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    21 April 2011, Volume 32 Issue 2
    特别关注
    The Detriment of Nuclear Radiation on Human and the Protection Principal
    ZHOU Ping-kun
    2011, 32(2):  171-176. 
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    Nuclear radiation is ionizing radiation, in terms of particulates and electromagnetic radiation emitted from atomic nuclei in various nuclear processes, e.g., nuclear fission, radioactive decay. Hit of nuclear radiation on matter results in the excitation and ionizing events. The major types of nuclear radiation include: α particle, β particle, γ-rays and neutrons. The major characteristics of physics and biophysics of nuclear radiation, detriment on human, and the protection principal were discussed. The severity and /or risk frequency of nuclear radiation detriment on living organisms are closely associated with radiation quality, absorbed dose and dose-rate.
    艾滋病基础与流行病学研究
    Restriction of HIV-1 and the Vif-deficiency strain by APOBEC3G
    LI Lan;ZENG Yi
    2011, 32(2):  177-181. 
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    Objective To study the restriction of HIV-1 and the Vif-deficient strain.
    Methods Viruses of HIV-1 wild type(BH10 WT) or the Vif-deficient strain(BH10 ΔVif) produced from transfected 293T cells were used to infect various cell lines, including MT4 and H9. The results were determined by reverse transcriptase(RTase) assay. Molecular-cloning technique was used to construct expression plasmid pEGFP-3G which express APOBEC3G with a C-terminal GFP tag. The virus strains of BH10 WT or BH10 ΔVif cotransfection with different dose of pEGFP-3G in 293T cells, and the expression of APOBEC3G-GFP was observed by live cell fluorescence microscopy. The infectivity of virus was determined by multinuclear activation of galactosidase indicator(MAGI) assay and RTase assay.
    Results BH10 WT replication in MT4 cells showed much faster replication kinetics than that in H9 cells, with peak RT values being observed as early as 4 days post-infection. RT activity of BH10 ΔVif in H9 cells was suppressed almost to the level of negative control and that in MT4 cells was observed on the 12th day. GFP-APOBEC3G and a GFP-only control localized to the cytoplasmic and cell-wide, respectively. The titer of BH10 ΔVif by MAGI assay is about 2.75×104 U/mL. The titer of viruses, produced in 293T cells by cotransfection with increasing amounts of APOBEC3G, was significantly reduced from 1.48×103 U/mL to 0.33×103 U/mL. The infectivity of BH10 ΔVif produced in the presence of 0.2 μg of co-transfected pEGFP-3G was twenty fold less infectious than BH10 ΔVif produced in the absence of APOBEC3G.
    Conclusion The anti-HIV activity of APOBEC3G was dose-dependent, and HIV-1 Vif has the essential role in the virus replication. Based on the results we would construct a new platform to accurately and promptly select efficacious drugs.
    Epidemiological Characteristics of HIV/AIDS Cases
    WANG Rui;ZHANG Mei;JI Yun-xia;SONG Ying-xue;WU Hao;JIAO Yan-mei
    2011, 32(2):  182-185. 
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    Objective To analyze the epidemic characteristics of HIV/AIDS cases.
    Methods The epidemiological data, including HIV transmission routes, HIV-1 co-infection with HBV/HCV/syphilis and the percentage of AIDS among HIV/AIDS cases diagnosed from Jan. 2003 to Dec. 2007 in AIDS clinic of Beijing You’an Hospital, Capital Medical University, were analyzed retrospectively.
    Results ① Data of randomly selected 330 HIV/AIDS cases from AIDS Clinic of Beijing were used. Among the cases, the rates of infection via sexual transmission and via blood transmission from 2003 to 2007 were 30% vs 60%, 24.8% vs 75%, 26.9% vs 70%, 67.9% vs 20%, 84.9% vs 10%, respectively. ② The percentage of HIV-1 co-infection with HBV were increasing from year2003 to 2007 were 4%, 10%, 14%, 21%, 22%, respectively; while co-infection with syphilis were 15%, 5%, 16%, 23%, 30%, respectively; the percentage of HIV-1 co-infection with HCV were 60%, 62%, 51%, 18%, 8%, respectively. ③ The percentage of AIDS among HIV/AIDS cases tended to be increasing year by year, It increased from 6.5%(year 2003) to 9.5%(year 2004) and 20%( year 2005), 25%(year 2006), 26.3%(year 2007).
    Conclusion ① Among the cases, there had been an alarming increase in the rates of infection via sexual transmission and co-infection with HBV/syphilis, while the rates of infection via blood transmission and co-infection with HCV had a rapid drop. Infection via sexual transmission tended to replace the blood transmission and is becoming the major route of transmission. ② The percentage of AIDS among HIV/AIDS cases tended to be increasing year by year.
    Treg Cells Depletion Can Lead to HIVspecific CD8+Cells Apoptosis in HIV Patients
    JIAO Yan-mei;FU Jun-liang;ZHANG Zheng;JI Yun-xia;WANG Rui;ZHANG Tong;CHEN De-xi;WU Hao
    2011, 32(2):  186-189. 
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    Objective To investigate whether the decrease of CD4+CD25+FoxP3+ cells(Treg) can lead to HIV-specific CD8+ cells apoptosis in HIV patients.
    Methods We characterized Tregs among 75 antiviral-naive HIV-1-infected individuals in this study. Treg and CD8+ cell activation and apoptosis were detected and its relevance was analyzed; Treg cells and CD8+ cells were simultaneously purified and then co-cultured in vitro, and HIV-specific CD8+ cell apoptosis was detected with pentamer and annexin V.
    Results Absolute counts of circulating Treg was inversely correlated with up-regulated activation and apoptosis of CD8+ T cells. In addition, Tregs were found to suppress HIV peptite-induced apoptosis of HIV-specific CD8+ T cells. ConclusionThese findings indicate that the decrease in Tregs closely correlate with increased apoptosis of CD8+ T cells, especially with HIV specific CD8+ T cells.
    Influences of HCV Infection on Incidence of Occult HBV Infection in Patients Infected with HIV-1
    LIANG Hong-xia;YU Zu-jiang;ZHANG Qian;LI Zhi-qin;PAN Yan-feng;LI Juan;JIANG He-qing;LI Jian-sheng;HE Yun
    2011, 32(2):  190-193. 
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    Objective To investigate the influences of HCV infection on occult HBV infection in patients infected with HIV-1.
    Methods The study enrolled 178 HBsAg-negative treatment-naive HIV-1 patients who transmitted by commercial blood donation in an HIV/AIDS CARE site in Henan province. Many parameters were detected, including liver function, HBV serologic markers(HBsAg, Anti-HBs, HBeAg, anti-HBe and anti-HBc), HCV antibody, HBV DNA and HCV RNA. Analyses were performed to compare the differences of HBV etiological markers in antiHCV+ group, anti-HCV- group and different HCV RNA groups.
    Results Among the 178 HBsAg-negative HIV-infected patients, 35 were HBV-M negative, 25 anti-HBs positive only, 25 anti-HBc positive only, 34 with both anti-HBs and anti-HBc positive. There were no significant differences in ages and gender between anti-HCV+ group and anti-HCV- group(P>0.05). There were significant differences in ALT and AST(P<0.05) but not total bilirubin between anti-HCV+ group and anti-HCV- group. There were no significant differences in HBV-M negative, isolated HBsAb positive only, isolated anti-HBc positive only, anti-HBs and anti-HBc double positive, HBV DNA positive between anti-HCV+ group and anti-HCV- group. There were no significant differences in HBV DNA positive rate between different HCV RNA groups(P>0.05).
    Conclusion In the HIV-infected patients who transmitted by commercial blood donation, HCV infection and viral loads of HCV RNA had no correlation with incidence of occult HBV infection, but HCV co-infection could aggravate the liver damage.
    Structural Characteristics and Biological Effects of HIV Negative Factor
    ZHANG Hong-wei;WU Hao
    2011, 32(2):  199-202. 
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    Negative factor(Nef) is one of the human immunodeficiency virus(HIV) accessory proteins with small molecular weight, large flexibility and spherical core. HIV Nef is an adaptor protein with no enzymatic activities. The protein has a central function in fighting against host immune response by downregulating CD4, MHC Ⅰ, MHC Ⅱ and CD28. The protein can enhance HIV replication and infectivity, and inhibit apoptosis of HIV-infected cells. The Nef protein plays a critical role in HIV pathogenesis.
    Vasoconstriction Effect of Thrombin on Pulmonary Artery and Thoracic Aorta ofRats and Its Possible Mechanism
    ZHANG Qian;;LIU Jie;;ZHANG Zhi-fei;;LIN Yuan;;WANG Jun;;WANG Chen;
    2011, 32(2):  203-207. 
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    Objective To explore the vasoconstrictive effect and mechanism of thrombin on pulmonary artery and thoracic aorta of rats.
    Methods The pulmonary artery and thoracic aorta ring of rat were isolated in vitro according to the procedures routinely used. Vascular constriction tension recordings were accomplished by using Powerlab biology signal collection system. RT-PCR method was used to detect the mRNA expression of thrombin receptor in vascular rings.
    Results The pulmonary artery and thoracic aorta rings were constricted by thrombin which was independent on the extracellular calcium. This effect was similar to that of the TRAP6, a thrombin receptor agonist. The mRNA expression of thrombin receptor-1 can be detected in the pulmonary artery and thoracic aorta.
    Conclusion Thrombin induced pulmonary artery and thoracic aorta ring constriction was independent of extracellular calcium and might be mediated by thrombin receptor 1(PAR1).

