Objective To investigate the anticoagulant activities of phlorotannins STP-1 and STP-2 from Sargassum thunbeergii Kuntze.Methods Coagulation time(CT)and bleeding time(BT)of mice were determined by in-vivo experiment to test the effect of phlorotannins by capillary method and cutting tail method respectively.Furthermore,activated partial thromboplastin time(APTT),prothrombin time(PT)and thrombin time(TT)were measured to evaluate the in-vitro anticoagulant activity of phlorotannins on New Zealand rabbits.Results Phlorotannins STP-1 had a significant effect on the prolongation of CT,BT,APTT,PT and TT,especially at the dosage of 1.0 mg/mL.Conclusion Phlorotannins STP-1 has both in-vivo and in-vitro anticoagulant activity.

The development of biodiesel and production feedstock in China was introduced, and the biodiesel production technologies as well as corresponding representative enterprises were reviewed. In addition, the development prospect of biodiesel industry in China was addressed.
Biofuels ; China

Objectiv e Renal ischemia-reperfusion ( I/R) may cause myocardial injury and dexmedetomidine ( DEX) is a new alpha-2 adrenergic agonist with the effects of antisympathia , seda-tion, and analgesia.This study was to investigate the effect of DEX on the myocardial tissue of rats at different time points after renal I/R. Methods Forty male Wistar rats were randomized into 5 groups of equal number,sham operation, 60 min renal ischemia and 3 h reperfusion (I/R1), 120 min ischemia and 3 h reperfusion (I/R2 ), 60 min ischemia and DEX+3 h reperfusion (D1), 120 min ischemia and DEX+3 h reperfusion ( D2) .Renal I/R was induced by removal of the right kidney and ligation of the left re-nal artery and vein followed by 3 hours of reperfusion.Meanwhile, intraperitoneal injection of DEX at 50μg/kg was given to the ani-mals in groups D1 and D2 at 60 at 120 min respectively after ischemia.After 3 hours of reperfusion, blood samples were collected for measurement of the concentrations of serum creatinine (Cr) and blood urea nitrogen (BUN), and renal and myocardial tissues harvested for observation of pathological changes under the light microscope and determination of the expressions of TNF-αand IL-10 by ELISA.Results Significant increases were observed in the concentrations of serum Cr and BUN , the expressions of TNF-αand IL-10 in the renal tissues and those in the myocardial tissues in groups I /R1([84.67 ±9.62] μmol/L, [8.55 ±1.08] mmol /L), I/R2 ([167.11 ±18.81] μmol/L, [13.42 ±1.25] mmol/L), D1 ([69.67 ±9.52] μmol/L, [7.56 ±0.70] mmol/L), and D2 ([114.29 ±12.50] μmol/L, [10.27 ±0.78] mmol/L), as compared with the sham operation group ([53.20 ±9.21] μmol/L, [3.75 ±0.78] mmol/L), (all P <0.05).Significant decreases was observed in the sham operation group as compared with other groups in the expressions of TNF-αand IL-10 (P<0.05).Significant decreases was observed in the D1 and D2 groups compared with other groups in the expressions of TNF-α, but increasing in IL-10.②Injury was reduced in the D1 and D2 groups compared with other groups.③The horizontal stripes of myocardial tissue disappeared in I/R1 and I/R2 decreases of inflammatory cells was observed in D1 and D2 groups compared with others. Conclusion Dexmedetomidine can attenuate myocar-dial injury induced by renal ischemia-reperfusion in rats and its inhibitory effect on inflammatory factors may be involved in the mechanism.

BACKGROUND:There are two types of epithelial stem cells in the ocular surface tissue:corneal epithelial stem cells and conjunctival epithelial stem cells. The corneal epithelial stem cells play an important role in renewal of corneal epithelial cells and maintenance of corneal transparency.
OBJECTIVE:To study the location of corneal epithelial stem cells using laser in vivo confocal microscopy and immunofluorescent staining.
METHODS:Patients with unilateral limbal stem celldeficiency who went to Henan Eye Institute from September 2009 to September 2012 were enrol ed in this study. Bilateral eyes were scanned by laser in vivo confocal microscopy, and the healthy eye was imaged as a control. The central cornea and limbus were scanned and images were recorded for statistical analysis. The eye bal s were obtained from Henan Eye Bank, China. Central cornea and limbus were dissected and embedded in the OCT compound for frozen section and the proper thickness of the section was 5-7μm. Immunofluorescent staining was used to detect the expression of p63, ABCG2, K3 and Connexin 43 in the epithelial layers of central cornea and limbus.
RESULTS AND CONCLUSION:Twenty-four patients diagnosed with unilateral limbal stem celldeficiency were recruited. Under confocal microscopy, in the affected eyes, the typical morphology of conjunctival epithelial cells and goblet cells was detected instead of corneal epithelial cells;in the limbus, a great amount of fiber scarring tissue was detected instead of Vogt palisade, rete pegs and pigment cells. Immunofluorescent staining showed the expression of p63, ABCG2 was mainly in the basal layer of limbal epithelium, especial y in the outer and middle parts, but the expression of p63 and ABCG2 was not detected in the epithelial celllayers of central cornea. K3 and Connexin43 were not expressed in suprabasal layers of limbal epithelium, but in central cornea, they were expressed highly in the whole epithelial celllayers. Laser in vivo confocal microscopy and immunofluorescent staining showed the corneal epithelial stem cells were located in the basal layer of outer and middle limbal epithelium, mainly in Vogt palisade and rete pegs.

