Objective To study the mechanism of β3 adrenoceptor (β3-AR) activation underlying cholesterol efflux by activating or inhibiting the β3-AR of HepG2 cells.Methods Cultured HepG2 cells were randomly divided into control group,β3-AR agonist group and β3-AR antagonist group.Serum levels of apoA-Ⅰ,apoA-Ⅱ,and β3-AR in supernatant fluid,and cholesterol,free cholesterol,cholesterol ester in HepG2 cells were measured by ELISA.Cholesterol efflux from macrophages was tested by 3H-labled cholesterol.Expressions of ABCA1 and LXRα mRNA and protein were detected by RT-PCR and Western blot respectively.Results The efflux rate of apoA-Ⅰ,cholesterol and cholesterol ester was significantly higher while the serum levels of cholesterol and cholesterol ester were significantly lower and the expression levels of ABCA1 and LXRα mRNA and protein were significantly higher in β3 AR agonist group than in control group.The serum levels of cholesterol and cholesterol ester were significantly higher while the efflux rate of cholesterol and cholesterol ester and the expression levels of ABCA1 and LXRα mRNA and protein were significantly lower in β3-AR antagonist groupt than in β3-AR agonist group (0.49±0.10 vs 1.24±0.02,0.85±0.05 vs 1.32±0.05,0.38±0.01 vs 1.45±0.20,0.08±0.01 vs 0.76±0.02,P＜0.01).Conclusion β3 AR promotes cholesterol efflux by upregulating the expression of apoA-Ⅰin HepG2 cells.

Objective To investigate the dynamic changes in expression of cell markers desmin, glial fibrillary acidic protein (GFAP), and (-smooth muscle actin (?-SMA) in primary cultures of human and rat pancreatic stellate cells (PSCs). Methods PSCs were isolated from human as well as rat pancreas using Nycodenz discontinuous density gradient centrifugation following digestion with combination of collagenase IV, Pronase E and DNase I, and purified by centrifugal elution techniques. Freshly isolated cells were examined by laser scanning confocal microscopy for vitamin A autofluorescence, by immunostaining for desmin, GFAP, and ?-SMA. Expression of ?-SMA was as well measured by Western analysis. Procollagen ?1(Ⅰ) mRNA expression was analyzed by Northern analysis. Results The purity of rat PSCs obtained by centrifugal elution were above 95%. More than 85% of either freshly-isolated human or rat PSCs displayed positive vitamin A autofluorescence. Rat PSCs stained positively for desmin and GFAP and negatively for ?-SMA, whereas human PSCs were negative for either desmin, GFAP or ?-SMA. During the process of primary culture, rat PSCs were positive for ?-SMA at 3d and completely transformed from quiescent state to myofibroblast-like phenotypes at 7d, which negatively or scarcely expressed desmin and GFAP, but fully expressed the ?-SMA protein and procollagen ?1(Ⅰ) mRNA, similarly to the settings of human PSCs. Conclusions Human and rat PSCs could be successively isolated in above 95% purity by combining gradient centrifugation with following centrifugal elution techniques. The results show some species differences in desmin and GFAP expression between freshly-isolated human and rat PSCs. Both of which, however, acquire a myofibroblast-like phenotype largely expressing ?-SMA protein and procollagen ?1(Ⅰ) gene in culture.

Objective To study Heat shock proteins (HSPs) induction and their association with Hantaan virus (HTNV) structural proteins in the brain tissues of mice experimentally infected with HTNV 76～118. Methods Newborn mice less than 72 h of age were experimentally infected with HTNV by intracerebral inoculation. The brains of mice at the 8th day post infection were removed and prepared for tissue extracts. The possible interaction or association of Hantaan virus nucleocapsid protein (HTNV NP) and glycoprotein G2 (HTNV G2) with GRP94, HSP70 and HSP27 were analyzed by double specific antibo dies sandwich ELISA and immunoprecipitation methods. Results HTNV infection induced the expression a set of HSPs including GRP94 and HSP70 in the brain tissues of mice experimentally infected with HTNV. HTNV NP was associated with GRP94, HSP70 and HSP27, which led to formation of HTNV NP HSP70 GRP94 HSP27 complexes. It was also observed that HTNV G2 was associated with NP and HSP27, which led to formation of HTNV G2 NP HSP27 complexes. Conclusions HTNV infection induces expression of a set of HSPs, which were associate with viral structural proteins in the brain tissues of mice experimentally infected with HTNV. This association may demonstrate the chaperone roles of HSPs in the synthesis, transportation and maturity of viral proteins during viral replication in the host cells.

