Journal of Capital Medical University

Medical Hierarchy Construction, Research and Practice on Medical Personnel Training in Rural Areas of the Capital of PR China——Introduction of the Grand Prize in 6^th National Higher Education and Teaching Achievements

PANG Wen-yun;FU Li

2009, 30(5): 577-579. doi:10.3969/j.issn.1006-7795.2009.05.001

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The task of health-care in rural areas is directly related to the peasants' health, the agricultural economic development and social stability. In this paper, based on the status of lacking medical personnel and training qualified medical personnel as the goal in rural areas of the Capital of PR China, the research and practice on cultivating medical personnel in rural areas were performed. With new ideas and new perspectives, the training bases, teaching resources, educating & teaching mode and training mechanisms of medical personnel in rural areas were investigated and actually established. A medical personnel training system in the rural areas that is innovative, suitable to the condition of national economy, and in favor of sustainable progress was formed.

Rhythmic Oscillation of Neurotransmitter-related Genes Expression in the Mouse Striatum

DING Hui;CAI Yan-ning;WANG Ling-ling;CHAI Yu-xin;ZHANG Yan-li;CHEN Biao

2009, 30(5): 580-583. doi:10.3969/j.issn.1006-7795.2009.05.002

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Objective The aims of this study were to determine whether striatal neuropeptides(dynorphin(DYN), enkephalin 1(ENK1), substance P(SP), cholecystokinin(CCK)) and dopamine receptor 1 and 2(D1r and D2r) are regulated by the molecular clock; whether their rhythmic changes depend on the matural molecular clock. Methods Twenty-four hour mRNA oscillations of these genes were evaluated in the mouse striatum at early postnatal(postnatal day 3), pre-weaning(postnatal day 14), and adult(postnatal day 60) stages. At P3, no daily oscillations were observed. Results A significant time effect was present for D2r, DYN, and ENK1 at P14, and for all genes except D1r, at P60. Conclusion Circadian expression of these neurotransmitter-related genes develops in the mouse striatum gradually.

Immunolocalization of α-Synuclein in the Rat Spinal Cord

YIN Juan-juan;Giorgio Vivaqua;LI Xin;LIU Guang-wei;LI Yao-hua

2009, 30(5): 584-589. doi:10.3969/j.issn.1006-7795.2009.05.003

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Objective To provide a systematic map of α-synuclein(α-Syn) localization in the normal rat spinal cord. Methods Two home-made monoclonal antibodies(2E3 and 3D5) against α-Syn were immumohistochemically used to reveal the localization of α-Syn in the spinal cord. Luxol Fast Blue staining was applied to identify the unmyelinated nerve fibres. Double immunofluorescence was used to observe the co-localization of α-Syn with neuropeptide Y(NPY). Results α-Syn was localized in the neuronal nuclei and the unmyelinated nerve fibers within the gray matter. In both 2E3-and 3D5-stained sections, intensely stained nerve fibers were observed in laminae Ⅰ and Ⅱ. However, the immunopositive neuronal nuclei were only found in 3D5-stained sections, which were localized within the gray matter, especially in laminae Ⅶ, Ⅷ, Ⅸ, Ⅹ. Double immunofluorescent staining showed that some of the α-Syn-immunoreactive fibers and terminals in laminae Ⅰ and Ⅱ stained also for neuropeptide Y(NPY) in the dorsal horns, whereas in the anterior horns, a few motor neurons expressing α-Syn in the nuclei had cytoplasmic NPY. Conclusion α-Syn is localized in the neuronal nuclei and the unmayelinated nerve fibers within the gray matter of the spinal cord. Some of the α-Syn-positive neurons and fibers are NPY-positive.

Enhanced Oligomerization of α-Synuclein in Sera of Patients with Parkinson's Disease

LI Yan;YIN Juan-juan;LI Xin;WU Yan-chuan;LIU Guang-wei;LI Yao-hua;YU Shun

2009, 30(5): 590-592. doi:10.3969/j.issn.1006-7795.2009.05.004

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Objective To study the oligomerization of α-Synuclein (α-Syn) in the sera of patients with Parkinson's disease (PD) and the potential mechanism for this oligomerization. Methods Recombinant α-Syn was incubated in the sera of PD patients and of normal subjects. Immunoblot analysis was used to identify the formation of α-Syn oligomers. A special ELISA method was used to evaluate the amount of α-Syn oligomers in PD and control sera. Results The oligomeric forms of α-Syn formed in the serum were dimers, trimers, tetramers, and fourteen-polymers. Compared with control sera, the sera of PD patients significantly enhanced the oligomerization of α-Syn. Reduced glutathione significantly decreased the amount of α-Syn oligomers in the sera of PD patients. Conclusion α-Syn can form different forms of oligomers in the serum. Compared with control, sera from PD patients enhance the oligomerization of α-Syn, which may be mediated by oxidization of α-Syn.

Isolation and Activity Analysis of a Minimal Promoter of the Human Tyrosine Hydroxylase Gene

LI Yao-hua;YE Yi-wen;GAO Nan;LI Xin;YU Shun;YANG Hui;CHEN Biao

2009, 30(5): 593-596. doi:10.3969/j.issn.1006-7795.2009.05.005

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Objective To investigate the regulatory functions of human tyrosine hydroxylase gene upstream region. Methods A human TH gene upstream fragment was amplified by PCR, and was inserted into plasmid pGL3-Basic, to build a luciferase reporter vector. The reporter vector was transient transfected into MES23.5, 293T and Cos7 cells, and the promoter activity was mensurated using Dual Luciferase Assay system. Results Length of the isolated DNA fragment is 520 bp, which located at -495/+25 region of the human tyrosine hydroxylase gene. By comparison with pGL3-Basic vector, the recombinant reporter vector could significantly expressed luciferase in three types of cells(P<0.01). Conclusion TH gene -493/+27 region possesses promoter activity, and the constructed reporter gene will help us study the regulation of TH gene expression.

