1.Effects of SAHA combined with PTX on lethality and autophagy of human ovarian cancer OC3 cells
Yue ZHAO ; Yuanlin LIU ; Zhaohui LIU ; Si ZHANG ; Ying TONG ; Yi ZHANG
Military Medical Sciences 2015;(7):504-507,513
Objective To evaluate the effect of suberoylanilide hydroxamic acid(SAHA) or/and paclitaxel(PTX) on lethality and autophagy of human ovarian cancer OC3 cells,and to explore whether the combination of the two drugs has a synergistic function.Methods The morphology of OC3 cells was treated with SAHA and/or PTX, and then the morphology of treated OC3 cells was observed under an inverted microscope, cell proliferation was detected by MTT assay and autoph-agy was analyzed by AO/EB double staining assay.The synergistic effect of SAHA and/or PTX was analyzed by factorial design and gold formula method.Results After treatment with SAHA and/or PTX, the morphology of OC3 cells in the combination group ( SAHA+PTX) displayed significant morphological changes.OC3 cells became less adherent and refrac-tive than in other groups.Cell proliferation by MTT assay demonstrated that the growth inhibition rate of the combination groups was higher than in groups treated with SAHA or PTX respectively( P<0.05) .Furthermore, the synergistic effect af-ter treatment with a combination of SAHA with PTX was proved by the factorial design and gold formula method.The auto-phagy rate of the combined groups was significantly higher than in single treatment groups (P<0.05) by AO/EB double staining.Conclusion SAHA and PTX can inhibit the survival of OC3 cells and induce its autophagy.The two drugs have synergistic antitumor effects.
2.Expression of MTSS1 in cervical cancer tissue and its clinical significance
Si ZHANG ; Yuanlin LIU ; Xiangdong ZHOU ; Yue ZHAO ; Ying TONG ; Yi ZHANG
Military Medical Sciences 2016;40(3):237-240
Objective To detect the expression of metastasis sappressor 1(MTSS1) gene in cervical cancer tissue and to clarify its association with cervical cancer.Methods Totally 103 cases of cervical tissue were collected between Dec 2011 and Dec 2014 and classified according to biopsy and stage .Q-PCR and Western blotting were used to detect the expression of MTSS1 in normal cervical tissue and in different clinical stages of cervical cancer tissue .Results The expression of MTSS1 inⅡB-Ⅳstages of cervical cancer tissue was significantly higher than that of normal tissue or Ⅰ-ⅡA stages through q-PCR (P=0.000).Western blotting results showed that MTSS1 was positively expressed in normal cervical tissue at a rate of 23.3% or 53.3% in cervical cancer tissue.Moreover, the expression of MTSS1 was poorly correlated with age, tumor differentiation and lymphnode metastasis in cervical cancer tissue (P>0.05).The protein level of MTSS1 expressed in ⅡB-Ⅳ stages was significantly higher than that ofⅠ-ⅡA stages(P=0.005).Conclusion The expression of MTSS1 indicates the clinical stage of cervical cancer , suggesting that MTSS1 may play an important role in the development of cervical cancer .
3.Effects of cyclopamine on endometrial carcinoma (HEC-1 A) and apoptosis
Pingping ZHANG ; Xue LI ; Yuanlin LIU ; Si ZHANG ; Xiangdong ZHOU ; Ying TONG ; Yi ZHANG
Military Medical Sciences 2017;41(1):48-52
Objective To investigate the effects of cyclopamine (CYP) on endometrial carcinoma (HEC-1A) cell survival and on induction of cell apoptosis .Methods HEC-1A cells were treated with various doses of CYP (0, 5,10, 20 and 40 μmol/L) for 24 h respectively .Then,the inverted microscope was used to observe cell morphology .Cell proliferation and apoptosis were tested by CCK-8 assay and AO/EB bi-labelling assay.The apoptosis rate of HEC-1A was analyzed using flow cytometric analysis , and the key gene expression of Bax and Bcl-2 was detected by quantitative PCR .Results The HEC-1A cells exhibited dramatic morphological changes after treatment with CYP and in a dose-dependent manner .CYP significantly inhibited HEC-1A cell proliferation using CCK8 assays(P<0.05), and induced cell death by AO/EB bi-labelling assay.Moreover,flow cytometry analysis showed that CYP treatment resulted in HEC-1A cell apoptosis, and that a higher concentration of CYP induced severer cell apoptosis (P<0.05).Meanwhile, CYP treated HEC-1A cells exhibited up-regulated expression of Bax and down-regulated expression of Bcl-2 according to Q-PCR.Conclusion Our findings indicatee that CYP can inhibit HEC-1A cell proliferation and induce cell apoptosis .
