1.Effects of curcumin on the AKT/mTOR signaling pathway in human melanoma cell lines A375 and C8161
Xiaodong HAN ; Youyou ZHOU ; Siwen ZHENG ; Zhen LI ; Zhiqi SONG
Chinese Journal of Dermatology 2015;48(6):382-386
Objective To explore molecular mechanisms underlying the in vitro counteracting effect of curcumin on malignant melanoma.Methods Cultured A375 and C8161 human melanoma cells were cultivated in vitro,and randomly divided into several test groups and a control group to be treated with different concentrations of curcumin and dimethyl sulfoxide respectively for different durations.Then,methyl thiazolyl tetrazolium (MTT) assay,Transwell assay,flow cytometry and Western blot were performed to evaluate the effect of curcumin on the proliferation,invasion and cell cycle of,as well as expressions of AKT/mTOR signaling pathway-related proteins in A375 and C8161 cells respectively.Statistical analysis was carried out by using t test.Results MTT assay showed that the treatment with curcumin of 5-35 mg/L for 24-96 hours significantly inhibited the proliferation of both A375 and C8161 cells compared with that with dimethyl sulfoxide (all P < 0.001),and the inhibitory effect was in a dose-dependent manner within the range of 5-15 mg/L for A375 cells and within the range of 5-10 mg/L for C8161 cells,and in a time-dependent manner from 0 to 48 hours for both cells.After treatment for 24 hours,the 50% inhibitory concentration (IC50) of curcumin against A375 cells and C 8161 cells was 10 mg/L and 5 mg/L respectively.Transwell assay demonstrated that the invasion of A375 and C8161 cells was significantly suppressed by 72-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively (both P < 0.001).Flow cytometry showed that the cell cycle of A375 and C8161 cells was arrested at G2/M phase after 24-hour treatment with curcumin at 10 mg/L and 5 mg/L respectively,with significant differences in the proportion of A375 cells and C8161 cells in G2/M phase between the test group and control group (A375 cells:35.00% ± 3.54% vs.120.80% ± 7.46%,P< 0.001;C8161 cells:19.33% ± 4.04% vs.85.00% ± 9.53%,P < 0.001).Western blot revealed that the expressions of AKT/mTOR signaling pathway-related proteins were decreased in A375 and C8161 cells after 24-hour treatment with 10 mg/L and 5 mg/L curcumin respectively.Conclusion Curcumin can inhibit the proliferation and invasion of A375 and C8161 cells,likely by blocking cell cycle and inhibiting activation of the AKT/mTOR signaling pathway.
2.PhaseⅡclinical trial of raltitrexed plus oxaliplatin combined with concurrent radiotherapy for advanced esophageal carcinoma
XIA Youyou ; WANG Lei ; SONG Da'an ; LI Shiqiu ; JIANG Xiaodong
Chinese Journal of Clinical Oncology 2014;45(11):716-719
Objective:To evaluate the efficacy and safety of raltitrexed plus oxaliplatin combined with concurrent radiotherapy for advanced esophageal carcinoma. Methods:A total of 54 patients with stageⅡ/Ⅲadvanced esophageal carcinoma according to the clinical staging of esophageal carcinoma nonsurgical methods were treated with raltitrexed plus oxaliplatin combined with concurrent radiotherapy. The patients were irradiated with a dose of 60 Gy in 30 fractions. Two cycles of concurrent chemotherapy were adminis-tered during radiotherapy, with 100 mg/m2 oxaliplatin and 2.6 mg/m2 raltitrexed on d1 and d22. Results:The complete response rate was 16.7%(9/54), and the partial response rate was 68.5%(37/54). The total response rate was 85.2%. The no response and progression rate was 14.8%(8/54). The one-and two-year local control rates and overall survival rates were 75.4%, 57.3%and 70.4%(95%CI, 0.6-0.8), 46.6%(95%CI, 0.3-0.6), respectively. The incidence rates of radiation-induced esophagitis, leucopenia, acute diarrhea, neuro-toxicity were 100%, 72.2%, 16.7%, and 44.4%, of which 7.4%, 7.4%, 1.9%, and 0%were≥grade 3, respectively. Conclusion:Ralti-trexed plus oxaliplatin combined with concurrent radiotherapy can enhance the response rate and prolong the survival of patients with advanced esophageal carcinoma. The regime has mild toxicity and is worthy of further study in PhaseⅢ.
