儿童系统性红斑狼疮患者外周血淋巴细胞亚群与疾病活动的研究

目的探讨儿童系统性红斑狼疮(SLE)患儿外周血中淋巴细胞亚群的变化及临床意义。方法采用流式细胞术检测59例SLE患儿及52名健康对照组患儿的外周血淋巴细胞亚群T细胞、B细胞、Th细胞、Tc细胞、NK细胞和总淋巴细胞数,并对结果进行比较,同时分析SLE患儿外周血淋巴细胞亚群与SLE疾病活动指数(SLEDAI)的相关性及其与狼疮肾炎、补体的相关关系。结果与健康对照组相比,SLE患儿Th细胞和Th百分比、NK细胞和NK细胞百分比及Th/Tc比值明显降低,而B细胞百分比、Tc细胞百分比增高,差异均有统计学意义(P<0.05);SLE患儿B细胞与SLEDAI评分呈正相关,NK细胞百分比与SLE评分呈负相关(P<0.05);狼疮肾炎组患儿的T细胞、Tc细胞计数和Tc细胞百分比均明显高于无狼疮肾炎的患儿,而狼疮肾炎组患儿Th细胞百分比、Th/Tc比值和NK细胞、NK细胞百分比明显低于无狼疮肾炎的患儿(P<0.05);补体降低组患儿的B细胞和B细胞百分比及SLEDAI评分均明显高于补体正常组(P<0.05)。结论 SLE患儿外周血淋巴细胞亚群存在异常,淋巴细胞亚群与SLEDAI评分、狼疮肾炎和补体之间存在显著的相关性,表明淋巴细胞亚群可能反映SLE患儿病情的严重程度。     

系统性红斑狼疮患者外周血B淋巴细胞亚群与病情活动的相关性

目的探讨系统性红斑狼疮(SLE)患者外周血B淋巴细胞亚群的变化及其与病情活动的相关性。方法用流式细胞仪检测了27例SLE患者及16名正常对照人群的外周血B淋巴细胞亚群。SLE活动性的判断采用SLEDAI评分方法,其中缓解期的患者11例,活动期的患者16例。分析以上SLE患者外周血B淋巴细胞亚群变化及其与病情活动的相关性。结果SLE患者外周血B1淋巴细胞亚群的百分比、B1淋巴细胞亚群/B2淋巴细胞亚群的比值显著高于正常对照,但未见其与病情活动相关。结论SLE患者外周血B1淋巴细胞亚群存在异常,可能在SLE的发病中具有重要作用。     

T细胞亚群与NK细胞在难治性多发性骨髓瘤患者表达水平的研究

目的:研究多发性骨髓瘤患者外周血T淋巴细胞亚群与自然杀伤(NK)细胞的表达水平,探讨其细胞免疫功能与临床疗效的关系,并为判断、预测容易转归为难治患者提供具有参考价值的指标。方法:采用流式细胞术检测64例初治骨髓瘤患者化疗前及化疗中和全疗程结束后外周血T淋巴细胞亚群与NK细胞的表达水平。按化疗结果分完全缓解组、部分缓解组和难治组,与健康对照组比较。结果:1完全缓解组Th细胞、Ts细胞和NK细胞与对照组的差别无统计学意义。2部分缓解组Th细胞下降与对照组的差别有统计学意义。3难治组Th细胞、Ts细胞和NK细胞均呈明显下降,与对照组相比差别明显。4在最终死亡的三个患者,NK细胞百分比均在3%以下。结论:治疗后Th细胞、Ts细胞和NK细胞如果下降不明显,则提示该患者预后相对乐观,如果上述指标均明显下降,则提示预后不良。对于NK细胞严重降低的患者,提示预后极差。     