4.Effects of SAHA combined with PTX on survival and apoptosis of human paclitaxel-resistant ovarian cancer cells
Zhaohui LIU ; Yuanlin LIU ; Junfeng LI ; Yue ZHAO ; Ying TONG ; Yi ZHANG
Military Medical Sciences 2014;(3):203-206
Objective To evaluate the effect of SAHA or/and PTX on survival and apoptosis of human paclitaxel-resist-ant ovarian cancer OC3/P cells, and explore whether the combination of two drugs has a synergistic effect .Methods The morphology of OC3/P cells in different drug-groups was observed by inverted microscope .Cell viability was evaluated by MTT assay.The apoptosis rate was analyzed by Annexin V-FITC/PI assay.Results The morphology change of OC 3/P cells treated with different drug was observed by inverted microscope , and the change in combination group was more signif-icant than one drug alone group .The result of cell survival measured by MTT assay showed that inhibition rate of combina -tion group was more higher than one drug alone group (P<0.05).The analysis of factorial design and gold formula method all proved that the two drugs had synergy .Further the result of flow cytometry showed that apoptosis rate in combination group was significantly higher than SAHA or PTX alone group (P<0.05).Conclusion SAHA and PTX can inhibit the survival and induce apoptosis of OC 3/P cells, and two drugs have synergistic antitumor effects .
5.Cisplatin inhibites HeLa cell proliferation by suppressing activation of metastasis suppressor gene 1-extracellular signal-regulated kinase/serine-threonine kinase
Si ZHANG ; Yuanlin LIU ; Xue LI ; Xiangdong ZHOU ; Yue ZHAO ; Pingping ZHANG ; Ying TONG ; Yi ZHANG
Chinese Journal of Pharmacology and Toxicology 2016;30(4):350-355
OBJECTIVE To study the molecular mechanism of cisplatin(DDP)by which HeLa cell growth and proliferation are inhibited. METHODS Cultured HeLa cells were treated with DDP 0.02-75 μmol · L-1 for 24 or 48 h. CCK-8 assay was used to determine the cell proliferation. The wound scratch assay was used to detect the cell migration and invasion. Flow cytometry was used to detect the cell cycle arresting. q-PCR was used to test the expression of metastasis suppressor gene 1 (MTSS1)mRNA. Western blot was used to determine protein levels of MTSS1,phosphorylated-extra?cellular signal-regulated kinase(p-ERK) and phosphorylated-serine-threonine kinase(p-AKT). RESULTS Following the treatment with DDP for 24 or 48 h,the proliferation of HeLa cells was inhibited significantly (P<0.05),the value of the half inhibitory concentration (IC50) of cells was 4.14 and 11.82 μmol · L-1. Migration and invasion activity of HeLa cells were reduced according to the wound scratch assay(P<0.05). Flow cytometry results showed that the cell cycle was arrested at S phase. q-PCR results showed that MTSS1 mRNA expression changed with DDP in a concentration-dependent manner (r24 h=-0.965,P<0.01;r48 h=-0.953,P<0.01). Western blot showed that the protein levels of MTSS1,p-ERK and p-AKT expression declined significantly with the increase in DDP concentrations(p-ERK:r24 h=-0.875,P<0.01;r48 h=-0.966,P<0.01. p-AKT:r24 h=-0.831,P<0.01;r48 h=-0.863,P<0.01. MTSS1:r24 h=-0.969,P<0.01;r48 h=-0.988,P<0.01). CONCLUSION DDP treatment inhibits HeLa growth and proliferation by interfering with the MTSS1 expression and disturbing the activation of ERK and AKT signaling pathways.