Differential Expression of Low Molecular Weight Proteins in Sera of Parkinson's Disease Patients and Normal Subjects

LI Yao-hua;YE Yi-wen;LI Xin;YU Shun;YANG Hui;CHEN Biao

2009, 30(5): 597-600. doi:10.3969/j.issn.1006-7795.2009.05.006

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Objective To study the differential expression of serum low molecular weight proteins in Parkinson's disease(PD) patients and normal subjects. Methods Serum samples from 35 idiopathic PD patients and 35 normal subjects were selected. Serum proteins were captured by weak cationic exchange(WCX) magnetic beads. Molecular weight of the proteins in beads-binding fraction was detected by MALDI-TOF-MS. Differential expression molecules in PD patients and normal subject were screened by statistics. A classification model was constructed by bioinformatics tools like Supervised Neural Network (SNN), and was validated by using 25 newly recruited samples. Results A total of 8 discriminating M/Z peaks related to PD were identified(P<0.05, nonparametric test, Z: -4.458~-3.059). The classification model based on SNN generated a separation between PD and healthy controls. The correct rate was 81.4% for training set, and was 70.0% for 25 newly recruited samples. Conclusion Protein expression in serum of PD patients is different from the normal controls. Serum proteomics data combined with bioinformatics approaches may contribute to the diagnosis of PD.

Establishment of a Novel Co-culture Model of Suprachiasmatic Nucleus Slices and NIH/3T3 Cells in a Serum-free Condition

ZUO Xiao-hong;CAI Yan-ning;LI Ning;YU Shun;ZHANG Yan-li;CHEN Biao

2009, 30(5): 601-606. doi:10.3969/j.issn.1006-7795.2009.05.007

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Objective To establish a novel co-cultures model of suprachiasmatic nucleus(SCN) slices and NIH/3T3 cells in a serum-free condition. Methods SCN or cortical slices prepared from 7-day-old male SD rats were cultured with NIH/3T3 cells for 6 d. 24 h expression profile of Per1 was examined in NIH/3T3 cells using real-time PCR. Results A significant daily oscillation of Per1 mRNA expression was observed in NIH/3T3 cells co-cultured with SCN slices. The peak time of Per1 was at CT11. No daily oscillation was found in NIH/3T3 cells induced with cortical slices. Conclusion A novel co-culture model of SCN slices and NIH/3T3 cells was established, which will facilitate the studies on the communications between SCN and peripheral tissues.

α-Synuclein Promotes Neurite Outgrowth of Primarily Cultured Rat Brain Neurons

LIU Guang-wei;WANG Peng;LI Yao-hua;LI Xin;HE Xin;YU Shun

2009, 30(5): 607-610. doi:10.3969/j.issn.1006-7795.2009.05.008

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Objective To investigate the effect of α-Synuclein(α-Syn) on the neurite outgrowth of brain neurons. Methods Neurons isolated from the neocortex of newborn rats were cultured and α-Syn was added into the culture medium to observe the effect of the protein on the neurite outgrowth of the neurons. In addition, the effect of different concentrations of α-Syn on the neurite outgrowth was studied. Results The mean neurite length of the neurons treated with α-Syn was significantly longer than that of control neurons at 1 h, 2 h and 4 h after adding α-Syn. The effect of α-Syn on neurite outgrowth was enhanced with the increase of α-Syn concentration and was reversed by anti-α-Syn monoclonal antibody. Conclusion α-Syn can promote neurite outgrowth of primary rat brain neurons in a dose-dependent manner.

T Lymphocytes from Copolymer-1 Immunized Mice Protect Dopaminergic Neurons in the MPTP Mouse Model of Parkinson's Disease

XU Sheng-li;XUAN Qi;ZHOU Ming

2009, 30(5): 611-615. doi:10.3969/j.issn.1006-7795.2009.05.009

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Objective To define the phenotype of Copolymer-1(Cop-1) antigen-specific lymphocytes to protect the dopaminergic neurons in 1-methy1-4-pheny1-1,2,3,6-tetrahydropyritine(MPTP)-intoxicated C57BL/6J mice. Methods The C57BL/6J mice were divided randomly into Cop-1/BSA antigen-specific lymphocyte adoptive transfer, Cop-1/BSA antigen-specific T lymphocyte adoptive transfer, Cop-1/BSA antigen-specific lymphocyte without T cells adoptive transfer, MPTP model control and normal control groups. All the mice except normal controls received four intraperitoneal injections at 2 hour intervals of MPTP(18 mg·kg^-1). After the last injection, the animals of transfer groups received adoptive transfers of different types of antigen-specific lymphocytes immediately. All mice were killed after 7 days of last MPTP injection. The dopaminergic neurons were analyzed quantitatively using immunohistochemistry of tyrosine hydroxylase(TH) and stereological counts. Results Stereological counts revealed that that MPTP caused a significant loss of TH-positive neurons in substantia nigra(SN) (37.78% of normal controls). Similar results were observed in MPTP-injected mice that received BSA-lymphocytes(38.15% of normal controls), BSA-T-lymphocytes(41.44% of normal controls), Cop-1/BSA lymphocytes without T cells transfer(Cop-1: 40.06% of normal controls; BSA: 34.81% of normal controls). In contrast, MPTP-injected mice that received Cop-1 lymphocytes or Cop-1-T-lymphocytes exhibited a much smaller reduction in the number of TH-positive neurons(74.38%, or 84.64% of normal controls). Conclusion Cop-1 antigen-specific T lymphocytes may protect dopaminergic neurons in SN of MPTP C57BL/6J mouse.