3.Application value of clinical typing in the treatment of Budd-Chiari syndrome
Xiaowei DANG ; Luhao LI ; Lin LI ; Hai LI ; Shaokai XU ; Youyou LIU ; Peiqin XU
Chinese Journal of Digestive Surgery 2016;15(7):696-701
Objective To investigate the application value of clinical typing in the treatment of BuddChiari syndrome (BCS).Methods The retrospective corss-sectional study was adopted.The clinical data of 95 patients with BCS who were admitted to the First Affiliated Hospital of Zhengzhou University from January 2012 to September 2015 were collected.Based on patients' compensation and clinical symptoms,3 clinical typing and 8 subtypes of BCS were proposed,and each subtype was treated with corresponding strategies.Observation indices included (1) the clinical typing of BCS,(2) selection of treatment,(3) treatment effect,(4) follow-up situations.Follow-up using telephone interview and outpatient examination was performed once within 3 months after the first treatment and then once every 6 months up to December 2015 or death,loss to follow-up and experienced decompensation.During follow-up,color Doppler ultrasound and blood bio-chemistry test were performed regularly,and CT angiography was also conducted when necessary.Count data were presented as the case or percentage.The survival rate was calculated using Kaplan-Meier method and the survival curve was drawn.Results (1) BCS clinical typing of 95 patients:4 were detected in type Ⅰ (3 in type Ⅰ a and 1 in type Ⅰ b),7 in typeⅡ (4 in type Ⅱa and 3 in type Ⅱb),and 84 in type Ⅲ(43 in type Ⅲa,4 in type Ⅲb,32 in type Ⅲc,and 5 in type Ⅲd).(2) Selection of treatment in 95 patients:① among the 3 patients with type Ⅰ a,2 of them received inferior vena cava balloon angioplasty while 1 patient had to give up the operation due to failure in opening the occlusion.This patient underwent close observation and follow-up afterwards.② The patient with type Ⅰ b underwent cavity-antrum artificial blood vessel bypass operation due to failure in opening the occlusion.③Among the 4 patients with type Ⅱ a,one of them underwent hepatic vein balloon angioplasty.The other 3 patients underwent close observation and follow-up because of failure in intervention therapy,such as segmental occlusion of hepatic vein or difficulty in finding the hepatic vein.④ Among the 3 patients with type Ⅱ b,due to the history of upper gastrointestinal bleeding,2 patients received modified spleen-lung fixation and intestine-cavity blood vessels bypass,respectively,and 1 patient received intestine-cavity artificial blood vessels bypass due to severe peritoneal effusion.⑤ Among the 43 patients with type Ⅲ a,35 patients underwent inferior vena cava balloon angioplasty due to failure in hepatic vein intervention therapy (6 of them received firstly thrombolysis treatment due to combined thrombosis.Four patients received inferior vena cava and hepatic vein balloon angioplasties.Another 4 patients received close observation and follow-up due to failure in both inferior vena cava and hepatic vein intervention therapy.⑥Among the 4 patients with type Ⅲ b,2 underwent inferior vena cava balloon angioplasty and intestine-cavity artificial blood vessel bypass.The other 2 patients only received modified spleen-lung fixation because of failure in inferior vena cava intervention therapy.