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目的 探讨先天性心脏病(CHD)患儿肺动脉高压(PH)形成的影响因素.方法 研究对象均为2003-06-2005-02于北京大学深圳医院收集病例,以健康者30名为对照组(A组),以肺动脉压正常和并发PH的左向右分流的CHD患儿各30例为观察组(B组、C组).以高效液相色谱法、硝酸还原法及放射免疫法测定其血清精氨酸(L-Arg)、一氧化氮(NO)、血浆内皮素(ET-1)的浓度.结果 血清L-Arg浓度对照组(A组)为(72.00±18.01)nmol/mL,肺动脉压正常的患儿(B组)为(30.74±8.97)nmol/mL,伴PH的患儿(C组)为(23.51±12.37)nmol/mL.血清NO浓度A组为(76.10±17.10)nmol/mL,B组(90.55±26.57)nmol/mL,C组(60.05±17.60)nmol/mL.血浆ET-1浓度A组(50.82±7.58)pg/mL,B组(64.90±16.28)pg/mL,C组(69.64±10.66)pg/mL.结论 血清NO浓度和血浆ET-1浓度及其之间的平衡关系共同影响PH的形成及其程度.血浆ET-1浓度的升高是肺动脉压升高的直接因素,血清NO浓度的降低是间接因素,而血清NO浓度降低是由血清L-Arg浓度的降低引起.     

Immune responses against SARS-coronavirus nucleocapsid protein induced by DNA vaccine

The nucleocapsid （N） protein of SARS-coronavirus （SARS-CoV） is the key protein for the formation of the helical nucleocapsid during virion assembly. This protein is believed to be more conserved than other proteins of the virus, such as spike and membrane glycoprotein. In this study, the N protein of SARS-CoV was expressed in Escherichia coli DHSalpha and identified with pooled sera from patients in the convalescence phase of SARS. A plasmid pCI-N, encoding the full-length N gene of SARS-CoV, was constructed. Expression of the N protein was observed in COS1 cells following transfection with pCI-N. The immune responses induced by intramuscular immunization with pCI-N were evaluated in a murine model. Serum anti-N immunoglobutins and splenocytes proliferative responses against N protein were observed in immunized BALB/c mice. The major immunoglobulin G subclass recognizing N protein was immunoglobulin G2a, and stimulated splenocytes secreted high levels of gamma interferon and IL-2 in response to N protein. More importantly, the immunized mice produced strong delayed-type hypersensitivity （DTH） and CD^8＋ CTL responses to N protein.     

过氧化氢对大鼠离体肺组织精细切片活性的影响

目的研究过氧化氢(H2O2)对离体肺组织精细切片活性的影响,建立肺组织氧化损伤的简易模型.方法制备离体肺组织精细切片,以0.5 mL Krebs-Henseleit缓冲液作为孵育液,在37℃培养箱内进行孵育.在孵育液中加人H2O2,使其浓度为2 mmol/L,分别孵育0(对照组)、5、10、15和20 min;然后在不含H2O2的孵育液中进行恢复孵育1 h.测定肺片的四甲基偶氮唑盐(MTT)染色、孵育液中乳酸脱氢酶(LDH)和肺片组织ATP含量.结果随H2O2损伤时间延长,MTT染色变浅,其吸光度逐渐减小,损伤时间大于10 min组与对照组比较有显著差异(P＜0.01).H2 O2损伤10 min组肺片的LDH释放多于对照组(P＜0.05),损伤时间更长者增加更明显(P＜0.01).H2O2损伤5 min组的肺片组织中ATP含量明显低于对照组(P＜0.05),损伤时间更长者下降更明显(P＜0.01).经偏相关分析发现,肺片MTT染色与LDH释放呈负相关(r=-0.866,P＜0.01),与ATP含量呈正相关(r=0.869,P＜0.01).结论离体肺组织切片在含H2O2的孵育液中进行孵育时,随孵育时间延长其活性降低.此模型简单可靠,可用于肺组织氧化损伤的研究.     

关于802.1X认证技术在校园网应用中问题的探讨

介绍了802.1X认证技术的体系结构和工作原理,分析了802.1X认证技术在校园网中应用的优势与不足.     

CNAL组织的水溶液pH值测定的能力验证

pH值的测量是许多实验室大量开展的检测项目之一,在污水处理、水质环境、监测、食品安全、医疗卫生、药品监督和化学化工等行业得到广泛应用。为了评价本实验室pH值测定的测试能力和测试水平,通过实验室检测能力的外部措施来补充实验的内部质量控制程序,本实验室参加了由中国实验室国家认可委员会(CNAL)组织的水溶液pH值测定的能力验证[1]。1测试样品本实验室收到A和B两种测试样品,均为缓冲溶液,为A样和B样。A样为邻苯二甲酸氢钾水溶液,B样为混合磷酸盐水溶液,共有4瓶100 ml装满待测缓冲溶液的塑料瓶。A样分为A1和A2(错层式设记),pH…     

Metabolic activation of fluoropyrrolidine dipeptidyl peptidase-IV inhibitors by rat liver microsomes.

