Objective To establish a high performance liquid chromatography (HPLC) method for determining the choles‐terol efflux from macrophage‐derived foam cells mediated by apolipoprotein A‐1(apoA‐1) .Methods Human THP‐1 monocytic cells ,pre‐treated with 160 nmol/L phorbol‐12‐myristate acetate (PMA) for 24 h to differentiate into adherence macrophages ,then incubated with 50 μg/mL acetylated low density lipoprotein (ac‐LDL) for 48 h to induce foam cells formation ,then added apoA‐1 for 24 h .THP‐1‐derived macrophage foam cells were identified by oil red O‐staining ,and the cellular cholesterol content by meas‐ured by HPLC method .Cholesterol efflux from macrophage foam cells was determined by HPLC analysis and liquid scintillation counting ,respectively .Results Oil red O‐stainable lipid droplet accumulation were observed in entire cytoplasm of THP‐1‐derived macrophage foam cells .Measuring cellular cholesterol content showed that free cholesterol ,total cholesterol and cholesterol ester content in macrophage foam cells were increased remarkably than PMA group macrophages (P<0 .01) .After treated with ac‐LDL for 48 h ,the macrophage foam cells accumulated 80 .25 μg/mg cell protein and 47 .65 μg/mg cell protein respectively ,and the cho‐lesterol ester accounted for 59 .38% of the cellular total cholesterol (P<0 .01) .The ratio of cholesterol efflux reached 5 .63% and 7 .08% respectively by HPLC analysis and liquid scintillation counting using apoA‐1 mediation (P<0 .01) .Conclusion Combina‐tion of an enzymatic catalysis and HPLC method for determining cholesterol efflux from foam cells is successfully established in this study , thus providing a technical foundation for the further study of cellular lipid homeostasis .

Cytokine-induced killer cells (CIK) is the fourth largest cancer treatment after surgery,chemotherapy and radiotherapy,and it is the development direction of cancer treatment.It is a new type of immune cell,and it is named after natural killer cell samples T lymphocytes as it express CD3 and CD56.Currently,CIK treatment has a broader range of clinical applications,and it has achieved the better clinical efficacy in the blood system cancer and solid tumors,The CIK adoptive immunotherapy is considered to be a new hope for the anticancer treatment.

Hepatocyte is the core raw materials of bioartificial liver support system, primary hepatocyte is lim-ited to application because of short survival and difficult cul-ture in vitro. Porcine hepatocyte which has been used re-cently exist the risks of endogenous retrovirus transmission.With the development of molecular biology, it has been pos-sible that hepatocyte is immortalized recently. Immortalized hepatocytes have greatly significant to drug toxicology, bio-artificial liver support system and tissue engineering of liver.Therefore, we will review the prospects for research and ap-plication of immortalized hepatocytes.

[Abstract ] Objective The purpose of this study was to construct a short hairpin RNA (shRNA) interference lentiviral vector targeting the humanβ-COP gene and to evaluate its inhibitory effect on β-COP in THP-1 cells. Methods We designed and synthesized 4 humanβ-COP-specific oligonucleotide sequences and inserted them into the pGMLV-SC1 vector to construct a recombinant vector fol-lowed by transfection of HEK 293T cells with the recombinant vector and Lenti-HG Mix to produce lentiviruses and detect the viral con-tent.After infecting the THP-1 cells with the packaged lentiviruses , we analyzed the inhibitory effect of β-COP-shRNA on the β-COP gene by quantitative PCR and Western blot . Results Sequencing confirmed that the β-COP-specific oligonucleotide sequences were in-serted into the lentiviral vector and the lentiviruses were packaged in the transfected HEK 293T cells, with the final viral content of 1 × 109 TU/mL.Quantitative PCR showed that the 4 β-COP-shRNA vectors significantly decreased the mRNA expression of β-COP (P<0.01), with interference rates of 16.9 %,32.5%, 74.0%, and 50.3%, respectively.Western blot also indicated their inhibitory effect on the protein expression of β-COP, with an inhibition rate of 76.4% onβ-COP-shRNA3. Conclusion Lentiviral shRNA interference vectors targeting human β-COP were constructed successfully , which could suppress the expression of the human β-COP gene.

AIM To study the effects of homoharringtonine(HHT) on inhibition of proliferation and induction of apoptosis of nasopharyngeal carcinoma cells CNE 2Z. METHODS The inhibitory rates of the proliferation and IC 50 were detected by MTT method. The apoptosis was analyzed by flow cytometry (FCM), agarose gel electrophoresis and Hoechst 33258/PI fluorescence staining. RESULTS After cells were treated with HHT of different concentrations for 24, 48 and 72 h,respectively,the inhibitory rates of the proliferation rised with concentration and time. The IC 50 values of 24, 48 and 72 h were (0 629?0 039), (0 483?0 011) and (0 389?0 027) mg?L -1 , respectively. The difference among IC 50 values was obvious ( P

Objective To study the common aetiology and the principle of treatment of neonatal hyponatremia.MethodsThe clinical data of 48 neonatal hyponatremia cases,admitted to our hospital from June 2006 to January 2010,were analyzed retrospectively. ResultsThe main manifestations of neonatal hyponatremia were poor response ( 37.5％ ) and anorexia ( 31.3％ ).The common causes included anoxic diseases during perinatal period,improper feeding,effects of specific diseases,premature birth et al.After active treatment,the serum sodium returned to normal in 47 neonate within three days,except for 1 premature infants died of serious respiratory distress syndrome.Conclusion Neonatal hyponatremia shows no special clinical manifestation.The aetiologies are complicated,which should be treated in different methods correspondingly.The inquiry of feeding history,the treatment of primary illness,the early detection of serum sodium levels and timely correction of the serum sodium levels are very important to improve the successful rate and reduce sequela.

