Objective To investigate the effect of long-term high-fat diet on insulin resistance and the morphology and function of islets in rats and the relationship between them.Methods Thirty normal male Wistar rats (8 weeks old) were divided into two groups and fed either with normal chow (NC,n=15),or high-caloric and high-fat diet (HF,n=15).Insulin resistance was assessed by euglycemic hyperinsulinemic clamp technique. The insulin secretory function of islets was evaluated by intravenous insulin releasing test.Morphological and quantitative analysis of pancreatic tissues was performed by double-label insulin and glucagon immunohistochemistry.Proinsulin mRNA was detected by RT-PCR.Results The glucose infusion rate (GIR) in HF rats was significandy lower than that in NC rats [(5.83?0.79)mg?kg~(-1)?min~(-1) vs (7.60?1.29)mg?kg~(-1)?min~(-1),P＜0.05].Immunohistochemistry showed that HF rats had larger islet size [(15168?1327)?m~2 vs (6264?1840)?m~2,P＜0.01] and significantly reduced insulin relative concentration of?cells[(-5.15?0.03) vs (-4.81?0.17),P＜0.01],as compared with NC rats.The islet relative?cell volume was decreased signifieandy (P＜0.01),whereas the relative?cell volume was increased (P＜0.01).So the ratio of?to?were lower in HF [(4.68?1.01) vs (11.84?3.82),P＜0.05].The peak of insulin secretion in intravenous insulin releasing test in HF was at 10 min,whereas that in NC rats was at 5 min.AUC (area under curve) 10-60 rain of insulin in HF was higher than that in NC rats [(152.51?34.53)mIU?L~(-1)?min~(-1) vs (86.40?21.21) mIU?L~(-1)?min~(-1),P＜0.01].There was no difference in proinsulin mRNA levels between two groups. Conclusion Long-term high-caloric and high-fat diet results in early impairment of islet morphology and function, as well as significant insulin resistance,suggesting that the compensation ability of islets has already been impaired in the early course of type 2 diabetes mellitus.

Objective To investigate the efficacy and toxicity of neoadjuvant regional arterial chemotherapy in the treatment of advanced gastric cancer. Methods The clinical data of 158 patients with advanced gastric cancer and with the same clinical stages who were admitted to Renji Hospital of Shanghai Jiaotong University from February 2002 to May 2005 were retrospectively analysed. Preoperative regional arterial chemotherapy was applied to 76 patients (test group) and the remaining 82 patients only received surgical treatment (control group). The chemotherapy regimen was epirubicin (50 mg/m2) + cisplatin (60 mg/m2) + 5-fluorouracil (1000 mg/m2).This regimen was modified to oxaliplatin (130 mg/m2) + 5-fluorouracil (1000 mg/m2) since 2003, and surgery was performed 6-11 days after the chemotherapy. All patients received postoperative intravenous chemotherapy.The clinical effects, radical resection rate, operative complications and long-term survival of the two treatment methods were evaluated. All data were analysed using the chi-square test and Kaplan-Meier analysis. Results The radical resection rate was significantly higher at 86% (65/76) in the test group compared with 71% (58/82)in the control group ( x2 = 5.01, P ＜ 0. 05 ). The toxicity of the chemotherapy in the test group was mild. The postoperative complication rate was 20% (15/76) in the test group and 16% (13/82) in the control group, with no significant difference between the two groups (x2 = 0.41, P＞0.05). The median survival time was 41 months in the test group and 23 months in the control group. The 5-year overall survival rate was higher in the test group (44.6%) than that in the control group (29.1%) (x2 =3.95, P＜0. 05). Conclusions Neoadjuvant regional arterial chemotherapy is well tolerated by patients with advanced gastric cancer. It is also effective for increasing the radical resection rate and improving the long-term survival.

Armillariella tabescens EJLY2098 was induced to produce ?-mannanase with konjac fine flour (Amorphopallus rivieri) as single carbon source. This induced enzyme was then purified using DEAE ion exchange chromatography and named atMAN47. Zymologic analysis showed that the molecular weight of this ?-mannanase was approximately 47 kD. The enzyme was stable when pH ranged from 5.0 to 6.5 and could be activated by Na+ and Ba2+. With an optimal temperature of 50?C. Action mode analysis of TLC revealed that the enzyme belonged to the endo-?-mannanase family. Being a meta-acid endo-?-mannanase, it was suitable to be applied to feed industry with a promising future as an enzyme preparation.

