1.Effects of camptothecin on the expression of hypoxia-inducible factor-1α in HaCaT cells
Liangyu ZHANG ; Xiang WANG ; Yaqi LU ; Xiaoyang ZHU ; Yang CHEN
Chinese Journal of Dermatology 2015;48(6):400-403
Objective To estimate the effects of camptothecin (CPT) on the expression of hypoxia-inducible factor-1α (HIF-1α) in HaCaT cells under hypoxic conditions (2% O2),and to explore the potential therapeutic mechanism of topical CPT for psoriasis.Methods Some HaCaT cells were classified into 6 groups:5 test groups cultured in Dulbecco's modified Eagle's medium (DMEM) with the presence of CPT at 12.5,25,50,100 and 200 nmol/L respectively,and 1 control group cultured in DMEM with the presence of dimethyl sulfoxide (DMSO).All the 6 groups of cells were cultured under normoxic conditions for 12,24,48 or 72 hours or under hypoxic conditions for 12 hours.Cell counting kit-8 (CCK-8) assay was conducted to estimate the proliferation of HaCaT cells after the normoxic culture,and Western blot to quantify the protein expression of HIF-1α after the hypoxic culture.Some HaCaT cells were classified into a normoxia group (21% O2) and a hypoxia group (2% O2),and each group was divided into a CPT (100 nmol/L)-treated subgroup and a non-intervention subgroup (treated with the vehicle).After 12-hour culture,real-time fluorescencebased quantitative PCR was performed to measure the mRNA expression of HIF-1α.Statistical analysis was carried out by using Levene'.s test,one-way analysis of variance,Dunnett-t test and factorial analysis with the SPSS16.0 software.Results After treatment with CPT at 12.5-200 nmol/L for 12-72 hours,the proliferation of HaCaT cells was inhibited in a concentration-and time-dependent manner.More concretely,the cell proliferation rates were inhibited by 17.66% ± 6.46%,33.11% ± 4.63% and 56.31% ± 1.69% respectively in HaCaT cells after 12-hour treatment with 200 nmol/L CPT as well as 24-hour treatment with 100 and 200 nmol/L CPT compared with the control group at the corresponding time points (all P < 0.05).The protein expression level of HIF-1 α was significantly decreased in HaCaT cells after 12-hour treatment with CPT at 12.5,25,50,100 and 200 nmol/L under hypoxic conditions compared with the control group (0.348 ± 0.065,0.261 ± 0.112,0.115 ± 0.043,0.045 ± 0.024 vs.1.445 ± 0.329,all P< 0.05).The mRNA expression level of HIF-1α (expressed as △Ct) in the CPT-treated subgroup and non-intervention subgroup was-5.575 ± 0.29 and -5.451 ± 0.21 respectively in the normoxia group,significantly higher than that in the hypoxia group (-6.543 ± 0.57 and -6.203 ± 0.31 respectively,F =29.856,P < 0.05),while there was no significant difference between the CPT-treated and non-intervention subgroups (F =1.667,P > 0.05).Conclusions CPT at 100 nmol/L could inhibit the expression of HIF-1α protein,but had no obvious effect on that of HIF-1α mRNA.