6.Study on the reliability of CardioChek PA for measuring lipid profile
Ying GAO ; Chenggang ZHU ; Naqiong WU ; Yuanlin GUO ; Geng LIU ; Qian DONG ; Jianjun LI
Journal of Peking University(Health Sciences) 2016;48(3):523-528
Objective:To evaluate the clinical correlation between the CardioChek PA analyzer (CCPA)and a clinical laboratory reference method to use for screening program purposes.Methods:Fasting blood samples were collected on 325 patients (age:23 -86 years).One venous sample was col-lected using a serum tube for the evaluation on a Beckman reference analyzer.A second venous sample was collected in a lithium heparin tube and was evaluated on the CCPA analyzer.Linear regression analy-ses and Bland-Altman method were performed for each measured analyte:total cholesterol (TC),high density lipoprotein-cholesterol (HDL-C),triglycerides (TG)and low density lipoprotein-cholesterol (LDL-C).Results:Our results demonstrated a good clinical agreement for TC,HDL-C,TG and LDL-C (97.0%,92.9%,92.4% and 83.7%)in comparison with the CCPA to the reference analyzer.The correlation coefficients were 0.875,0.81 3,0.91 0,0.864,respectively.P values all <0.001 .There was no significant difference in the detection rate of hyperlipidemia in TC,HDL-C and LDL-C.Conclu-sion:We have identified the pre-analytic phase as an important step to guarantee the quality of results and indicated that the CCPA is a reliable lipid point-of-care testing system that can be used for the appli-cation of clinical screening anywhere.
7.Comparision of non-fasting with fasting blood lipid testing in in-hospital patients
Ying GAO ; Yuanlin GUO ; Naqiong WU ; Chenggang ZHU ; Ping QING ; Geng LIU ; Qian DONG ; Jianjun LI
Chinese Journal of Laboratory Medicine 2017;40(6):431-435
Objective To explore the feasibility of the application of non fasting blood lipid in the hospitalized population.Methods Self-control study was used.608 patients(aged 20~86 years old) were enrolled from April 2015 to October 2016 in lipid center of FuWai hospital.Fasting sample and non-fasting sample(1~4 h after breakfast) were collected from every patient and lipid profile including TG (triglyceride), TC (total cholesterol), HDL-C (high density lipoprotein cholesterol) and LDL-C (low density lipoprotein cholesterol) were measured in clinical laboratory.The results of two tests were compared using the Wilcoxon signed-rank test.Results The differences between non-fasting and fasting lipid test were +0.47 mmol/l (+30%) for TG,-0.03 mmol/l (-2.8%) for HDL-C,-0.09 mmol/l (-3%) for LDL-C and-0.24 mmol/l (-8.7%) for calculated LDL-C (P<0.001 respectively).The differenceswere +0.01 mmol/l for TC and +0.02 mmol/l for non-HDL-C,therefore no statistical difference was observed.When the TG level was stratified,the level of non-fasting LDL-C using directing test method was not significantly different between TG> 4.5 mmol/L and the whole (0.07 vs.0.09),but the level of non-fasting LDL-C using formula method wassignificantly different between TG> 4.5 mmol/L and the whole (0.66 Vs.0.24),andthe drops were 34.9% vs.8.7%.Conclusion Non-fasting lipid test could be an effective routine method for lipid evaluation in the hospitalized population.
8.Expension In Vitro of T Cells from Cord Blood CD34(+) Cells Stimulated with SCF and IL-2
Yuanlin LIU ; Yongjun SUI ; Shuangxi ZHANG ; Zikuan GUO ; Ying WU ; Ning MAO
Journal of Experimental Hematology 2000;8(1):48-51
The generation of large quantities of novel human T cell clones ex vivo would make a wide range of gene-and immuno-therapies for tumor and AIDS possibly. Although it is well established that T cells are derived from CD34(+) cells, the involvement of thymic fragments from either human or murine fetus makes the in vitro T cell perliferation very cumbersome. In this report, cord blood mononuclear cells were used as accessory cells to support the differentiation of CD34(+) cells into naive T cells stimulated with SCF and IL-2. CD4(+) and CD8(+) T cells, under the cultural conditions, were continuously produced in vitro at least over a period of 3 weeks and their ratios changed gradually. CD4/CD8 double positive T cells and RAG-2 gene were existed, and RAG-2 gene, reponsible for TCR rearrangement, was expressed during the cell proliferation. Our study presents a simple culture system in vitro to acquire large quantities of naive T cell clones.
