brain natriuretic peptide in chronic pulmonary heart disease
Jinghua XIAO ; Shengming LIU ; Dongbo TIAN ; Yun CHEN ; Weiping CHEN ; Zhiping FU ; Haie WANG
Chinese Journal of Primary Medicine and Pharmacy 2016;(4):603-606,607
Objective To investigate the diagnostic value of serum brain natriuretic peptide(BNP)in chron-ic cor pulmonale of different period.Methods According to the inclusion criteria,we recruited 216 cases from heart and respiratory medicine(including outpatients and inpatients)of Qingyuan People 's Hosptial from April 2013 to December 2014.All the cases were divided into healthy control group(group A,n =48),cor pulmonale heart function compensatory period group(group B,n =43),cor pulmonale right heart failure group(group C,n =45),the simple left heart failure group(D group,n =40)and the whole heart failure group(group E,n =40).The serum BNP value,pul-monary function,echocardiography were detected.We compared the differences amomg them with correlation analysis, and drew the ROC curves to obtain the best cutoff point.Results The BNP value was higher in group C(495.44 ± 219.90)ng/L than group B[(182.44 ±69.71)ng/L,P <0.001],while the value was higher in group D(882.57 ± 288.56)ng/L and E(891.78 ±256.45)ng/L than group C(P <0.001).In cor pulmonale groups,BNP was positive-ly correlated with RV,RVOT,and PASP,negatively correlated with FEV1 ,and not correlated with LVEF %.In group D and E,BNP was negatively correlated with LVEF %.The best cutoff point of BNP was 285.3ng/L between cor pul-monale heart function compensatory period group(group B)and cor pulmonale right heart failure group(group C). The best cutoff point of BNP was 764.2ng/L between cor pulmonale right heart failure group(group C)and the whole heart failure group(group E).Conclusion There is certain correlation between serum BNP level and the progress of chronic cor pulmonale.Dynamic monitoring of serum BNP level in the judgement of treating cor pulmonale is of certain reference significance.
10.The effect of the photon therapeutic apparatus on type 2 diabetic peripheral neuropathy
Xiao LIANG ; Zhiping LIU ; Qingfeng CHENG ; Juan YUAN ; Chengyun WU ; Qinghua ZHAO ; Qifu LI
Chongqing Medicine 2014;(6):656-657,660
Objective To observe the clinical efficacy of the photon therapeutic apparatus on type 2 diabetic patients with periph-eral neuropathy .Methods While giving conventional nerve nourishing ,improving microcirculation treatment and nursing ,adjuvant therapy with Carnation-11 photon therapeutic apparatus was applied to one lower limb only of the 20 diabetes mellitus patients with peripheral neuropathy and compare the curative effects by paired observation of the two lower limbs .Results The evaluation score of peripheral neuropathy dropped obviously with ankle reflex and thumb vibration sensation improved greatly after 10 days of treat-ment and remained an unchanged follow -up test 30 days later ,which means the clinical therapeutic effect of the treated limb was significantly superior to the comparison limb .The difference of the curative effects is statistically significant ( P < 0 .05 ) . Conclusion Conventional therapy accompanied by adjuvant treatment of photon therapy can relieve the clinical symptoms and phys-ical signs of diabetic patients with peripheral neuropathy ,which could improve curative effect on diabetic peripheral neuropathy .