2.Clinical Observation of Salmeterol Fluticasone Combined with Tiotropium Bromide in the Treatment of COPD via Different Inhalation Devices
Huilan ZHU ; Xiumei XIANG ; Limei ZHOU ; Lu SHI ; Feifei WANG
China Pharmacy 2017;28(8):1084-1087
OBJECTIVE:To observe clinical efficacy and safety of salmeterol fluticasone combined with tiotropium bromide in the treatment of COPD via different inhalation devices.METHODS:Eighty COPD patients were selected from our hospital during Jan.2014 to Jan.2015,and then divided into trial group and control group according to random number table,with 40 cases in each group.Both groups were given Salmeterol fluticasone inhalant 500 μg,bid+Tiotropium bromide inhalant 18 μg,qd.Control group was given medicine via inhalation device coming with medicine,while trial group was given medicine via gas compression type ultrasonic spray inhalator.Both groups were treated for 1 year.Blood concentration of medicine 0.5 h after medication,mMRC score and COPD acsessment test (CAT) score 3,6,9 months after treatment,the times of acute exacerbation during treatment,FEV1% before and af ter treatment were all observed in 2 groups.The occurrence of ADR was recorded.RESULTS:Four cases withdrew from trial group and 1 case from control group.After medication,there was no statistical significance in blood concentration of fluticasone,salmeterol and tiotropium bromide between 2 groups (P>0.05).0.5 h after medication,mMRC score of trial group was slightly lower than that of control group,without statistical significance (P>0.05);CAT score of it was significantly lower than that of control group,with statistical significance (P<0.05).The times of acute exacerbation in trial group during treatment was significantly less than control group,with statistical significance (P<0.05).The decrease of FEV1% in trial group was slightly lower than control group,without statistical significance (P>0.05).The incidence of ADR in trial group was significantly lower than control group,with statistical significance (P<0.05).CONCLUSIONS:For COPD patients,salmeterol fluticasone combined with tiotropium bromide via gas compression type ultrasonic spray inhalator is better than inhalation device coming with medicine in clinical efficacy and safety.
3.Benign glandular schwannoma
Xiang DU ; Xiongzeng ZHU ; Hongfen LU ; Al ET
China Oncology 2001;0(03):-
Purpose:To report a seldom case of benign glandular schwannoma and review its pathological characteristic. Methods:The tumor was observed by histology, histochemistry and immunohistochemistry. Results:Histologically, the tumor is composed of spindle cell component showing typical schwannoma and collections of glandular component with the expressions of Leu 7,GFAP,MBP,CEA,AE1/AE3 and 34?E12. Conclusions:The histogenesis of the glandular schwannoma remains uncertain. In our opinion, the glands within the tumor, when examined immunohistochemically, probably represent entrapped dermal appendageal glands with proliferation. [
4.Effects of liraglutide on eNOS and IRS-1 expressions in human umbilical vein endothelial cells cultured with high glucose
Ling YUE ; Jing DONG ; Guangda XIANG ; Linshuang ZHAO ; Junxia ZHANG ; Guangping ZHU ; Lin XIANG ; Min LIU ; Junyan LU
Chinese Journal of Endocrinology and Metabolism 2016;32(4):327-329
Humanumbilicalveinendothelialcells(HUVECs)weretreatedwith3nmol/Lliraglutidefor10, 15, 30, 45, 60, 90, 120, 150, 180, 210, 240, and 270 minutes at the concentrations of 5. 5 or 30 mmol/L glucose. Western blot analysis was used to detected protein expression and phosphorylation of insulin receptor substrates-1 ( IRS-1 ) and endothelial nitric oxide synthase ( eNOS ) . The results showed that the baseline level of phosphorylated-eNOS/eNOS was lower in high glucose group than that in normal group(0. 239 ± 0. 016 vs 0. 400 ± 0. 02,P<0. 05). Liraglutide time-dependently increased phosphorylated-eNOS/eNOS and phosphorylated-IRS-1/IRS-1 levels at 5. 5 or 30 mmol/L glucose.