Molecular Epidemiologic Study on Human Immunodeficiency Virus Type 1 Infection in Partial Men Who have Sex with Men(MSM) in Beijing

LIU Zhi-ying;LI Hai-ying;ZHANG Tong;HUANG Xiao-jie;WEI Fei-li;XU Meng;JIAO Yan-mei;XU Xiao-ning;CHEN De-xi;WU Hao

2009, 30(5): 616-620. doi:10.3969/j.issn.1006-7795.2009.05.010

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Objective To investigate the molecular epidemiology of human immunodeficiency virus 1(HIV-1) infection among men who have sex with men(MSM) in Beijing and survey the up to date HIV-1 prevalence in this region. Methods Seventeen anti-HIV-1 positive samples of MSMs from 2007 2008 were collected, the genomic DNA were isolated directly from the whole blood, HIV-1 gag/env genes were amplified by nested PCR from the genomie of MSMs. The c2-v5 region of env gene and p17-p24 region of gag gene were sequenced for analyzing the subtype directly. Phylogenetic analyses were performed based on the nucleotide sequence data. Results There were three HIV-1 strains including CRF01-AE, B and CRF07-BC among seventeen MSMs in Beijing, genotyping showed that 52.94%(9/17) were CRF01-AE, 29.41%(5/17) were B and 17.65%(3/17) were CRF07-BC. Conclusion There are three HIV-1 subtypes among MSMs in Beijing from the small samples epidemiologic survey, and the CRF01-AE has replaced the B and became the most prevalent HIV-1 subtype in Beijing, and the CRF07-BC is increasing fast among MSMs in Beijing recently.

Investigation on Proliferation and Death of CD8⁺ T-cell and Its Subsets in Chronic HIV-1 Infected Persons

REN Yi;LIU Yi;LI Hai-ying;ZHANG Tong;WU Hao;CHEN Xin-yue

2009, 30(5): 621-625. doi:10.3969/j.issn.1006-7795.2009.05.011

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Objective To observe the changes associated with proliferation and death of CD8⁺ T cell and its subsets including naive CD8⁺ T-cell (T_N), central memory CD8⁺ T-cell (T_CM),effection memory CD8⁺ T-cell (T_EM) and effection memory CD8⁺ T-cell RA (T_EMRA) in chronic HIV-1 infected patients and healthy controls. Same observations were also carried out according to different CD4⁺ T-cell count in chronic HIV-1 infected patients. Methods Sixteen healthy controls and 23 untreated HIV-1 infected patients whose disease course was over 12 months were enrolled in observation. Flow cytometry tests were carried out after routine staining. CD8⁺ T-cells were divided into four subsets based on CD45RO and CD27 expression. They were T_N(CD45RO-CD27+), T_CM (CD45RO+CD27+), T_EM (CD45RO+CD27-) and T_EMRA(CD45RO- CD27-)respectively. Cell proliferation was studied by measuring expression of the Ki-67 antigen. Cell death was studied by annexin V staining. Cell proliferation and death ratio were compared between HIV-1 chronic infection group and healthy group. Same investigations were conducted among three HIV infected groups which were divided according to CD4⁺ T-cell count at 200/mm³ and 350/mm³ levels. Results The distribution of CD8⁺ T-cell subsets in normal control(n=16) were T_N 34.92%±12.68, T_CM 25.44%±10.36%, T_EM 14.64%±10.58%, and T_EMRA 25.00%±12.59%,respectively. The counterparts in HIV-1 infected group(n=23) were T_N 17.14%±8.03%(t=5.368, P=0.000), T_CM 31.40%±14.02%(t=-1.448, P=0.156), T_EM 20.17%±13.17%(t=-1.393, P=0.172), and T_EMRA 31.48%±15.16%(t=-1.405, P=0.168) respectively. The ratio of proliferating cells in CD8⁺ T-cells were 0.15%±0.09% in healthy control and 1.33%±0.90% in infection group(z=-4.655, P=0.000). The ratio of dying cells in CD8⁺ T-cells were 8.74%±4.73% in healthy control and 24.08%±13.72% in infection group(z=-4.169, P<0.001). There was a statistically significant difference in T_N subset among CD4≤200/mm³ group(n=5), CD4 200~350/mm³ group(n=12) and CD4>350/mm³ group(n=6). But there was no significant difference in proliferation and death ratios among these three groups. Conclusion 1 Compared with healthy control, death and proliferation ratios in CD8⁺ T-cells and its subsets increased dramatically. CD8⁺ T-cells were shifted into high flow-ratio cell cycling. 2 The composite ratio of CD8⁺ T_N subset was dramatically down regulated especially in the CD4<200/mm³ group. The resource of CD8⁺ T-cells T_N subset was exhausted following disease progress.