AIM To investigate the effects of homoharringtonine (HHT) on the expression of caspase-3 pro-tein and mRNA in nasopharyngeal carcinoma cells CNE-2Z. METHODS After CNE-2Z cells were treated with HHT [0(control),0.125,0.25,0.5,1 mg?L -1] for 8 h, the expression of pro-caspase-3 protein was analyzed by Western Blot and the expression of caspase-3 mRNA was detected by semi-quantitative RT-PCR. RESULTS As HHT-concentration increased, the expression of pro-caspase-3 protein decreased significantly (P

Objective To evaluate the clinical value of adenosine triphosphate (ATP) determination in CD4+ cells in cytomegalovirus pneumonia after renal transplantation.Methods The ATP level of CD4+ T cells was measured by ImmuKnowTM kit.The ATP levels were determined in 187 renal transplant recipients before and 30,60,90,180 days after operation,and at the time of CMV pneumonia and 4 weeks after treatment of CMV pneumonia.The associations between ATP levels and CMV pneumonia were analyzed.Analysis of variance (ANOVA),Pearson-Spearman and relative risks were used for data analysis.Results 17 cases out of 187 renal transplant recipients were diagnosed as CMV pneumonia (9.1％),and the onset of CMV pneumonia started on the (2.8 ±1.2)month after renal transplantation.ATP concentrations in CD4+ T cells were significantly lower after operation than those before operation (P＜0.01).ATP concentrations reached the lowest on the about postoperative day 90 (P＜0.05),then increased gradually.In 17 recipients with CMV pneumonia,the ATP levels before and 30,90 days after operation,at the time of CMV pneumonia and 4th week after treatment of CMV pneumonia were (376 ±182),(283 ± 146),(196 ± 112),(145 ± 102) and (236 ± 117) μg/L respectively.ATP levels at the time of CMV pneumonia were significantly lower than any other time points (P＜0.05).There was close correlation between ATP levels and CMV pneumonia.Conclusion The determination of ATP in CD4+ cells could reflect the status of cell-mediated immunity in renal transplant recipients,and could evaluate the severity and prognosis of CMV pneumonia and guide the clinical treatment.

Background:Fenretinide,which is capable of generating reactive oxygen species( ROS ),has emerged as a promising antineoplastic agent based on numerous in vitro and in vivo studies and clinical chemoprevention trials. Preliminary studies showed that fenretinide could induce apoptosis in human hepatocellular carcinoma( HCC)cells in vitro, however,the precise mechanism was not clarified. Aims:To elucidate the effect of ROS on apoptosis of human HCC cells induced by fenretinide and the underlying mechanism. Methods:Human HCC cell line Huh-7 was treated with antioxidant vitamin E,fenretinide or their combination,respectively. ROS in live cells was evaluated by confocal microscopy and flow cytometry;cell viability and apoptosis were assessed by CellTiter-Glo Luminescent Cell Viability Assay Kit and Caspase-Glo3/7 Assay Kit;expression and intracellular localization of nuclear receptor Nur77,as well as expression of stress-induced transcription factor GADD153 were measured by immunofluorescence staining and Western blotting,respectively. Results:Vitamin E pretreatment fully blocked the fenretinide-induced ROS production. In Huh-7 cells pretreated with vitamin E,cell apoptosis induced by fenretinide was significantly reduced(P<0. 05). Furthermore,effect of vitamin E pretreatment was noteworthy on reducing fenretinide-induced GADD153 expression, while no significant impact on fenretinide-induced Nur77 expression and translocation was observed. Conclusions:Elimination of ROS by vitamin E can abrogate the pro-apoptotic effect of fenretinide on Huh-7 cells,which indicates the participation of ROS in fenretinide-induced apoptosis of human HCC cells. Its mechanism might be associated with induction of GADD153 protein expression.

This study established a population pharmacokinetics-pharmacodynamics model of clopidogrel in patients with acute coronary syndrome. Fifty-nine patients were enrolled. The plasma concentration of clopidogrel active metabolite and vasodilator stimulated phosphoprotein platelet reactivity index (VASP-PRI) were selected as the pharmacokinetics index and the pharmacodynamics index, respectively. The covariates including demographic characteristics, laboratory indexes, combined medication, complications and genetic polymorphisms of related enzymes were screened for their influence on the pharmacokinetic and pharmacodynamics parameters. Population pharmacokinetic and pharmacodynamics data analysis was performed using NONMEM software. The general linear model and the indirectly effect model-turnover model for pharmacokinetic and pharmacodynamic analysis were selected as the basic model, respectively. The population typical values of K12, CL/F, V/F, EC50, K(in), and E(max) were 0.259 h(-1), 179 L x h(-1), 632 L, 1.57 ng x mL(-1), 4.29 and 0.664, respectively. CYP2C19 was the covariate in the final pharmacokinetic model, and the model was to design a prior dosage regimen.