5.Association between endometrial cancer and metabolic syndrome
Huafeng SHOU ; Juan NI ; Tao ZHU ; Jianhong CHEN ; Xiang ZHANG ; Xiaoxian XU ; Lu CHEN ; Hua YU
Chinese Journal of Obstetrics and Gynecology 2010;45(2):128-131
Objective To study the association between endometrioid uterine carcinomas and metabolic syndrome (MS). Methods A retrospective study was conducted on 123 patients who were admitted in Department of Gynecology Oncology, Zhejiang Cancer Hospital (study group) and 90 healthy women (control group) with matching age from Jan. 2005 to Mar. 2009. The general conditions[including age, whether menopausal, body mass index (BMI)];the risk factors for MS [including waist circumference,fasting plasma glucose, triglycerides(TG), high-density lipoprotein (HDL) and systolic and diastolic blood pressure]were analyzed. The clinical stage, histological type, and pathology differentiated degree of study group with or without MS were also analyzed by univariate analysis and Cox proportional hazards models.Results (1) The univariate survival analysis shown that there were no significant difference with age in two groups[(54.3±0.6) vs. (54.2±0.9) years;P>0.05], while the rate of menopausal, BMI(≥25 kg/m~2), the cases coupled with MS, the size of waist circumference (> 80 cm), the level of fasting plasma glucose (≥5.6 mmol/L),TG(> 1.7 mmol/L)and abnormal systolic and diastolic blood pressure in study group were higher than those in control group (67.5% vs. 48. 9%, 45.5% vs. 23.3%, 43.9% vs.18.9%, 50.4% vs. 27.8%, 53.7% vs. 21.1%, 40.7% vs. 21.1% and 40.7% vs. 25.6%,respectively, all P <0.05). The percentage of HDL(< 1.30 mmol/L) was higher in study group than that in control granp(63. 4% vs. 32. 2%, P <0.05). (2) There were not significant difference for the clinical stage, pathological type, grades between patients with or without MS in study group (P > 0.05). (3) The Logistic multivariate survival analysis shown that central obesity, higher TG, lower HDL and abnormal plasma glucose were independent risk factors for endometrioid uterine carcinomas coupled with MS (P< 0.05). Conclusion Metabolic syndrome is marginally associated with an increased risk of endometrioid uterine carcinomas, which may be the new point to screen, prevention and treatment endometrioid uterine carcinomas.
6.Identification of constituents in Suanzaoren tang by LC-Q-TOF-MS and LC-IT-MS.
Yun-Xiang ZHU ; Lu-Lin CHEN ; Jing-Ru GONG ; Shu-Fang WANG
China Journal of Chinese Materia Medica 2014;39(3):457-465
LC-Q-TOF-MS and LC-IT-MS in positive and negative ion mode were applied to simultaneously characterize the constituents in Suanzaoren tang. Analysis was performed on an Agilent Zorbax SB-C18, Rapid Resolution HT column(4.6 mmx 50 mm, 1. 8 micro m) with gradient elution of acetonitrile(A) -aqueous solution containing 0. 05% formic acid(B) at a flow rate of 0. 6 mL min(-1) and the column temperature was 30 degreesC. By comparing MS fragmentation, accurate molecular weight, literature date and standard compounds information, a total of48 compounds were successfully identified or speculated. The origins of these compounds were assigned to the corresponding Chinese medicine. Thirty-one compounds were reported in Suanzaoren tang for the first time. LC-Q-TOF-MS combined with LC-IT-MS is a simple and rapid tool for the identification of constituents of Suanzaoren tang, and the results could provide evidence for the research on quality combined and effective constituents of Suanzaoren tang.
Chromatography, Liquid
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Drugs, Chinese Herbal
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chemistry
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Mass Spectrometry
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Organic Chemicals
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analysis
8.Effects of camptothecin on the autophagy of HaCaT cells
Yangyang HAO ; Liangyu ZHANG ; Xiang WANG ; Yaqi LU ; Xiaoyang ZHU ; Yang CHEN
Chinese Journal of Dermatology 2017;50(2):86-90
Objective To evaluate effects of camptothecin on the autophagy of HaCaT cells.Methods Some cultured HaCaT cells were divided into several groups to be treated with camptothecin at concentrations of 5,10,25,50,100 and 200 nmol/L,and 0.1% dimethyl sulfoxide (DMSO) (control group),respectively.Cell counting kit-8 (CCK-8) assay was conducted to estimate the proliferative activity of HaCaT cells after 24-and 48-hour treatment,flow cytometry to evaluate cell apoptosis after 24-hour treatment,and Western blot analysis to measure the expression of autophagy-related proteins microtubuleassociated protein 1 light chain 3 (LC3) and p62.Some HaCaT cells were divided into 2 groups to be treated with 10 nmol/L camptothecin and 0.1% DMSO