    Changes of Membrane Potential in Pulmonary Smooth Muscle Cell Isolated from Chronic Thromboembolic Pulmonary Hypertension Patients
    LI Jing;WANG Chen;LIU Jie;LI Ji-feng;;LIU Yan;GU Song;YANG Yuan-hua;ZHAI Zhen-guo;GAN Hui-li;WANG Jun;
    2011, 32(2):  208-213. 
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    Objective To investigate the method of isolating and culturing the pulmonary artery smooth muscle cells(PASMCs) from the endarterectomized specimens of CTEPH patients and to observe their electrophysiological characteristics.
    Methods The endarterectomized tissues of CTEPH patients undergoing pulmonary thromboendarterectomy(CTEPH-PAMSCs) and normal lung tissues of bronchogenic carcinoma or bullae patients(normal-PASMCs) were collected, the PASMCs were isolated by enzymatic method and identified by SM-α-actin. Patch clamp technique was used to measure the cell’s membrane potential and action potential.
    Results Human PASMCs from normal lung tissues and CTEPH patients’ tissues were successfully isolated and cultured that was confirmed by SM-α-actin staining. Compared with PASMCs of normal control, the membrane potential in CTEPH-PASMCs was more depolarized(-38.12mV±2.28 mV, n=10 vs -21.05 mV±2.20 mV, n=11, P<0.001) and the action potential duration at 50%, 75%, 90% were all prolonged(APD50: 0.100 s±0.016 s vs 0.185 s±0.035 s, APD75: 0.150 s±0.024 s vs 0.277 s±0.053 s, APD90: 0.180 s±0.028 s vs 0.333 s±0.064 s, P<0.05).
    Conclusion The results suggested that the PASMCs in CTEPH patients has a more depolarized resting membrane potential and a prolonged action potential duration, which might be important factors determining the calcium influx via the voltage-gated calcium channel in pulmonary vascular remodeling.

