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• 2013 Volume 23 Issue 4
  Published: 29 July 2013

    

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  Construction and identification of recombinant adenovirus encoding TCF4 and ΔTCF4

  HU Jun, YE Rong, WANG Chun-yan, et al

  2013, 23(4): 277-283.

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  [Abstract] Objective: To construct recombinant adenovirus encoding full-length T cell factor 4(TCF4) and a mutant TCF4 (ΔTCF4), which delete the binding sites of β-catenin, and to investigate the expression and activity of these recombinant adenoviruses in MSCs, thereby providing the foundation for further study about the role of Wnt/β-catenin signaling in MSCs and sheding a light on the usefulness of MSCs in cell transplantation and gene therapies. Methods: The DNA sequences of TCF4 and ΔTCF4 were amplified from cDNA of MSCs by RT-PCR, and were subcloned into the shuttle plasmid pAdtrack-CMV, forming transfer vectors pAdtrack-CMVTCF4 and pAdtrack-CMV-ΔTCF4. The plasmids were recombined in BJ5183 that has transformed by adenoviral backbone plasmid pAdEasy-1. Subsequently, both recombinant plasmids of Ad-TCF4 and Ad-ΔTCF4 were confirmed by RT-PCR, PacⅠ digesting analysis, and DNA sequencing. The identified Ad-TCF4 and Ad-ΔTCF4 were transfected to QBI-293A cells to produce the adenoviruses. To purify and titrate of recombinant adenoviral vectors, and to discuss Ad-TCF4 and Ad-ΔTCF4 vectors infected MSCs. Western blotting was applied to detect the expression of TCF4 in MSCs infected with Ad-TCF4. Luciferase activity assay was used to measure the activity of Ad-ΔTCF4 in MSCs.Results: The adenovirus vectors encoding target genes were proved to be recombined successfully in QBI 293A; and their titer virus were 1.1×107 pfu／mL and 1.3×107 pfu／mL respectively, The optimal virus of Ad-TCF4 and Ad-ΔTCF4 were 150 MOI, in which the efficiency of infection reached more than 90％. Western blot analysed the high expression of TCF4 in Ad-TCF4-infected MSCs, while luciferase activity was effectively suppressed in MSCs infected with Ad-ΔTCF4. MSCs infected with Ad-ΔTCF4 displayed decreased migration as compared with the control, however, infection with Ad-TCF4 showed no obvious differences. Conclusion: The recombinant adenoviruses encoding TCF4 and ΔTCF4 were successfully constructed and could infect MSCs efficiently.
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  RNA interference in pseudogene Cripto-3   on proliferation and invasion of human colon cancer cell

  ZHU Ling, JIANG Peng-cheng, MAN Chang-feng,et al

  2013, 23(4): 284-287.

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  [Abstract] Objective: To study the effects of RNA interference-mediated silencing of  pseudogene Cripto-3(Cr-3) on proliferation and invasion of colon cancer cells. Methods: The cells were divided into five groups, blank control group (Con-A group), empty control group (Con-B group), siRNA transfection group (5 nmol/L group, 10 nmol/L group and 20 nmol/L group). After human colon cancer cell line SW480 were transfected by Cr-3 small interfering RNA (siRNA), the levels of Cr-3 were determined by quantitative real-time PCR. The cell growth and invasion ability was examined by clone formation in soft agar, and  Transwell, respectively. Results: Compared with Con-A group and Con-B group, siRNA transfection group (5 nmol/L group, 10 nmol/L group and 20 nmol/L group) had lower levels of Cr-3 mRNA expression with lower invasion ability of colon cancer cells and in a dosedependent manner(both P＜0.05). Conclusion：Cr-3 might play an important role in the development of human colon cancer, and down-regulation of Cr-3 by its siRNA could inhibit the invasion of human colon cancer cells.
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    Effect of microRNA-21 transfection on  PDCD4  expression andcell proliferation in cervical cancer cells

  DAI Gui-hong1, LI Guo-li2, QIAN Hua1, YUAN Dong-lan3, JIAO Xia1, MEI Jia4, ZHU Xiao-wei1, YU Hong1

  2013, 23(4): 288-291.

