急性髓系白血病患者外周血单个核细胞Toll样受体9以及血清肿瘤坏死因子和凋亡基因FAS的表达

背景：Toll样受体9（TLR9）、肿瘤坏死因子a、Fas可能共同参与白血病的发生发展过程。目的：测定TLR9在急性髓系白血病患者外周血单个核细胞中的表达及血清肿瘤坏死因子α、Fas水平。方法：从急性髓系白血病患者与正常对照组中分离出外周血单个核细胞,采用反转录-聚合酶链反应法检测外周血单个核细胞中TLR9mRNA的表达水平,采用酶联免疫吸附试验法检测血清肿瘤坏死因子α、Fas水平。结果与结论：在急性髓系白血病患者初治组和难治复发组中,外周血单个核细胞中TLR9mRNA表达高于正常对照组（P〈0.01）,化疗后完全缓解组与正常对照组差异无显著性意义（P〉0.05）;各病例组血清肿瘤坏死因子α、Fas水平显著高于正常对照组（P〈0.01）。TLR9mRNA的表达与血清肿瘤坏死因子α、Fas水平均呈正相关。     

急性白血病骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D的表达(英文)

背景:据作者查新检索,国内外有关急性白血病骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D活性及mRNA表达与白血病类型、患者肝脾淋巴结肿大关系的报道罕见.目的:探讨糖基化磷脂酰肌醇特异性磷脂酶D的表达与急性白血病类型、急性髓系白血病患者肝脾淋巴结肿大的关系.方法:急性髓系白血病组骨髓标本43例,急性淋巴细胞白血病组骨髓标本28例,以21例健康志愿者作为正常对照组.密度梯度离心法获得骨髓单个核细胞,用GPI锚定的胎盘碱性磷酸酶作为底物,Triton-X114分相法检测糖基化磷脂酰肌醇特异性磷脂酶D的活性,半定量RT-PCR法检测糖基化磷脂酰肌醇特异性磷脂酶D mRNA的表达,并分析糖基化磷脂酰肌醇特异性磷脂酶D活性和mRNA表达水平与急性白血病类型、急性髓系白血病患者肝脾淋巴结肿大的关系.结果与结论:与正常对照组比较,急性髓系白血病组骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D活性和mRNA表达均明显升高(P < 0.01);急性淋巴细胞白血病组骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D活性和mRNA表达均明显降低(P < 0.01).在43例急性髓系白血病患者中,13例患者检查发现有肝脾淋巴结肿大,30例患者检查无肝脾淋巴结肿大,无肝脾淋巴结肿大标本骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D活性和mRNA表达均明显高于有肝脾淋巴结肿大标本(P < 0.05).结果证实急性白血病骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D的活性与mRNA变化一致,其表达水平与急性白血病类型及急性髓系白血病患者有无肝脾和/或淋巴结肿大有关.     

急性髓细胞性白血病患者外周血单个核细胞CD28mRNA的表达

背景:大量研究显示,肿瘤患者外周血T细胞表面共刺激分子CD28蛋白表达存在差异,提示共刺激通路异常可能与恶性肿瘤的发生进展有关.目的:观察急性髓细胞性白血病外周血单个核细胞共刺激信号分子CD28 mRNA在中的表达.方法:急性髓细胞性白血病患者80例,其中M0型7例,M1型6例,M2型18例,M3型15例,M4型17例,M5型9例,M6型8例.并根据急性白血病疗效标准将80例患者分为完全治愈组、缓解组、未缓解组.采用Taqman探针实时荧光定量PCR检测80例患者及76名健康人群外周血单个核细胞CD28 mRNA的表达.结果与结论:急性髓细胞性白血病外周血单个核细胞M1,M3和M4亚型中的CD28 mRNA表达量低于健康人群 (P < 0.05);急性髓细胞性白血病未缓解组中CD28 mRNA低于健康人群 (P < 0.05),完全治愈组和缓解组中CD28 mRNA表达与健康人群差异无显著性意义.说明急性髓细胞白血病患者外周血单个核细胞存在CD28 mRNA表达缺陷,并与临床分期、病情进展及预后有关.     