    Differential Expression of NKCC1 and NKCC2 in Rat Gastric Mucosa
    DOU Zhang-feng;ZHANG Yue;JI Tuo;XUE Hong;ZHU Jin-xia
    2011, 32(2):  214-217. 
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    Objective To investigate differential distribution and gene content of two isoforms of Na+-K+-2Cl- co-transporter(NKCC), NKCC1 and NKCC2 in rat gastric mucosa.
    Methods Immunofluorescence and realtime PCR were used to investigate the distribution and expression of NKCC1 and NKCC2 in different parts of stomach(gastric fundus, corpus and antrum).
    Results ① NKCC1 immunoactivity in the gastric mucosa was found in the bottom part of gastric fundus and antrum, and middle and bottom mucosa of gastric corpus of rats, and cell membrane showed strong NKCC1 labeling. NKCC2 distributed across the full-thickness mucosa of gastric corpus and antrum but in the upper 1/3 mucosa of gastric fundus of rat, and it was dispersed in the cytoplasm and cell membrane. ② Real-time PCR indicated that the mRNA of NKCC1 expressed mostly in the gastric corpus, however, the mRNA of NKCC2 expressed mostly in the gastric antrum.
    Conclusion In normal rats, the two isoforms of NKCC showed differential distribution and mRNA expression in the gastric mucosa; NKCC1 distributed mostly in the bottom or middle and bottom part of gastric mucosa, NKCC2 distributed mostly in the upper or full-thickness of gastric mucosa.

    Blocking Effects of Benzamil and Ruthenium Red onCarotid Baroreceptor Discharges
    LIU Ting-hui;WEI Hua;ZHAO Hai-yan;LIU Ping;HUANG Hai-xia;WANG Wei;FU Xiao-suo;NIU Wei-zhen
    2011, 32(2):  218-223. 
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    Objective To investigate the effects of benzamil, amiloride and ruthenium red(RR) on the activity of carotid sinus baroreceptor and to probe into the properties of the mechanosensitive ion channels in arterial baroreceptor.
    Methods The carotid sinus nerve discharge(CSND) was recorded from rabbit carotid sinus-nerve preparations in vitro with doubly perfusing the sinus internally and externally while the intrasinus pressure was clamped. The effects of the above-mentioned chemicals on the baroreceptor discharge were investigated.
    Results Benzamil blocked CSND dose-dependently with IC50=18.5 μmol/L. The effect was reversible after washing out. Amiloride(2 mmol/L) inhibited the nerve activity reversibly by 95%. RR(100 μmol/L) blocked CSND irreversibly by 89%.
    Conclusion Benzamil, amiloride and RR could block the baroreceptor activities of both type A and type C nerve endings, implying that similar mechanosensitive ion channels might locate in the two types of receptors. However, the relevant mechano-electrical transductive molecule remains to be clarified.