Objective To investigate the influence and the possible mechanism of tea polyphenols on maintenance hemodialysis (MHD)patients’ oxidative stress and micro inflammatory condition.Methods 60 cases of MHD patients were randomly divided into routine group and tea polyphenols group.30 healthy subjects were chosen as normal control group.After 3 months,MDA,AOPP,SOD,TNF-α,CRP,IL-6 and NF-κB of routine group and tea polyphenols group were detected.Results Before treatment,compared with normal control group,MDA and AOPP of routine group and tea polyphenols group increased (P <0.01 ),SOD decreased (P <0.01 );after treatment,compared with routine group,MDA and AOPP of tea polyphenols group decreased (P<0.01),SOD increased (P<0.01).After treatment,compared with normal control group,NF-κB,TNF-α,CRP, IL-6 and oxidative index of routine group and tea polyphenols group increased,and the difference was statistically significant (P<0.01);compared with routine group,oxidative index of tea polyphenols group decreased,and the difference was statistically significant (P <0.01 ).Conclusion Tea polyphenol can improve MHD patients’oxidative stress and micro inflammatory state.

OBJECTIVE Topreparealipidderivativeofgemcitabine(Gem)anditspolymericmi-celles to overcome the disadvantages of Gem.METHODS N-benzyl-3′-acetyl-gemcitabine(BAG)was synthesized.A BAG-loaded poloxamer polymeric micelle (BAG∶poloxamer 188 =10∶1 ,mol/mol)was prepared using an injection method.The micelles were characterized with a laser particle size and elec-tric charge instru ment and negatively-stained trans mission electron microscopy.Hu man breast cancer cells MCF-7 were cultured with Gem or BAG polymeric micelles of 5,10,20,30,50,70,90 μmol·L-1 for 24,48 and 72 h,respectively.The inhibitory rate of cells was measured with an MTT method.The MCF-7 cytotoxicity of BAG polymeric micelles was investigated.A pharmacodynamic study was per-formed on the mice bearing mouse hepatocellular cancer cells H22.Intravenous (iv)and oral (ig)ad-ministration was used at the dose of Ge m 40 mg·kg -1 or BAG polymeric micelles 62 mg·kg -1 .The mice were administered on the 1 st,4th and 7th day and sacrificed on the 8th day.Tumor inhibitory rates were measured.RESULTS TheBAGstructurewasidentifiedbythinlayerchromatograph,1Hand13C NMR,infrared ray chromatograph and mass spectrum.The appearance of BAG micelles was a slightly blue suspension.The micelles were spheres according to the electron microscopic observation.Their size was 62.82 nm and the zeta potential was -18.8 mV.The half inhibition concentration (IC50)of Gem and BAG polymeric micelles was 40.6 and 90.0 μmol·L-1 ,5.0 and 14.9 μmol·L-1 ,5.0 and 1 3.6 μmol·L-1 at 24,48 and 72 h,respectively according to the MTT results.According to the in vivo results,compared with the tumor model group,Gem (ig),Gem (iv)and BAG polymeric micelles (iv and ig)had significant effect on the tumor weight of H22 cell xenograft mice (P<0.01 ).As for anti-tumor efficiency,BAG polymeric micelles (ig)were better than Gem (ig)(P<0.05);BAG polymeric micelles (iv)were better than BAG polymeric micelles (ig)(P<0.05),and BAG polymeric micelles (iv)were almostequaltoGem(iv).CONCLUSION ThelipidderivativeofGemcanbeloadedinthepoloxamer 1 88 polymeric micelles.BAG polymeric micelles show in vitro MCF-7 cell inhibition and in vivo inhibition of mouse H22 xerografts;iv or ig.BAG polymeric micelles (ig)show better anti-tumor effect than Gem (ig),indicating that BAG polymeric micelles are a promising novel anti-tumor oral preparation.