Analysis of Drug Resistance Results in 20 Cases of HIV/AIDS Patients

WEI Fei-li;WU Hao;ZHANG Tong;LI Qing;LIU Zhi-ying;WANG Rui;JIAO Yan-mei;CHEN De-xi

2009, 30(5): 626-630. doi:10.3969/j.issn.1006-7795.2009.05.012

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Objective To analyze drug resistance status in 20 HIV/AIDS patients who had received highly active antiretroviral therapy(HAART) with treatment failure and to evaluate the significance of drug resistance test. Methods Plasma viral load, CD4⁺ T lymphocyte counts were determined by Versant HIV-1 RNA 3.0 and TriTEST CD4 FITC/CD8 PE/CD3 PerCP Reagent, respectively. Using the Trugene HIV-1 Genotyping kit and OpenGene DNA Sequencing System, we sequenced HIV-1 target genes and analyzed drug-related mutations. We used molecular biology software(such as Blast) to identify HIV-1 subtypes. Results Of the 20 patients, 17 had pre-treatment plasma samples, and 1 case showed an non-nucleoside reverse transcriptase inhibitors(NNRTIs)-related resistance mutation before HAART. When antiviral treatment initiated, 3/20 cases with no drug resistanceǒrelated mutations were found at any time point; in 17/20 cases drug resistanceǒrelated mutation was detected. Thirteen of 17 patients showed NNRTI-related mutations, mostly in K103N, Y188L, Y181C, V106M and G190A. Eleven of 17 patients showed Nucleoside/Nucleotide Reverse Transcriptase Inhibitors(NRTIs)-related mutations, mostly in M184V, T/215Y/F, D67N, K70R and K219E/Q. Three of 17 patients were showed Protease Inhibitors(PIs)-related mutations, with primary mutation site at M46I/L and V82F/A. In secondary mutation of PI, M36I and L63P were the most common sites. Of the 20 patients, had type B infection, had 5(25%, 5/20) CRF01_AE recombinants another, had 5(25%, 5/20) CRF07_BC recombinants, had CRF06_CPX1 recombinants is 1(5%, 1/20). Conclusion For patients with treatment failure and drug-resistance, mutation detection can help clinicians determine the cause of treatment failure and optimize the medication combinations. Termination or change of treatment programs could significantly affect drug resistance test results, and therefore, clinicians should be fully aware of the treatment situation of patients before drug resistance test. The subtypes revealed that the HIV subtypes have an association with routes of infection.

Optimization of Alu-long Terminal Repeat Polymerase Chain Reaction Approach to Detect Integrated HIV-1

LI Juan;JIAO Yan-mei;HUA Wei;ZHANG Tong;WU Hao;GAO Yan-qing

2009, 30(5): 631-634. doi:10.3969/j.issn.1006-7795.2009.05.013

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Objective To optimize Alu-long terminal repeat(LTR) polymerase chain reaction(PCR) approach to detect integrated HIV-1. Methods The genomic DNA from purified CD4⁺T cells of 20 HIV-1 patients and 10 healthy control subjects was obtained. A pair of primers were designed according to the characteristics of human genome and HIV-1 genome. The first PCR was carried out by using the nested 5' primers from conserved sequences of human Alu and 3' primer from conserved HIV-1 long terminal repeat(LTR) sequences. The product of the first PCR was representative of both cellular DNA upstream of the integration site and integrated HIV-1 LTR. An aliquot of the diluted first PCR product was further subjected to the second round of PCR by using nested HIV-1 LTR-specific primers and then detect the DNA sequence from the product of Alu-LTR PCR. Results After optimized Alu-LTR PCR, integrated HIV-1 was detected in each of the 20 cases of HIV/AIDS. Conclusion After optimization, the sensitivity of Alu-LTR PCR for detection of integrated HIV-1 was markedly improved.

Progresses in the Mechanism of HIV-1 Latency Formation

JIAO Yan-mei;WU Hao

2009, 30(5): 639-642. doi:10.3969/j.issn.1006-7795.2009.05.015

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Although HIV-1 can replicate continuously in untreated patients, they can also be hidden intracellularly to evade the host immune clearance, that is latent virus. As the viral latency is a challenge to antiviral treatment, great concern has arisen. This article reviews literature on the mechanism of HIV-1 latency formation.

Experimental Studies on Hypoxic Preconditioning-induced Neuroprotection against Focal Ischemic Injury of Mice and Its p38 MAPK Mechanism

LIU Xu;FENG Su-juan;ZHANG Cai-yan;BU Xiang-ning;ZHANG Nan;YAN Rui;WANG Qian-yun;LI Jun-fa

2009, 30(5): 643-647. doi:10.3969/j.issn.1006-7795.2009.05.016

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Objective To observe the effect of hypoxic preconditioning(HPC) on middle cerebral artery occlusion(MCAO)-induced brain injury of mice and explore the change of p38 mitogen activated protein kinase(p38 MAPK) in neuroprotection of HPC. Methods Male BALB/c mice(18 g-22 g, 8 weeks-10 weeks) were randomly divided into H0 sham, H0, H4 sham and H4 groups. Using our unique hypoxic preconditioned mouse model and middle cerebral artery occlusion mouse model, combined with triphenyltetrazolium chloride(TTC) staining, neurological deficits evaluation, immunohistochemistry and Western blotting, the authors observed the changes of neurological score, brain infarct volume, neuronal injury, p38 MAPK phosphorylation and protein expression levels in the cerebral cortex of mice. Results HPC significantly decreased the neurological score, reduced infarction size, density of infarct area and edema ratio, and attenuated neuronal loss. Compared with the sham control, ischemia increased the phosphorylation levels of p38 MAPK significantly in penumbra of mouse brain. Furthermore, compared with the normoxic control, the phosphorylation levels of p38 MAPK increased more significantly in penumbra of HPC ischemic group. Conclusion p38 MAPK signal pathway might be involved in neuroprotective effect of HPC on focal ischemic brain of mice.

