Objective To investigate the relationship between insulin level and mild cognitive impairment(MCI) of type 2 diabetes and analyze the risk factors for MCI. Methods We collected 101 type 2 diabetes subjects who were divided into two groups:MCI and normal cognition(NC).All subjects were evaluated with cognition scores of MMSE,GDS,CMS,ADL.Insulin,glycosylated hemoglobin,blood routine test,blood pressure,ECG,brain CT were detected.Independent t-test,correlation and multiple regression analysis were performed. Results 1. The diabetic patients with MCI showed the higher levels of insulin at fasting, 2hr and 3hr after meal than did NC(P

Objective:To investigate whether the IgG from patients with antiphospholipid syndrome(APS) is capable of stimulating blood monocytes to express tissue factor(TF) activity and to explore the mechanisms possibly concerned.Methods:Freshly isolated peripheral blood monocytes were cultured in media containing purified APS IgG or normal IgG or RP-1 [a monoclonal anti-?_2 glucoprotein I(?-?_2GPI)antibody] and/or other agonists and the TF activity on monocytes was investigated by measuring factor Ⅶa-dependent generation of factor Xa.Results:APS IgG as well as RP-1 significantly increased monocytes TF activity,comparing to normal IgG;exogenous ?_2GPI enhanced the effects as the intact IgG molecule.Conclusion:That certain antiphospholipid antibodies,mainly anti-?_2GPI antibodies,induce monocytes TF activity in an antigen-specific manner,not via Fc receptor pathway,is contributed to the thrombotic diathesis in APS.

Objective:To establish bacterial endotoxin test for brozopine. Methods: Interference pre-test and interfering factors test were conduced on 3 batches of samples from 2 manufacturers to confirm the applicability of bacterial endotoxin test and the non-in-terfering concentration. The bacterial endotoxin test was carried out based on the method described in the second part of Chinese Phar-macopeia (2010 edition) and relevant standards and guidelines. Results: The three batches of brozopine showed no interference in bacterial endotoxin test at the concentration less than or equal to 2. 5 mg·ml-1 . The bacterial endotoxin test of the three bathes of samples all met the requirements. Conclusion:Bacterial endotoxin test can be used for the quality control of brozopine.

Objective:Making single chain antibody(scFv)-alkaline phosphatase(Ap)fusion protein.Methods:An expression vector pSTE2-C66-Ap was constructed by sequentially inserting the Ap coding region into plasmid pSTE2-8E5 and replacing the VH-VL fragment of 8E5 by VH-VL fragment of C66.The fusion protein scFvC66-Ap was expressed in E.coli.and analysed by SDS-PAGE and immunoblotting.Results:Have obtained the scFvC66-Ap fusion protein with a molecular weight of 75 kD.It bound a 60 kD molecule from KG1a cell proteins on immunoblotting membrane detected directly by Ap enzymatic activity.Conclusion:A method permits the production of scFv-Ap conjugates in E.coli.which can replace conventionally prepared Ap-labeled antibodies in immunoassays.

AIM:To investigate the effect of activation of retinoid X receptor (RXR) on transforming growth factor β1 (TGF-β1) induced collagen synthesis under hypoxic environment in rat cardiac fibroblasts (CFs) and underlying molecular mechanisms .METHODS: CFs were cultured using myocardial tissue with dry method .Hypoxic environment was established for CFs by continuous nitrogen supplement .Type I and type III collagens in supernatants were detected by ELISA.Nuclear and cytoplasmic extractions were prepared using NE-PER nuclear and cytoplasmic extraction reagents .The protein levels of Smad2 and p-Smad2 were determined by Western blot and immunocytochemical staining .RESULTS:Un-der hypoxic condition , TGF-β1 (0.01~10 μg/L) increased the synthesis of type I and type III collagens in a dose-de-pendent manner in the CFs .At the concentration of 5μg/L, the synthesis of collagen I and III was significantly increased as compared with control group (P<0.01).RXR agonist 9-cis-retinoic acid (9-cis-RA;10 -9 ~10 -6 mol/L) decreased TGF-β1 (5μg/L)-induced synthesis of type I and III collagens in a dose-dependent manner in the CFs under hypoxic con-dition.The synthesis of type I and type III collagens was significantly inhibited by 9-cis-RA (P<0.01).Smad2 inhibitor ( 20 nmol/L) showed similar inhibitory effect on the synthesis of type I and III collagens induced by TGF -β1 under hypoxic condition.Compared with TGF-β1 intervention group, the cytoplasmic level of p-Smad2 in the CFs was significantly in-creased in TGF-β1+9-cis-RA group, but the nuclear p-Smad2 level was significantly decreased (P<0.05).CONCLU-SION:Retinoid X receptor agonist 9-cis-RA inhibits TGF-β1-induced synthesis of type I and type III collagens in the CFs by repressing p-Smad2 nuclear translocation under hypoxic condition .

