1.Effect of IGF-1 on expressions of VEGF under suppression of HIF-1α by shRNA in cultured human RPE cells
Chinese Journal of Experimental Ophthalmology 2012;30(4):316-319
BackgroundProliferative vitreo-retinal disease (PVD)is one group of ocular complications marked by the enhanced proliferation of various cells included retinal pigment epithelial (RPE) cells.Insulin-like growth factor-1 (IGF-1)and vascular endothelial growth factor(VEGF) are implicated in the aberrant cell proliferation and pathological neovascularization that characterizes PVD,but the signaling mechanism is unclear now. Objective This study was to explore the effect of IGF-1 on VEGF in cultured human RPE cells under the small hairpin loop RNA (shRNA) keeping hypoxia-inducible factor-1 α ( HIF-1 α) silencing. Methods Human retinas were isolated from 4 healthy male donors,and the RPE cells were harvested and cultured.The ceils were identified using anti-human keratin antibody.The third to fifth generation of human RPE cells were used in the experiment.One target site of HIF-1α mRNA was chosen by certain design principle,and shRNA was designed and synthesized by the target site and transferred into the cells in vitro,and then the cells were cultivated with 50 μg/L IGF-1 for 24 hours.The mRNA and protein expressions of HIF-1α and VEGF were detected by RT-PCR and Western blot respectively. Results Cultured human RPE cells showed the flat irregularly multangular shape,and 97%cells appeared the positive response for keratin.HIF-1α mRNA expression in human RPE cells was significantly lower in 50 μg/L IGF-1 group than the 0 pg/L IGF-1 group ( 1.49±0.18 vs 1.46±0.17 ) ( t =0.335,P =0.743 ),however,the expressing levels of HIF-1α protein( 1049.86±172.54 vs 0.00±0.00) and VEGF mRNA(0.95±0.15 vs 0.35±0.07) and VEGF protein (391.98±56.77 vs 214.36±37.15)were raised in the 50 μg/L IGF-I group compared with 0 μg/L IGF-1 group (t=16.098,9.935,6.928,P<0.05).After the HIF-1α-specific shRNA was transferred into cultured RPE cells,the expressions of both HIF-1α mRNA and its protein significantly decreased in RPE cells under 50 μg/L IGF-1 concentration condition( F=68.679,89.904,P=0.000),moreover,the expression of VEGF mRNA and its protein were significantly lowed(F=21.770,6.205,P<0.05). ConclusionsIGF-1 promotes the accumulation of HIF-1α protein and induce the expression of VEGF in human RPE cells,which probably play a pivotal role in the development of PVD.
2.Left ventricular functional changes of stunned myocardium during perioperation of coronary artery bypass graft surgery
Liang-liang, ZHAO ; Hang, L(U) ; Hong-yu, LIU ; Bai-chun, WANG ; Guo-wei, ZHANG ; Li-guo, YANG ; Chao, CHI
Chinese Journal of Endemiology 2012;31(3):283-286
ObjectiveTo evaluate the functional changes of stunned myocardium before and after coronary artery bypass graft(CABG) treatment,and clear the meaning of revascularization which CABG has brought to patients with diffused vascular changes.MethodsA total of 36 patients with 99% diffused coronary artery stenosis in left anterior descending branch underwent non-pump CABG treatment in the Department of Cardiothoracic Surgery the First affiliated hospital of Harbin Medical University.Real-time three-dimensional echocardiography (RT3DE) was repeatedly performed 1 week before operation and 10 days,1 month,and 3 months after CABG.Regional diastolic volumes,systolic volumes,ejection fractions,regional stroke volume to global diastolic volume and the values of abnormal segments before and after CABG were studied.ResultsOne week before operation and 10 days,1 month and 3 months after CABG,the differences of volumes between groups in the last phases of diastole and systolic were statistically significant in anterior wall basement segment,anterior septal basement segment,anterior wall intercalary segment,anterior septal intercalary segment,anterior wall of apex cordis and septation of apex cordis(F =3.51,3.55,4.08,4.05,2.98,3.01,all P < 0.05; F =4.51,4.55,4.08,3.00,2.96,2.99,all P < 0.05).The values of the six segments mentioned above,3 months after operation[(6.74 ± 1.23),(6.64 ± 1.21),(6.02 ± 1.10),(5.95 ± 1.09),(5.82 ± 1.06),(5.10 ± 0.93)ml; (2.74 ± 0.50),(2.69 ± 0.49),(2.51 ± 0.46),(2.32 ± 0.42),(2.36 ± 0.43),(2.03 ± 0.37)ml] were compared with those of 1 week before operation[(8.33 ± 1.52),(8.20 ± 1.50),(7.43 ± 1.36),(7.36 ± 1.34),(7.19 ± 1.31),(6.29 ± 1.15)ml; (4.94 ± 0.90),(4.85 ± 0.88),(4.53 ± 0.83),(4.18 ± 0.76),(4.25 ± 0.78 ),(3.65 ± 0.67)ml],the differences were statistically significant (all P < 0.05); the differences between groups in regional ejection fractions,regional-global ejection fractions were statistically significant(F =4.56,4.88,4.28,3.15,2.93,2.88,P < 0.01 or < 0.05; F =5.56,5.28,4.98,5.15,3.03,2.78,P < 0.01 or < 0.05).Compared with 1 week before the operation, 1 month after the operation in regional ejection fractions,10 days,1 month in regionalglobal ejection fractions after the operation,4 segments of them were significantly improved(all P < 0.05) and 3 months after operation,all the 6 segments had been improved significantly(all P < 0.05).The maximum volume of the sum of group difference of the 6 segments and the 4 segments in the last phase of diastole was statistically significant(F =2.58,5.81,P < 0.05 or < 0.01 ),and the summation began to decrease 10 days after the operation.The values of 3 months after operation[ (36.27 ± 1.10),(25.35 ± 1.16)ml] were compared with that of 1 week before operation[ (44.80 ± 1.36),(31.32 ± 1.43)ml ] the difference was statistically significant (all P< 0.05).The maximum volume summafion comparisons of 6 segments and 4 segments in the last phase of systolic had statistical significance(F =5.77,5.57,all P < 0.01 ),and 10 days after the operation,the summation began to decrease.The values of 1 month[(16.4 0 ± 0.48),(11.58 ±0.51 )ml],and 3 months after operation[ (14.65 ± 0.45),(10.26 ± 0.46)ml],were compared with those of 1 week before operation[ (26.40 ± 0.80),(18.50 ± 0.84)ml],the differences were statistically significant (all P < 0.05).ConclusionsStunned myocardium can be improved through CABG in myocardium systolic,diastole function and ejection fractions of the relevant segments and all of this have proved that patients undergoing CABG revascularization can improve the heart function of the ischemic area.
3.Effect of Thumb Pressing at Yangming Meridian on Function Rehabilitation After Total Knee Arthroplasty
Jun-Qing GU ; Yong-Ying LIANG ; Ying L(U) ; Lian-Bo XIAO
Shanghai Journal of Acupuncture and Moxibustion 2017;36(11):1348-1351
Objective To observe the effect of thumb pressing at Yangming Meridian on the function rehabilitation after total knee arthroplasty (TKA) for knee osteoarthritis (KOA).Method Sixty KOA patients after TKA were randomized into a treatment group (thumb pressing at Yangming Meridian plus conventional rehabilitation) and a control group (single conventional rehabilitation),30 cases each.The treatment group was intervened by thumb pressing and kneading manipulation at the acupoints of Foot Yangming Meridian and Foot Taiyin Meridian plus rehabilitation training of passive flexion and extension by CPM machine,while the control group was intervened by rehabilitation training of passive flexion and extension by CPM machine.The American Hospital for Special Surgery (HSS) for knee joint,the American Knee Society score (KSS) and the Western Ontario and the McMaster Universities Osteoarthritis Index (WOMAC) of KOA were used to evaluate the function.Result After the treatment,the HSS,KSS and WOMAC scores were changed significantly in both groups (P<0.05).The improvements of HSS,KSS and WOMAC scores in the treatment group were superior to those in the control group,and the differences were statistically significant (P<0.05).Conclusion The efficacy of thumb pressing at Yangming Meridian plus conventional rehabilitation for post-TKA KOA is superior to single conventional rehabilitation.