⑦ Among the 32 patients with type Ⅲ c,3 underwent inferior vena cava and hepatic vein balloon angioplasties,and 27 patients underwent only inferior vena cava balloon angioplasty due to failure in hepatic vein intervention therapy (7 of them received balloon angioplasty following thrombolysis treatment due to combined thrombosis).On account of failure in both inferior vena cava and hepatic vein intervention therapy,2 patients underwent resection of lesion membranes and cavity-antrum artificial blood vessel bypass,respectively.⑧ Among the 5 patients with type Ⅲ d,1 underwent inferior vena cava balloon angioplasty and intestine-cavity artificial blood vessel bypass,and 4 underwent only modified spleen-lung fixation due to failure ininferior vena cava intervention therapy.(3) Treatment efficacy:of 95 patients,8 received followup observation,and 87 patients recovered to varied extent after interventional therapies and operations,with symptomatic relief of leg edema,ulcer,peritoneal effusion and esophageal varicosity.Eighty-seven patients went through the perioperative period safely,and no death occurred.The incidence of postoperative complications was 10.3% (9/87).The complications mainly include venous thrombosis in lower limbs during catheter-directed thrombolysis therapy,pleural effusion,pneumatosis,and peritoneal effusion after surgery,all of which were cured after symptomatic treatment.(4) Follow-up results:87 were followed up for 3-42 months with an average time of 19 months.During the follow-up,5 patients (1 in type Ⅰ a and 4 in type Ⅲa) received recanalization surgery because of the reocclusion after the inferior vena cava balloon angioplasty,and no decompensation occurred.However,decompensation was found in 11 patients (disease progression in 4 patients and symptom relapse in 7 patients).The survival rates of patients without decompensation at 0.5,1.0,2.0 and 3.0 years after the first treatment were 96.5%,95.0%,83.4% and 80.5%,respectively.Conclusion According to patients' compensation and clinical symptoms,clinical typing of BCS and treatment strategiesis are determined,and it will provide a satisfactory clinical efficacy.
4.CT diagnosis of criss-cross heart
Youyou YANG ; Ruping DAI ; Yi LI ; Xiangmin LI ; Xuhui ZHOU ; Quanfei MENG
Chinese Journal of Radiology 2010;44(4):379-382
Objective To probe the clinical application of CT in the diagnosis of criss-cross heart(CCH).Methods Five patients with CCH confirmed by operation were retrospectively analyzed.Enhanced 64-slice spiral CT was performed in 2 patients and enhanced single-slice electron beam CT was performed in 3 patients.Three dimensional reconstructions were applied for the fully display of anatomic malformations,and the results were compared with that of echocardiogram and angiocardiogram with Chi-square test Results(1)Visceroatrial situs solitus,twisted and concordant atrioventricular connection,horizontally oriented ventricular septum,ventricular septum defect and pulmonary stenosis were found in all patients on CT.The ventriculoarterial connection was discordant,including double-outlet right ventricle in 4 patients and complete transposition of great arteries in 1 patient In addition,associated anomalies including persistent left superior vena cava(n=2),coarctation of the aorta(n=1)and right aortic arch with right descending aorta(n=1)were detected as well.(2)Total 33 anomalies in 5 cases were found during operation.The diagnostic accuracy of CT,angiocardiogram and echocardiogram was 93.9%(31/33),81.8%(27/33)and 54.5%(18/33)respectively.There was a significant difference between CT and echocardiogram(X~2=13.39,P<0.01),and no significant difference between CT and angiocardiogram(X~2=1.29,P>0.05).Conclusion CT is an excellent imaging technique for the diagnosis of CCH.