Case teaching method was used to undergraduates’specialized courses of clinical laboratory medicine such as the clinical laboratory hemotology and basic clinical laboratory medicine. This teaching method achieved satisfactory effect in clinical ability culture and obtained the positive opinion of students.

Objective To investigate the impaired characteristics of working memory in Parkinson's disease patients with depression and the recovery status of working memory after antidepressant treatment.methods Totally 60 cases were enrolled in this study and divided into three groups:30 Parkinson's disease patients with depression (PD-D),15 Parkinson's disease patients without depression (PD) and 15 persons without Parkinson's disease as normal controls.All subjects were evaluated with Wechsler Memory Scale (WMS-R) audio-visual span,Tower of Hanoi (TOH) and Wisconsin Card Sorting Test (WCST).30 PD-D patients were randomly divided into two subgroups:routine treatment subgroup (8 weeks levodopa therapy) and antidepressant combined treatment subgroup (8 weeks levodopa + citalopram therapy).The evaluation of Hamilton Depression Scale (HAMD),WMS-R audio-visual span,TOH and WCST were performed on these two groups before and after treatment.Result s① There were significant differences comparing PD-D and PD groups with the normal control group in scores of WMS-R,TOH and WCST indicators (P<0.05).There were significant differences in WMS-R visual span,accuracy rate and speed of TOH,as well as the percent errors and percent perseverative responses of WCST comparing PD-D group(11.88±5.91,0.420±0.345,0.408±0.334,0.882±0.253,0.565±0.229) with PD group (15.87±5.21,0.768±0.167,0.634±0.232,0.493±0.161,0.327±0.122) (P<0.05).② Before and after treatment in PD-D routine treatment subgroup,there were significant differences in digital memory and audio-visual memory of WMS-R((6.73±3.72,5.95±3.13) vs (3.77±2.16,1.91±1.58)),accuracy rate of TOH(0.45±0.26 vs 0.23±0.13),as well as percent errors((-0.58±0.17) vs (-0.37±0.14)),percent perseverative responses((-0.32±0.15) vs (-0.14±0.09)),percent conceptual level responses(0.38±0.09 vs 0.13±0.07) and number of categories completed(3.27±1.56 vs 1.06±0.91) of WCST(P<0.05).③ After 4-week treatment and 8-week treatment,there were significant differences in HAMD score comparing PD-D antidepressant combined treatment group(16.33±2.72,10.27±2.66) with PD-D routine treatment group(21.73±2.28,18.4±2.47) (P<0.05).Conclusion Comparing Parkinson's disease patients without depression,the impaired working memory is more serious and extensive in Parkinson's disease patients with depression.Antidepressant treatment can improve the working memory of Parkinson's disease patients with depression.

Objective To explore the effect and mechanism of magnesium on calcification induced by hyperphosphate.Methods Vascular smooth muscle cells (VSMCs) were primarily cultured in vitro and induced calcification by β-glycerophosphate (β-GP).VSMCs were randomly divided into control group,high phosphorus group (10 mmol/L β-GP),magnesium intervèntion group(10 mmol/L β-GP + 3 mmol/L MgSO4) and 2-aminoethoxy-diphenylborate (2-APB,an inhibitor of magnesium transporter) intervention group(10 mmol/L β-GP+3 mmol/L MgSO4+ 10-4 mol/L 2-APB).Calcium deposition and alkaline phosphatase (ALP) activity were measured by alizarin red staining,quantification of calcium and euzyme linked immunosorbent assay.RT-PCR and Western blotting were used to observe the expression of core binding factor α-1 (Cbfα-1) mRNA and protein,respectively.In vivo,male Sprague-Dawley rats (n=24) were randomly divided into control group (methylcellulose+high phosphorous diet),vascular calcification group (adenine suspension + high phosphorous diet),high magnesium intervention group(adenine suspension+high phosphorous and magnesium diet).The aortic pulse wave velocity (PWV) was measured,and vascular calcification was determined by von Kossa stain and quantification of calcium.Cbfα-1 in aortic was measured by immunohistochemistry.Results In vitro,compared with high phosphorus group,calcification,ALP activity (P ＜ 0.05) and Cbfα-1expression in VSMCs were significantly decreased in magnesium intervention group after incubation for 14 days,but the addition of 2-APB might inhibit the protective effect of magnesium on VSMCs.Dynamic observation of Cbfα-1 showed that magnesium significantly inhibited the expression of Cbfα-1 (P ＜ 0.05) on the third day and the inhibitory role was obviously increased in a time-dependent manner.Consistent with the findings in vitro,the aortic PWV,calcification were all significantly reduced (P ＜ 0.05) in high magnesium intervention group with high serum magnesium level,when compared with vascular calcification group.Immunohistochemistry showed that hypermagnesemia downregulated obviously the expression of Cbfα-1 induced by hyperphosphatemia(P ＜ 0.05).Conclusion Magnesium protects against vascular calcification by inhibiting osteogenic differentiation of VSMCs.