4.Clinicopathological differences in laterally spreading tumors between rectum and colon
Meili XU ; Jie WU ; Chunlian WANG ; Jirong HUO ; Liang L(U)
Journal of Central South University(Medical Sciences) 2018;43(2):192-197
Objective:To investigate the clinicopathological differences in laterally spreading tumor (LST) from the rectum and colon.Methods:Clinicopathological records of 198 patients with LST (116 cases in rectum,82 cases in colon) from the Second Xiangya Hospital of Central South University between January 2012 and January 2017 were evaluated.Results:A total of 198 colorectal LST were included.According to the endoscopic classification,nodular mixed type (LST-GM),homogeneous type (LST-GH),flat elevated type(LST-FE) and pseudodepressed type (LST-PD) were 127(64.1%),13(6.6%),41(20.7%) and 17(8.6%),respectively.LST-GM was predominant in the rectum (71.7%),while LST-FE was predominant in the colon (78.0%),with significant difference (P<0.01).The mean size of LST was (52.03±35.62) mm or (25.37±11.56) mm in the rectum or the colon,with significant difference between them (P<0.01).High grade intraepithelial neoplasia frequency was higher in the rectum than that in the colon (31.0% vs 18.3%),while the low grade intraepithelial neoplasia frequency was lower in the rectum than that in the colon (61.2% vs 75.6%) (both P<0.05).The mean size of LSTGM and LST-GH diameter were larger in the rectum than that in the colon,and the malignant potential of LST-GM was higher in the rectum than that in the colon.The percentage of high grade intraepithelial neoplasia + invasive carcinoma was 41.8% and 22.2%,respectively (both P<0.05).LST in colon was mostly treated with endoscopic mucosal resection,while LST in rectum was treated by endoscopic submucosal dissection predominantly.Conclu sion:LSTs from the rectum and colon show different clinicopathological characteristics to some extent.LST-GM is predominant in the rectum,while LST-FE is predominant in the colon.The malignant potential of LST-GM is higher in the rectum than that in the colon.
5.Paired determination and analysis of 303 mercury samples of cord blood, fetal hair, placenta and maternal scalp hair
Hong-Dao L(U) ; Jun-Liang GUO ; Mei-Zhu CHEN ; Hai-Lan SU
Chinese Journal of Applied Clinical Pediatrics 2013;28(11):820-822
Objective To determine mercury contents in pair in cord blood,fetal hair,placenta and maternal scalp hair,and to analyze the correlation among them and the effect of mercury exposure risk factors in the mercury contents of samples.Methods Puerperants in the hospital,who lived in the local area without history of major diseases during the whole pregnancy,were investigated by questionnaire composed to several health factors.Cord blood,fetal hair,placenta and maternal scalp hair of these puerperal and their neonates (polyembryony and birth defects excluded) were collected to determine mercury content in pairs.Results The mean mercury contents of cord blood,fetal hair,placenta and maternal scalp hair in 303 samples were 1.65 μg/kg,234.58 μg/kg,3.85 μg/kg,497.62 μg/kg,respectively.Fifty percentile of them were 1.72 μg/kg,252.24 μg/kg,3.98 μg/kg,508.88 μg/kg,respectively.There were direct correlations between mercury in cord blood and that in fetal hair,mercury in cord blood and that in placenta,mercury in cord blood and that in maternal scalp hair,mercury in fetal hair and that in placenta,mercury in fetal hair and that in maternal scalp hair,mercury in maternal scalp hair and that in placenta(all P < 0.05).The mercury contents in cord blood,fetal hair and maternal scalp hair of those living nearby factories involved in mercury,higher intake fish during the pregnancy,firing coal and consumption of whiting cosmetics and smoking were determined as (2.24 ± 0.20) μg/kg,(315.65 ± 35.31) μg/kg and (663.53 ± 71.83) μg/kg.The mercury in those without the high risk factors mentioned above were(1.62 ± 0.16) μg/kg,(245.79 ± 28.21) μg/kg and (499.39 ± 47.72) μg/kg.There was a significant difference between 2 groups(all P < 0.01).Conclusions In addition to control industrial pollution,pregnant women should avoid the above-mentioned high risk factors and pay more attention to health care during pregnancy.The mercury content in cord blood is highly correlated with the mercury in fetal hair,and that in placenta and in maternal scalp hair.The 2 kinds of detection both can achieve the purpose of monitoring mercury intrauterine exposure conditions.
