1.Effect of CD147 monoclonal antibody on paclitaxel resistance in HCE1 multicellular spheroids
Journal of Central South University(Medical Sciences) 2011;36(3):192-202
Objective To investigate the effect of CD147 monoclonal antibody (mAb) on the natural resistance to paclitaxel (TAX) in the human cervical cancer line (HCE1) multicellular spheroid (HCE1/MCS) model and if CD147 mAb can reverse the HCE1/MCS resistance to TAX. Methods HCE1/MCS was obtained by liquid overlay and rotating technique. HCE1/MCS morphological changes were observed before or after the interference of CD147 mAb. The effects of TAX on HCE1/MCS (including inhibition ratio, IC50 and index of multicellular resistance) before or after CD147 mAb treatment were determined by the method of WST-1 and the inhibition ratio curve was mapped. Cell cycle and apoptosis were detected by flow cytometer (FCM). The expression of CD147 and P-gp of both HCE1/MC and HCE1/MCS was detected by immunocytochemistry. Results HCE1/MCS was established successfully. CD147 mAb could inhibit HCE1/MCS from forming spheroids. CD147 mAb could enhance the sensitivity of HCE1/MCS to TAX. IC50 in different concentrations of CD147 mAb (5,10,20 μg/mL) HCE1/MCS group were (40.31±3.73), (32.43±1.56), and (30.69±1.01) μg/mL. CD147 mAb resulted in G1/G0 arrest in HCE1/MCS. CD147 mAb of low concentrations (0-10 μg/mL) caused a dose-dependent inhibition of HCE1/MCS (P<0.05). Combined with TAX, CD147 mAb could also induce HCE1/MCS cell cycle arrest in both G1/S and G2/M stage. The expression of CD147 and P-gp was consistent in HCE1/MCS groups. Conclusion CD147 plays an important role in muliticellular resistance of cervical cancer and inhibition of CD147 can synergistically reverse the multicellular drug resistance (MCR) in cervical cancer. The MCR of HCE1/MCS mediated by CD147 is related to P-gp.
2.Effects of pseudolaric acid B on the metastatic activity and invasiveness of HeLa cells and the expression of matrix metalloproteinase
Yun HU ; Keshen LI ; Xiaoke WU
Medical Journal of Chinese People's Liberation Army 2001;0(10):-
Objective To investigate the effects of pseudolaric acid B(PAB) on the invasiveness and metastatic activity of cervical cancer HeLa cells in vitro and the possible mechanism.Methods HeLa cells were divided into 5 groups: blank control group,DDP 3.125?g/ml group,20?mol/L PAB group,10?mol/L PAB group and 5?mol/L PAB group.The effect of PAB on invasive ability of HeLa cells was observed with a Transwell cell culture chamber.The mRNA expression of MMP22 and MMP29 in HeLa cells was detected by RT-PCR and the protein expression was detected by Western blotting.Results The average number of invaded HeLa cells was 15.60?2.45 in 20?mol/L PAB group and 20.14?2.48 in 10?mol/L PAB group,which was significantly lower than those in control group(46.50?4.70) and PAB 5?mol/L group(42.44?5.80,P
5.Pharmacodynamics Study of Different Lactone Ratios of Hydroxycamptothecin on Mice Models of H22 Hepatoma
Rong WANG ; Wei HU ; Song WU ; Yun FANG
Herald of Medicine 2015;(4):471-474,475
Objective To investigate the anti-tumor effects of hydroxycamptothecin ( HCPT ) with different lactone ratios on the mice models of H22 hepatoma. Methods Mice models of H22 hepatoma were established. Tumor inhibiting rates of HCPT with different lactone ratios ( 0%, 25%, 50%, 75%, 100%) and the growth status of model mice before and after chemotherapy were observed. Serum biochemical indices were determined to investigate the effects of HCPT with different lactone ratios on hepatic and renal function of the mice. Results Positive control drug and HCPT with different lactone ratios all inhibited the tumor in mice with H22 hepatoma, the inhibition rate was 65. 30%, 12. 57%, 49. 23%, 75. 47%, 90. 06% and 93. 22%, respectively. Compared with the model control group, the living conditions of the mice in HCPT groups were improved. With increasing of lactone ratios, the hepatic injury was alleviated markedly, but the renal injury was aggravated. Conclusion There is a positive correlation between lactone ratios and its anti-tumor effect, and HCPT with 75% lactone can achieve preferable anti-tumor effect with less toxicity as compared with that with 100% lactone ratio.