Effect of BDNF on Degeneration of Dopaminergic Neurons in the Substantia Nigra of a Model of Intra-ventricular Injection of Lipopolysaccharide in the Rat

ZHAO Yong-mei;LV Feng-yue;XU Qun-yuan

2009, 30(5): 648-652. doi:10.3969/j.issn.1006-7795.2009.05.017

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Objective To investigate neurotoxic effects of intra-ventricularly injected lipopolysaccharide(LPS) on dopaminergic(DA) neurons in the substantia nigra and to explore the role of brain-derived neurotrophic factor(BDNF) in the long-term degeneration of dopaminergic neurons in rats. Methods Altogether 48 healthy male SD rats were assigned into LPS-injected group and saline-injected group randomly. All injections were made intra-ventricularly on right side with LPS 20 μL(1.25 g/L)or saline 20 μL. The changes of dopaminergic neurons in the substantia nigra were studied by using tyrosine-hydroxylase(TH) immunohistochemical staining. The changes of BDNF were detected by immunohistochemical staining and enzyme immunoassay system(ELISA). Results The number of TH-positive neurons in the substantia nigra in LPS-injected rats decreased at 24 weeks after LPS injection. The number of TH-positive neurons in LPS-injected group was 75.4% of that in saline-injected group(P<0.01). The BDNF-positive cells in the substantia nigra in LPS-injected group increased markedly compared with saline-injected group at 4 weeks, while at 12 weeks and 24 weeks, the BDNF-positive cells in the substantia nigra in LPS-injected group decreased obviously compared with those in saline-injected group. The results of ELISA showed that BDNF protein levels in LPS-injected group were 186.3% of those of saline-injected group at 4 weeks(P<0.01). While at 12 weeks and 24 weeks post LPS injection, BDNF protein levels in the substantia nigra and corpus striatum of LPS-injected group were 42.3%(P<0.01) and 61.0%(P<0.05) of those in saline-injected group, respectively. Conclusion A single intra-ventricular administration of LPS in rats may result in a significant and a delayed loss of dopaminergic neurons in the substantia nigra. After LPS injection, a decrease of BDNF protein level in the substantia nigra occurs before the degeneration of dopaminergic neurons, suggesting that the decrease of BDNF may be one of the reasons that induced the loss of dopaminergic neurons induced by cerebral inflammation.

Experimental Study on the Effects of Glucocorticoids on Syndrome Evaluation in Rat Asthma Model at Different Stage of Intervention

LIU Ren-hui;ZHAO Hui;MU Yang;WANG Pei;YANG Jing;ZNANG Yi;LIU Su;WANG Xiu-juan

2009, 30(5): 653-657. doi:10.3969/j.issn.1006-7795.2009.05.018

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Objective To study the change of Syndrome of asthmatic rats at different treatment stage with glucocorticoid(GC) intervention(before withdrawal, during the withdrawal, and after withdrawal). Methods Ninety SD rats were randomized into 3 groups, normal control group, model control group, and GC group. Dexamethasone was injected into asthmatic model of rats and the syndrome was observed at different stage of GC intervention, including weight, armpit temperature, claw temperature and digital photos analysis of the color of claw and tail. Results Before the stage of withdrawing GC, the syndrome of GC group at first was consistent with the characteristics of Yin deficiency syndrome, and later not only had the Yin deficiency syndrome, but also had the trend of Yang deficiency syndrome. During the period of hormone withdrawal, and after the stage of hormone withdrawal, GC group was in line with the syndrome of Yang deficiency. Conclusion This study showed that GC group had the different syndrome characteristics at different stage of the treatment with dexamethasone.

Cluster Analysis on Carotid Sinus Baroreceptor Discharge

ZHAO Hai-yan;WEI Hua;LIU Ping;LIU Ting-hui;WANG Wei;HUANG Hai-xia;NIU Wei-zhen

2009, 30(5): 658-663. doi:10.3969/j.issn.1006-7795.2009.05.019

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Objective To investigate if it is possible to extract the information of single nerve fiber discharges from multiple fiber discharges of carotid sinus baroreceptors with Spike 2 software and to establish a detailed method. Methods To record the pressure in the carotid sinus and in vitro few fiber discharges of the sinus nerve simultaneously and then to sort all the action potentials by means of Spike 2 software. Results 1 It is necessary to define several template parameters according to the baroreceptors properties. 2 Reliable unit discharges could be extracted with Spike 2 program from the sinus nerve discharges with a high signal-to-noise ratio containing 2-3 units or more 5 units. 3 It should be prudent when analyzing the baroreceptor discharges containing much more units with this program. Conclusion Spike 2 software is suitable for cluster analysis on carotid sinus baroreceptor discharges containing units not more than 5. For an accurate sorting it is a prerequisite to obtain raw records with a fine signal-to-noise ratio and to set up the template parameters pertinently.

Change in Microenvironment for Nerve Regeneration in Rats with Streptozotocin-induced Experimental Diabetes Following Sciatic Nerve Crush Injury

ZHANG Zai-qiang;CAO Shi-jian;WANG Yong-jun

2009, 30(5): 664-670. doi:10.3969/j.issn.1006-7795.2009.05.020

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Objective To evaluate the significance of molecular microenvironment in neurons for nerve regeneration and repair by investigating the dynamic changes of nerve regrowth-associated proteins following sciatic nerves crush in rats with streptozotocin-induced experimental diabetes. Methods Bilateral sciatic nerve crush was performed 4 weeks after induction of streptozotocin-induced experimental diabetes. the mean percentage of TUNEL positive cells in dorsal root ganglion(DRG) following bilateral sciatic nerves crush for 7 days, 14 days, 21 days, and 28 days, were evaluated. Immunohistochemical staining and Western-blot techniques were used to investigate the expression of nerve growth associated protein(GAP-43) and nerve growth factor receptor(trk A) in different duration following sciatic nerve crush injury. Results The percentage of TUNEL positive neurons in DRG significantly increased in all stages after sciatic nerve crush, reaching a peak at 14~21 days. The expression of GAP-43 and trk A in DRG were upregulated at all time points after nerve injury in streptozotocin-induced experimental diabetes, but the overall level was lower than that of non-diabetic rats. Conclusion In streptozotocin-induced experimental diabetes, neurons in DRG undergo survival crisis, the nerve regrowth-associated gene expression system was disintegrated, the capacity to regenerate their axons declines after nerve injury.