Objective To explore the expression of HCK gene during the cardiomyocyte differentiation of mouse embryonic stem cells and analyze the role of HCK gene in maintenance of pluripotency of embryonic stem cells.Methods Mouse embryonic stem cells were cultured,then induced to differentiate into cardiomyocytes.Total RNAs were isolated from mouse embryonic stem cells in the differentiation days:0 day(D0),the second day(D2),the fourth day(D4),the sixth day(D6),the eighth day(D8),respectively.The levels of HCK mRNAs were assessed by the method of semi-quantitive reverse transcriptase-polymerase chain reaction(RT-PCR).In the meanwhile,Total proteins were also isolated from mouse embryonic stem cells in the differentiation D0,D2,D4,D6,D8,and the levels of HCK proteins were evaluated by Western-blot.Results HCK mRNAs could be detected in the mouse embryonic stem cells in D0 and D2,however,they were undetectable from D4 to D8.The expression of HCK mRNAs was rapidly down-regulated during cardiomyocyte differentiation of mouse embryonic stem cells.Expression of HCK proteins,which coincided with HCK mRNAs,down-regulated during differentiation and couldn't be detected in D4.Conclusions With the cardiomyocyte differentiation of mouse embryonic stem cells,the expression of HCK in the levels of mRNA and proteins are sharply down-regulated;HCK may play an important role in maintaining the pluripotency of embryonic stem cell.

OBJECTIVETo explore the capability of human periodontal ligament stem cells (PDLSCs) differentiating into adipose cells in vitro and to determine their changes in cell morphology, structure and function during differentiation.

METHODSPDLSCs isolated by magnetic-activated cell selection were treated continuously with adipogenic medium for 21 d. Then the cell morphology, ultrastructure, adipose specific markers of low density lipoprotein (LPL) and peroxisome proliferator activated receptor-gamma (PPAR-gamma) were analyzed by inverted contrast microscope, trans mission electron microscope (TEM), flow cytometry, immunofluorescence, RT-PCR and Western blot, respectively. These adipose-like cells were also identified by oil red O staining to determine the formation of lipid droplet, and the non-induced cells were used as control.

RESULTSAfter continuous induction, the treated cells differentiated into adipose-like cells with round shape, and large amount of lipid drop in cytoplasm. 96.54% of the PDLSCs were found to differentiate into adipose cells as showed by flow cytometry, the specific markers of LPL mRNA and PPAR-gamma mRNA, and oil red O staining, respectively. Further, PPAR-gamma protein was detected in the induced cells in a time-dependent manner.

CONCLUSIONHuman PDLSCs have the potential of differentiating into adipose cells under appropriate condition, and the differentiated cells exhibited characteristics of adipose cells both from cell morphology and from their functions.

Adipocytes ; Cell Differentiation ; Humans ; PPAR gamma ; Periodontal Ligament ; Stem Cells

The purpose of this work was to investigate the effects of niflumic acid (NFA), a chloride channel blocker, on the proliferation of human hepatoma cell line (HHCC). Cell proliferation was analyzed by cell count and MTT assay. Cell cycle analysis was carried out by flow cytometry. [Ca(2+)](i) was determined by laser scanning confocal system. It was found that NFA decreased significantly the cell number and the MTT optical density (OD) of HHCC cells, and that the OD value was reversed after washout of NFA. Compared with control, NFA blocked cell cycle progression in G(1) phase. Extracellular application of NFA (100 micromol/L) induced a rapid decrease in [Ca(2+)](i). These findings demonstrate that blockage of chloride channels by NFA induces growth arrest of HHCC in G(1) phase, which may be due to the inhibition of Ca(2+)/CaM-dependent signaling pathways.
Calcium ; metabolism ; Calmodulin ; metabolism ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chloride Channels ; antagonists & inhibitors ; Humans ; Liver Neoplasms ; pathology ; Niflumic Acid ; pharmacology

OBJECTIVEAmbulatory arterial stiffness index (AASI) has been recently proposed to reflect the dynamic relation between diastolic and systolic blood pressure throughout the whole day. The aim of our study was to investigate the change in AASI with advancing age and the correlation with 24 hours pulse pressure (24 h PP) in healthy individuals.

METHODS246 healthy subjects [mean age (59.7 +/- 14.6) years, women 38.6%] underwent 24 hours ambulatory blood pressure monitoring (ABPM) in normal life style. The blood pressure recordings, heart rate (HR), mean arterial pressure (MAP) and pulse pressure (PP) were analyzed simultaneously by computer for every 30 minutes during 6:00 am-22:00 pm and every 60 minutes during 22:00 pm-6:00 am. Using all the blood pressure recordings, we plotted diastolic against systolic blood pressure from each individuals and calculated the regression slope. AASI was derived from 1 minus this regression slope.

RESULTSIn 246 healthy individuals, AASI increased with age. Among the healthy individuals, the 95th percentile of AASI was 0.56, the upper boundary of the 95% prediction interval of AASI in relation to age were 0.49 at 20 - 39 years, 0.59 at 40 - 59 years, 0.69 at 60 - 79 years, 0.79 at > or = 80 years. The correlation coefficient between AASI and 24 h PP was 0.497 (P < 0.01). AASI linearly increased with age in healthy individuals, whereas the relation between pulse pressure and age was curvilinear.

CONCLUSIONSAASI as a index reflecting blood pressure relationship, manifested the corresponding change with advancing age. The correlation between AASI and traditional index 24 h PP indicated AASI as a new measure of arterial stiffness.

