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.

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 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

[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 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 role of Lactobacillus plantarum (LP) in the treatment of colitis in interleukin (IL)-10 knockout (IL-10-/-) mice and to explore its possible mechanisms.Methods Eight weeks old female wildtype (WT) mice and IL-10-/- mice, twenty mice of each type,were randomly assigned to four groups, WT group, WT+ LP group, IL-10-/- group and IL-I0-/- +LP group. The WT and IL-10-/- mice were gavaged with 0.5 ml saline, WT+Lp and IL-10-/- +Lp groups were gavaged with Lp 0.02 g (0.5 ml) ,took Lp 1 × 109 cfu everyday,continued for 4 weeks and then the experiment finished. The fresh mice faeces was collected once every week before (week 0) and during the experiment. The mice were executed at the end of experiments, the change of mice weight Was recorded, the length and the wet weight of colon were measured, fresh colon tissue specimens were taken for biological slices and proinflammatory cytokines TNF-a and IFN-γ were measured in colon mucosa. The fresh faeces were selectively cultured. The colonization of Lp in normal and colitis mice and its regulation role in intestinal flora were observed. Results Compared with WT mice, IL-10-/- mice demonstrated severe diarrhea, significantly decreased in body weight (P ＜0.05)and serious malnutrition. After Lp treatment, diarrhea relieved in IL-10-/- mice and the body weight increased significantly (P＜0.05). Pathological examination suggested that 100％ of IL-10-/-mice had intestinal inflammation, however after Lp treatment intestinal inflammation improved significantly. Mucosal ulcer, epithelial hyperplasia, the infiltration of neutrophils and lymphocytes in the lamina propria were also significantly reduced.The histopathological score was significantly lowered (P＜0.01). After Lp treatment, colon wet weight and the ratio of wet weight to length of IL-10-/- mice changed significantly (P＜0.01). Colon edema and thickening improved remarkably. The TNF-a and IFN-γ concentration of colon in IL-10-/- mice were 377.4±84.4 μg/g and 602.6±108.1 μg/g,which increased obviously than WT group (139.2 ± 32. 7 μg/g and 173.0± 52.4 μg/g, P＜0.05). After treated with Lp for four weeks, the TNF-α and IFN-γ concentration of colon in IL-10-/-+Lp group mice were 207.2±65.7 μg/g and 442.1 ± 138.4 μg/g, both were lower than that of IL-10-/- group mice (P＜0.05). The intestinal flora was disrupted in IL-10-/- mice. Conclusion Lp can effectively reduce intestinal inflammation in IL-10-/- mice, which take certain part in treatment in colitis. This treatment effect is associated with intestinal flora regulation and the inhibition of proinflammation cytokines expression.

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.

ObjectiveTo investigate the efficacy and toxicity of postoperative radiochemotherapy compared with chemotherapy alone in the treatment of locally advanced gastric cancer.MethodsA total of 83 patients with resected adenocarcinoma of the stomach were randomly assigned to postoperative radiochemotherapy group (RCT) ( n =43 ) or chemotherapy alone group (CT) ( n =40 ).Patients in RCT group received radiotherapy concurrent with capecitabine chemotherapy then followed by 4 - 6 cycles of FOLFOX4 chemotherapy.The total dose of radiation was 45 Gy.The dose of capecitabine was 1600 mg/m2per day.In the CT group,patients received 6 - 8 cycles FOLFOX4 chemotherapy.Survival was analyzed using Kaplan-Meier method and Logrank test. Results The follow-up rate was 96％. The number of patients who had a minimum of 2-,3-year follow-up time were 37,12 in the RCT group and 31,10 in the CT group.The 1-,2-,3-year local control rates for RCT and CT groups were 100％,97％,94％ and 95％,87％,73％ (x2 =4.54,P =0.033),respectively.The 1-,2-,3-year survival rates were 98％,86％,81％ in the RCT group,with 93％,80％,64％ in the CT group ( x2 =3.96,P =0.047 ).The incidence of grade 3hematological toxicity in the RCT and CT group was 23％ vs 15％ ( x2 =0.93,P =0.630 ),and grade 3gastrointestinal toxicity was 16％ vs 10％ ( x2 =0.95,P =0.624 ). Conclusions Compared with chemotherapy alone,postoperative radiochemotherapy can improve survival of locally advanced gastric cancer patients with acceptable toxicities.