for 24 hours,respectively.Then,indirect immunofluorescence assay (IFA) was performed to determine the LC3 expression.Results Camptothecin at low concentrations of 5 and 10 nmol/L had no significant effects on the proliferation and apoptosis of HaCaT cells.Compared with the control group,the cellular proliferative rates were significantly inhibited by (31.23 ± 1.00)%,(54.21 ± 8.10)% and (66.75 ± 10.70)% in the 50-,100-and 200-nmol/L camptothecin groups after 24-hour treatment respectively,and by (25.81 ± 5.99)%,(44.35 ± 5.32)%,(65.81 ± 8.28)% and (73.23 ± 9.59)% in the 25-,50-,100-and 200-nmol/L camptothecin groups after 48-hour treatment respectively (all P < 0.001).After 24-hour treatment,the apoptosis rates were significantly higher in the 50-,100-and 200-nnol/L camptothecin groups (14.46% ± 2.38%,19.15% ± 1.59%,29.88% ± 1.37%,respectively) than in the control group (3.80% ± 0.13%,all P < 0.001).After 24-hour treatment with 5 and 10 nmol/L camptothecin,the protein expression of LC3 Ⅱ was significantly up-regulated,while p62 protein expression was significantly down-regulated:IFA showed that the percentage of autophagosome-positive cells was significantly higher in the 10-nmol/L camptothecin group than in the control group after 24-hour treatment (36.67% ± 4.55% vs.6.23% ± 0.92%,t =6.546,P =0.003).Conclusions Camptothecin at low concentrations of 5 and 10 nmol/L can induce autophagy of HaCaT cells,but has no obvious effects on cell proliferation and apoptosis.Camptothecin at concentrations of 50,100 and 200 nmol/L can inhibit cell proliferation,promote cell apoptosis,and decrease autophagy levels.
9.Impact of medical staff loyalty on patient satisfaction:a study based on structural equation model
Yuezhou ZHU ; Pei WAN ; Xiang LU ; Guozhong JI ; Hui FU ; Wanwen DAI
Chinese Journal of Hospital Administration 2017;33(3):190-193
Objective To learn the relationship and operating mechanism between medical staff's loyalty and patient satisfaction.Methods Medical staff and inpatients of a tertiary hospital in Nanjing were sampled at a 1:2 ratio for study,with 840 questionnaires released and 269 valid questionnaires recovered;medical staff service quality as perceived by inpatients served as the intermediate variable,with Mplus7.0 used to build a structural formula model for an empirical study of the relationship between staff loyalty and patient satisfaction.Results Staff loyalty exerts certain positive influence on patient satisfaction and staff service quality ( P < 0.001 ); service quality also exerts some positive influence on patient satisfaction (P<0.001);staff loyalty indirectly exerts positive influence(P<0.001)via the service quality as perceived by patients(mediation effect value 0.364 > direct effect value 0.143).Conclusions Structural formula model can reveal the relationship and extent of the influence of staff loyalty on patient satisfaction.This indicates that hospitals should enhance the loyalty,medical competence and communication skills of their medical staff for higher patient satisfaction.
10.A pilot study of the effect of aerobic training with bicycle ergometer on hemiplegic stroke patients without ambulation ability
Zun WANG ; Hongjuan FAN ; Xiao LU ; Xiang WANG ; Xiaojun ZHU ; Lei WANG ; Yihuang GU ; Tong WANG
Chinese Journal of Physical Medicine and Rehabilitation 2011;33(12):924-927
Objective To study the effectiveness and feasibility of low intensity of aerobic bicycle ergometer training on hemiplegic stroke patients unable to perform assistive or independent ambulation.Methods Thirty stroke patients unable of assistive or independent ambulation were randomly divided into a control group and a aerobic training group.Patients in the aerobic training group performed low intensity of aerobic bicycle ergometer training for 30 min,3 times a week for 6 weeks in addition to a routine rehabilitation regime.The training duration and course of routine rehabilitation training were the same in both groups.The related cardiovascular response in aerobic training and changes in motor performance as well as the risk factors for stroke were observed in both groups.Results Twenty-four patients(12 patients in each group)completed the experiment.Patients in aerobic training group performed all training regime safely without any adverse response.Their motor function(including FMA score,Rivermead index,BI ADL ability),exercise endurance duration in exercise test,glucose level at 2 h in oral glucose tolerance test,fasting insulin level and HOMA-IR index all improved greatly as compared to the control group(P < 0.05).Conclusion Low intensity of aerobic bicycle ergometer training can be safely and effectively administered in hemiplegic stroke patients unable of assistive ambulation or independent ambulation.