    Splicing Variant and Expression of BKCa in Mice Colonic Smooth Muscle
    HOU Dong-yan;LIU Ping;HUANG Hai-xia;WEI Hua;FU Xiao-suo;WANG Wei;NIU Wei-zhen
    2011, 32(2):  224-229. 
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    Objective To investigate the whole length and the splicing variants of the large conductance calcium activated potassium channels(BKCa) mRNA in mice colonic smooth muscle, and the semi-quantify at BKCa α-subunit in various segments of gastrointestinal tract.
    Methods To observe the expression of BKCa α-subunit mRNA and protein in smooth muscle from various segments of gastrointestinal tract by RTPCR and Western blotting, and to examine the splice forms of all the splicing sites of BKCa mRNA in colonic smooth muscle by RT-PCR.
    Results The expressions of BKCa both at the mRNA and the protein levels, in the smooth muscles of mouse stomach and colon were more abundant than those in duodenum and ileum. Exon 19(stress-regulated exon, STREX) was spliced in the mRNA of BKCa α subunit in colonic smooth muscle while no splicing on other exons was detected.
    Conclusion The abundance of BKCa in colonic smooth muscle is high among the smooth muscles of the various gastrointestinal segments. The splicing variant BKCa in colonic smooth muscle was STREXlacking type, namely ZERO type.

    Changes of Monophasic Action Potential in Rabbit Left Ventricles Induced by Ventricular PressureClamping Method
    WEI Hua;LIU Ping;HUANG Hai-xia;WANG Wei;ZHAO Hai-yan;NIU Wei-zhen
    2011, 32(2):  230-233. 
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    Objective To establish a ventricular pressure-clamping method for isolatedperfused hearts of small animals, observe the effect of mechanical stimuli on cardiac electrical activities and explore the phenomena and mechanisms of cardiac mechano-electric feedback.
    Methods Using the ventricular pressure-clamping method to stretch the left ventricle(LV) during the end of action potential, the effect on monophasic action potential duration(MAPD) of ventricles was observed.
    Results LVMAPD90 was lengthened by 15.4%, 26.1%, 44.4%(P<0.05) comparing to the control when the left ventricle was clamped by extra stretch which was 130, 140, 150 mmHg, respectively, while LVMAPD20 and LVMAPD50 had no changes; There was no significant difference in RVMAPD20, RVMAPD50 and RVMAPD90 between before and after extra inflations were applied.
    Conclusion This ventricular pressure-clamping method developed by our lab can exactly mimic the changes of load in hearts under the physiological/pathophysiological conditions and can freely deliver mechanical stimuli to the ventricle, which improved the method of pressure-controlling on researching cardiac MEF at organ level; stretching the left ventricle during the end of the action potential could lengthen MAPD, slower repolarization, while the right ventricle had not the same changes, which suggested that the inflations caused a kind of inward current in the left ventricle.

    The Effects of Cyp2j3 Gene Transfection on Myocardial Ischemia-reperfusion Injury in Rats in vivo
    LU Ling-qiao;YU Gang-gang;ZHANG Dong-mei;ZENG Xiang-jun;WANG Hong-xia;ZHANG Li-ke
    2011, 32(2):  234-238. 
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    Objective To observe the expression of cyp2j3 after intramyocardial injection of plasmid pcDNA3.1-2J3 and examine the effects of cyp2j3 gene transfer on myocardial ischemia-reperfusion injury.
    Methods Fifty-four male Wistar rats were randomly divided into three groups as follows: normal saline NS group, pcDNA3.1 group and pcDNA3.1-2J3 group, 80 μL normal saline or 80 μL pcDNA3.1 or 80 μL pcDNA3.1-2J3(1 g/L) was injected into 4 sites of left myocardium respectively. Two weeks after gene transfection, six rats from each group were sacrificed to determine the expressions of CYP2J3 mRNA and CYP2J3 protein in myocardial tissue by RTPCR and Western blotting respectively. The other rats were subjected to 45 min ischemia and 120 min reperfusion. Cardiac function was recorded by the use of BL-420F instruments. Infarction size was measured by triphenyltetrazolium chloride(TTC) staining.
    Results Compared with the control group, CYP2J3 mRNA and CYP2J3 protein were overexpressed in pcDNA3.1-2J3 group after two weeks gene transfer via direct injection. Left ventricular end-systolic pressure(LVESP) and left ventricular maximal velocities of contraction and relaxation(±LV dp/dtmax) were significantly increased in pcDNA3.1-2J3 group after 45 min ischemia and 120 min reperfusion. Myocardial infarction size also reduce in gene transfer pcDNA3.1-2J3 group compared with pcDNA3.1 group in I/R model(P<0.05).
    Conclusion Cyp2j3 can be effectively transferred into the myocardial tissue and express via direct injection pcDNA3.1-2J3 in vivo. Intramyocardial injection of plasmid pcDNA3.1-2J3 can attenuate the myocardial injury induced by ischemia or reperfusion.