T淋巴细胞亚群检测对SLE的临床意义

目的探讨SLE患者T淋巴细胞亚群水平及其对SLE活动评估提供依据。方法采用流式细胞仪检测SLE活动期组患者30例,稳定期组患者30例及正常对照组人群30例外周血T淋巴细胞亚群水平,并对T淋巴细胞亚群的变化情况进行统计分析。结果与正常对照组相比,SLE活动期组及稳定期组患者CD4＋细胞的百分率均明显降低（P〈0.01）,CD8＋细胞的百分率均明显增高（P〈0.01）,CD4＋/CD8＋比值也明显降低（P〈0.01）。SLE活动期组与稳定期组比较,CD4＋细胞的百分率稍低于稳定期组（P〈0.05）,CD8＋细胞的百分率明显高于稳定期组（P〈0.01）,CD4＋/CD8＋明显低于稳定期组（P〈0.01）。结论 SLE患者的T淋巴细胞表达是异常的,而且活动期和稳定期患者的表达水平有所不同,外周血T淋巴细胞亚群水平的检测,对SLE病情判断和临床治疗和观察是有一定价值的参考指标。     

系统性红斑狼疮患者淋巴细胞增殖活性分析

目的探讨系统性红斑狼疮(SLE)患者外周血单核细胞(PBMC)中T细胞仁嗜银蛋白(AgNORs)和T细胞亚群及辅助性T细胞亚群(Th)的关系及其临床意义,指导临床诊断与治疗。方法PBMC经多克隆T细胞活化剂刺激,以图像分析法检测AgNORs区面积与细胞核仁面积的百分比(I.S%);以微量细胞毒法检测T淋巴细胞亚群水平;以ELISA法检测血清中干扰素(IEN)-γ、白细胞介素(IL)-4水平来评价辅助性T细胞亚群活性。结果①13例稳定期和17例活动期SLE患者PBMC中活化T细胞AgNORs表达分别为(5.14±1.06)I.S%和(3.21±1.40)I.S%,与正常对照组(6.69±1.30)I.S%相比,均显著下降(P<0.05,P<0.01);活动期显著高于稳定期(P<0.01);②与稳定组和正常对照组相比,活动期SLE患者CD4-T细胞计数(23.5±7.8%)和CD4/CD8比值(0.58±0.31)显著降低(P<0.01),CD8计灵敏(37.4±12.1%)明显升高(P<0.05);③与稳定组和正常对照组相比,活动期SLE患者血清IL-4(37.2±2.7pg/ml)水平明显升高(P<0.05),IFN-γ(17.9±2.7pg/ml)分泌较正常对照组明显降低(P<0.05),与稳定期患者无显著差异(P>0.05)。结论①SLE患者细胞免疫活性低于正常水平;②活动期患者血清IL-4分泌水平增高,INF-γ水平降低,提示Th1类活性降低,在SLE中存在Th1/Th2的不平衡。     

系统性红斑狼疮患者T细胞亚群的变化

目的:探讨T细胞亚群在系统性红斑狼疮(SLE)发病机制中的作用及其能否作为SLE病情评估和指导治疗的参考指标.方法:收集SLE高活动期患者40例,低活动期患者26例,稳定期患者15例,健康查体者30例.采用流式细胞术检测外周血样本的CD4+、CD8+细胞百分率及CD4+/CD8+比值;记录SLE患者的临床表现、疾病活动性指数(SLEDAI)评分、一般实验室检查以及血清免疫学指标,并对所得数据进行比较和相关性分析.结果:SLE患者不同活动期之间及和正常对照组之间外周血T细胞亚群存在差异.高活动期SLE患者CD4+/CD8+比值与SLDAI评分及尿蛋白负相关,与补体C3呈正相关;高活动期SLE患者经糖皮质激素和免疫抑制剂等治疗2周后,SLEDAI评分下降,T细胞亚群较治疗前无明显变化.结论:T细胞亚群SLE的发病机制中具有一定作用,并与疾病的严重程度有关,但不能作为短期内治疗是否有效的参考指标.     