乙型肝炎肝硬化患者外周血中单个核细胞Toll样受体4的表达及其与血清内毒素的关系

目的:通过检测乙型肝炎肝硬化患者外周血单个核细胞(PMBCs)Toll样受体4(TLR4) mRNA的表达、血清内毒素水平及肿瘤坏死因子α(TNF-α);探索TLR4 mRNA表达与血清内毒素及TNF-α的关系.方法:抽取29例乙型肝炎肝硬化患者外周静脉血,分离单个核细胞,用RT-PCR半定量分析TLR4 mRNA水平;同时用鲎试剂测定血清内毒素水平和用放免法测定血清TNF-α水平.结果:正常对照组和肝硬化患者血清内毒素水平分别为0.050±0.020 EUmL-1和0.107±0.080 EUmL-1(P<0.05),血清TNF-α水平分别为1.282 pgmL-1±0.249 pgmL-1和2.076±0.358 pgmL-1(P<0.05),后者均明显升高,而且肝功能损害越严重内毒素水平升高越明显;肝硬化患者PMBCs中TLR4 mRNA的表达(0.900±0.263)与正常对照(0.923±0.461)无显著差异(P>0.05).结论:在肝硬化患者,尤其主要是Child-Pugh C级患者,血清内毒素及TNF-α水平较正常对照组明显升高,但PMBC的TLR4 mRNA的表达水平无变化,这可能与肝硬化患者同时存在内毒素耐受有关.     

急性髓系白血病骨髓单个核细胞糖基化磷脂酰肌醇特异性磷脂酶D mRNA的表达与意义

本研究探讨糖基化磷脂酰肌醇特异性磷脂酶D(glycosylphosphatidilinoditol-specific phospholipase D,GPI-PLD)在急性髓系白血病病人骨髓单个核细胞中的表达及其意义。采用半定量RT-PCR和酶转化底物的百分率的方法检测急性髓系白血病初发组、难治复发组、缓解组与正常对照组骨髓单个核细胞GPI-PLD酶活性和mRNA表达,并比较它们之间的差异。结果显示:急性髓系白血病初治组骨髓单个核细胞GPI-PLD mRNA表达和GPI-PLD活性分别为1.86±0.32、(46.96±7.15)%;缓解组分别为1.26±0.29和(33.36±5.13)%;难治复发组分别为1.79±0.19和(44.31±7.22)%;而正常对照组分别为1.27±0.23和(35.38±5.15)%。初治组和难治复发组的GPI-PLD活性与mRNA表达均明显高于正常对照组,差异具有统计学意义(p0.01);而缓解组与正常对照组之间比较,差异无统计学意义(p0.05)。结论:GPI-PLD活性和mRNA表达水平变化一致;初治组和难治复发组急性髓系白血病病人骨髓单个核细胞GPI-PLD活性与mRNA表达水平明显高于正常组。GPI-PLD是否在白血病的发生、发展过程中发挥一定作用,有待进一步研究。     

急性髓细胞性白血病患者外周血单个核细胞CD28 mRNA的表达

背景：大量研究显示,肿瘤患者外周血T细胞表面共刺激分子CD28蛋白表达存在差异,提示共刺激通路异常可能与恶性肿瘤的发生进展有关。目的：观察急性髓细胞性白血病外周血单个核细胞共刺激信号分子CD28 mRNA在中的表达。方法：急性髓细胞性白血病患者80例,其中M0型7例,M1型6例,M2型18例,M3型15例,M4型17例,M5型9例,M6型8例。并根据急性白血病疗效标准将80例患者分为完全治愈组、缓解组、未缓解组。采用Taqman探针实时荧光定量PCR检测80例患者及76名健康人群外周血单个核细胞CD28 mRNA的表达。结果与结论：急性髓细胞性白血病外周血单个核细胞M1,M3和M4亚型中的CD28 mRNA表达量低于健康人群（P〈0.05）;急性髓细胞性白血病未缓解组中CD28 mRNA低于健康人群（P〈0.05）,完全治愈组和缓解组中CD28 mRNA表达与健康人群差异无显著性意义。说明急性髓细胞白血病患者外周血单个核细胞存在CD28 mRNA表达缺陷,并与临床分期、病情进展及预后有关。     