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  [Abstract] Objective: To investigate the effect of miR-21 transfection on the expression of programmed cell death 4(PDCD4) and cell proliferation in cervical cancer cell line HeLa cells, and to provide experimental foundation for miR-21 gene therapy of cervical cancer. Methods: Lipid-mediated transfection was used to transfect pre-miR-21，pre-miR-21 negative control and anti-miR-21,anti-miR-21 negative control into HeLa cervical cancer cells. The miR-21 expression was measured by quantitative real-time PCR(qRT-PCR). The cellular growth activity was determined by MTT assay. The protein expression of PDCD4 was measured by western blotting. Detection of the target sites of miR-21 was analyzed by luciferase report gene assays. Results: In pre-miR-21 transfected cells, the cellular growth activity increased, and the expression of PDCD4 protein decreased. Conversely, in anti-miR-21 transfected cells, the cellular growth activity decreased, and the expression of PDCD4 protein increased. pMIR-Luc-PDCD4-3′UTR co-transfected with pre-miR-21, luciferase activity decreased; while it co-transfected with anti-miR-21, luciferase activity increased. Conclusion: PDCD4 gene was the target gene of miR-21, which suggested the potential possibility of miR-21 as a target gene for cervical cancer therapy.
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  Effects of siRNA-mediated HDAC2 gene silence on proliferation and apoptosis of human pancreatic cell line

  YAN Yan, ZHANG You-li, XU Min

  2013, 23(4): 292-295.

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  [Abstract] Objective: To investigate the effects of siRNA-mediated histone deacetylase 2(HDAC2) gene silence on the proliferation and apoptosis of human pancreatic cell line PaTu8988. Methods: Human pancreatic cancer cell line PaTu8988 cells were cultured and divided into five groups: control group, 20 nmol／L negative siRNA group, 40 nmol／L negative siRNA group, 20 nmol／L HDAC2 siRNA group, and 40 nmol／L HDAC2 siRNA group. Transient transfection of small interfering RNA (siRNA) against HDAC2 on human pancreatic cancer cell line PaTu8988 cells were performed by Lipofectamine 2000. Expression of HDAC2 gene was detected by quantitative real-time PCR and western blotting, respectively. The cell proliferation was determined by MTT assay. Flow cytometry was used to examine the cells apoptosis. Results: Compared with control group and negative siRNA groups, HDAC2 siRNA groups had markedly lower levels of HDAC2 expressions and lower cell growth rate, but higher cell apoptosis rate(P<0.05 or P<0.01). Conclusion: Transfection of HDAC2 siRNA could effectively inhibit the expression of HDAC2 in PaTu8988 cell lines, which suppressed the proliferation and induced apotosis of PaTu8988 cells.
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  Influence of hydrogenrich saline on senescence in aging rat kidney

  YUAN Xiao-liang, HE Xiao-zhou, XU Xian-lin, FAN Min

  2013, 23(4): 296-299，303.

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  [Abstract] Objective: To observe the impact of hydrogen-rich saline on the senescence of kidney in aging rats. Methods: Ten healthy adult male Sprague-Dawley rats aged 3-4 months were selected in the study as the adult group; meanwhile, twenty healthy male SD rats aged 1820 months were randomized into aging group and hydrogen-rich saline group. Adult group and aging group rats received intragastric administration of saline(5 mL·kg-1·d-1), while hydrogenrich saline group rats received hydrogen-rich saline(5 mL·kg-1·d-1).The other conditions were the same in the three groups. After eight weeks, the superoxide dismutase(SOD), malondialdehyde(MDA), 8-hydroxy-2′-deoxyguanosine(8-OHdG) contents in kidney tissues from all the rats were determined; and the mRNA levels of p53, p16, SIRT1 were measured by RT-PCR. Results: Aging group had greatly lower SOD contents, higher MDA and 8-OHdG concentrations than adult group; while hydrogen-rich saline group had significantly higher SOD contents, lower MDA and 8-OHdG concentrations than aging group(P<0.01 or P<0.05); Compared with adult group, aging group had significantly higher p53 and p16 mRNA levels with lower SIRT1 mRNA levels (P<0.01 or P<0.05); on the contrary, hydrogen-rich saline group had markedly lower p53 and p16 mRNA levels with higher SIRT1 mRNA levels, compared with aging group(both P<0.01). Conclusion: Hydrogen-ich saline could play a role in delaying kidney senescence, which was probably related to the inhibition of p53 and p16 expression and the improvement of SIRT1 expression.
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  Toxicological study on the security of calcium titanate biological materials

  WU Zhan-ao, XUAN Zhi-gang, SUN Min, JIANG Tao

  2013, 23(4): 300-303.