系统性红斑狼疮患者P-糖蛋白与淋巴细胞亚群的相关性研究

目的探讨多药耐药蛋白P-糖蛋白(P-gp)的表达水平与淋巴细胞亚群计数在系统性红斑狼疮(SLE)患者中的关系及在预测疾病耐药性方面可能的机制。方法选择初发SLE患者20例和健康对照组15名。通过流式细胞术测定外周血淋巴细胞P-gp的表达水平和淋巴细胞亚群的计数。分析SLE患者外周血淋巴细胞P-gp的表达水平与淋巴细胞亚群中CD4+T细胞,CD8+T细胞、自然杀伤细胞、总T细胞和总B细胞等的相关性。采用t检验和χ2检验、Pearson相关性分析进行统计学分析。结果①健康对照组与SLE患者外周血淋巴细胞的相对荧光强度(RFI)平均值分别为2.2±0.7、3.3±2.0,差异有统计学意义(t=18.8,P<0.01)。②SLE患者外周血淋巴细胞P-gp的表达与淋巴细胞亚群中总T细胞,总B细胞,NK细胞的计数均无相关性,但P-gp的表达水平与CD4+T细胞、CD4+/CD8+比值有相关性(r=0.602,P=0.002;r=0.561,P=0.005)。③根据SLE患者外周血淋巴细胞亚群计数分为增高组和降低组,分别与相应的RFI值比较,发现CD4+T细胞及CD4+/CD8+比值的增高组与相应P-gp的表达呈正相关(r=0.713,P=0.025;r=0.628,P=0.014)。结论外周血淋巴细胞的P-gp表达是SLE患者多药耐药性的指标,SLE患者药物跨膜转运蛋白P-gp表达水平尚与其存在免疫功能异常亢进淋巴细胞亚群中CD4+T细胞及CD4+/CD8+比值密切联系,提示P-gp可能是药物治疗疗效的指标,对改善预后和联合治疗减少耐药有重要的意义。     

系统性红斑狼疮患儿外周血淋巴细胞亚群变化

目的：探讨系统性红斑狼疮（SLE）患儿外周血淋巴细胞亚群变化特征。方法选择24例SLE病儿和20例健康儿童为研究对象,应用流式细胞术（FCM）检测外周血的淋巴细胞亚群,对比分析两组检测结果。结果SLE患儿外周血CD4＋CD25＋T细胞、自然杀伤（NK）细胞百分率较对照组明显降低（t=2.480,3.773;P＜0.05）,CD8＋T细胞、活化T细胞和B细胞百分率明显升高（t=3.470,4.779,2.480;P＜0.05或P＜0.01）。结论SLE患儿外周血淋巴细胞亚群紊乱,调节性T细胞和NK细胞减少、B细胞增多和T细胞活化增强,在SLE发病机制中可能起重要作用。     

B细胞淋巴瘤患者外周血T淋巴细胞亚群的检测及与Ki67的关系

目的:了解B细胞淋巴瘤患者外周血T淋巴细胞亚群、NK细胞的表达以及与Ki67的关系。方法:选择B细胞淋巴瘤患者32例,男18例,女14例,年龄36~62岁。应用流式细胞仪(FCM)检测总T细胞,Ts,Th,NK细胞绝对值计数,并计算出Th/Ts值,对各种数据进行比较。同时用免疫速率比浊法测定血清中免疫球蛋白。据Ki67表达阳性率分为低表达组和高表达组,了解Ki67与免疫功能的关系。结果:非霍奇金淋巴瘤(NHL)患者Th明显低于正常对照组,P<0.05;Ts低于正常对照组,P>0.05;总T细胞明显低于正常对照组,P<0.01;Th/Ts,NK均明显低于正常对照组,P<0.01。Ki67高表达组Th,总T细胞,Th/Ts,NK细胞明显低于正常对照组,P<0.05;Ts明显高于正常对照组,P<0.05;Ki67低表达组Th,总T细胞,Th/Ts,NK细胞低于正常对照组,P<0.01;Ts高于正常对照组,但差异无统计学意义(P>0.05)。结论:B细胞淋巴瘤患者存在细胞免疫抑制与紊乱,且基础免疫低下。Ki67表达越高,免疫功能越低;反之,免疫功能越强。     

