检测外周血特异性结核分枝杆菌抗体对活动性结核病的诊断价值研究

目的研究外周血浆细胞分泌特异性结核分枝杆菌抗体对活动性结核病的诊断价值。方法纳入2013年4月至7月期间深圳市第三人民医院肺科门诊活动性结核病患者;健康对照组人员来自体检科门诊志愿者,分为3个组：活动性结核病组（104例）、结核分枝杆菌（Mtb）潜伏感染组（26例）和健康对照组（33名）。分离其外周血单个核细胞（PBMCs）。体外培养4d后收集培养上清液,用ELISA法和蛋白免疫印迹法,ELISA和蛋白免疫印迹法检测外周血浆细胞分泌的特异性结核分枝杆菌抗体;检测各组收集的PBMCs培养上清液中的特异性结核分枝杆菌抗体,比较各组间的差异。所有数据均使用GraphPadPrism5．0进行统计学分析,用受试者工作特征（ROC）曲线评价ELISA法检测的诊断价值,得出敏感度、特异度;多组间的比较采用Kruskal—Wallis检验;两组间比较采用Dunnmultiplecomparison检验;蛋白免疫印迹法用四格表的卡方检验;以P〈0．05为差异有统计学意义。结果ELISA法检测活动性结核病组特异性结核分枝杆菌抗体的A450nm值（0．593±0．206）高于Mtb潜伏感染组（0．342±0．152）和健康对照组（0．246±0．121）,差异有统计学意义（H=77．27,P〈0．001）。特异性结核分枝杆菌抗体区分活动性结核病患者和Mtb潜伏感染者、健康对照者曲线下面积（AUC）分别为0．857、0．944,当诊断界限值为0．42时,其区分活动性结核病和Mtb潜伏感染的敏感度为77．9％（81／104）,特异度为80．8％（21／26）;区分活动性结核病和健康人的敏感度为77．9％（81／104）,特异度为93．9％（31／33）。蛋白免疫印迹法检测活动性结核病和健康人的敏感度、特异度和准确性分别为79．2％（61／77）、100．0％（11／11）、81．8％（72／88）（x2=24．8,P〈0．001）。结论ELISA法和蛋白免疫印迹法检测外周血浆细胞分泌特异性结核分枝杆菌抗体诊断活动性结核病特异度高,可以作为临床结核病实验诊断的辅助方法。     

Acute sleep deprivation in healthy young men: Impact on population diversity and function of circulating neutrophils

Lack of sleep greatly affects our immune system. The present study investigates the acute effects of total sleep deprivation on blood neutrophils, the most abundant immune cell in our circulation and the first cell type recruited to sites of infection. Thus, the population diversity and function of circulating neutrophils were compared in healthy young men following one night of total sleep deprivation (TSD) or after 8 h regular sleep. We found that neutrophil counts were elevated after nocturnal wakefulness (2.0 ± 0.2 × 10⁹/l vs. 2.6 ± 0.2 × 10⁹/l, sleep vs. TSD, respectively) and the population contained more immature CD16^dim/CD62L^bright cells (0.11 ± 0.040 × 10⁹/l [5.5 ± 1.1%] vs. 0.26 ± 0.020 × 10⁹/l [9.9 ± 1.4%]). As the rise in numbers of circulating mature CD16^bright/CD62L^bright neutrophils was less pronounced, the fraction of this subpopulation showed a significant decrease (1.8 ± 0.15 × 10⁹/l [88 ± 1.8%] vs. 2.1 ± 0.12 × 10⁹/l [82 ± 2.8%]). The surface expression of receptors regulating mobilization of neutrophils from bone marrow was decreased (CXCR4 and CD49d on immature neutrophils; CXCR2 on mature neutrophils). The receptor CXCR2 is also involved in the production of reactive oxygen species (ROS), and in line with this, total neutrophils produced less ROS. In addition, following sleep loss, circulating neutrophils exhibited enhanced surface levels of CD11b, which indicates enhanced granular fusion and concomitant protein translocation to the membrane. Our findings demonstrate that sleep loss exerts significant effects on population diversity and function of circulating neutrophils in healthy men. To which extent these changes could explain as to why people with poor sleep patterns are more susceptible to infections warrants further investigation.     