慢性阻塞性肺病外周血单个核细胞表面TLR2、TLR4的表达与血清TNF-α、IL-6、IL-8相关性的研究

目的:探讨慢性阻塞性肺病(chronic obstructive pulmonary disease,COPD)患者外周血单个核细胞表面Toll样受体(Toll-like receptors)TLR2和TLR4的表达情况及其与血清肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)的关系。方法:本研究涉及的3组研究对象分别为:吸烟COPD组(n=68)、吸烟非COPD组(n=42)和不吸烟非COPD组(n=43)。将68例吸烟COPD患者进一步分为急性发作期亚组(n=32)和稳定期亚组(n=36)。采用实时荧光定量PCR法检测外周血单个核细胞表面TLR2、TLR4的表达水平,同时采用酶联免疫吸附试验(ELISA)试剂盒测定血清细胞因子的浓度。结果:吸烟COPD组外周血单个核细胞表面TLR2、TLR4的表达水平及血清TNF-α、IL-6、IL-8的浓度均显著高于吸烟非COPD组(TLR2:1.87±2.04比0.47±0.24;TLR4:4.08±3.84比0.41±0.18;TNF-α:68.20±56.26比26.20±5.43;IL-6:201.62±174.69比72.99±8.03;IL-8:235.42±158.22比77.50±8.38)。吸烟非COPD组外周血单个核细胞上TLR2、TLR4的表达水平均显著低于不吸烟非COPD组(TLR2:0.47±0.24比2.96±2.71;TLR4:0.41±0.18比2.67±1.74),而吸烟非COPD组血清TNF-α、IL-6、IL-8的浓度均显著高于不吸烟非COPD组(TNF-α:26.60±5.43比8.93±6.41;IL-6:72.99±8.03比25.61±8.63;IL-8:77.50±8.38比47.10±11.11)。COPD急性发作期单个核细胞表面TLR2、TLR4的表达水平及血清TNF-α、IL-6、IL-8的浓度均显著高于COPD稳定期(TLR2:3.56±1.80比0.368±0.222;TLR4:6.97±3.76比1.51±0.943;TNF-α:106.17±61.94比90.01±19.66;IL-6:327.18±183.56比105.36±13.45;IL-8:381.74±108.51比34.44±10.45)。吸烟COPD组外周血单个核细胞表面TLR2、TLR4的表达水平与血清TNF-α、IL-6、IL-8的浓度成正相关。结论:COPD患者尤其是COPD急性发作期患者外周血单个核细胞表面TLR2、TLR4的表达水平及血清TNF-α、IL-6、IL-8的浓度均显著升高;并且COPD患者外周血单个核细胞表面TLR2、TLR4的表达水平与血清TNF-α、IL-6、IL-8的浓度成正相关。提示COPD的发生、发展与机体天然免疫密切相关。     

急性髓系白血病中uPA和uPAR的表达及意义

目的观察急性髓系白血病人骨髓单个核细胞uPAR（CD87）的表达及血浆uPA的水平,并探讨其临床意义。方法用ELISA法测定22例急性髓系白血病患者（观察组）及34例良性血液病患者（对照组）血浆中uPA表达水平,用流式细胞学技术测定骨髓单个核细胞uPAR水平,比较两组之间的表达差异。结果血浆中uPA水平和骨髓单个核细胞uPAR水平急性髓系白血病组均高于良性血液病组,差异具有显著性（P〈0.05）;pearson相关分析显示uPA表达水平及uPAR水平之间存在正相关性（观察组r1=0.614,对照组r2=0.921;P〈0.05）。结论急性髓系白血病患者血浆uPA水平及骨髓单个核细胞uPAR表达均升高,二者具有协同表达关系,联合检测对于急性髓系白血病的病情监控和预后分析有临床指导意义。     

肿瘤坏死因子相关凋亡诱导配体TRAIL及其受体在急性髓系白血病细胞中的表达及意义

为了检测肿瘤坏死因子相关凋亡诱导配体(TRAIL)及其受体在急性髓系白血病细胞中的表达，并探讨其在白血病治疗中的意义，采用RT-PCR方法及流式细胞术，对39例急性髓系白血病细胞(患者组)、18例完全缓解白血病细胞(缓解组)和21例正常人骨髓或外周血单个核细胞(对照组)表面TRAIL及其受体的表达进行检测。结果表明：①患者组和缓解组TRAIL、DR4和DR5表达高，而DcR1和DcR2表达低。②缓解组DR5的表达高于患者组。③患者组和缓解组DR5的表达均高于DR4。④患者组AML不同亚型表达TRAIL及其受体相似。结论：TRAIL及其受体在急性髓系白血病细胞中的表达具有明显的差异性：DR5在TRAIL介导的急性髓系白血病细胞凋亡中起重要的作用。     