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  [Abstract] Objective: To evaluate the biological safety of calcium titanate biological materials. Methods: Using genetic toxicity test (mouse bone marrow nuclear test) and mice hemolysis test in vitro to evaluate the biological safety and biological compatibility of the biological materials, calcium titanate,which was prepared by hydrothermal synthesis technique at 120 ℃ for 6 h with the modifier such as polyethylene glycol,sixteen alkyl three methyl bromide (CTAB), and sodium citrate. Results:  The results of mouse bone marrow micronucleus test were <4‰ and hemolytic rate was <5％. Conclusion: Under this experimental condition, calcium titanate biological materials modified by the polyethylene glycol, sixteen alkyl three methyl bromide, sodium citrate and showed nothing were toxic  and had no mutagenicity and hemolysis.
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  Growth inhibition of 5-Aza-2′-deoxycytidine on human lung adenocarcinoma cell line A549

  LI Jing, GU Ying-chao, CHEN Yuan-yuan, DUAN Li-rong

  2013, 23(4): 304-307.

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  [Abstract] Objective: To investigate the inhibition of 5-Aza-2′ deoxycytidine (5-Aza-dC) on human lung adenocarcinoma cell line A549 and its possible mechanism.Methods: Use MTT method to find the differences of the effect on A549 cell growth; use Coomassie Brilliant Blue G-250 staining to determine the relative protein content of the tested A549 cell; use Hoechst 33258 fluorescence staining to observe the apoptosis of A549 cell and its morphological variations. Results：When the 5-Aza-dC was given in a certain range of concentration(40-100 μg／mL),the A549 cell proliferation could be significantly inhibited in a dose and time manner, compared with the control group(P<0.05, P<0.01). Meanwhile, the relative content of protein in the tested A549 cell was significantly decreased. When the concentration was in the range of 60-100 μg／mL, the apoptosis of A549 cell and its morphological variations could be observed. Conclusion： 5-Aza-dC can inhibit the growth of human lung adenocarcinoma A549 cells, induce the apoptosis and decrease the relative content of the protein. Its mechanism can be related to the block of cell cycle, alternation of the protein function and apoptosis induction.
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  Construction of prokaryotic expression vector carrying HIV Tat gene and the expression, purification and identification of recombinant Tat in E.coli

  QIU Hui-ping1, CHENG Xiao-dong2, LU Chun2

  2013, 23(4): 308-312.

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  [Abstract] Objective: To construct the prokaryotic expression vector carrying HIV-1 Tat gene and to purify the fusion protein in E.coli. Methods: The DNA fragment of the Tat gene from pcDNA3-1（+）／Tat101 was cloned into the prokaryotic expression vector pET-28a(+),named pET-28a(+)Tat. Then the recombinant vector was transformed into E.coli BL21 (DE3). The expression of the histidinetagged (His-Tag) fusion protein was induced with isopropyl-β-D-thiogalactopyranoside (IPTG). The fusion protein was detected and purified by Western blot and immobilized Ni2+absorption chromatographic column, respectively. Results: The prokaryotic expression vector carrying Tat gene was successfully constructed. The expression of the His-Tag fusion protein in E.coli BL21 (DE3) was detectable. Finally, the fusion protein with the relative molecular weight of 18×103 was gained after purified using the affinity chromatographic column. Conclusion: Recombinant Tat gene can be expressed in E. coli BL21 (DE3) and the fusion protein obtained can be functionally active.
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  Mitochondrial mechanism of cardioprotective effect of hydroxysafflor yellow A against anoxia／reoxygenation injury in rats

  LIU Yi-na, ZHU Jian-hua, WU Xiang, WANG Zheng-hua

  2013, 23(4): 313-316.

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  Objective: To determine whether the cardioprotection of hydroxysafflor yellow A (HSYA) against anoxia／reoxygenation injury is mediated by mitochondrial transmembrane pores. Method: Cardiomyocytes were isolated from male Sprague-Dawley rats. Cell viability was assessed by trypan blue exclusion and mitochondrial membrane potential was measured by loading with TMRE. The opening of mitochondrial permeability transition pore was determined spectrophotometrically. Result: Pretreatment with HSYA at 0.1 mmol／L for 5 min increased the cell viability against anoxia／reoxygenation injury and attenuated mitochondrial depolarization induced by anoxia／reoxygenation. In mitochondrial isolated from hearts pretreated with 0.1 mmol／L HSYA for 5 min, a significant inhibition of Ca2+ induced swelling was observed, while NG-nitro-L-argininemethyl ester (L-NAME), an inhibitor of NOS, abrogated the protective effects of HSYA. Conclusion: These findings indicate that HSYA protected cardiomyocytes against anoxia／reoxygenation injury via inhibited mitochondrial permeability transition pore opening, while nitric oxide may take a part in it.
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  Changes and clinical significance of ischemia modified albumin, high-sensitivity C-reactive protein and CXC ligand 16 in early  acute coronary syndrome patients

  GU Zhi-miao1, YANG Zhen-yu2

  2013, 23(4): 317-319.