Expression of Bactericidal/Permeability-Increasing Protein in Murine Tissues and Cells

GUO Ling;XU Qing;Lü Zhe;LIU Zhen-long;FAN Yi-qiang;WANG Wei;KONG Qing-li;AN Yun-qing

2009, 30(5): 671-676. doi:10.3969/j.issn.1006-7795.2009.05.021

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Objective The bactericidal/permeability-increasing(BPI) protein is an important protein expressed in some cells, which has bactericidal activity on gram-negative organisms. It is involved in the defense against gram-negative bacterial infections and binds to LPS with high affinity. Its expression profile in human tissues has been revealed by previous studies in our laboratory and other laboratories. But no reports about its expression profile in mouse, the most popularly used animal model were published. This study aimed to reveal the expression profile of BPI protein in various tissues from mice in physiological condition and by LPS stimulation. Methods BPI protein expression was detected with immunohistochemical staining using anti-muBPI polyclonal anti-serum. Pre-bleeding rabbit serum was served as negative control. Different tissues from mice treated with LPS(550 ng per mouse) and untreated mice were examined, including testis, small intestine, heart, liver, lung, spleen, kidney, bone marrow, peritoneal enterocoelia clysis(PECs) and bone marrow-derived dendritic cells(BMDCs). Results 1 Murine BPI protein is weakly expressed in testis, small intestine, kidney, bone marrow, PECs and BMDCs in control group(untreated with LPS), but its expression increased dramatically after LPS stimulation; 2 The expression of BPI protein could not be detected in liver and lung in control mice, but could be detected after LPS stimulation; 3 Expression of BPI protein could not be detected in the other examined tissues, no matter with or without LPS stimulation. Conclusion This study has successfully revealed the expression profile of BPI protein in some tissues from mice in physiological condition or by LPS stimulation. Our results showed that BPI protein expressed physiologically in some tissues and cells in mice, and its expression level could be increased or induced by LPS stimulation.

Effect of Low-intensity Pulsed Ultrasound on Schwann Cell Proliferation and Expression of Neurotrophin-3 Protein in Vitro

LI Jia-mou;ZHANG Hua;LIN Xin;WAN Hong;TIAN Bao-peng

2009, 30(5): 677-681. doi:10.3969/j.issn.1006-7795.2009.05.022

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Objective It is generally known that low-intensity pulsed ultrasound(LIPUS) accelerates peripheral nerve tissue regeneration. However, the precise cellular mechanism is still unclear. The purpose of this study was to determine how the Schwann cells respond directly to LIPUS stimuli. Methods Schwann cells were enzymatically isolated from postnatal 1-3 day rat sciatic nerve tissue and cultured in a 6-well plate. The ultrasound was applied at a frequency of 1 MHz and an intensity of 100mW/cm² spatial average temporal average(ISATA) for 5 min/day. The control group was cultured in the same way but without an administration of ultrasound. We investigated the effect of LIPUS on cell proliferation, neurotrophin-3(NT-3) and brain-derived neurotrophic factor(BDNF) mRNA expression by the methods of BrdU uptake, immunohistochemistry and RT-PCR. Results Immunohistochemistry demonstrated that more than 98% of the cultured cells were positive in S-100, NT-3 and BDNF. Stimulated cells also revealed an increased rate of cell proliferation by using 5-bromo-2'-deoxyuridine(BrdU) assay. The mRNA expression of NT-3 by RT-PCR assays was significantly up-regulated at day 14 after the LIPUS stimulation, whereas, the mRNA expression of BDNF was significantly down-regulated. Conclusion These results suggested that the application of LIPUS can promote cell proliferation and NT-3 gene expression, and down-regulates BDNF gene expression in Schwann cells. The signal-regulated mechanisms involved in this process remains unclear and requires further research.

A Murine Model of Cryptococcus Neoformans Infection and PCR Rapid Diagnosis

YI En-tong;YIN Cheng-hong;LI Min

2009, 30(5): 682-686. doi:10.3969/j.issn.1006-7795.2009.05.023

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Objective The authors established a cryptococcus neoformans-inoculated murine model which can be used to explore the diagnostic effect of cryptococcosis by PCR method. Methods The murine model for PCR diagnostic use was made by inoculating cryptococcus agents through tracheal intubation in BALB/c mice. PCR diagnosis was tested on DNA samples extracted from lung tissue of challenged mice. Results A pair of primers(ITS1-CN4) from eight pairs of candidate primers were screened out, which targeted the ribosomal DNA gene cluster and ITS1, ITS2 regions and could detect cryptococcus from murine tissue sensitively and specifically. Conclusion The murine model of cryptococcus neoformans infection were successfully established. One pair of primers(ITS1-CN4) could detect cryptococcus from murine tissue with high sensitivity and specificity.