COVID-19患者实验室指标变化的观察研究

目的总结新型冠状病毒肺炎(corona virus disease 2019,COVID-19)患者治疗前后实验室指标变化趋势。方法选择2020年1-2月宝鸡市中心医院感染科确诊的13例COVID-19患者为研究对象,回顾分析实验室指标在治疗前后的变化情况。结果治疗前后白细胞总数(WBC)、血小板(PLT)、C反应蛋白(CRP)、红细胞沉降率(ESR)、肌酸激酶(CK)、α羟丁酸脱氢酶(α-HBDH)、白细胞介素6(IL-6)、淀粉样蛋白A(SAA)、肌红蛋白(Mb)水平存在明显差异(P<0.05或P<0.01)。WBC、淋巴细胞计数(LYMP)、PLT、CRP、ESR、CK、α-HBDH、直接胆红素(DBIL)、IL-6、SAA、便潜血试验异常率在治疗前后存在显著差异(P<0.05或P<0.01)。结论WBC、PLT、CRP、ESR、CK、α-HBDH、IL-6、SAA、Mb指标变化可用来评估COVID-19患者治疗的有效性,可作为疾病预后的评估指标;而WBC、LYMP、PLT、CRP、ESR、CK、α-HBDH、DBIL、IL-6、SAA、便潜血试验可作为其早期诊断及辅助诊断指标。     

Intermittent fasting during Ramadan attenuates proinflammatory cytokines and immune cells in healthy subjects

Intermittent fasting and caloric restriction have been shown to extend life expectancy and reduce inflammation and cancer promotion in animal models. It was hypothesized that intermittent prolonged fasting practiced during the month of Ramadan (RIF) could positively affect the inflammatory state. To investigate this hypothesis, a cross-sectional study was designed to investigate the impact of RIF on selected inflammatory cytokines and immune biomarkers in healthy subjects. Fifty (21 men and 29 women) healthy volunteers who practiced Ramadan fasting were recruited for the investigation of circulating proinflammatory cytokines (interleukin [IL]-1β, IL-6, and tumor necrosis factor α), immune cells (total leukocytes, monocytes, granulocytes, and lymphocytes), and anthropometric and dietary assessments. The investigations were conducted 1 week before Ramadan fasting, at the end of the third week of Ramadan, and 1 month after the cessation of Ramadan month. The proinflammatory cytokines IL-1β, IL-6, and tumor necrosis factor α; systolic and diastolic blood pressures; body weight; and body fat percentage were significantly lower (P < .05) during Ramadan as compared with before Ramadan or after the cessation of Ramadan fasting. Immune cells significantly decreased during Ramadan but still remained within the reference ranges. These results indicate that RIF attenuates inflammatory status of the body by suppressing proinflammatory cytokine expression and decreasing body fat and circulating levels of leukocytes.     

窒息新生儿外周血白细胞计数与血小板参数的动态变化分析

目的 :探讨窒息新生儿外周血白细胞计数与血小板参数的动态变化及其临床意义。方法 :将 5 8例窒息新生儿分为轻度窒息组 ( 2 0例 )和重度窒息组 ( 38例 ) ;另外选择 2 0例正常新生儿作为对照组 ,检测各组外周血白细胞计数、血小板参数 ,比较各组间白细胞数及血小板参数新生儿窒息程度的关系。结果 :外周血白细胞数及血小板计数 ( PC)、血小板平均体积 ( MPV)、血小板分布宽度 ( PDW) ,在窒息组和对照组及重度窒息组和轻度窒息组比较 ,外周血 WBC、MPV、PDW均显著升高 ( P<0 .0 1 ) ,血小板数显著减少 ( P<0 .0 5 )。窒息组治疗后与正常组比较 ,外周血白细胞数、血小板参数无显著性差异 ( P均 >0 .0 5 )。结论 :外周血白细胞数、血小板参数变化与新生儿窒息程度密切相关 ,在临床上可作为窒息后多脏器损伤的一个预警参数。     

多药耐药基因转染脐血单个核细胞及其表达与功能的体外研究

目的：为克服白血病化疗中的骨髓抑制，采用含人多药耐药（MDR1）基因全长cDNA逆转录病毒载体转染入脐血单个核细胞，探索一种MDR1基因导入脐血造血细胞稳定、有效的方法，评价MDR1基因转染脐血细胞及其功能的表达，为体内转染MDR1基因保护骨髓免受大剂量化疗药物损伤提供条件。方法：采用含有MDR1基因表达质粒pHaMDR1/A的包装细胞PA317传代培养产病毒法，通过浓缩病毒上清液将MDR1基因导入人脐血单个核细胞，应用聚合酶链反应（PCR）方法、免疫组化法、流式细胞学等方法从基因、蛋白质、功能及细胞生物学特性等不同水平检测MDR1基因在脐血单个核细胞的表达、转染率及其与转染时间的关系和功能的表达；转染行为是否影响脐血细胞生物学特性。结果：（1）建立了一种安全可行、稳定高效的MDR1基因体外转染脐血造血细胞的体系和方法；（2）PCR方法证实外源性MDR1基因可有效性地体外整合到脐血单个核细胞中；（3）免疫组化法测得2日、4日、6日转染率分别为18.0%、29.8％、34.7%;(4)柔红霉素排出试验证实导入的MDR1基因表达产物P-gp有正常的生物学功能；（5）转染后的脐血细胞周期生物学特异性无异常。结论：MDR1基因体外转染脐血造血细胞是安全可行的，且在体外有稳定、有效的表达。这一方法为进一步在化疗骨髓保护的体内实验提供了有利的依据。     