5.A clinical comparative study of levosimendan on patients with acutely heart failure
Rui YAO ; Tongwen SUN ; Youyou DU ; Yapeng LI ; Yanzhou ZHANG ; Ling LI
Chinese Journal of Emergency Medicine 2015;24(8):893-896
Objective To investigate the efficacy and safety of levosimendan on patients with acutely decompensated heart failure (ADHF).Methods A prospective randomized and controlled study was carried out from June 2013 to June 2014.Patients were randomly divided into levosimendan group and dobutamine group,with 60 patients in each group.All patients received an intravenous infusion of levosimendan or dobutamine for 24 hours.Brain natriuretic peptide (BNP),stroke volume (SV) and left ventricular ejection fraction (LVEF) were measured.The cardiovascular mortality,rehospitalization rate,the composite endpoint differences and the incidence of adverse events were compared between two groups in 1,3,6 months after treatment.Comparisons between two groups were made using Student t-test or one-way ANOVA.Statistical analysis was performed using SPSS 17.0 software and a P value of < 0.05 was considered statistically significant.Results There was no significant difference in the basic characteristics between two groups.Compared with baseline level,the plasma BNP levels,SV and LVEF were improved at 24 h in both groups (P < 0.05).The cardiac function indexes were better in levosimendan group than in dobutamine group at 24 h [BNP (1147±407) pg/mL vs.(1 502±501) pg/mL,SV (60.9±9.6) mL vs.(57.3±10.3) mL,LVEF (31.6±6.0)% vs.(28.8±5.1)%,P<0.05].One month later,the cardiac function indexes were still better in levosimendan group than baseline [BNP (796 ± 296) pg/mL vs.(1 951 ±692) pg/mL,SV (64.6±9.5) mL vs.(52.2±9.1) mL,LVEF (33.4 ±5.8)% vs.(25.7 ± 6.1) %,P < 0.05].After l months of treatment,the composite endpoint in levosimendan group was significantly lower than dobutamine group (5% vs.16.3%,P =0.043).There was a downward trend of mortality and rehospitalization rate in levosimendan group in six months follow-up (P > 0.05).The incidence of side effects was no statistically significant between groups (both were 13.3%).Conclusions Levosimendan is superior to that of dobutamine in improving the hemodynamic status and prognosis in ADHF patients,and the adverse reaction of levosimendan is less.
6.Rat ADSCs transfected by lentivirus vector-mediated enhanced green fluorescent protein
Shaolei LI ; Youyou YANG ; Yunjiang LIU ; Li JIANG ; Xiaofeng NIU ; Yinfeng XU ; Jianhua YI
Chinese Journal of Microsurgery 2014;37(2):147-151
Objective To explore the labeling method of rat adipose-derived stromal cells,and observe the stem cell characteristics and the activities of EGFP-positive adipose-derived stromal cells (EGFP-ADSCs) in vitro and in vivo.Methods ADSCs were transfected for 12 h with enhanced green fluorescent protein gene (EGFP) carried by lentivirus(Lv-EGFP) vector at different value of MOI (0,5,10,25,50,100,respectively).The rate of EGFP expression and fluorescence intensity were evaluated by flow cytometric analysis and fluorescence microscopy,and cell viability was detected by MTT-test after transfection.Secondly,cells were exposed either to adipogenic medium or osteogenic medium,then stained with Oil Red O and Alizarin Red S.Cell growth was investigated on frozen longitudinal sections when EGFP-ADSCs were injected into acellular nerves to build tissue-engineered peripheral nerves repairing sciatic nerve defects in rats for 1 week in vivo.Results EGFP-positive rate and fluorescence intensity peak at 4 days after transfection.The rate of EGFP expression was 0.13%,31.09%,75.33%,92.66%,96.70%,98.38% for MOI =0,1,5,25,50,100,respectively.The positive rate between the experimental group and control (MOI =0) existed significantly difference (P < 0.05) ; the difference between MOI =1,5 groups and MOI =25,50,100 groups were also observed (P < 0.05).There was no statistical difference in EGFP-positive rate and cell proliferation activity among MOI =25,50,100 groups (P > 0.05).MOI =25 was chosen as best scheme to transfect ADSCs for subsequent experiments.Osteogenic and adipogenic differentiation for 20 days,orange calcium deposits,orange-red lipid droplets were seen in EGFP-ADSCs after Alizarin red and oil red O staining.At 1 week in vivo,EGFP-ADSCs evenly distributed and became fusiform on frozen longitudinal sections.Conclusion Lv-EGFP transfection does not affect the ADSCs activity and their osteogenic and adipogenic differentiation,so could be as a tracing method for ADSCs-tissue-engineered peripheral nerves repairing nerve defects.
7.STUDIES ON THE CONSTITUENTS OF ARTEMISIA ANNUA L.
Tu YOUYOU ; Ni MUYUN ; Zhong YURONG ; Li LANNA ; Gui SHULIAN ; Zhang MUQUN ; Wang XIUZHEN ; Liang XIAOTIAN
Acta Pharmaceutica Sinica 2015;50(10):366-370
Six crystalline components were isolated from the lipophilic fraction of Artemisia annua L. They have been identified as four sesquiterpenes, one flavonol and one coumarin. Qinghaosu I and III are new sesquiterpenes. Five main constituents, camphene, iso-artemisia ketone, 1-camphor, β-carophyllene, and β-pinene were identified from the volatile oil of this herb.