Role of Adhesion Molecules in the Relationship between Obstructive Sleep Apnea-hypopnea Syndrome and Coronary Heart Disease

FAN Xin;Wei Xin;Du Feng-he

2009, 30(5): 687-690. doi:10.3969/j.issn.1006-7795.2009.05.024

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Objective To explore the role of adhesion molecules in the relationship between obstructive sleep apnea-hypopnea syndrome(OSAHS) and coronary heart disease(CHD). Methods The levels of serum soluble intercellular adhesion molecule-1(ICAM-1) and vascular cell adhesion molecule-1(VCAM-1) in 31 OSAHS patients with CHD, 31 OSAHS patients without CHD and 28 healthy controls were measured by enzymed-linked immunosorbent assay(ELISA). The relationship of the concentration of ICAM-1, VCAM-1 with the polysomnogram parameters was analyzed. Results The levels of serum ICAM-1 in OSAHS patients with CHD and OSAHS patients without CHD were significantly higher compared with the healthy controls(P<0.05). The level of serum VCAM-1 in OSAHS patients was significantly higher than that in the healthy controls(P<0.05), but the level of serum VCAM-1 did not significantly differ between OSAHS patients with CHD and the healthy controls(P>0.05). ICAM-1 in OSAHS patients positively correlated with apnea-hypopnea index(AHI)(r=0.544, P<0.01). ICAM-1 and VCAM-1 also showed significantly positive correlation with body mass index(BMI)(r=0.280, P<0.05). Conclusion The high levels of serum ICAM-1 is an important risk factor for the development of CHD in OSAHS patients.

Clinical Characteristics of the Elderly with Metabolic Syndrome

HONG Xu;MA Qing;CHEN Hai-ping;YU Heng-chi;YIN Jie;HAO Ya-bin

2009, 30(5): 691-695. doi:10.3969/j.issn.1006-7795.2009.05.025

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Objective To investigate clinical characteristics of the elderly with metabolic syndrome and type 2 diabetes and the influence of metabolic syndrome on coronary heart disease and chronic renal disease. Methods The study was conducted in 257 patients(age range 60-85 years) with type 2 diabetes mellitus. The patients were divided into two groups which were with or without MS according to the Adult Treatment Panel Ⅲ criteria:120 patients with MS and 137 patients without MS.The clinical features and morbidity of coronary heart disease, arteriosclerosis of the lower extremity, and cerebral infarction. Results Compared with patients without MS, BMI, HOMA-β, total cholesterol, triglyceride, uric acid and microalbuminuria of patients with MS were significantly higher(all P<0.01). Compared with patients without MS, more patients with MS suffered from CHD(P<0.01). Conclusion Obesity,dyslipidemia, insulin resistance, hyperuricacidemia, microalbuminuria and CHD were more common in the elderly with MS and type 2 diabetes.

Comparison between Propofol and Sevoflurane during Operation with Suspension Laryngoscopy

LI Li;YANG Wen-jing;LI Tian-zuo

2009, 30(5): 696-698. doi:10.3969/j.issn.1006-7795.2009.05.026

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Objective To compare propofol with sevoflurane with remifentanil target-controlled infusion(TCI) respectively during operation of suspension laryngoscopy. Methods Forty ASA Ⅰ-Ⅱ patients scheduled for the vocal cord surgery through suspension laryngoscopy were randomly divided into two groups: Propofol group(P group) and Sevoflurane group (S group), 20 cases in each. Both groups were induced with intravenous midazolam 1.0~1.5 mg, succinylcholine 1.5~2.0 mg/kg and remifentanil 4 μg/L in TCI. P group was induced with Propofol 3 μg/ml in TCI and S group with intravenous propofol 1.5~2.0 mg/kg. After intubation, patients in S group inhaled 2%~4% sevoflurane to get sevoflurane concentration to 1 MAC, and combined with remifentanil in TCI. Patients in P group were maintained with remifentanil and propofol in TCI. HR and MAP at different time point, waking time, extubation time and side effects were recorded and compared. Results In both groups after induction, the HR at T₁(1 min after induction) declined as compared with T₀(before operation)(P<0.05), and MAP at T₁ and T₂(1 min after intubation) declined as compared with T₀(P<0.05). But HR in P group at T₁, T₂ and T₇(operation finished) declined more significantly than that in S group(P<0.05). While waking time and extubation time in S group were shorter than in P group(P<0.05). Conclusion Both methods of anaethesia can achieve good effect in the suspension laryngoscopy, but the patients maintained with sevoflurane had more stable hemodynamics and recovered sooner.

Expression and Clinical Significance of Lung Resistance Protein in Breast Cancer

ZHAO Ning;ZHANG Zhong-tao;QU Xiang;WANG Yu

2009, 30(5): 699-702. doi:10.3969/j.issn.1006-7795.2009.05.027

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Objective To investigate the expression of lung resistance protein(LRP) genes in breast cancer and to explore the role of multi-drug resistance mechanism in breast cancer and its clinical significance. Methods Totally 81 female cases of breast cancer, who were treated in the department of general surgery of Beijing Friendship Hospital in 2000, aged 30~83 years (average 53.2 years), accepted the comprehensive treatment with surgery without neoadjuvant chemotherapy. Pathological types included invasive ductal carcinoma, invasive lobular carcinoma, the only cancer, mucus cancer and early invasive cancer. According to standards of International Union Against Cancer(UICC) 2003 version for clinicopathological staging of breast cancer, 17 cases were at stage Ⅰ, 43 cases stage Ⅱ, and 21 cases stage Ⅲ. Thirty-six cases had five-year follow-up results, 30 cases had disease-free survival for 5 years, 6 cases had metastasis(1 case survived with tumor and 5 cases died). Tissue chips were prepared. LRP expression in breast cancer tissue was determined by using immunohistochemistry and in situ hybridization, and other detection technology. Results According to manual reagents criteria of Maixin company, the results were separated into negative(-/+) and positive(++/+++). The positive rate of LRP in breast cancer was 70.4%(57/81). The 81 cases were divided into two groups: young and middle-aged group(age<50 years, 36 cases) and elderly group(≥50 years, 45 cases). It was found that positive correlation was not only found between LRP and ER but also between LRP and PR. The correlation coefficient were 0.327, 0.264, P＜0.05. But there was no correlation between LRP and p53, Her-2 and Ki-67, P> 0.05. Conclusion The expression of LRP increased in breast cancer evidently, there was correlation between the LRP expression and age, expression of ER and PR, which may provide basis for selecting the drug of chemotherapy.