9.Application of Lipoprotein Apheresis in Patients With Familial Hypercholesterolemia
Chenggang ZHU ; Geng LIU ; Naqiong WU ; Yuanlin GUO ; Ruixia XU ; Qian DONG ; Ying GAO ; Yan ZHANG ; Sha LI ; Jianjun LI
Chinese Circulation Journal 2016;31(12):1175-1178
Objective: To explore the safety and efficacy of lipoprotein apheresis (LA) in treating the patients with familial hypercholesterolemia (FH).
Methods: A total of 12 FH patients treated in our hospital from 2015-02 to 2016-10 were retrospectively studied. Based on intensive cholesterol lowering therapy with rosuvastatin (10-20) mg Qd and Ezetimibe 10 mg Qd, the patients received LA by double ifltration plasma pheresis (DFPP) via bilateral elbow central vein or femoral vein. The changes of lipid level were compared at before and after LA treatment.
Results: For pre- and immediately after LA treatment, the average total cholesterol (TC) was (9.42±3.65) mmol/L vs (2.84±0.83) mmol/L, low density lipoprotein cholesterol (LDL-C) was (7.31±3.46) mmol/L vs (1.95±0.82) mmol/L; at 1, 3, 7 and 30 days after treatment, TC and LDL-C levels showed increasing trend, while they were still lower than they were before treatment, allP<0.01. For pre- and immediately, 1 day, 3 days after treatment, the average HDL-C level was (0.96±0.31) mmol/L, (0.63±0.17) mmol/L, (0.56±0.15) mmol/L and (0.68±0.22) mmol/L respectively,P<0.05-0.01. For pre- and immediately after LA treatment, the average TG level was (1.90±0.86) mmol/L vs (0.88±0.38) mmol/L,P<0.05. Only 1 patient had the symptoms of hypotension, nausea and sweat, the patient was relieved by expectant treatment.
Conclusion: LA therapy may decrease blood levels of TC and LDL-C at short term in FH patients with good tolerance;even TC and LDL-C could slowly increase after treatment, while combining with lipid lowering therapy, it has been a safe and effective method for treating relevant patients.
10.Relationship Between ABO Blood Type and Spontaneous Re-canalization in Patients With Acute Myocardial Infarction
Xianliang LIN ; Jing SUN ; Sha LI ; Chenggang ZHU ; Yuanlin GUO ; Naqiong WU ; Ruixia XU ; Ying GAO ; Chuanjue CUI ; Xiaolin LI ; Ping QING ; Yan ZHANG ; Geng LIU ; Qian DONG ; Zhurong LUO ; Jianjun LI
Chinese Circulation Journal 2017;32(6):564-568
Objective: To study the relationship between ABO blood type and spontaneous re-canalization (SR) in patients with acute myocardial infarction (AMI). Methods: A total of 1209 consecutive AMI patients were enrolled. Based on TIMI grade, the patients were divided into 2 groups: Non-SR group, the patients with TIMI grade 0-1,n=442 and SR group, the patients with TIMI grade 2-3,n=767. The relationship between ABO blood type and SR was investigated. Results: Compared with Non-SR group, SR group had more patients with blood type O (32.3% vs 24.7%) and less blood type A (31.7% vs 24.9%). Meanwhile, we found that a lower cholesterol level was related to patients with O blood type and SR occurrence, bothP<0.05. Multi regression analysis indicated that with adjusted age, gender, BMI, hypertension, diabetes, smoking, LDL-C and C-reactive protein, ESR, fibrinogen, D-dimmer, endothelial cardiac function, blood type O may independently predict SR occurrence in AMI patients (OR=1.49, 95% CI 1.10-2.05), while blood type A may have disadvantage for SR (OR=0.65, 95% CI 0.48-0.80). Conclusion: ABO blood type has been related to SR in AMI patients, blood type O is in favor of SR, while blood type A has disadvantage for SR occurrence.