Objective To explore the effect of vitamin K2 on β-glycerophosphate(β-GP)-induced rat vascular smooth muscle cells (VSMCs) calcification and and the mechanism.Methods VSMCs were obtained from rat aortic,and identified by immunocytochemistry,then randomly divided into control group,high phosphorus group,vitamin K2 group (the group was settled three subgroups according to the concentration of vitamin K2 based on the high phosphorus medium,namely 10 μmol/L,25 μmol/L,50 μmol/L) and noggin (bone morphogenetic protein pathway inhibitor) group.Calcification was visualized by Alizarin red staining,calcium load in cells was quantified by o-cresolphthalein complexone method and alkaline phosphatase (ALP) activity was measured after stimulating 14 days,gene expressions of bone morphogenetic protein-2 (BMP-2),SMAD1,SMAD7 and Runx2 mRNA were detected by RT-PCR,Runx2 protein levels was detected by Western blotting after stimulating 3 days.Results Compared with the cells in control group,high phosphorus induced cell calcification,increased ALP activity,up-regulated the expression of BMP-2,SMAD1,Runx2 mRNA (P ＜ 0.05) and down-regulated the expression of SMAD7 (P ＜ 0.01),while compared with high phosphorus group,the calcium deposition,ALP activity and the expression of BMP-2,SMAD1,Runx2 mRNA were remarkably reduced in a dose-dependent manner by treatment with vitamin K2 (P ＜ 0.05) and the expression of SMAD7 was increased (P ＜ 0.01).Compared with high phosphorus group,SMAD1 and Runx2 expression in noggin group were remarkably reduced(P ＜ 0.01).Conclusion Vitamin K2 inhibits β-glycerophosphate-induced VSMCs calcification which correlates with the suppression of the expression of osteoblast markers through the down-regulation of bone morphogenetic protein pathway.

Objective To explore the genetic mechanism of Yang-deficiency constitution by detecting genomic copy number variations (CNVs). Methods Thirty cases of Yang-deficiency constitution and 30 cases of balanced constitution were included according to the standards of Classification and Determination of Constitution in Traditional Chinese Medicine. DNA was extracted from white blood cells in peripheral blood. A genome-wide association study was conducted by using Affymetrix SNP 6.0 platform. CNVs of each sample were analyzed using PennyCNV software. The Yang-deficiency constitution-specific copy number variation regions (CNVRs) of each autosome were identified. CNVR-related genes and their annotations were searched at online Human Genome Browser. Results The mean number of CNVs in balanced constitution group was 12.63±3.39, ranging from 8 to 20. After stepwise elimination of two Yang-deficiency constitution subjects, the mean number of CNVs in Yang-deficiency constitution group was 15.04±8.95, ranging from 2 to 38. A total of 26 CNVRs were identified from 28 Yang-deficiency constitution subjects, including 19 duplicated CNVRs, 6 deleted CNVRs, and 1 mixed type CNVR. Most CNVRs were shared by a few Yang-deficiency constitution subjects, and only 7 CNVRs were shared by more than 5 Yang-deficiency constitution subjects. The functions of representative genes in Yang-deficiency constitution-specific CNVRs were related with extracellular and intracellular signal transduction, metabolic regulation, and immune response, etc. Conclusion Yang-deficiency constitution subjects have some specific genomic CNVs, which might result in Yang-deficiency constitution phenotypes by influencing the expression of genes associated with extracellular and intracellular signal transduction, material metabolism (energy metabolism), and immune response, etc.