CT Imaging Features of Specific Pulmonary Infection in Kidney Transplantation Patients

DING Yi;ZHANG Lei;QIAN Xiao-jun;ZHAI Ren-you;WANG Wei

2009, 30(5): 703-709. doi:10.3969/j.issn.1006-7795.2009.05.028

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Objective To investigate the CT findings in specific pulmonary infection in kidney transplantation patients. And thereby to improve the knowledge of CT imaging features of these disease. Methods A retrospective analysis was carried out in 49 patients with specific pulmonary infection after kidney transplantation. There were 31 males and 18 females, the age ranged from 21 to 60 years(mean 42 years). The time duration from renal transplantation to onset of illness was 12 days to 7 years. Forty-nine patients underwent plain CT and high resolution(HR) CT scans of the chest. In helical CT mode at 1∶ 1 pitch, by using 5 mm beam collimation. HRCT mode used 1mm or 0.625 mm beam collimation, the interval was 10mm.The spectrum of infections included invasive pulmonary aspergillosis in 29 patients, pneumocystis carinii pneumonia in 12 patients, cytomegalovirus pneumonia in 4 patients and pulmonary tuberculosis in 4 patients. Final diagnosis was obtained by sputum culture in 19 patients, bronchoalveolar lavage(BAL) and transbronchial biopsy in 20 patients, blood culture in 8 patients, and on autopsy 2 patients. The findings on chest CT were interpreted by three radiologists. Results The 49 patients showed the typical findings on chest CT. Twenty-two of 29 invasive pulmonary aspergillosis on CT typically showed multiple nodules, 17 of 22 typically showed nodules surrounded by a halo of ground-glass haze(halo sign), 15 of 22 showed peribronchovascular nodules, 23 of 29 showed ground-glass opacity, 18 of 29 showed consolidation; 11 of 29 showed wedge-shaped consolidation. Twelve cases with pneumocystis carinii pneumonia typically showed diffuse ground-glass opacity on CT, 4 of cytomegalovirus pneumonia cases typically showed interstitial pneumonia on CT, ground-glass opacity with thick interlobular septum, 3 of 4 showed ground-glass opacity with small multiple nodules; 2 of 4 showed pleural fluid. CT of 4 cases with pulmonary tuberculosis showed miliary shadows, 1 of 4 showed tree-in-bud sign. Conclusion There are some characteristics in the CT films of patients with different specific pulmonary infection after kidney transplantation. CT findings helped in promp diagnosis of the underlying diseases in these patients.

Progresses in Studies on Immune Reconstitution Inflammatory Syndrome in HIV/AIDS Patients

WU Wei;HUANG Shi-jing;WANG Jie

2009, 30(5): 710-713. doi:10.3969/j.issn.1006-7795.2009.05.029

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Immune reconstitution inflammatory syndrome (IRIS) occurred in the process of immune reconstitution disease after highly active antiretroviral therapy. In this paper, the concept, diagnostic criteria, clinical manifestations and treatment of IRIS were reviewed. It is important to improve diagnosis and differentiation of the IRIS and reduce delays. One of the characteristics of immune reconstitution is restoration of immune response to opportunistic infections . Therefore, unless there is a threat to patient's life or a risk that may lead to a very high rate of disability, the highly active antiretroviral therapy should be continued.

Discovery and Significance of Telomere and Telomerase——Introduction of Nobel Prize in Physiology or Medicine 2009

ZHANG Ting;Wang Xiao-min

2009, 30(5): 718-722. doi:10.3969/j.issn.1006-7795.2009.05.031

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This year’s Nobel Prize in Physiology or Medicine is awarded to three American scientists who have solved a major problem in biology: how the chromosomes can be copied in a complete way during cell divisions and how they are protected against degradation. Elizabeth H. Blackburn and Jack W. Szostak discovered that a unique DNA sequence in the telomeres protects the chromosomes from degradation. Carol W. Greider and Elizabeth H. Blackburn identified telomerase, the enzyme that makes telomere DNA. These discoveries made great contributions to promote the investigations on aging and tumor.

What You See Is What You Learn, What You Learn Guides What You Do——Introduction to the Contribution of the Awardees of Nobel Prize in Chemistry 2009

LU Zhao;DING Wei

2009, 30(5): 723-726. doi:10.3969/j.issn.1006-7795.2009.05.032

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"What you see is what you learn" is believed to be the essence of the structural biology research. In the 2009 Nobel Prize in Chemistry awarded work, three outstanding scientists from the United States and Israel have vividly demonstrated that how high-resolution crystallography not only helped to solve long-standing problems in the structural-functional studies for the roles of ribosomes in the process of protein biosynthesis, but also revealed the underlying mechanisms. Their contributions have provided far-reaching implications for basic biological and medical sciences, and meanwhile inspired and expanded the important applications for related drug development, such as the designs of antibiotics.

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