6.Effects of limb ischemic preconditioning on myocardial ischemia-reperfusion injury in a rat model of severe hemorrhagic shock and resuscitation
Xianwen HU ; Lingling JIANG ; Xiaofen LIU ; Yun WU ; Yun LI ; Ye ZHANG
Chinese Journal of Anesthesiology 2014;34(1):116-119
Objective To evaluate the effects of limb ischemic preconditioning on myocardial ischemiareperfusion injury in a rat model of severe hemorrhagic shock and resuscitation.Methods Twenty-four male Sprague-Dawley rats,weighing 300-350 g,were randomized into 3 groups (n =8 each) using a random number table:control group (group C); severe hemorrhagic shock and resuscitation group (HSR group); limb ischemic preconditioning group (group LIP).Hemorrhagic shock and resuscitation was induced by withdrawing blood (50% of the total blood volume) from the left common carotid artery over an interval of 1 h,and 30 min later the animals were then resuscitated by infusion of the shed blood via the jugular vein over 30 min.Limb ischemic preconditioning was induced by 4 cycles of 5 min limb ischemia followed by 5 min reperfusion at 40 min before ischemia in LIP group.Before withdrawing blood (T0),immediately after the end of withdrawing blood (T1),before infusion of the shed blood (T2),and at 0,1 and 2 h after infusion of the shed blood (T3-5),mean artery pressure (MAP) was measured,the cardiac output (CO),left ventricular ejection fraction (LVEF) and myocardial performance index (MPI) were detected using color Vivid flow imaging,and total vessel density (TVD),perfusing vessel density (PVD),proportion of perfused vessels (PPV),microvascular flow index (MFI) of sublingual microcirculation were measured using sidestream dark-field imaging.The survival rates within 72 h after hemorrhagic shock and resuscitation were recorded.Results Compared with C group,MAP,CO,LVEF,TVD,PVD,PPV and MFI were significantly decreased and MPI was increased at T1-5 in HSR group and at T1 and T2 in LIP group (P < 0.01).Compared with HSR group,MAP,CO,LVEF,TVD,PVD,PPV and MFI were significantly increased and MPI was decreased at T3-5,and the survival rate within 72 h was increased in LIP group (P < 0.01).Conclusion Limb ischemic preconditioning can significantly attenuate myocardial ischemiareperfusion injury induced by severe hemorrhagic shock and resuscitation in rats and is helpful for prognosis.
7.Anti-tumor mechanisms of lymphopenic mice transferred with tumor-specific lymphocytes and immunized with mIL-21 tumor vaccine
Yun WU ; Chunsheng LIU ; Jun DOU ; Fengshu ZHAO ; Weihua HU ; Ping WEN ; Kai HU ; Xiangfeng HE
Chinese Journal of Microbiology and Immunology 2008;28(12):1143-1148
Objective To explore the mechanism of anti-tumor effects of transferring tumor-specif-ic lymphocytes obtained from pre-immunized BALB/c mice with inactive rolL-21 tumor vaccine (mIL-21-Sp2/0)to syngeneic mice, associated with mIL-21 tumor vaccine immunization, in the condition of cyclo-phosphamide (Cy)-induced lymphopenia. Methods Activated lymphocytes of spleen and lymph nodes ob-tained from pre-immunlzed syngeneic mice with irradiated mIL-21-Sp2/0 cells were infused into BALB/cmice treated with Cy 2 days before, subsequently vaccinated with mlL-21 tumor vaccine, after 7 days, chal-lenged with Sp2/0 tumor cells, observed the growth of tumor of mice. T lymphocyte subsets differentiation was measured by flow cytometry (FCM) analysis. The proliferation and cytotoxie activities of activated lym-phocytes were analyzed by FCM, respectively, staining with CFSE and 7-AAD. The number of IFN-γ-secre-ting cells was evaluated by ELISPOT. Results The lymphopenic mice were transferred with activated lym-phocytes and inoculated with raiL-21 tumor vaccine might provide superior anti-tumor immunoprotection, re-tard tumor growth of the mice. The proliferating capabilities and killing rate of transferred tumor Ag-specific lymphocytes enhanced obviously, the number of IFN-γ-secreting cells was significantly higher compared with the control groups. Conclusion Under Cy-induced lymphopenia condition, tumor Ag-specific lymphocytes sensitized by raiL-21 tumor vaccine were transferred to mice and immunized with mlL-21 tumor vaccine at the same time, benefit the proliferation of transferred effective cells and immune cells itself, assist to form and sustain special anti-tumor effects.