同种异体骨髓单个核细胞移植急性心肌梗死大鼠:心肌细胞凋亡及相关基因的表达

背景:细胞凋亡增加所导致的进行性心肌细胞丢失是引起急性心肌梗死后充血性心力衰竭的主要原因之一,通过降低肿瘤坏死因子α浓度及Bax蛋白、PDCD5 mRNA表达可以抑制急性心肌梗死后心肌细胞凋亡.目的:探讨同种异体骨髓单个核细胞移植对急性心肌梗死大鼠心肌细胞凋亡及相关基因表达的影响.设计、时间及地点:随机对照动物实验,于2006-01/11在华中科技大学附属协和医院完成.材料:体质量为(240±20)g的Wistar大鼠60只,随机分为假手术组、模型对照组、细胞移植组,20只/组.另取体质量为100～150 g的Wistar大鼠60只用于制备骨髓单个核细胞.方法:模型对照组、细胞移植组大鼠结扎冠状动脉左前降支,建立急性心肌梗死模型;假手术组大鼠仅开胸,不结扎冠状动脉左前降支.造模后,细胞移植组大鼠在梗死心肌周围分4点于心外膜下植入骨髓单个核细胞悬液,每点100 μL(107个细胞);假手术组、模型对照组同法注射等量DMEM培养基,培养4周.主要观察指标:血清及非梗死区心肌组织中肿瘤坏死因子α的水平,左室非梗死区心肌细胞凋亡指数、Bax蛋白和PDCD5mRNA的表达.结果:移植后4周,细胞移植组在发生凝固坏死的宿主心肌内可以看到局灶BrdU标记阳性的骨髓单个核细胞.与假手术组比较,移植后4周模型对照组、细胞移植组血清和心肌组织中肿瘤坏死因子α水平均显著升高(P<0.05),但细胞移植组升高幅度明显小于模型对照组(P<0.05).与假手术组比较,模型对照组和细胞移植组左心室非梗死区心肌细胞凋亡指数、Bax蛋白和PDCD5 mRNA(PDCD5/GAPDH)均明显升高(P<0.05),但细胞移植组升高幅度明显小于模型对照组(P<0.05).结论:同种异体骨髓单个核细胞移植可以抑制大鼠心肌梗死后心肌细胞凋亡的发生,其机制可能与其抑制肿瘤坏死因子α的表达、降低Bax蛋白和PDCD5基因的促凋亡作用有关.     

Measurement of L-carnitine and acylcarnitines in body fluids and tissues in children and in adults

We have developed a reliable and validated radio-enzymatic method for the assay of L-carnitine and acylcarnitines, using a modification of existing methods. The sensitivity of the assay is 10 mumol/l using 10 microliters of plasma or urine. It is also suitable for measurements of carnitine in a 10 mg sample of liver or muscle obtained by percutaneous biopsy. The use of N-ethylmaleimide in the reaction mixture together with an excess of [1-14C]acetyl CoA ensures that the reaction proceeds to completion and a linear response is obtained. Using this method control ranges have been established for plasma and urine carnitine concentrations in healthy children and adults, and for the carnitine content of liver and muscle in adults. No significant difference was found between fasting and post-prandial plasma carnitine levels. An age-related increase was found in urinary total carnitine and acylcarnitine concentration throughout childhood. These data provide a reliable basis for studies of patients with abnormal carnitine and acylcarnitine metabolism, distribution and excretion.     

Rosamicin in Urethral and Vaginal Secretions and Tissues in Dogs and Rats

In animal studies we investigated the distribution of rosamicin in plasma and urethral and vaginal tissues in rats as well as in urethral and vaginal secretions in dogs. We found concentration ratios between urethral secretion and plasma of 1.9 and between vaginal secretion and plasma of 2.4. The rosamicin concentrations in urethral and vaginal tissue significantly exceeded the levels of all other tissues investigated. Because rosamicin could be valuable for the treatment of bacterial urethritis and the colonization of the vaginal introitus with fecal bacteria in women, it should be investigated clinically in this respect.     