Objective To investigate the diagnostic value of determination of the genotypes in codon 12 and 13 of K-ras oncogene in blood samples of patients with pancreatic carcinoma(PC).Methods Blood samples were obtained from 54 patients with pathologically confirmed PC,and 33 healthy controls.The DNAs were obtained in these samples.and then genotype of K-ras mutation was detected by using the PNA-clamping real-time quantitative PCR.Then the correlation between the K-ras genotypes of blood DNA and the clinical characteristics was analyzed.Results K-ras mutations were found in 74.1%(40/54)of patients with PC.There was no such mutation in control samples.The mutations of K-ras was associated with age,lymph node and vessel invasion.poorly differentiated tumor,CA19-9,while it was not associated with sex,tumor location,size of tumor,clinical staging and pathological type.Conclusions The one-step method was highly sensitive for detecting K-ras mutation in blood samples.Detection of circular blood cells harboring K-ras mutation suggested the tumor was highly invasive with poor prognosis.

To identify the key elements and available tools of regional health information resources plamling,the study introduced an innovative I-beam map,aiming to solve the two key questions of what and how to plan The paper covered such issues of planning as what is the present situation of regional health information,how to divide functional categories of regional health information resources,what are included in the business subclasses,which core data are used in the business process,which data to share and exchange,how to regulate such data,how to manage the regulations and make continuous improvement,and how to evolve towards planning standardization discriminatively.The study also drew a midfield line with such seven fields as medical service,maternity and child care,disease control and Drevention,health administration,emergency management,blood management and primary healtheare.This midfield line is used,along with stakeholders,planning process,information flow and planning output make the four quadrants,describing the whole process in the I-beam form.This practice can provide the methodology ofhow to plan,in the hope of guiding regional health information resource planning from theory to practice.

Regional health planning is a leading and scientific management philosophy for healthcare development in the international community.Information resources planning paves the way for scientific development.In the ongoing health reform in China,planning of regional health resources will play a key role by leveraging information technology.From the viewpoints of health information resources and information resources planning,the paper defines the concept of regional health information resources planning in microcosmic perspective.It summarized the five features of public welfare,specificity,imbalance,small channel and low noise,and non-symmetry.Authors compared the five planning modes,namely the community medical information resources mode,the general hospitals as the center of information resource mode,the regional PACS mode,the regional health data resource mode,and the regional medical coordination resources mode.In the paper,the planning is rounded up as four key points,that is,the basic steps of situation assessment and analysis,routine work of health information standardization,the planning should both comply with general rules of information resource planning and characteristics of the health sector,in addition to guidelines of forward-looking concepts.

AIM:To study the chemical constituents of Fissistigma oldhamii(Hemsl.)Merr.METHODS:Silica gel column chromatography was used in the isolation procedure,the structures of the isolated compounds were elucidated by spectral data.Meanwhile cytotoxic activity of compound Ⅰ was made according to the cell experiment in vitro.RESULTS:Five compounds were isolated from Radix of Fissistigma oldhamii(Hemsl.)Merr..On the basis of physical-chemical constants and spectral data(EI-MS,1H-NMR,13C-NMR),these compounds were identified as:Aristolactam A Ⅱ(Ⅰ),Aristolactam B(Ⅱ),Physcion(Ⅲ),?-sitosterol(Ⅳ),Stigmastan-7-one(Ⅴ).Compound Ⅰ showed cytotoxic activities on GLC-82 and HL_ 60 cell strains.CONCLUSION:Compounds Ⅲ,Ⅳ and Ⅴ are firstly isolated from Fissistigma oldhamii(Hemsl.)Merr..The IC_ 50 of compound Ⅰ on GLC-82 and HL_ 60 cell strain are:234.45,101.17 ?M.

