GlyoxalaseⅠthat presents in all human tissues detoxifies α-oxoaldehydes and prevents the formation of advanced glycation end products(AGEs).AGEs and their main precursor namely methylglyoxal(MG)produce cytotoxicity and have extensive relationship with diabetic complications.Therefore,to elevate glyoxalaseⅠactivity may be a new path to release such complications.

AimThe aim of this study was to establish a rodent model with similar characters of human metabolic syndrome(MS).Methods Three species mice and Wistar rats were fed with high energy chows(HEC)for 6 to 23 weeks.Animals were weighted every week.Fasting blood glucose(FBG)together with total cholesterol(TC)and low density lipoprotein-cholesterol(LDL-C)were investigated by oxidase test every two week.And fasting blood insulin(FINS)was determined by radioimmunoassay.Homeostasis model assessment of insulin resistance(HOMA-IR)was calculated as FBG×FINS/22.5.At the end of the experiment,oral glucose tolerance test(OGTT)was performed.Then animals were decapitated,and coel-fat and orchio-fat were collected and weighted to calculate the visceral fat coefficient(VFC).Results FBG,serum TC and LDL-C significantly increased(P<0.01)after 6 weeks feed of HEC in KM mice.The mice also formed abdominal obesity and insulin resistant together with impairment of glucose tolerance(P<0.05 or P<0.01).Though similar to the KM mice,C57BL/6 and BALB/c mice couldn't form abdominal obesity while the latter had increased body weight(P<0.05 or P<0.01).Wistar rats formed hyperlipidemia from 1 to 10 week and hyperglycemia from 10 to 23 week together with insulin resistance and impaired glucose tolerance(P<0.05 or P<0.01).Conclusion KM mice feed with HEC for 6 weeks could successfully establish metabolic syndrome mice model which might be suitable for drug-screening,the major characters includes the formation of abdominal obesity(increase of VFC),the increase of serum TC,LDL-C,FBG and HOMA-IR,and the decrease of OGTT.

AimPurpose-The aim of this study is utilizing the highthrough genechip data to Compare the difference of the pharmacological pathways among the Qingkailing effective components Baicalin(BA),Jasminoidin(JA),cholic acid(CA) and Concha margaritiferausta(CM)in the treatment process of cerebral ischemia.Methods The focal cerebral ischemia-reperfusion model mice were randomly divided into groups of Baicalin(BA),Jasminoidin(JA),cholic acid(CA),Concha margaritiferausta(CM)and model group(M),15 mice for each group,24 hours later total RNA were abstracted from the hippocampus,we selected 374 gene expression profile related to cerebral ischemia,made cDNA chip marked by Cy3/Cy5,detect the variation of different components,Then apply Arraytrack software to select differentiate expressed genes between BA and M,JA and M,CA and M,CM and M by T-tests,select genes with P<0.05,Fold change>1.5,according GeneGO software to find the top two pathways of each components.Results the number of differentiate expressed genes between BA,JA,CA,CM and M is separately 46,50,54 and 30,according to the top two pathways of GeneGo display JA,CA,CM all participate Apoptosis and survival_TNFR1 signaling pathway,besides BA participate in regulating G-protein signaling and Development_A2A receptor signaling while CA in Neurophysiological process_NMDA-dependent postsynaptic long-term potentiation in CA1 hippocampal.Conclusion Qingkailing effective components take diversity Pharmacological characteristics,BA mainly for anti-apoptosis,JA mainly for inhibit apoptosis and promote ischemic brain protection,etc,CA focused on inhibiting calcium influx,and anti-neuron variability.But CM has no good results on this.