    Difference of Reaction Time in the Cognition of Different Gender Name
    GAO Ying-xiao;ANDREW C.N. Chen;ZHANG Ming-xia
    2011, 32(2):  239-243. 
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    Objective To investigate the difference in the recognition speed while subjects of the different gender viewed Chinese names(male, female and neutral) under separated, mixed, and reversed conditions, and further study the “opposites attraction” in the recognition of different gender names.
    Methods Forty healthy adults(20 male and 20 female, age: 20~24 years) were enrolled in this study. The visual Chinese test stimuli consisted of male names, female names, and neutral names(50 trials each), respectively under separated, mixed, and updown conditions. The visual presentation and reaction time of the behavior task were quantified with the eevoke software.
    Results Comparison of the 3 name stimuli: fastest to female name〔(750.94±104.53)ms〕, next to male name〔(879.07ms±126.44)ms〕, and slowest to neutral name〔(1109.36±158.98) ms〕. Post-hoc t-tests of these 3 name classes also exhibited significant differences among them in R.T.(all at P<0.001 level). Between male subjects and female subjects under three conditions〔name separated: F(1,19)=0.059, P=0.811; name mixed: F(1,19)=0.139, P=0.713; name reversed: F(1,19)=3.592, P=0.073〕, the reaction time on the three types of name had no significant difference.
    Conclusion Female name elicits fastest R.T. than male name, and the slowest for neutral name. No “oppositesex attraction” affect in name recognition. Gender of subjects makes no difference in the reaction speed of name classes. Apparently, the stimulus conditions affect the cognitive speed, but they did not change the discipline that reaction speeds of female names are fastest and neutral names are slowest.

    Effects of Hypoxia on PTEN Phosphorylation and Nuclear Localization
    ZHOU Yun-ying;LIU Hua;GUO Rui-han;YANG Long-yan;BIAN Wei-hua;HE Jun-qi
    2011, 32(2):  244-248. 
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    Objective To investigate the effects of hypoxia on endogenous phosphatase and tension homolog deleted on chromosome ten(PTEN) phosphorylation and nuclear localization.
    Methods COS-7 cells were cultured in hypoxic condition(1% O2) and harvested at 0, 1, 8, 24, 32 h. Each lysate was used to detect p-PTEN and PTEN downstream molecule p-Akt expression by Western blotting. Subcellular localization of PTEN was observed at 0, 24 h with hypoxic treatment by nucleus and plasma separation method and confocal microscopy respectively.
    Results Upon hypoxic treatment for 24 h, phosphorylated PTEN level decreased significantly,accompanied by p-Akt expression level decrease; and nuclear PTEN level increase.
    Conclusion In response to hypoxia treatment, PTEN was activated due to decrease of phosphorylated PTEN level, then PTEN inhibited downstream Akt activation. Under hypoxic condition, PTEN was promoted to enter nucleus.

    Enrichment of egfr Gene Promotor in Human Genome Wide by RNA Capture
    YANG Jin;ZOU Bin-bin;ZHANG Yu-xiang;WANG Ze-sheng
    2011, 32(2):  249-253. 
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    Objective We took epidermal growth factor receptor(egfr) gene promotor DNA as target fragment and established RNA capture assay which is very efficient for targeted genomic DNA enrichment and can be used in deep sequencing of target regions with second-generation sequencing.
    Methods HeLa S3 cervical cancer cells were used as a model in the study. The genomes were extracted, digested with Mbo Ⅰ, filled ends, added with PE adapters, then 400 bp~1 000 bp size fragments were selected as DNA library. egfr promotor RNA was used to hybridize with the library. Hybridized fragments were selected by magnetic beads, PCR amplified for 15 cycles, and sequenced with Solexa second generation sequencing system.
    Results The sequenced tags were analyzed by bioinformatics. There were 1 889 of 106 reads mapping to egfr gene promotor and only 2, 3, 3 reads mapping to each notch 1, platelet derived growth factor-B(pdgf-B) and src gene loci. We selected 106 the same long reads randomly from human genome as control group. The results showed that egfr fragments were enriched to 630 times by the RNA capture assay.
    Conclusion RNA capture of target genome sequence is an efficient assay for high-throughput re-sequencing of targeted regions of genome. RNA capture and high-throughput sequencing technology can be applied to re-sequencing of target regions, including regions related to alternative splicing, nucleosome distribution, single nucleotide polymorphisms(SNPs) and loci of complex disease-related genes.

