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 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.

Objective To sunomanize the experience of consecutive 353 patients and mean 2-years follow-up following Mei mini maze procedure for atrial fibrillation.Methods Between June 2010 and May 2015,353 patients(240 males, 113 females) of atrial fibrillation received this therapy.The age of these patients were(59.7 ± 8.5) years.Among them, 186 were with paroxysmal and 167 were with non-paroxysmal.The procedure, through three ports on left chest wall, included pulmonary vein isolation and ablations of the roof and posterior wall of left atrium which were achieved by bipolar radiofrequency ablation.Ganglionic plexus ablation was made by the ablation pen.Left atrial appendage was excluded.Results Durations of their procedures were(92.3 ± 19.1) mins.No conversion to sternotomy or pacemaker implantation occurred and none of the patients died.The hospital stay was(8.5 ± 2.1) days.The mean follow-up duration was (25.0 ± 15.1) months.308 (90.1％) patients were in sinus rhythm.34 patients could not maintain sinus rhythm.Stroke, thrombus in the left atrium and stenosis of pulmonary vein were not found after their procedures.Conclusion Mei mini maze procedure is a safe, effective, and appropiiate treatment for AF, which restores sinus rhythm and may be associated with the prevention of AF-related stroke.It deserved to be promoted in future.

Objective:The aim of the current study was to explore the alterations of TLR4 after traumatic brain injury. Methods: 20 Male ICR rats were randomly divided into four groups(5 rats each group)including control group without brain injury and traumatic brain injury groups(4,24 hours and 7 days).All rats were decapitated at corresponding time point and mid-jejunum samples were taken.The intestinal mucosa structure was detected by histopathological examination.Immunohistochemistry and realtime RNA were used for detection of TLR4 expression in ileum samples.Results: After traumatic brain injury,the histopatholocal alterations of gut mucosa occurred at 4 hours and progressed to a serious state.Compared to control group,TLR4 was significantly up-regulated on the endothelia of microvessels in villous interstitium and lamina propria by 4 h following TBI and maximally expressed at 24 h post-injury.The endothelial TLR4 immunoreactivity in ileum mucosa still remained strong on 7 d post-injury.There was a very low mRNA expression of TLR4 in the gut of control rats and significantly increased at 4 h following TBI(P