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  Objective: To explore the values of the serological screening for the ischemia modified albumin(IMA), high-sensitivity C-reactive protein(hs-CRP) and CXC ligand 16(CXCL16) in early acute coronary syndrome (ACS) patients. Methods: A total of 93 patients who were admitted to hospital in 3 hours after chest pain occurred were recruited and divided into control group(n=30) and acute coronary syndrome group(ACS n=63), IMA, hs-CRP and CXCL16 were detected by intravenous haemospasia and compared between the two groups. Results: The levels of serum IMA, hs-CRP and CXCL16 in ACS group were increased significantly compared with the control group(all P<0.01). The results of correlation analysis showed that IMA had a positive correlation with hs-CRP and CXCL16(r=0.286,P<0.05; r=0.301, P<0.05). Conclusion: IMA, hs-CRP and CXCL16 were correlated with ACS, and CXCL16 was probably one of the responsible factors of the high serum IMA and hsCRP levels. They could be the earliest sensitive indicators to the diagnosis and risk estimation of ACS.
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  Enforced expression and verification of sall4 in HGC27 cell line

  ZHANG Ling1, GAO Shuo1, ZHANG Bin1, WANG Mei1, XU Xiao1, QIAN Hui1, XU Wen-rong1,2

  2013, 23(4): 320-324.

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  Objective: To establish the stable sall4-overexpressing HGC27 cell line, so as to lay a foundation for exploring the role of sall4 in the development of gastric cancer. Methods: pCMV6-entry-sall4 and pCMV6-entry plasmids were transfected into HGC27 cells via Lipofectamine 2000. At 48 h after transfection, cells were then cultured with G418 at a final concentration of 600 mg／L. Finally, stably transfected sublines of HGC27 cells were obtained. The expression of sall4 in stable cell line was determined by RT-PCR and western blotting. Results: The HGC27-sall4 cell line was obtained. The expression level of SALL4 in HGC27-sall4 cells was significantly higher than untransfected HGC27 cells and HGC27-vector cells. Conclusion: We successfully screened out the stable sall4-overexpressing HGC27 cell line.
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  Expression and correlation of plasma interleukin-17 and C-reactive  protein in patients with chronic obstructive pulmonary disease

  TANG Hai-cheng, BAI Hong-jian, JIANG Yong-qian, ZHANG Qun

  2013, 23(4): 325-327.

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  Objective: To investigate the changes of plasma interleukin-17 and C-reactive protein(CRP) with acute exacerbation and remission of chronic obstructive pulmonary disease (COPD), and analyze its clinical significance. Methods: Thirty-five patients with acute exacerbation of COPD(COPD group) and 23 healthy subjects (control group) were enrolled in the study. The plasma CRP and IL-17 levels in two groups were measured and analysed. Results: The levels of plasma CRP and IL-17 from patients with acute exacerbation and remission of COPD were higher than that in healthy control group(both P<0.01). Analysis results indicated that plasma IL-17 was positively correlated with CRP in patients with acute exacerbation of COPD before treatment(r=0.402,P=0.001). Conclusion: IL-17 and CRP both involved in the acute inflammation of COPD, and combination of  them had certain clinical significance in determining the acute exacerbation of COPD.
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  Pharmacognosy studies on a Li nationality medicine: Cymbopogoni citrati herba

  WANG Yong, HUANG Min-qin, WEI Na

  2013, 23(4): 328-331,336.

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  Objective: The pharmacognosy studies of the herb of Cymbopogon citratus (DC.) were systematically carried out. Methods： Macroscopical,microscopical and  thin-layer chromatography（TLC） identification methods as well as moisture, total ash and acidinsoluble ash assay, determination of ethanol and ether extracts methods and assay of citral by HPLC were used for the identification and assay of the crude drug. Results： The main microscopical characteristics were bicollateral peltate vascular bundles, secretory cavity containing oil, nonglandular hair with single cell, small glandular hair, diacytic type stoma, beaded incrassate xylem parenchyma cells. The colored spot of citral was obviously shown in the TLC chromatogram. The moisture content in lemongrass is about 11.9％, the total ash is about 7.39％ and the acid-insoluble ash is about 1.54％. The content of ethanolsoluable extract with hot maceration and the ether-soluable extract of volatile components are 9.04％ and 2.20％, respectively. The average content of citral in lemongrass is about 1.32％. Conclusion： The research provided scientific basis for the identification, quality control and further exploitation of C. citratus.
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  Aqueous two-phase system coupled with heat reflux for the extraction of tetrandrine from Stephania tetrandra S. Moore

  LUO Kai-wen, QI Xue-yong, GE Yan-ru, JIANG Xue-yin, ZHU Jing-jing, FU Hai-zhen

  2013, 23(4): 332-336.