    Cloning, Expression and Purification of Human Synuclein Gamma
    LIU Xiao-juan△;ZHANG Li-qing△;LI Jian;ZHAI Fang-li;LI Shen-tao;
    2011, 32(2):  254-258. 
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    Objective To study cloning, prokaryotic expression and purification of human synuclein gamma.
    Methods A DNA fragment encoding human synuclein gamma was amplified by polymerase chain reaction from a human cDNA library and was cloned into vector pGEX-6p-1. The construct carrying the coding DNA sequence of synuclein gamma fused with GST was transformed into E.coli BL21(DE3), and the fusion protein of synuclein gamma and GST was expressed by induction with IPTG. The product was purified by affinity chromatography, ion exchange chromatography and gel filtration.
    Results Human synuclein gamma was successfully expressed in E.coli in soluble form. The product was identified to be human synuclein gamma by SDS-PAGE and mass spectrography. After purification, the purity of the recombinant protein was more than 98%.
    Conclusion An effective purification protocol was established. Recombinant synuclein gamma with a purity more than 98% was obtained, which provided a basis for the further studies of the crystal structure, biological activities and functions of synuclein gamma.

    Comparison of the Number of Decidual Natural Killer Cells between Decidua Basalis and Decidua Parietalis in Human First Trimester
    WANG Wei;WANG Jing;WENG Jing
    2011, 32(2):  259-262. 
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    Objective To investigate the human decidual natural killer(dNK) cells in decidua basalis(DB) and decidua parietalis(DP) during early pregnancy and its possible functions.
    Methods DB and DP of the same early pregnant individual were discriminated by HE staining and expression of Keratin 7 by immunohistochemistry. The CD56+ dNK cells were shown by immunohistochemistry. The number of dNK cells in DB or DP was counted.
    Results Cytokeratin 7-positive extravillous trophoblast cells was a marker to distinguish DB and DP because they only present in DB. The number of CD56+ dNK cells in DB was significantly larger than that in DP(P<0.01).
    Conclusion The differential distribution of dNK cells in early pregnant decidua may be related to trophoblast invasion.

    Models for Antitumor Evaluation and Intercalation Mechanism Investigation
    XU Yan-xia;ZHAO Ming;ZHANG Xiao-yi;WANG Yu-ji;WU Jian-hui;HUANG Fei;ZHANG Jian-wei;PENG Shi-qi
    2011, 32(2):  263-271. 
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    For improving the cancer chemotherapy the discovery of new DNA intercalator is one of the effective approaches. The investigations have demonstrated the utility of the models in both evaluation of anti-tumor activity and identification of intercalation mechanism. This paper describes the DOCK score, ultraviolet(UV), fluorescence, circular dichroism(CD), relative viscosity, melting curve, kinetics in calf thymus(CT) DNA stacking of small molecule intercalator, kinetics in CT DNA intercalation of small molecule intercalator, interaction stabilizing the stacking complex of small molecule intercalator and CT DNA and the gel electrophoresis based models as well as their use in the identification of small molecule intercalating towards DNA.

    Expression of Estrogen Receptor mRNA and Genes Related to Regulation of Bile acid Metabolism in Fetal Rat Liver of Pregnant Rats with Intrahepatic Cholestasis
    SHI Qing-yun;ZHAO Jin;LIN Yun-geng;YAN Shi;ZHOU Xin;LIN Ying-qi
    2011, 32(2):  272-275. 
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    Objective To study the expression of estrogen receptor(ER) and gene related to regulation of bile acid metabolism in fetal rat liver of pregnant rats with intrahepatic cholestasis.
    Methods Sixty clean SD pregnant rats were selected and divided into two groups at random. ① Since the 13th day of pregnancy after taking blood, the rats in control group were injected subcutaneously with refined vegetable oil 2.5 mL·kg-1·d-1, study group was injected subcutaneously with the 17-α-ethynylestradiol(EE) 1.25 mg·kg-1·d-1 till the 17th day. ② On the 21st day, all rats were killed. Then the fetus livers were collected for study. ③ The levels of serum total bile acid(TBA) were examined by ELISA. The expression of ER, CYP7A1, CYP27A1, CYP8B1(mRNA) in fetal rat liver was examined by real-time PCR.
    Results ① The levels of TBA were significantly higher in study group mother rats(58.7±3.2)μmol/L than that of control group(24.6±1.3)μmol/L on the 17th day(P<0.05); The levels of TBA were higher in study group mother rats(66.4±2.7)μmol/L than that of control group(26.5±3.1)μmol/L on the 21st day(P<0.05). The levels of TBA were higher in study group fetus rats(28.4±2.6)μmol/L than that of control group(10.5±3.1)μmol/L on the 21st day. ② The expression of CYP7A1 mRNA(1.25±0.01), CYP27A1 mRNA(2.05±0.03), CYP8B1 mRNA(1.85±0.01) in study group fetus liver increased as compared to control group(0.35±0.02, 0.75±0.03, 0.85±0.02, respectively)(P<0.05). 3) The expression of ERα mRNA in study group(0.75±0.02)was higher than that of control group(0.45±0.01)(P<0.05). Conclusion The expression of ERα, CYP7A1, CYP27A1, CYP8B1(mRNA ) increased in the fetal rat liver of pregnant rats with intrahepatic cholestasis, resulting in increased synthesis of bile acids, which may be one of the mechanisms of perinatal death of the fetal rats.

