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.

[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

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 evaluate the value of combined hepatectomy in radical resection for hilar cholangiocarinoma. Methods The clinical data and follow-up data of 67 patients of resection for hilar cholangiocarinoma in Henan Tumor Hospital from June 2005 to october 2008 were retrospectively analyzed. Results According to intraoperative exploration situation and bismuth types, tumor resection was combined performed with hepatectomy (n＝38)or non-hepatectomy (n＝29). The rate of R0 resection was 55.3% in hepatectomy group(n＝21) and 34.5% in non-hepatectomy group(n＝10), and the difference was significant（P＝0.024）. The incidence of complications were 39.5% in hepatectomy group(n＝15) and 13.4% in non-hepatectomy group(n＝4), and one patient with liver and kidney failure died in hospital. The 1, 3, 5 years of survival rate were 89.3%,53.6% and 32.1% respectively in R0 group (n＝31) and 69.7%,30% and 10% respectively in R1～R2 group(n＝36), there were significant differences in the postoperative survival rate between both groups（P＝0.018). The 1, 3, 5 years of survival rate were 81.8%,48.5% and 24.2% in hepatectomy group and 75%,32% and 16% in non-hepatectomy group respectively, and the differences were significant（P＝0.037). Conclusions Aggressive resection including combined hepatectomy for hilar cholangiocarcinoma can play an important role for curative effect and long term survival rate.

The feasibility and the clinical value of the enzyme-multiplied immunoassay technique （EMIT） monitoring of blood concentrations of cyclosporine A （CsA） in patients treated with CsA were investigated after kidney transplantation. The validation method was performed to the EMIT determination of CsA blood concentration, the CsA whole blood trough concentrations （Co） of patients in different time periods after renal transplantation were monitored, and combined with the clinical complications, the statistical results were analyzed and compared. EMIT was precise, accurate and stable, also with a high quality control. The mean postoperative blood concentration of CsA was as follows： 〈1 month, （281.4± 57.9）ng/mL; 2 - 3 months, （264.5 ± 41.2） ng/mL; 4 - 5 months, （236.4 ± 38.9） ng/mL; 6 - 12 months, （206.5± 32.6）ng/mL; 〉12 months, （185.6± 28.1）ng/mL. The toxic reaction rate of CsA blood concentration within the recommended therapeutic concentration was 14.1%, significantly lower than that of the none-recommended dose group （37.2%） （P〈0.05）; the transplantation rejection rate was 4.4%, significantly lower than that of the none- recommended dose group （22.5%） （P〈0.05）. Using EMIT to monitor the blood concentration of CsA as the routine laboratory method is feasible, and is able to reduce the CsA toxicity and rejection significantly, leading to achieving the desired therapeutic effect.

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.