Adult ; Age Factors ; Aged ; Aged, 80 and over ; Arteries ; physiology ; Blood Pressure ; physiology ; Blood Pressure Monitoring, Ambulatory ; Female ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo develop an extraction and concentration method for the detection of hepatitis A virus (HAV) in shellfish, water, serum and saliva samples by nested RT-PCR.

METHODSHAV were artificially inoculated into the above samples, calm sample was extracted using glycine buffer pH9.5, PEG precipitation; water sample was PEG precipitated directly; then all the samples including serum and saliva samples were extracted using Trizol regent, followed by nested RT-PCR detection using primers from HAV VP1-2A region.

RESULTSThe detection limit for HAV in cultured cell lysis was 0.1TCID50; in water, serum or salva sample was 1TCID50 respectively, in calm sample was 1-10 TCID50. HAV RNA was detected in water and sera samples collected from the HAV outbreak region, sequenced and analysis.

CONCLUSIONThe method developed here is convenient, specific and capable of detecting low levels of HAV in different samples, would be useful for diagnostic laboratories in order to perform HAV analysis in cases of foodborne infections or for molecular epidemiology investigation of HAV outbreaks.

Animals ; Chemical Fractionation ; methods ; Fresh Water ; virology ; Genetic Techniques ; Hepatitis A ; diagnosis ; virology ; Hepatitis A virus ; genetics ; isolation & purification ; Humans ; RNA, Viral ; chemistry ; genetics ; isolation & purification ; Saliva ; virology ; Serum ; virology ; Shellfish ; virology

AIMTo study the mechanism of the therapeutic angiogenesis effect of bone marrow stromal cells (BMSCs) implantation on rat acute myocardial infarction models.

METHODSThe rat acute myocardial infarction models were made by coronary artery ligation and divided into 2 groups at random. In the experiment group, twice passaged BMSCs were labeled with BrdU and then implanted into the infarction region of the recipients in 4 weeks. The control group was the model rats received only DMEM injection. In control group, the hearts were harvested on the day 3, 7, 14, 28, 42 and 56. The infarction regions were examined to identify the angiogenesis and the expression of the VEGF and bFGF. In experiment group, the hearts were examined on the day 42 and 56 after AMI (the day 14 and 28 after cells implantation).

RESULTSViable cells labeled with BrdU could be identified in host hearts. Histologic examination found most donor cells within the infarction region expressed fibroblastic and endothelial phenotype. The transplantation regions had a greater capillary density than the control regions did (14 +/- 4.7/HPF vs 6 +/- 2.4/HPF, P < 0.05). In the control group, the expression of VEGF and bFGF within the infarction regions peaked on day 7, and then decreased over time. In the experiment groups, the expression of bFGF and VEGF on the day 42 and 56 had a higher level than the control group did.

CONCLUSIONThe expression of VEGF and bFGF is significantly increased after cells therapy during the late phase of AMI. It indicates that BMSCs implantation promoted the angiogenesis is mediated by its differentiation into endothelium and the increased release of VEGF and bFGF.

Animals ; Bone Marrow Cells ; cytology ; Bone Marrow Transplantation ; Female ; Fibroblast Growth Factor 2 ; metabolism ; Myocardial Infarction ; surgery ; Neovascularization, Physiologic ; physiology ; Rats ; Rats, Wistar ; Stromal Cells ; transplantation ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo evaluate the effect of the aluminum hydroxide (Al-OH) adjuvant on the 2009 pandemic influenza A/H1N1 (pH1N1) vaccine.

METHODSIn a multicenter, double-blind, randomized, placebo-controlled trial, participants received two doses of split-virion formulation containing 15 μg hemagglutinin antigen, with or without aluminum hydroxide (Al-OH). We classified the participants into six age categories (>61 years, 41-60 years, 19-40 years, 13-18 years, 8-12 years, and 3-7 years) and obtained four blood samples from each participant on days 0, 21, 35, and 42 following the first dose of immunization. We assessed vaccine immunogenicity by measuring the geometric mean titer (GMT) of hemagglutination inhibiting antibody. We used a two-level model to evaluate the fixed effect of aluminum Al-OH and other factors, accounting for repeated measures.

RESULTSThe predictions of repeated measurement on GMTs of formulations with or without Al-OH, were 80.35 and 112.72, respectively. Al-OH significantly reduced immunogenicity after controlling for time post immunization, age-group and gender.

CONCLUSIONThe Al-OH adjuvant does not increase but actually reduces the immunogenicity of the split-virion pH1N1 vaccine.

Adjuvants, Pharmaceutic ; chemistry ; Adolescent ; Adult ; Aluminum Hydroxide ; chemistry ; Antibodies, Viral ; blood ; Child ; Child, Preschool ; China ; Data Interpretation, Statistical ; Double-Blind Method ; Female ; Hemagglutination Inhibition Tests ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza Vaccines ; adverse effects ; chemistry ; immunology ; Influenza, Human ; epidemiology ; immunology ; prevention & control ; virology ; Male ; Middle Aged ; Models, Statistical ; Pandemics ; Young Adult