Similarities and differences in clients treated and in medications prescribed by APRNs and psychiatrists in a CMHC

This study is part of a larger study comparing prescribing practices of psychiatrists and advanced practice psychiatric nurses (APRNs) using the following three groups of patients: patients treated by psychiatrists, those treated by APRNs, and those treated by both APRNs and psychiatrists at different times in 1 year. Demographics for 5507 patients were examined. A subsample of APRNs and psychiatrists prescribed similar total numbers of medications. Psychiatrists prescribed more types of antidepressant medications other than the SSRI antidepressants, and they prescribed more than twice the number of benzodiazepines. APRNs prescribed more SSRIs and spent more time with clients during medication visits.     

Racial differences in oxidative stress and inflammation: in vitro and in vivo

African American race is an independent risk factor for enhanced oxidative stress and inflammation. We sought to examine whether oxidative-stress and inflammatory markers that are typically measured in humans also differ by race in cell culture. We compared levels between African American and Caucasian young adults and then separately in human umbilical vein endothelial cells (HUVECs) from both races. We found heightened oxidative stress and inflammation in the African Americans both in vitro and in vivo. African American HUVECs showed higher nitric oxide (NO) levels (10.8 ± 0.4 vs. 8.8 ± 0.7 μmol/L/mg, p = 0.03), Interleukin-6 (IL-6) levels (61.7 ± 4.2 vs. 23.9 ± 9.0 pg/mg, p = 0.02), and lower superoxide dismutase activity (15.6 ± 3.3 vs. 25.4 ± 2.8 U/mg, p = 0.04), and also higher protein expression (p < 0.05) of NADPH oxidase subunit p47phox, isoforms NOX2 and NOX4, endothelial nitric oxide synthase (NOS), inducible NOS, as well as IL-6. African American adults had higher plasma protein carbonyls (1.1 ± 0.1 vs. 0.8 ± 0.1 nmol/mg, p = 0.01) and antioxidant capacity (2.3 ± 0.2 vs. 1.1 ± 0.3 mM, p = 0.01). These preliminary translational data demonstrate a racial difference in HUVECs much like that in humans, but should be interpreted with caution given its preliminary nature. It is known that racial differences exist in how humans respond to development and progression of disease, therefore these data suggest that ethnicity of cell model may be important to consider with in vitro clinical research.     

Postural strategies and falls in elderly and in parkinsonism]

OBJECTIVE: To use a posture analysis to show the evolution of postural pattern connected with falls.MATERIAL AND METHOD: It is a prospective study on two groups of 16 persons of more than 60 years. A group concerns 16 small disability off drug parkinsonian patients, a group concerns 16 healthy witnesses. All the persons benefited from a posture recording by means of a force platform and were followed during 1 year. RESULTS: Data analysis underlines three groups of persons corresponding to three postural patterns, independently of the presence of Parkinson disease. A group (n = 18) did not contain fallers, the second (n = 10 ) contained 20% of fallers, the third (n = 4) contained 100% of fallers. Differences between the groups were identified on 16 posturographic parameters. DISCUSSION: A group has a good functional value and one does not record any fall. Its characteristics, which correspond to a category of persons who compensate well for the phenomena of ageing, are found in the literature. A group has an intermediate functional value and regrets 20% of fallers. Kinetic profile reveals a tendency to the stiffness of the posture. This group is going to operate rather ankle strategies. A group has an inferior functional value and regrets 100% of fallers. Kinetic profile seems disrupted and not to be able to adapt itself in a satisfactory way to the situation otherwise than by stereotypical reactions. This group is going to operate systematically much less stabilizing hip strategies. CONCLUSION: A close determinism between physiological neuromotor ageing and Parkinson disease does exist. We showed with a prospective follow-up, the arisen of fall and showed the evolution of postural patterns related to fall. It appears as well that evolution mainly follows three stages leading from a small risk of fall gait pattern to a major risk of fall gait pattern.