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  Objective: To improve the method of extraction and separation of tetrandrine from Stephania tetrandra S. Moore in order to reduce environment pollution and cut down the cost of production. Methods: The ethanol/(NH4)2SO4 as a two aqueous phase system for the extraction and separation of tetrandrine was studied.The effect of operating parameters, including the concentration of ethanol and (NH4)2SO4,pH and temperature on extraction was studied in depth. Results: The extraction rate of tetrandrine in the aqueous two phase system could reach 84.50%. The ratio of solid:solvent was 1∶8, the time of heat reflux were 2 h, the salinity of (NH4)2SO4 was 35%, the density of ethanol was 20% (w, %), the pH level was 9, accordingly. The purity and yield of tetrandrine crude were 20.30% and 19.80 mg/g accordingly after recycling ethanol.Conclusion: The method of aqueous two phases could be applied to extraction and separation of tetrandrine in industry due to conveniently of procedure, highly of extraction rate and repeatability.
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  Properties of molecularly imprinted polymers prepared with chondroitin sulfate based on nano-titanium dioxide

  HUANG Qing1, XIA Sheng-yue2, GE Yan-ru1, QI Xue-yong1

  2013, 23(4): 337-340,359.

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  Objective: To synthesize molecularly imprinted polymers(MIPs) for chondroitin sulfate(Chs) by using molecular imprinting technique(MIT) and study the biosorption of Chs onto Chs-MIPs. Methods: The MIPs with specific performances of recognition and adsorption towards to Chs were prepared in the presence of Chs as the template molecule, nano-titanium dioxide (TiO2) as the carrier and acrylamide (AM) as the functional monomer, the ethyleneglycol dimethacrylate(EGDMA) as the crosslinker, azodiisobutyronitrile as the initiator. The structure of Chs-MIPs was characterized by scanning electronic microscope(SEM). The synthesis conditions and the adsorption characteristics of Chs-MIPs were also studied. The fits of pseudo-first-order and pseduo-second-order were investigated to state the sorption kinetics. The Langmuir, Freundich were applied to model the biosorption equilibrium. Results: The optimum mass ratio between TiO2 and AM was 6∶1; the optimum molar ratio between EGDMA and AM was 7∶1. Compared with pseudo-first-order, pseudo-second-order kinetic model demonstrated a better correlation for the experimental data determined. Moreover, the Langmuir isothermal equation showed a good fit to the experimental data. Conclusion: The MIPs made by MIT presented better specific adsorption ability to Chs and durability performances, which were confirmed by repeated durability experiments. It might be used as an effective biosorbent to extract Chs from solution.
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  Association of serum chemerin with high sensitive C-reactive protein in pe 2 diabetes with metabolic syndrome

  YUAN Guo-yue, DONG Si-jing, ZHU Tian-yi,et al

  2013, 23(4): 341-344.

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  Objective: To investigate serum levels of chemerin in patients with type 2 diabetes mellitus(T2DM)with metabolic syndrome(MS), and to study the association between serum levels of chemerin and highly sensitive C-reactive protein(hs-CRP). Methods: A total of 88 cases were selected, including 28 healthy subjects and 60 T2DM patients which were divided into MS group and non-MS group according to the diagnostic criteria of MS. Oral glucose tolerance test and insulin releasing test were performed, meanwhile, blood lipid, serum chemerin, hs-CRP and other items were measured. Results: Compared with normal control group and nonMS group, the serum chemerin was significantly increased in MS group［(86.47±23.61） vs （60.09±15.32） ng／mL, （86.47±23.61） vs （61.16±17.90） ng／mL, P<0.01,respectively］; The serum hs-CRP in MS group was higher than in non-MS group ［(2.57±1.64） vs （0.98±0.90） mg／L,P<0.01］. Serum chemerin level was positively correlated with body mass index(BMI), waist circumference(WC), waist hip ratio(WHR), diastolic blood pressure(DBP), Fasting insulin(FINS), postprandial serum insulin(PSI), HOMA-IR, HOMA-β, triglyceride and hs-CRP, and negatively correlated with fasting plasma glucose(FPG), postprandial plasma glucose(2hPG), HDL-C. The multiple regression analysis showed hs-CRP, WHR, DBP and HOMA-β were independent related factors in the influence of the serum chemerin levels. Conclusion: T2DM patients with MS tend to have higher serum chemerin and the level of serum chemerin was associated with hs-CRP.
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  Dual-source CT coronary angiography using the high-pitch mode in diagnosing the coronary artery stenosis