Objective To explore the effect of ?-elemece on the reverse of drug resistance and the expression of P-glycoprotein(P-gp) in MDR K562/ADM cells. Methods MTT method was used to detect the sensitivity of cell and its reverse of drug resistance.Immunohistochemistry and immunoelectron microscopy were for observing the expression of P-gp.It's quantitative analysis was determined by flow cytometry.All experimental data were dealt with by SPSS(10.0 production pacility) soft ware. Results 1.Non-cytotoxic dose of ?-elemece(4.0?mg/L) could obviously decrease the IC-(50) value of K562/ADM cells to ADM.The reversing fold was 2.18;2.The level of P-gp expression was higher in K562/ADM cells than that in its parent K562 cells.Our studies also showed that the distribution of P-gp was on membrane of K562/ADM cells;3.Non-cytotoxic dose of ?-elemece(4.0?mg/L) could remarkably decrease the P-gp expression in MDR K562/ADM cells.Conclusion\ ?-elemece could evidently reverse drug resistance of K562/ADM to ADM.Decreasing the P-gp expression in drug resistance cell is one of the main mechanism of reversion.

Objective To investigate whether determination of CEA and CA19-9 levels in EUS-FNA pancreatic samples can be useful in detecting pancreatic adenocarcinoma and differentiating pancreatic adenocarcinoma from chronic pancreatitis.Methods Levels of CEA,CA19-9 were examined by chemiluminescence immunoassay analysis in EUS-FNA specimens obtained from 25 patients with chronic pancreatitis and 65 patients with pancreatic adenocarcinoma,and compared with those of their peripheral serum.Twelve patients with suspected pancreatic adenocarcinoma while with negative EUS-FNA pathological findings were followed up.Results First,the levels of CEA,CA19-9 in EUS-FNA specimens were higher than those in serum obtained from same patient with pancreatic adenoearcinoma(P＜0.01),but there was no difference in these variables of EUS-FNA specimens and serum obtained from patients with chronic pancreatitis.Second,in the EUS-FNA samples,the levels of CEA,CA19-9 in pancreatic adenocarcinoma were higher than those in chronic pancreatitis(P＜0.01).On the contrary,in serum samples,there was no significant difference in CEA level between pancreatic adenocarcinoma and chronic pancreatitis(P=0.079).CA19-9 level in serum of Dancreatic adenocarcinoma was higher than that of chronic pancreatitis(P＜0.01). Finally,during the follow-up,of all the 12 patients with suspected pancreatic adenocarcinoma,10 patients were diagnosed as having pancreatic adenocarcinoma and 2 patients as having chronic pancreatitis.Diagnostic accuracy of serum CEA and CA19-9were 30%and 70%respectively,while sensitivity of CEA and CA10-9 determined by EUS-FNA was both above 90%.Conclusion The method of measuring CEA and CA19-9 levels in samples obtained by EUS-FNAcan be useful in detection of pancreatic adenocarcinoma and differentiation of malignant pancreatic tissue from chronic pancreatitis.

Objective To investigate the expression of peroxisome proliferator-activated receptor-γ(PPAR-γ) and angiotensin receptor (AT2R) in chronic pancreatitis tissue. Methods Between April 2006 and February 2009, 24 patients were pathologically diagnosed as chronic pancreatitis were enrolled and 12 samples of normal pancreatic tissues were treated as controls. Immunohistochemical staining was used to detect PPAR-γ and AT1R, AT2R expression in pancreatic tissues. Results PPAR-γ and AT1R were mostly negatively expressed in normal pancreatic tissues, the positive scores were 0.33±0.49 and 0.42 ± 0. 51, respectively, while AT2R were weakly positively or negatively expressed in acinus and islet cell, and it was weakly positively expressed in ductal epithelial cells, the positive score was 2. 33 ± 1. 37. In chronic pancreatitis tissue, PPAR-γ, AT1R and AT2R were positively expressed in acinus, ductal epithelial cells, and islet cell, the positive scores were 3.28 ± 2.46, 4.36 ± 2.80 and 4.61 ± 2.89, which were significantly higher than those in the normal group (P<0.01, P <0.01, P<0.05). Conclusions PPAR-γ, AT1R and AT2R were positively expressed in chronic pancreatitis tissue, and they may be the treatment target of chronic pancreatitis.