皮肌炎合并间质性肺疾病与外周血淋巴细胞亚群的相关性研究

目的探讨皮肌炎(DM)患者外周血淋巴细胞亚群计数与其合并间质性肺疾病(ILD)的关系。方法选择皮肌炎患者59例,用流式细胞仪检测外周血淋巴细胞标记分子的绝对表达情况,分析皮肌炎合并间质性肺疾病的患者与外周血淋巴细胞亚群计数,如T淋巴细胞(CD3+),CD4+T细胞(即辅助性T细胞,Th),CD8+T细胞(即抑制性/细胞毒性T细胞,Ts/CTL),B淋巴细胞(CD3-/CD19+)、自然杀伤(NK)细胞(CD3-/CD16+CD56+)及淋巴细胞总数(T+B+NK)计数及CD4+/CD8+比值的相关性。结果胸部高分辨率计算机断层扫描(HRCT)显示ILD的皮肌炎患者CD4+T细胞数目(652±394)高于未合并ILD患者(428±331),且差异有统计学意义(P<0.05)。皮肌炎患者CD4+T细胞数目升高与间质性肺疾病呈正相关(r=0.269,P<0.05)。结论 CD4+T细胞异常增高是皮肌炎患者合并间质性肺疾病的危险因素,对评估皮肌炎预后亦有指导意义。     

Synthesis,Cytotoxicity and Antileishmanial Activity of Some N-(2-(indol-3-yl)ethyl)-7-chloroquinolin-4-amines

We report herein the condensation of 4,7-dichloroquinoline (1) with tryptamine (2) and D-tryptophan methyl ester (3) . Hydrolysis of the methyl ester adduct (5) yielded the free acid (6) . The compounds were evaluated in vitro for activity against four different species of Leishmania promastigote forms and for cytotoxic activity against Kb and Vero cells. Compound (5) showed good activity against the Leishmania species tested, while all three compounds displayed moderate activity in both Kb and Vero cells.     

Lung disease and PKCs

The lung offers a rich opportunity for development of therapeutic strategies focused on isozymes of protein kinase C (PKCs). PKCs are important in many cellular responses in the lung, and existing therapies for pulmonary disorders are inadequate. The lung poses unique challenges as it interfaces with air and blood, contains a pulmonary and systemic circulation, and consists of many cell types. Key structures are bronchial and pulmonary vessels, branching airways, and distal air sacs defined by alveolar walls containing capillaries and interstitial space. The cellular composition of each vessel, airway, and alveolar wall is heterogeneous. Injurious environmental stimuli signal through PKCs and cause a variety of disorders. Edema formation and pulmonary hypertension (PHTN) result from derangements in endothelial, smooth muscle (SM), and/or adventitial fibroblast cell phenotype. Asthma, chronic obstructive pulmonary disease (COPD), and lung cancer are characterized by distinctive pathological changes in airway epithelial, SM, and mucous-generating cells. Acute and chronic pneumonitis and fibrosis occur in the alveolar space and interstitium with type 2 pneumocytes and interstitial fibroblasts/myofibroblasts playing a prominent role. At each site, inflammatory, immune, and vascular progenitor cells contribute to the injury and repair process. Many strategies have been used to investigate PKCs in lung injury. Isolated organ preparations and whole animal studies are powerful approaches especially when genetically engineered mice are used. More analysis of PKC isozymes in normal and diseased human lung tissue and cells is needed to complement this work. Since opposing or counter-regulatory effects of selected PKCs in the same cell or tissue have been found, it may be desirable to target more than one PKC isozyme and potentially in different directions. Because multiple signaling pathways contribute to the key cellular responses important in lung biology, therapeutic strategies targeting PKCs may be more effective if combined with inhibitors of other pathways for additive or synergistic effect. Mechanisms that regulate PKC activity, including phosphorylation and interaction with isozyme-specific binding proteins, are also potential therapeutic targets. Key isotypes of PKC involved in lung pathophysiology are summarized and current and evolving therapeutic approaches to target them are identified.     