  WU Chun-hua, XUE Yue-jun, XU Yi-qun, RONG Wei-liang, PAN Chang-jie, QIAN Nong

  2013, 23(4): 345-349.

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  Objective: To evaluate the image quality, diagnostic accuracy and radiation doses for coronary artery imaging with second-generation 128-slice dual-source CT (DSCT) using the highpitch mode. Methods: A total of 123 cases patients with suspected or known coronary artery disease were randomly divided into two groups. Group A (n=58) underwent the adaptive prospective ECG-triggered sequence scan (CorAdpSeq) and group B (n=65) underwent the prospective high-pitch mode (pitch 3.4, Flash mode). Two experienced radiologists assessed the image quality and significant coronary stenosis (≥50％) with a double blinded fashion in a five score scale. Interobserver agreement for image quality was calculated with Cohen kappa statistics. The radiation doses values were measured. The diagnostic performance presented by DSCT in detecting stenosis ≥50％  were compared with the gold standard DSA. Results: There were 1 968 segments of coronary arteries in 123 patients included in this analysis. Diagnostic image quality (score≥3) was obtained in 1920 segments(97.6％) of the coronary artery segments, which was obtained in 98.1％(910／928)of segments in group A and 97.1％(1 010／1 040)in group B respectively with no significant difference (χ²=0.030,P=0.867). The mean image quality score for the whole coronary segments was (4.7±0.8) in 123 patients.There was no statistical difference between mean image quality of group A (4.7±0.7) and group B (4.6±0.9) (t=0.603,P=0.548). The kappa value between two radiologists was 0.799(P<0.01)on image quality. The effective radiation dose in group B (1.08±0.11) mSv was significantly (t=11.642,P=0.000) lower than that in group A (2.26±0.49) mSv. Sensitivity and specificity were 97.6％ and 97.5％ pervessel in group A; 98.1％,96.5％ per-vessel in group B. Conclusion: Compared with DSA both the high-pitch and CorAdpSeq mode for CTCA provide high image quality and accuracy of significant coronary stenosis, but the high-pitch mode can significantly reduce the radiation dose in the lower heart rate and regular heart rhythm.
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  Study on correlation between human urinary kallikrein treatment and influencing factors for acute cerebral infarction

  2013, 23(4): 350-353,359.

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  Objective: To investigate the effect of human urinary kallikrein(HUK) on acute cerebral infarction and the relationship between the treatment result and influential factors. Methods: One hundred and twenty patients with acute cerebral infarction were enrolled in the randomized control trail.All patients were randomly divided into treatment group(n=60) and control group(n=60).They were both treated by identical basis therapy.The patients in the treatment group were treated by syringe pumps of 0.15 PNA HUK diluted in 50 mL saline once per day for 21  days.The national institutes of health stroke scale (NIHSS) scores of two groups were analyzed before and after the treatment. All patients were classified according to the trial of org 10 172 in acute stroke treatment (TOAST) classification and oxforshire community stroke project (OCSP) classification. Results: Before treatment, there was no significant difference in the NIHSS scores between the two groups(P>0.05). At the end of treatment, the NIHSS scores in both groups were significantly decreased(P<0.05). Compared with the control group, the treatment group had lower NIHSS scores(P<0.05). The efficacy in the treatment group(81.67％)was better than that in the control group(51.67％). The differences between diabetes, TOAST classification and clinical results were significant(P<0.05). Age, sex, hypertension, heart disease, hyperlipidemia, smoking, drinking alcohol, transient ischemic attack(TIA), progressive cerebral infarction, and OCSP classification had no significant effects on clinical results(P>0.05).The efficacy using HUK to patients without diabetes and non cardiac cerebral infarction was better than the patients with diabetes and cardiac cerebral infarction. Conclusion: HUK is an effective treatment for acute cerebral infarction, especially for acute cerebral infarction patients without diabetes and non cardiac cerebral infarction patients.