Artemisia annua
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chemistry
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Artemisinins
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chemistry
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isolation & purification
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Bridged Bicyclo Compounds
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chemistry
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isolation & purification
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Camphor
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chemistry
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isolation & purification
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Monoterpenes
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chemistry
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isolation & purification
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Oils, Volatile
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chemistry
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Sesquiterpenes
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chemistry
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isolation & purification
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Terpenes
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chemistry
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isolation & purification
8.Electron-beam CT diagnosis of congenital cardiovascular diverticula
Youyou YANG ; Ruling DAI ; Lili ZHENG ; Xiangmin LI ; Xuhui ZHOU ; Qian PENG ; Quanfei MENG
Chinese Journal of Radiology 2008;42(9):919-922
Objective To investigate the clinical application of electron-beam CT(EBCT)in the diagnosis of congenital cardiovascular diverticula. Methotis Retrospective analysis of 9 patients with congenital cardiovascular diverticula confirmed by operation and pathology was done.Of them,enhanced continuous volume scan was performed on 8 patients and enhanced single slice scan was performed on one patient with an Imatron C-150 scanner.Results The group of 9 pailents included one patient with diverticulum of the left ventricle.3 patients with diverticulum of the atria and 5 patients with diverticulum of the aorta.EBCT scan and three dimensional reconstruction could demonstrate not only the origin,size,shape,Location and adjacent structure of diverticula,but also other important complicated abnormalities such as ventrieuloarterial connection disorder,cardiac sepud defect,aortic coarctation and even dissection.Conclusion EBCT is an ideal noninvasive technique in the diagnosis of congenital cardiovascular diverticula
9.The heart rate turbulence in patients with isolated systolic hypertension
Hongwei PAN ; Shenghua ZHOU ; Qiming LIU ; Xuping LI ; Tao ZHOU ; Hua SU ; Youyou DU
Clinical Medicine of China 2008;24(5):456-458
Objective To observe the changes of the heart rate turbulence(HRT)in patients with isolated systolic hypertension(ISH).Methods 85 patients with ISH,80 patients with common hypertension and 58 control subjects were investigated.Turbulence onset(TO)and turbulence slope(TS)were recorded of 24 hours dynamic electrocardiogram.Results The difference of HRT in the three groups was significant(P<0.01).The values of TO and TS was significantly higher in common hypertension group and ISH group than in control group(P<0.05);TS was significantly lower in ISH group than in common hypertension group(P<0.05),but the difference of TO between ISH group with common hypertension group was not obvious.Conclusion The HRT in patients with primary hypertension is significantly lower,which is more significant in patients with ISH than in patients with common hypertension.It suggests that the cardiac autonomic nerves dysfunction perhaps has an important influence on development of hypertension and its prognosis.
10.CT diagnosis of cardiac lipoma
Youyou YANG ; Ruping DAI ; Miao FAN ; Jifei WANG ; Xiangmin LI ; Xiaomei LUO ; Quanfei MENG
Chinese Journal of Radiology 2010;44(8):803-806
Objective To investigate the application of CT in the diagnosis of cardiac lipoma.Methods Retrospective analysis of 6 patients with cardiac lipoma confirmed by operation and pathology was done. Four patients had singles slice electron beam CT plain and contrast and movie scan. Two patients had 64-slice CT plain and enhanced scan. Results (1) One patient was isolated intracavitary lipoma in the right artium, 1 patient was isolated intrapericardial lipoma and 4 patients were intramural lipomas. Of the 4 intramural lipoma, 2 were infiltrative lipomas located in the left ventricle wall or the right ventricle and septum, 2 patients were isolated in the atrio-ventricular septums. (2) CT and three-dimensional reconstruction could depict the location, shape, size, margin and characteristic fat density of lipoma,indicating the diagnosis and classifications. The displacement of coronary artery, pulmonary inflammation and effusions of pericardium and pleural cavity could also be revealed. Conclusion Cardiac lipoma can be accurately diagnosed and classified by CT.