The current study evaluated the potential for two dipeptidyl peptidase-IV (DPP-IV) inhibitor analogs (1S)-1-(trans-4-([(4-trifluoromethoxyphenyl)sulfonyl]amino)cyclohexyl)-2-[(3S)-3-fluoropyrrolidin-1-yl]-2-oxoethanaminium chloride and (1S)-1-(trans-4-([(2,4-difluorophenyl)sulfonyl]amino)cyclohexyl)-2-[(3S)-3-fluoropyrrolidin-1-yl]-2-oxoethanaminium chloride (MRL-A and MRL-B), containing a fluoropyrrolidine moiety in the structure, to undergo metabolic activation. The irreversible binding of these tritium-labeled compounds to rat liver microsomal protein was time- and NADPH-dependent and was attenuated by the addition of reduced glutathione (GSH) or N-acetylcysteine (NAC) to the incubation, indicating that chemically reactive intermediates were formed and trapped by these nucleophiles. Mass spectrometric analyses and further trapping experiments with semicarbazide indicated that the fluoropyrrolidine ring had undergone sequential oxidation and defluorination events resulting in the formation of GSH or NAC conjugates of the pyrrolidine moiety. The bioactivation of MRL-A was catalyzed primarily by rat recombinant CYP3A1 and CYP3A2. Pretreatment of rats with prototypic CYP3A1 and 3A2 inducers (pregnenolone-16alpha-carbonitrile and dexamethasone) enhanced the extent of bioactivation which, in turn, led to a higher degree of in vitro irreversible binding to microsomal proteins (5- and 9-fold increase, respectively). Herein, we describe studies that demonstrate that the fluoropyrrolidine ring is prone to metabolic activation and that GSH or NAC can trap the reactive intermediates to form adducts that provide insight into the mechanisms of bioactivation.     

Enhancement of manumycin A-induced apoptosis by methoxyamine in myeloid leukemia cells.

Farnesyltransferase inhibitors (FTIs) are currently under investigation for leukemia treatment. We evaluated the FTI manumycin A (manumycin) in two myeloid leukemia cell lines (U937 and HL-60). Manumycin induced nitric oxide production and apoptosis of the leukemia cells. Nitric oxide or other reactive oxygen species may induce oxidative DNA damage, and the number of apurinic sites increased after manumycin treatment, which was reversed by concurrent treatment with N-acetyl-L-cysteine. Since repair of DNA damage is important to cell survival, we hypothesized that methoxyamine, an inhibitor of base-excision repair, would enhance the antineoplastic effect of manumycin. The combination of manumycin and methoxyamine resulted in enhanced apoptosis by six criteria increased annexin V binding, release of mitochondrial cytochrome c into the cytosol, activation of caspase-9, activation of caspase-3, specific cleavage of poly-adenosyl ribose polymerase, and increase in the sub-G1 cell cycle fraction. The drug combination enhanced inhibition on the soft agar clonogenic assay and on the formazan dye cell viability assay. The effects of manumycin or manumycin plus methoxyamine on apoptosis were blocked by N-acetyl-L-cysteine, and partially by nitric oxide synthase inhibitors or scavenger of peroxide. We conclude that methoxyamine enhances manumycin-induced apoptosis in myeloid leukemia cells.     

The association of XRCC1 haplotypes and chromosomal damage levels in peripheral blood lymphocyte among coke-oven workers.

Theoretically, a haplotype has a higher level of heterozygosity than individual single nucleotide polymorphism (SNP) and the association study based on the haplotype may have an increased power for detecting disease associations compared with SNP-based analysis. In this study, we investigated the effects of four haplotype-tagging SNPs (htSNP) and the inferred haplotype pairs of the X-ray cross-complementing group 1 (XRCC1) gene on chromosome damage detected by the cytokinesis-block micronucleus assay. The study included 141 coke-oven workers with exposure to a high level of polycyclic aromatic hydrocarbons and 66 nonexposed controls. The frequencies of total MN and MNed cells were borderline associated with the Arg(194)Trp polymorphism (P = 0.053 and P = 0.050, respectively) but not associated with the Arg(280)His, Arg(399)Gln and Gln(632)Gln polymorphisms among coke-oven workers. Five haplotypes, including CGGG, TGGG, CAGG, CGAG, and CGGA, were inferred based on the four htSNPs of XRCC1 gene. The haplotype CGGG was associated with the decreased frequencies of total MN and MNed cells, and the haplotypes TGGG and CGAG were associated with the increased frequencies of total MN and MNed cells with adjustment for covariates among coke-oven workers. This study showed that the haplotypes derived from htSNPs in the XRCC1 gene were more likely than single SNPs to correlate with the polycyclic aromatic hydrocarbon-induced chromosome damage among coke-oven workers.     

黄芪注射液对刀豆蛋白诱导的小鼠肝脏损伤的保护作用

目的：观察免疫损伤模型组与实验组小鼠外周血中T细胞亚群的变化情况，探讨黄芪注射液对小鼠肝脏损伤的保护作用。方法实验组小鼠每天腹腔注射黄芪注射液400μl，正常组与模型组小鼠每天给予等剂量的生理盐水。第7天在给药1小时后，模型组与实验组小鼠分别经尾静脉注射刀豆蛋白，正常组小鼠则给予等剂量的生理盐水。8小时后检测各组动物外周血中CD4^＋T、CD8^＋T细胞以及CD4^＋／CD25^＋T细胞所占比例，检测血浆转氨酶水平，制备肝脏病理标本，进行HE染色。结果尾静脉注射8小时后，实验组小鼠外周血的CD4^＋所占T细胞比例较模型组有所升高（P〈0．05），但仍低于正常组水平（P〉0．05）：而CD8^＋T细胞的比例则无明显变化（P〉0．05）。实验组ALT、AST水平明显低于模型组（P〈0．05）。实验组汇管区淋巴细胞浸润明显减少。结论黄芪注射液对刀豆蛋白诱导的小鼠肝损伤有确切的保护作用。