乙型肝炎病毒宫内感染对新生儿外周血细胞因子干扰素γ和白介素4的影响

目的 探讨乙型肝炎病毒(HBV)侵犯新生儿外周血单个核细胞(PBMC)后,对新生儿免疫功能的影响,了解其免疫失败发生的机理.方法 聚合酶链反应法(PCR)检测67对乙型肝炎表面抗原(HBsAg)、乙型肝炎e抗原(HBeAg)阳性孕妇及其新生儿血清和PBMC的HBV DNA.根据新生儿PBMC中HBV DNA分成阴性和阳性组,将新生儿的PBMC分别在植物血凝素(PHA)和纯化HBsAg刺激下进行体外细胞培养,检测培养上清液中干扰素-γ(IFN-γ)、白介素-4(IL-=4)的分泌含量.结果 (1)35例母亲PBMC HBV DNA阳性者其新生儿15例为阳性,母亲与患儿PBMC HBV DNA阳性,差异有显著统计学意义(P＜0.01).(2)新生儿PBMC内HBV DNA阳性组与阴性组比较,纯化HBsAg刺激时,阳性组IFN-γ的分泌量较阴性组低(P＜0.01),IL-4含量显示PBMC HBV DNA阳性组高于阴性组(P＜0.05),PHA刺激时,两组IFN-γ、IL-4含量差异无统计学意义(P＞0.05).结论 PBMC内的HBV DNA可能是HBV母婴垂直传播的一条重要途径;宫内新生儿PBMC感染HBV,IFN-γ特异性反应低下,而IL-4特异性反应增强,细胞调节失衡,可能是新生儿免疫失败和易于免疫耐受的一个重要原因.     

Formyl-methionyl-leucyl-phenylalanine and a calcium ionophore A23187 reverse the inhibition of phorbol myristate acetate-induced oxidative burst by linoleic and oleic acid anilides

Linoleic and oleic acid anilides profoundly inhibited the production of reactive oxygen metabolites (ROM) in human polymorphonuclear leukocytes (PMNL) induced by a tumor promoter, phorbol myristate acetate (PMA). The addition of a Ca²⁺ ionophore, A23187, or a chemotactic peptide, formyl-methionyl-leucyl-phenylalanine (fMLP), readily reversed linoleic and oleic acid anilide-induced inhibition of PMA-evoked respiratory burst in PMNL without affecting PMA-induced respiratory burst. fMLP or A23187 caused a marked increase in the production of ROM in PMNL that did not produce ROM after their co-exposure to PMA and cis-fatty acid anilides. This suggests a role for Ca²⁺ in this restoration of respiratory burst activity in PMNL. Oleic and linoleic acid anilides enhanced also respiratory burst in PMNL subsequent to their stimulation with fMLP. Interestingly, corresponding fatty acids, linoleic and oleic acid, also inhibited PMA-induced production of ROM in PMNL, but this inhibition was not reversed by A23187 or fMLP. These findings suggest that the aniline moiety of cis-fatty acids significantly modifies the effects of linoleic and oleic acids in the production of ROM in PMNL. Moreover, free intracellular Ca²⁺ may play a critical role in the activation of PMNL to produce ROM, and in the modulation of the effects of cis-fatty acid anilides.     

激素敏感型单纯性肾病外周血白细胞乙酰肝素酶基因表达的研究

目的　研究激素敏感型单纯性肾病 (SRSNS)外周血白细胞中乙酰肝素酶 (Hpa)基因表达水平及其与尿蛋白定量之间的关系 ,探讨肾小球基底膜电荷屏障破坏的机制。方法　采用逆转录 聚合酶链反应的方法 ,分别对 4 3例SRSNS患儿 (活动期 2 3例、恢复期 10例、缓解期 10例 )、2 3例肾炎性肾病、15例紫癜性肾炎 (肾病型 )和 15例正常对照儿童外周血白细胞中Hpa基因表达水平进行半定量研究。结果　 (1)SRSNS活动期Hpa表达量 (1 2 7± 0 36 )高于肾炎性肾病、紫癜性肾炎 (肾病型 )和正常对照组 (0 36± 0 13) ,差异有显著意义 (P <0 0 0 1)。 (2 )不同阶段的SRSNS患者之间Hpa表达量活动期 >恢复期 >缓解期 ,差异有显著意义 (P <0 0 5 ) ;缓解期 (0 34± 0 18)与正常对照组之间差异无显著意义。 (3)SRSNS组活动期、恢复期和缓解期Hpa表达水平与尿蛋白定量之间呈正相关 (rs=0 75 1,P <0 0 0 1)。结论　外周血白细胞Hpa表达的增高可能与SRSNS大量蛋白尿的产生有关。