    Establishment of Reference Method for Serum Urea Detection and Comparison with Routine Urea Kits
    ZHANG Ya-jun;ZHANG Jie
    2011, 32(2):  276-282. 
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    Objective To establish and validate the coupledenzyme equilibrium reference method as a reference method for urea detection and compare it with other 4 routine urea kits.
    Methods The reference method was verified by testing the two levels of urea in SRM909b and participating in RELA-2009. The EP9-A2 protocol was used to compare the results in forty serum samples in 4 routine kits methods with coupled-enzyme equilibrium reference method.
    Results The deviation of SRM909b level 1 was 2.19% and SRM 909b level 2 were 0.24% respectively through determining by the reference method. The results for RELA-2009 were within the accepted range. Comparing the confidence interval of the predicted bias with the definition acceptable error at the medical decision point Xc(Xc=9.64 mmol/L) for the 4 routine kits, the results were all within the range of defined acceptable error(4.5%) and the range of biological variation bias(5.5%).
    Conclusion The coupled-enzyme equilibrium reference method for serum urea was established in our lab. The urea results measured with 4 routine kits were different from results with the reference method, but the deviation was acceptable, and it will not affect the detection results.

    Use of Expandable Intramedullary Nail System in the Treatment ofLimb Fractures
    LU Meng;ZHAO Er-hong;QIANG Hua
    2011, 32(2):  283-286. 
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    Objective To investigate and assess the effectiveness and methods of using expandable intramedullary nail system(EINS) in the clinical treatment of limb fractures.
    Methods From June, 2006 to June, 2008, a total of 31 cases with limb fracture were treated with EINS. In total, 32 nails were used for 20 male patients and 11 female patients, ranging from ages of 19 to 85; femur, 9 cases; tibiofibular, 4 cases(both tibiofibular, 1 case; compound fracture, 1 case); humerus, 6 cases; proximal femur, 12 cases.
    Results In all 31 cases, the wounds healed within the first stage, time of surgery ranged from 45 to 150 minutes, averaging 97.5 minutes per surgery. Single side limb blood lose ranged from 50 to 500 mL, on average, 275 mL. Thirty-one patients were followed up post surgically. Follow-up periods ranged from 8 to 32 weeks, averaging 20 weeks. The average healing time of treated limb fractures: 10 weeks for proximal femur, 12.8 weeks for femur, 12.4 weeks for tibiofibular, 13.5 weeks for humerus.
    Conclusion EINS has a unique design that is advantageous in the clinical treatment of limb fractures. Its surgical procedures are relatively simple, less invasive, and the duration of surgery short. In addition, EINS significantly decreases the amount of X-ray exposure, thus, decreasing the potentially harmful effects of X-ray on both doctors and patients, especially for young patients that have not had children. The axial flexibility is stable and accelerates the healing of fracture. PF expandable stud extrudes cancellous bone and does not pose the problem of bone cutting used in internal bone fixation methods. This is especially suitable for osteoporotic patients. There is no need for remote locking and is therefore suitable for both distal limb patients and patients with skin conditions. EINS provides an innovative alternative method for the treatment of long bone fracture.

    Applicability of GlideScope Video Laryngoscope for Tracheal Intubation in Clinical Anaesthesia
    ZHAO Zhi-dan
    2011, 32(2):  287-289. 
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    Objective To explore applicability and safety of GlideScope video laryngoscope for tracheal intubation in clinical anesthesia.
    Methods One hundred ASAⅠ~Ⅱ patients, aged(45±15) years, were randomly assigned into 2 groups(n=50 each), scheduled for gynaecological laparoscopic surgery under general anesthesia(group G) with tracheal intubation. Anaesthesia was induced with fentanyl 2~3 μg·kg-1, propofol 2 mg·kg-1, vecuronium 0.1 mg·kg-1 and maintained with 1% isoflurane. Orotracheal intubation with GlideScope video laryngoscope was performed for group Ⅰ patients at 3 min after intravenous vecuronium; Group Ⅱ patients were intubated by direct laryngoscopy using Macintosh(group M). The glottic exposure time and the tracheal intubation time were recorded. The BP and HR were recorded before and after induction of anesthesia during tracheal intubation.
    Results The glottic exposure time was(15.38±5.21)s in the group G and(14.21±3.11)s in the group M. The mean tracheal intubation time was(30.22±11.31)s in the group G and (30.11±12.23)s in the group M. The BP was significantly decreased after induction of anesthesia, but HR was significantly increased during tracheal intubation in both groups.
    Conclusion GlideScope video laryngoscope achieved better glottic exposure with less tracheal irritation and damage. The method may reduce the risk of cross-infection to doctors.

