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.

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

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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.

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.

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.

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.

ObjectiveTo investigate the effects of morphine preconditioning on myocardial ischemiareperfusion (I/R) injury and the expression of phosphorylated extracellular signal-regulated kinase 1/2 (p-ERK1/2)in rats with chronic heart failure.MethodsForty-eight healthy male SD rats weighing 220-250 g were randomly divided into 6 groups ( n =8 each):control group (group C),sham operation group (group S),I/R group and preconditioning with low,median and high doses of morphine groups (groups MP1-3 ).Chronic heart failure was induced by iv edriamycin 2.0 mg/kg once a week for 6 weeks in groups S,I/R and MP1-3.Left ventricular end-diastolic diameter (LVEDD) and left ventricular end-systolic diameter (LVESD) were measured using ultrasound,and left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) were calculated at the end of 14th day after the end of adriamycin administration.Blood samples from the carotid artery were collected after ultrasonography for determination of the plasma brain natriuretic peptide (BNP) concentration.Myocardial I/R was induced by 30 min occlusion of left anterior descending branch of coronary artery followed by 120 min reperfusion at 2 day after ultrasonography in groups I/R and MP1-3.In groups MP1-3,iv morphine 0.015,0.030 and 0.050 mg/kg were repeated 3 times at 5 min interval at 30 min before ischemia respectively,while normal saline 5 ml/kg was given in group I/R.The animals were sacrificed at the end of reperfusion in groups S,I/R and MP1-3,and the hearts were removed to measure the area at risk (AAR),infarct size (IS),and IS/AAR ratio was calculated.The p-ERK1/2 expression in myocardium was assessed by Western blot.ResultsThe LVESD and plasma BNP concentration were significantly higher,while the LVEF and LVFS lower in the other 5 groups than in group C (P ＜0.01).No myocardial infarction was found in group S.The p-ERK1/2 expression was significantly lower in groups I/R and MP1 than in group S (P ＜ 0.05).IS and IS/AAR ratio were significantly lower,and p-ERK1/2expression was significantly higher in groups MP2.3 than in group I/R ( P ＜ 0.05).There were no significant differences in IS,IS/AAR ratio and p-ERK1/2 expression between groups MP1 and I/R (P ＞ 0.05).IS and IS/AAR ratio were decreased gradually,and the p-ERK1/2 expression was up-regulated gradually in groups MP1-3 ( P ＜0.05).ConclusionMorphine preconditioning can confer cardioprotection against myocardial I/R in a dose-dependent manner in rats with chronic heart failure.Up-regulation of p-ERK1/2 expression is involved in the underlying mechamism.

Objective To investigate the effect of the ubiquitin-proteasome pathway inhibitor MG132 on the natural resistance to cisplatin in the human cervical cancer line (HCE1) muhicellular spheroid (HCE1/MCS) model and to probe it if MG132 could reverse the HCE1/MCS resistance to cisplatin, as well as the possible mechanism of drug resistance.Methods (1) HCE1/MCS was obtained using liquid overlay and rotating technique.(2)Four groups were established (MG132 group, cisplatin group, MG132 + cisplatin group, the control group).Cell viability were measured by trypan blue exclusion assay.Cell cycle and apoptosis were detected by flow cytometry.(3) The expression of nuclear factor (NF) kB of both HCE1 monolayer cells (HCEI/MC) and HCE1/MCS was detected by western blot, and the expression of B cell lymphoma/leukemia-2 (bcl-2) was detected by immunohistochemistry.Results (1) HCE1/MCS was established successfully.(2) Cell inhibited rate of HCE1/MC and HCE1/MCS was: in MG132 group, (11.67 ± 2.34) % vs (10.78 ± 1.17) % (P > 0.05) ; in MG132 + cisplatin group, (92.67 ± 2.52)% vs (91.33 ±2.18)% (P>0.05); in cisplatin group, (45.01±7.44)% vs (9.45±5.98)% (P<0.05).(3)The rate of apoptosis of HCE1/MC and HCE1/MCS were: in MG132 group, 8.14% and 5.97% ; in MG132 + cisplatin group, 99.01% and 95.22% ; in cisplatin group, 33.61% and 0.88%.(4)The expression level of NF-kB and the high expression rate of bcl-2 were: in HCE1/MCS of control group, 0.67 and 60% ; in HCE1/MCS of cisplatin group, 0.85 and 83% ; in HCE1/MCS of MG132 group, 0.39 and 20% ; in HCE1/MCS of MG132 + cisplatin group, 0.47 and 33%.Conclusions (1) HCE1/ MCS present natural resistance to cisplatin and may become a good model for the study of cervical cancer drug resistance in vitro.(2) MG132 could induce the inhibition and apoptosis of HCE1/MCS cells and partially reverse the natural resistance of HCE1/MCS to cisplatin, of which partially reverse the natural resistance may be in relation to the down-regulation of NF-kB and bcl-2 expression.