7.Effect of intensity modulated radiation therapy on oral mucosa and immune function in patients with nasopharyngeal carcinoma
Shaoqiang LIANG ; Ning ZHANG ; Lusi CHEN ; Yang ZHANG ; Zhenhe ZHENG ; Weijun LUO ; Tao XU ; Zhiqian L(U) ; Shao'en LI
Journal of Central South University(Medical Sciences) 2018;43(5):505-510
Objective:To study the potential effects of intensity modulated radiation therapy (IMRT) on clinical efficacy,oral mucosa reaction and immunological foundation;and to explore the effect of immunological changes on clinical efficacy and oral mucosa reaction in patients with nasopharyngeal carcinoma.Methods:A total of 200 patients with nasopharyngeal carcinoma,who came from First Department of Nasopharyngeal Radiotherapy,the First People's Hospital of Foshan from October 2008 to November 2011,were selected.The patients were treated with nasopharyngeal radiotherapy,and divided into an observation group and a control group (n=100 in each group).The control group underwent common conventional two-dimensional radiotherapy treatment,while the observation group underwent IMRT.The 5-year survival rates and recurrence rates were recorded at follow-up.After the radiotherapy,the oral mucosa in the patients were evaluated by the classification standard of acute radioactive mucositis by American Radiotherapy Oncology Group (RTOG),and the number of T lymphocyte subsets before and after treatment was detected.Results:There were significant difference in non-regional-recurrence survival rate,disease-free survival rate,local recurrence rate between the above 2 groups (all P<0.05),but no significant difference in the distant metastasis-free survival rate (P>0.05).The acute oral mucosa reactions of grade 1,2,3,4 in the control group were 8.00%,20.00%,12.00%,7.00%,respectively,and those were 7.00%,22.00%,15.00%,1.00% respectively.There was no significant difference in the acute response of oral mucosa in grade 1,2 and 3 in the 2 groups (all P>0.05),but there was significant difference in the grade 4 (P<0.05).There were significantly difference in CD8+,CD4+/CD8+ and CD4+ T lymphocyte subsets before and after treatment in the above 2 groups (all P<0.01);there were also significantly difference after treatment between the observation group and the control group (all P<0.01).Conclusion:In the process of treatment in patients with nasopharyngeal carcinoma,the use of IMRT on the basis of chemotherapy is more effective than the conventional two-dimensional radiotherapy,which can reduce the proportion of grade 4 (severe) acute oral mucosa reaction.It may be related to the protective effect of IMRT on immune function in the patients.
8.Effect of HeLa cells infected with Coxsackie virus B3 on mTOR and p70S6K mRNA expression under different nutritional conditions
Liang CHENG ; Chun-Yuan CHEN ; Yong-Gui ZHU ; Li-Hua HUANG ; Hong-Wei L(U) ; Zuo-Cheng YANG
Chinese Journal of Applied Clinical Pediatrics 2013;28(10):733-736
Objective To explore the effect of HeLa cells infected with Coxsackie virus B3 (CVB3) on the changes of mTOR signal pathway under different nutritional conditions.Methods The HeLa cells were cultured under conventional culture and serum starvation culture.(1) For the conventional method,the medium with 10 g/L fetal bovine serum was added for 24 h after the Hela cells were fused into 40% to 50%,and the medium was changed on the next day,then the virus group was infected with CVB3 of 50% tissue culture infective dose (TCID50).However,the control group was cultured by 2 g/L fetal bovine serum.(2) For the serum starvation method,HeLa cells were cultured with the medium without fetal bovine serum for 24 h.Then the virus group was infected with CVB3 of TCID50.The cells in control group were cultured by 2 g/L fetal bovine serum.Cell morphology changes were observed by inverted microscope,and the expressions of the mTOR,p70S6K mRNA were detected with Real-time PCR at 3 h,6 h,9 h,12 h,24 h respectively in both conventional culture and serum starvation groups.Results The expressions of mTOR and p70S6K mRNA were lower in the virus group than those in control group at 12 h and the 24 h (all P <0.05) in the conventional culture group.And the expressions of mTOR and p70S6K mRNA in the virus group were lower than those in the control group at every time points (all P < 0.05) in serum starvation group.The expressions of mTOR and p70S6K mRNA in group with serum starvation virus and the control groups were higher than those in conventional culture group in all time points,but only the expressions of mTOR mRNA were significantly different between the 2 groups (all P <0.05),however,the expressions of p70S6K mRNA had no significant difference between the 2 groups (all P > 0.05).Conclusion CVB3 may be able to down-regulate the expressions of mTOR and p70S6K mRNA.