Aim To investigate the effects of retinoic acid (RA) on induction osteroporosis model rats andpreventive effects of Danshengubao.Methods 4-month-old female Sprague-Dawley rats were given RA at 70 mg·kg~(-1)·d~(-1) and were given Danshengubao at different doses at the same time.All rats were treated by oral gavaged for 28 days.The static and dynamic parameters in cancellous bone of the fifth lumbar vertebrae (LV5) were examined and the dynamic changes of the tibial shaft (Tx) were observed with histomorphometrical analyses; the forth lumbar vertebrae (LV4) was used to perform the compression test.Results Compared with control group, biomechanical properties of LV4, the static parameters ( total tissue area, trabecular area, trabecular perimeter) and the dynamic parameters of LV5 were significantly decreased in RA group.Compared with control group, bone formation of Tx was decreased in periosteal surfaces but enhanced in endocortical surfaces in RA group.Compared with RA group, the biomechanical properties of LV4 were increased significantly in low and medium dose of Danshengubao groups.Conclusion sRA can decrease the size and the biomechanical properties of LV, but it can not change the percentage trabecular area. The mechanism may be related to the act that RA can inhibit cancellous bone formation, decrease the modeling of cortical bone in periosteal surfaces and enhance the remolding of cortical bone in endocortical surfaces. Danshengubao can improve biomechanics of LV induced by RA in rats.

Aim To explore the effects of cannabinoid (WIN55,212-2) on mRNA and protein expressions of PKAC-β,c-fos and BDNF in cerebral cortex after intracerebral hemorrhage(ICH)in rats.Methods ICH model of rats were made by Ⅶ Collagenase,which were injected by the intraperitoneal route,thirty minutes after the operations.The rats were killed for brain tissue as specimens with 24 hours.The localizations of PKAC-β,c-fos and BDNF were assessed by immunohistochemistry;The expressions of PKAC-β,c-fos and BDNF mRNA were detected by RT-PCR;The expressions of PKAC-β and BDNF protein were revealed by Western blot.Results WIN55,212-2 increased not only the levels of BDNF mRNA and protein,but also c-fos mRNA in ipsilateral cerebral cortex.However,it decreased the levels of PKAC-β mRNA and protein.PKAC-β,c-fos,and BDNF proteins were expressed on membrane of neurons,nucleus of neurons or the cytoplasm of glial cells respectively.Conclusion WIN55,212-2 induces the expression of BDNF in the cerebral cortex,which provides a theoretical basis for the treatment of cerebrovascular disease.

Aim To demonstrate the effects and mechanism of adenosine A1 receptor agonist R(-)-N6-(2-phenylisopropyl) adenosine(R-PIA) on high glucose(HG)-induced myocardial hypertrophy by in vitro cultured myocardial cells from neonatal rats.Methods The protein content was assayed by the method of Lowry. The expression of p-ERK1/2 and ERK1/2 was determined by Western blot.The [Ca~(2+)]I transient changes of cell loaded Fura-2/AM were measured by Till image system.Results 1 μmol·L~(-1) R-PIA and U0126 inhibited similarly HG-induced increase of the protein content and [Ca~(2+)]I transient along with the relative expression of p-ERK1/2.These responses were completely abolished by adenosine A1 receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine(CPDPX).Conclusion Adenosine A1 receptor stimulation significantly inhibits HG-induced myocardial hypertrophy by mediating ERK1/2 pathway and Ca~(2+).

Aim To explore the effect of survivin in PC12 cells against injuries induced by cobalt chloride(CoCl_2).Methods PC12 cells were exposed to CoCl_2 at different doses in different time to set up the chemical hypoxia induced PC12 cells injuries model.Cell viability was tested by using cell counter kit-8.Dose-effect(200～1 000 μmol·L~(-1))and time-effect(0～48 h)relationship between hypoxia induced by CoCl_2 and the expression of survivin was evaluated by western blot.Results PC12 cells viability was inhibited significantly by CoCl_2 in a dose and time dependent manners;At the concentrations from 200 to 600 μmol·L~(-1) CoCl_2 for 24 h,survivin expression was upregulated in a dose dependent manner,peaking at 600 μmol·L~(-1) CoCl_2 treatment,exceeding this concentration of CoCl_2,with dose increasing,survivin expression decreased.At the dose of CoCl_2 up to 1 000 μmol·L~(-1),survivin did not express basically;Treatment with 600 μmol·L~(-1) CoCl_2 in different time,within the range of 0～36 h,the expression of survivin enhanced in time dependent manner.But with the extension of time,survivin expression was declining; 17-allylamino-17-demethoxygeldanamycin (2 μmol·L~(-1)),an inhibitor of Hsp90,not only decreased survivin overexpression but also obviously enhanced the injuries of PC12 cells induced by CoCl_2,which didn't damage PC12 cells alone.Conclusion Upregulation of survivin expression may be one of the endogenous defense mechanisms for PC12 cells against chemical hypoxia.