Objective To investigate the relationship between with or without non?alcoholic fatty liver disease(NAFLD) and diabetic retinopathy(DR) in type 2 diabetes mellitus(T2DM)?Methods Clinical and laboratorial data of 517 cases T2DM hospitalized patients were collected,and the patients were divided into two groups according to if the NAFLD was complicated or not?Group A was T2DM with NAFLD and group B was T2DM without NAFLD?The general information and the laboratorial checking results were Compared, then various index were used as the independent variable, DR was used as the dependent variable for Logistic regression analysis?Results (1)In the 517 cases of T2DM patients,the incorporative rate of the NAFLD was 65?7%(349/517)?(2)Compared with group B,the levels of body mass index(BMI),insulin resistance index (HOMA?IR),glutamyltransferase(GGT),alanine aminotransferase(ALT),aspartate aminotransferase(AST), triglyceride(TG),total cholesterol(TC),low?density lipoprotein cholesterol(LDL?C) and uric acid(UA) for group A were significantly increased, while the high?density lipoprotein cholesterol ( HDL?C ) level was significantly decreased?All the differences were statistically significant( P<0?05)?( 3) Logistic analysis showed that the duration of the extension,hypertension,the increasing level of NAFLD,LDL?C were the risk factors of DR?Even though excluded the influence of duration,high blood pressure and LDL?C level,NAFLD was still the risk factor for T2DM complicated by the DR( OR=2?176,95% CI ( 1?354,3?199) )?Conclusion NAFLD and DR are closely related, so early diagnosis and intervention of NAFLD may prevent the occurrence and development of DR.

Objective A retrospective cohort study was carried out to observe the long-term effect of ESD in treating early gastrointestinal cancer or precancerous lesions. Methods The clinical and follow-up data of 73 patients were collected. Kaplan-Meier, Log-rank and Breslow test and Cox's proportional hazards regression model were used to analyze the data. Results The median survival time in the gastric and colo-rectal early cancer or precancerous lesions is longer than 65 months in our study, respectively. For esophagus, the median survival time was 44.5 months; the disease free survival time (DFS) after ESD was significantly reduced in the esophagus, compared to the stomach and colo-rectum (χ2 = 12.61, P = 0.000; χ2 = 7.09, P = 0.008); the degree of atypia (or infiltration), and lesion size were considered to be two factors to influence the DFS after ESD (P = 0.027, OR^ =2.38, 95%CI:1.10 ~ 5.12, P = 0.074, 95%CI; OR^ =0.90, 95%CI: 0.80 ~ 1.01). Conclusion ESD is an effective curative treatment in the resection of early upper gastrointestinal cancer and precancerous lesions. The degree of atypia (or infiltration) was concluded as an independent risk factor for the DFS post-ESD, and the size of lesion was a valuable parameter with regard to the recurrence after ESD procedure.

Insects exhibit a particular resistance to infections.The activation of the innate immune response of insects is involved in the recognition of the infectious nonself and subsequent activation of cellular and humoral reactions.In humoral reactions,insect antibacterial peptides and lysozyme are very important in resistance to infections.Housefly(Musca domestica) is one of the most important kinds of insects and it has strong ability to adapt to the adverse circumstances.It is of momentous theoretical and practical significance to research the immunity system of housefly.The methods of inducement of housefly larvae were firstly studied.Then an antibacterial protein,whose molecular weight is 28kDa,was purified from housefly larvae,induced by 30% H2O2,through saltout,Sephadex G25 column,Sephadex G75 column and CMSepharose Fast Flow column.This antibacterial peptide had activities against most of Gram positive and Gram negative bacteria.

BACKGROUND: Apelin/APJ system has a wide range of physiological functions,but its intracellular signal transduction,in particular,apelin receptor desensitization,internalization,resensitization degradation,have still no consistent opinion.OBJECTIVE: To construct eukaryotic expression vector expressing human apelin receptor(APJ)tagged to red fluorescent protein(pDsRED-express-C1),and to determine the expression in human embryo kidney 293 cells.METHODS: The plasmid pcDNA3.1-hAPJ was used as a template for PCR amplification of human APJ.Following PCR amplification the PCR product were removed and enzymatic digestion with EcoR I and BamH I.Same enzymes were used to cut vector pDsRED-express-C1.The digestive product was ligated by conventional methods of connection,then transfected into Competent E.coli TOP10.Single clones were picked plasmid extraction,followed by restriction enzyme digestion and finally DNA sequencing.The recombinant plasmid with correct sequencing was transfected into human embryonic kidney cells,PI staining,followed by the observation under a confocal microscope.RESULTS AND CONCLUSION: PCR amplified a 1.2-kb fragment,which was consistent with the expected size of the human APJ.The pDsRed-hAPJ recombinant plasmid was cut into two fragments,one corresponded to the pDsRED-express-C1 vector size,and the other fragment corresponded to APJ target fragment.Confocal microscopy analysis showed that,APJ was expressed mainly in the membrane of human embryo kidney 293 cells.The pDeRed-hAPJ eukaryotic plasmid expression vector was successfully constructed and effective expression of this fusion protein is achieved,which might be instrumental in the study of displacement and intracellular localization of human APJ.