    Observation of the Clinic Bleaching Effect of the BeyondTooth Whitening System
    ZANG Tao;FANG Chao;LI Ruo-xuan;LV Ya-lin;LIU Ying;WANG Jie
    2011, 32(2):  290-292. 
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    Objective To observe the bleaching effect and side effects of BEYOND Tooth Whitening System in different light time.
    Methods Postoperative tooth color change and pulp gum stimulated situation of 54 patients who were given different light time of bleaching treatment were recorded.
    Results The differences in color of the 3 experimental groups were statistically significant(P<0.05), in gum and pulp reaction to the drug stimulation was statistically significant(P<0.05), all these differences were in group 3(12 minutes)> in group 2(10 minutes) and >group 1(8 minutes), P<0.05 between 12 minutes and 8 minutes.
    Conclusion Increased light exposure time was associated with increased whitening effect, but also associated with the increased stimulation of dental pulp and increased degree of pulp reaction, therefore it is suggested that clinical operation with light exposure should be less than 12 minute each time.

    Value of Contrast-enhanced Ultrasonography in Distinguishing Hysteromyoma from Adenomyosis
    LI Guo-zheng;YUAN Min;WU Zi-fang;LIU Zeng-qiang;LIU Li-yan
    2011, 32(2):  293-296. 
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    Objective To investigate the value of contrast-enhanced ultrasonography(CEUS) in distinguishing hysteromyoma(HM) from adenomyosis(AM).
    Methods A total of 103 cases with hysteromyoma and 76 cases with adenomyosis were retrospectively studied with CEUS.
    Results The enhancement patterns of hysteromyoma on CEUS were peripheral pseudocapsule type in 89 cases(86.4%), synchronous type in 14 cases(13.6%), and no case had centripetal type enhancement. The enhancement patterns of adenomyosis on CEUS were synchronous type in 47 cases(82.5%), centripetal type in 10 cases(17.5%) and no case with peripheral pseudocapsule enhancement. The most of hysteromyoma are peripheral pseudocapsule type enhancement(Z=-10.502, P<0.01). While in adenomyosis, synchromous and centripetal enhancement(respectively Z=-8.562, P<0.01 and Z=-4.377, P<0.01) were the common type.
    Conclusion The enhancement pattern of hysteromyoma on CEUS was significantly different from adenomyosis. As a new non-invasive method, contrast-enhanced ultrasonography has important clinical value on the differential diagnosis of hysteromyoma and adenomyosis.

    Investigation on the Knowledge and Attitude of Wuhan Citizens to Hospice
    ZHAO Xian-da;WU Xin;LIU Dong-ying;HE Yu-yu
    2011, 32(2):  297-300. 
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    Objective To assess the situation of Wuhan citizens in the knowledge and the attitude to hospice.
    Methods A questionnaire survey using Multistage sampling survey method was applied.
    Results Totally 921 responded, including 468 male and 453 female;13.6% of the citizens heard of hospice frequently, 49.5% heard of hospice rarely and 36.9% never heard of; 26.70% would like to accept hospice care, 55.00% expressed that hospice should be thought over solemnly, 8.30% showed unwillingness to hospice; there are differences among age, education and the awareness of hospice. Most responders expect that hospice can show respect for life and relieve patients. Many of them worried about the outlay of hospice and the service quality.
    Conclusion The degree of awareness of hospice is not high in Wuhan citizens. Much should be done to strengthen publicity, improve the practitioner’s qualification and learn advanced concepts.The government is expected to support the development of hospice and reduce the expense.

    The Role of IL-17 in Ischemia Reperfusion Injury
    ZHU Wei-wei;WANG Peng-cheng;
    2011, 32(2):  304-308. 
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    In ischemia reperfusion, blood supply to tissues and organs was blocked, and blood flowing can aggravate the situation of tissue injury during reperfusion. In the past, studies were focused on the oxygen free radical damage, energy metabolism, intracellular calcium overload, leukocyte adhesion, endothelial cell injury and other theories about ischemiareperfusion injury(IRI) mechanism. However, the role of cytokines in IRI is increasingly reported recently. The latest research shows that IL-17secreting Th17 cells play an important role in the process of IRI. In this article, the roles of Thl7 cells and the cytokine IL-17 in ischemia-reperfusion injury are summarized.