Gender-related symptoms and correlates of alcohol dependence among men and women with a lifetime diagnosis of alcohol use disorders

This study explored gender-related symptoms and correlates of alcohol dependence in a crosssectional study of 150 men and 150 women with a lifetime diagnosis of alcohol use disorders (AUD). Participants were recruited in equal numbers from treatment settings, correctional centres and the general community. Standardized measures were used to determine participants' use of substances, history of psychiatric disorders and psychosocial stress, their sensation seeking and family history of substance use and mental health disorders. Multivariate analyses were used to detect patterns of variables associated with gender and the lifetime severity of AUD. Men had a longer history of severe AUD than women. Women had similar levels of alcohol dependence and medical and psychological sequelae as men, despite 6 fewer years of AUD. More women than men had a history of severe psychosocial stress, severe dependence on other substances and antecedent mental health problems, especially mood and anxiety disorders. There were differences in family history of alcohol-related problems approximating same-gender aggregation. The severity of a lifetime AUD was predicted by its earlier age at onset and the occurrence of other disorders, especially anxiety, among both men and women. The limitations in the generalizability of these findings due to sample idiosyncrasies are discussed.     

Biochemical and molecular characterisation of cubozoan protein toxins

Class Cubozoa includes several species of box jellyfish that are harmful to humans. The venoms of box jellyfish are stored and discharged by nematocysts and contain a variety of bioactive proteins that are cytolytic, cytotoxic, inflammatory or lethal. Although cubozoan venoms generally share similar biological activities, the diverse range and severity of effects caused by different species indicate that their venoms vary in protein composition, activity and potency. To date, few individual venom proteins have been thoroughly characterised, however, accumulating evidence suggests that cubozoan jellyfish produce at least one group of homologous bioactive proteins that are labile, basic, haemolytic and similar in molecular mass (42-46 kDa). The novel box jellyfish toxins are also potentially lethal and the cause of cutaneous pain, inflammation and necrosis, similar to that observed in envenomed humans. Secondary structure analysis and remote protein homology predictions suggest that the box jellyfish toxins may act as α-pore-forming toxins. However, more research is required to elucidate their structures and investigate their mechanism(s) of action. The biological, biochemical and molecular characteristics of cubozoan venoms and their bioactive protein components are reviewed, with particular focus on cubozoan cytolysins and the newly emerging family of box jellyfish toxins.     

Dose tolerability of chronically inhaled voriconazole solution in rodents

Invasive pulmonary aspergillosis (IPA) is a fungal disease of the lung associated with high mortality rates in immunosuppressed patients despite treatment. Targeted drug delivery of aqueous voriconazole solutions has been shown in previous studies to produce high tissue and plasma drug concentrations as well as improved survival in a murine model of IPA. In the present study, rats were exposed to 20 min nebulizations of normal saline (control group) or aerosolized aqueous solutions of voriconazole at 15.625 mg (low dose group) or 31.25 mg (high dose group). Peak voriconazole concentrations in rat lung tissue and plasma after 3 days of twice daily dosing in the high dose group were 0.85 ± 0.63 μg/g wet lung weight and 0.58 ± 0.30 μg/mL, with low dose group lung and plasma concentrations of 0.38 ± 0.01 μg/g wet lung weight and 0.09 ± 0.06 μg/mL, respectively. Trough plasma concentrations were low but demonstrated some drug accumulation over 21 days of inhaled voriconazole administered twice daily. Following multiple inhaled doses, statistically significant but clinically irrelevant abnormalities in laboratory values were observed. Histopathology also revealed an increase in the number of alveolar macrophages but without inflammation or ulceration of the airway, interstitial changes, or edema. Inhaled voriconazole was well tolerated in a rat model of drug inhalation.