Aim To investigate the molecular mechanisms of differentiation in human leukemia HL-60 cells induced by diallyl disulfide(DADS)using suppression subtractive hybridization(SSH).Methods In our privious study,the subtractive cDNA library was constructed successfully and efficiently. 30 clones were randomly analyed with restriction enzyme.The inserts of cDNAs were analyzed by restrictive enzyme EcoR I.Positive clones were sequenced and the homology of resulting cDNA sequences were analyzed through bioinformatics software Blastn.Results 18 clones contained 100～600 bp cDNA inserts.10 differantiation genes were obtained and involved in cell cycle,signal transduction,metabolism and RNA binding.And 3 of 10 genes,p21,STAT1 and CAMTA1 were up-regulated and detected by RT-PCR,the results matched with SSH.Conclusion sThere are tight correlation between the differentiation induced by DADS and three-upregulated gene:p21,STAT1 and CAMTA1.

Aim To investigate the effect of catalpol from Radix rehmanniae on the proliferation,differentiation and matrix mineralization of MC3T3-E1 cells in vitro.Methods Catalpol from Radix rehmanniae of different concentration preparations were extracted with ethanol(catalpol ethanol-extract),respectively.A mouse osteoblast cell line MC3T3-E1 was used to determine the potency of catalpol.MTT were applied to determine proliferation of the cell treated by catalpol at different concentrations.Differentiating effects of the catalpol with different concentrations in the cell were evaluated through the examinations of alkaline phosphatase(ALP)activities and bone gla protein(BGP)levels.Von kossa staining method was used to demonstrate the effects of the catalpol on calcification of the cells.Results Catalpol at concentration from 1×10~(-7) to 1×10~(-9) mol·L~(-1) for 24 hours and 48 hours effective promoted the proliferation of osteoblasts of MC3T3-E1 cells line.Catalpol at concentration from 1×10~(-5) to 1×10~(-6) mol·L~(-1) for 48 hours and 72 hours effective stimulated the activity of ALP of osteoblasts of MC3T3-E1 cells line.Catalpol at concentration from 1×10~(-5) to 1×10~(-6) mol·L~(-1) for 8 days and 12 days effective increased the synthesis and secretion of bone gla protein(BGP) of osteoblasts of MC3T3-E1 cells line.Catalpol at concentration from 1×10~(-5) to 1×10~(-6) mol·L~(-1) for 19 days effective increased the mineralized bone nodular structure number of osteoblasts of MC3T3-E1 cells line.Conclusion Catalpol could promote proliferation and differentiation of MC3T3-E1 cells in vitro.Catalpol may be one of effective monomer of Radix rehmanniae on treatment of osteoporosis.

Aim To investigate the anti-tumor effect of nitidine chloride(NC)on human HepG2 hepatocellular transplanted tumor in nude mice and its effect on topoisomerase.Methods The subcutaneous transplantable tumor model of human liver cancer in nude mice was established and the anti-tumor effect of NC was calculated.The effects of NC on TopoⅠ/Ⅱ mediated-pBR322 DNA relaxation were measured by using agarose gel electrophoresis.Results NC inhibited significantly the growth of hepatoma,The inhibitory rate at the dose of 2.5,5,10 mg·kg~(-1) was 12.06%,35.63% and 60.91% respectively.At the concentration of 6.25 μmol·L~(-1),NC completely inhibited the pBR322 DNA cleavage mediated by TopoⅠ;at the concentration of 25 μmol·L~(-1),NC completely inhibited the pBR322 DNA cleavage mediated by Topo Ⅱ.Conclusion Nitidine Chloride can inhibit hepatic carcinoma growth in nude mice,The anti-tumor mechanism is probably related to the inhibitory effect on Topo.