8.Evaluate of transesophageal echocardiography during lung transplantation
Yafeng WU ; Shengcai HOU ; Hui LI ; Anshi WU ; Huaping DAI ; Bin HU ; Yun YUE ; Yidan LI
Chinese Journal of Thoracic and Cardiovascular Surgery 2011;27(1):24-26
Objective To investigate the clinical value of transesophageal echocardiography during the lung transplanta tion. Methods From August 2005 to August 2009, 19 patients with advanced lung diseases received lung transplantation.The average age was(48.35±13.04) years. The echocardiographic probe was placed in patient's esophagus before surgery.The left and right pulmonary venous openings, artery blood flow velocity, right ventricular wall motion, left and right ventricular volume, right ventricular ejection fraction were recorded at different time intervals during lung transplantation, especially at the break and after completion of bronchus, pulmonary veins, and pulmonary artery anastomosis. Results The procedure included sequential-type lung transplantation in 6 cases and single lung transplantation in 13. The blood flow disappeared when blocking pulmonary artery and vein and right ventricular volume increased slightly. The right ventricular volume restored after completion of trachea, pulmonary veins, pulmonary artery anastomosis. TEE detected that the blood flow velocity of pulmonary veins, pulmonay artery anastomosis increased slightly. In 1 case the opening of the right pulmonsry artery blood flow velocity increased significantly and blood flow velocity decresed and blood oxygen partial pressure resumed after re-anastomosis of pulmonary artery. Conclusion TEE play an important role in monitoring pulmonary artery and vein anastomosis diameter and blood flow velocity and right ventricular function and predicting complications during lung transplantation.
9.Role of asymmetric dimethylarginine in acute lung injury induced by cerebral ischemia/reperfusion injury in rats.
Yun-hu WU ; Xuan ZHANG ; Dian-hua WANG
Journal of Southern Medical University 2011;31(8):1289-1294
OBJECTIVETo determine the role of asymmetric dimethylarginine (ADMA) in acute lung injury induced by cerebral ischemia/reperfusion (I/R) injury in rats.
METHODSAdult male SD rats were randomly divided into 4 groups, namely the sham-operated group (S), cerebral I/R model group, ADMA+I/R group, and dimethylarginine dimethylaminohydrolase (DDAH)+I/R group. In the latter 3 groups, acute lung injury was induced by left middle cerebral artery occlusion for 120 min. After a 24-h reperfusion, the rats were sacrificed and the activities of nitric oxide synthase (NOS) and contents of nitric oxide (NO) were measured using reductase and colorimetric assay. The mRNA and protein expressions of protein kinase C (PKC) and myosin light chain kinase (MLCK) in the lung tissues were detected with RT-PCR and Western blotting, respectively. The contents of ADMA in the bronchoalveolar lavage fluid (BALF) and blood flowing into and out of the lungs were measured by ELISA.
RESULTSCerebral I/R injury caused significantly elevated ADMA levels in the BALF and blood flowing into the lungs, and obviously lowered the NO concentration and NOS activity in the lung tissues (P<0.05). Following cerebral I/R injury, MLCK and PKC mRNA and protein expressions were significantly upregualted in the lung tissues (P<0.05). Exogenous DDAH obviously decreased the levels of ADMA in the BALF and blood flowing into the lungs, increased NO concentration and NOS activity, and down-regulated MLCK and PKC mRNA and protein expressions in lung tissues of rats with cerebral I/R injury (P<0.05).
CONCLUSIONADMA contributes to the development of acute lung injury following cerebral I/R injury in rats by upregulating MLCK and PKC expression. ADMA may serve as a novel therapeutic biomarker and a potential therapeutic target for acute lung injury induced by cerebral I/R injury.
Acute Lung Injury ; etiology ; physiopathology ; Animals ; Arginine ; analogs & derivatives ; metabolism ; pharmacology ; Brain Ischemia ; complications ; Male ; Myosin-Light-Chain Kinase ; genetics ; metabolism ; Nitric Oxide Synthase ; antagonists & inhibitors ; Protein Kinase C ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; complications ; physiopathology ; Up-Regulation ; drug effects
10.Optimization of spraying dry technology of Biqiu ranules ethanol extract by Box-Behnken response surface method.
Yan-jun ZHANG ; Li-li LIU ; Jun-hua HU ; Yun WU ; En-xiang CHAO ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(18):3585-3589
With inlet temperature, specific gravity, feeding speed as independent variables, the comprehensive evaluating indexes of content of schisandrin and arctiin as dependent variable, the experimental data were fitted to a second order polynomial equation. Based on establishing the mathematical relationship between the comprehensive evaluating indexes and respective variables, Box-Benhnken central composite test and response surface analysis method was employed to optimize the spray drying technology of Biqiu granules ethanol extract. The optimal drying parameter was as follows: the inlet temperature was 175 degrees C, the specific gravity was 1.10, feeding speed was 32 r x min(-1). Under these conditions, the comprehensive evaluating indexes of spraying dry processes was 92.68, which was close to the model prediction. The spraying dry technology of Biqiu granules ethanol extract optimized by response surface methodology was accurate and feasible, which provided theoretical experiment basis for the industrialization production.
Chemistry, Pharmaceutical
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methods
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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chemistry
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isolation & purification
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Ethanol