9.Effect of alloxan intraventricular injection on learning and memory abilities of mice
Ran-Liang CUI ; Kai RONG ; Pu L(U) ; Hai-Yan HU ; Yu-Jing CHU ; Nan DING ; Yan-Qiu DENG
Chinese Journal of Neuromedicine 2011;10(4):346-350
Objective To investigate the effect of glucose metabolism alteration induced by alloxan intraventricular injection on learning and memory abilities of mice, and its role in the development of AD. Methods Mice were randomly divided into high-dose alloxan intraventricular injection group (n=7, 4 mg/kg) and low-dose alloxan intraventricular injection group (n=7, 1.5 mg/kg)and control group (n=7, physiological saline); intraventricular injection of alloxan, the O-GLcNAc transferase inhibitor, was performed in the high-dose and low-dose alloxan intraventricular injection groups to interfere the brain glucose metabolism. Morris water maze was used to detect the learning and memory abilities of mice. Western blotting and immunohistochemistry were used to analyze the alterations of phosphorylation and O-Glycosylation of neurofilament in mice brain induced by alloxan intraventricular injection. Results In the located navigation tests, the swimming time and distance to find the platform in the mice of alloxan administration were significantly increased as compared with those in the control group (P< 0.05); in space exploration experiments, compared with those in the control mice, the number of crossing the hidden platform was decreased and the initial angle of entry to water was increased in the mice of alloxan administration (P<0.05). Western blotting and immunohistochemistry displayed that phosphorylation was obviously increased and the O-Glycosylation was significantly reduced in the cytoskletal neurofilament of the mice with alloxan administration as compared with those in the control group (P<0.05), which was similar to the alteration of neurofilament's modification in AD brain. Conclusion The inventricular injection of alloxan could impair the learning and memory of mice, which might have a relation with the dysregulation of phosphorylation and O-Glycosylation in neurofilament caused by the impaired glucose metabolism, which is similar to the alteration of phosphorylation and O-Glycosylation in neurofilament in AD brain.
10.Analysis of local recurrence pattern for limited stage small cell lung cancer after IMRT plus chemotherapy
Wenbin YAN ; Xuan LIU ; Zongmei ZHOU ; Yuxia WANG ; Zefen XIAO ; Qinfu FENG ; Dongfu CHEN ; Jima L(U) ; Jun LIANG ; Lei DENG ; Tao ZHANG ; Wenqing WANG ; Nan BI ; Xin WANG ; Xiaozhen WANG ; Zhouguang HUI ; Luhua WANG
Chinese Journal of Radiation Oncology 2020;29(3):175-178
Objective To investigate localized regional recurrence after chemotherapy and chest radiotherapy in limited stage small cell lung cancer (LS-SCLC),and explore the relationship between recurrence location and radiotherapy and chemotherapy and its influencing factors.Methods From 2006 to 2014,pathological LS-SCLC treated in CAMS,125 patients had local recurrence,Kaplan-Meier statistical method was used to analyze the survival rate and PFS of each recurrence site.Log-rank was used to compare the survival rate of each group.Univariate analysis includes Chi-squareand t-test for the factors for the recurrence site.Multivariate analysis using Logistic regression.Results The 1-,2-and 5-year overall survival rates were 92.0%,46.4% and 14.7%,respectively.The median progression time was 12.96 months,The median survival time after progression was 1 1.5 months,and the 1-,2-,and 5-year overall survival rates were 45.0%,23.0%,and 10.0%,respectively.The recurrence sites include intrapulmonary recurrence (67 patients),regional lymph nodes (21 patients),simultaneous intrapulmonary and regional lymph nodes (28 patients),and contralateral or supraclavicular lymph nodes (9 patients).The median survival time were 23.96 months,24.76 months,23.23 months,and 18.66 months,and the 2-year survival rates were 49%,52%,46%,and1 1%,respectively (P=0.000,0.004,0.008).In 6 patients (4.0%),5 patients were located in the supraclavicular region,and 1 patient (0.8%) in the field.Conclusions For LS-SCLC undergoing IMRT and chemotherapy,the local failure location is mainly located in the pulmonary,and further treatment of the split dose and targets requires further clinical exploration.