系统性红斑狼疮患者血清及骨髓基质细胞分泌转化生长因子β1水平

背景：转化生长因子β1是重要的免疫调节因子已被证实，但有关其与系统性红斑狼疮之间的关系还不很清楚。 目的：检测转化生长因子β1在系统性红斑狼疮患者血清及骨髓基质细胞培养液中的水平。 设计：病例-对照分析。 对象：选择2006-02/2007-02中山大学附属第五医院门诊及住院的系统性红斑狼疮患者55例，其中5例同意行骨髓像检查的重度系统性红斑狼疮患者合并中度贫血，经实验室检查符合慢性病贫血；应用SLEDAI评分标准对患者活动度进行评估，轻度13例、中度22例、重度20例。以同期自愿献血的健康者24例作为对照组，其中4例经临床表现及实验室诊断为缺铁性贫血。 方法：所有受试者晨起空腹抽取静脉血2 mL，离心后取上清液，-20℃保存待测。采用密度梯度离心和贴壁筛选法对5例贫血系统性红斑狼疮患者和4例缺铁性贫血患者骨髓基质细胞进行分离培养，制备骨髓基质细胞培养上清液待测。 主要观察指标：采用双抗体酶联免疫吸附法检测各组血清标本、骨髓基质细胞培养上清液标本中的转化生长因子β1水平，及其与SLEDAI积分、血沉、补体C3的相关性。 结果：与对照组比较，系统性红斑狼疮组血清转化生长因子β1水平明显降低（P < 0.01）；且轻、中、重度系统性红斑狼疮患者血清转化生长因子β1水平逐渐降低（P < 0.05）。与对照组比较，系统性红斑狼疮组骨髓基质细胞培养上清液中转化生长因子β1水平明显降低（P < 0.01）。系统性红斑狼疮患者血清转化生长因子β1水平与SLEDAI积分呈负相关（r=-0.862，P < 0.01），与疾病同期活动指标血沉呈负相关（r = -0.56，P < 0.05），与补体C3呈正相关（r=0.78，P < 0.05）。 结论：系统性红斑狼疮骨髓基质细胞分泌转化生长因子β1存在异常，并且转化生长因子β1可能参与了系统性红斑狼疮的发病及转归，对判断其病情活动度及研究系统性红斑狼疮的发病机制有一定临床意义。     

多细胞因子诱导成年鼠骨髓基质细胞向神经细胞分化的实验研究

目的观察大鼠骨髓基质细胞(rBMSCs)的生长特点及诱导条件下分化成神经细胞的能力,并对其机制进行初步探讨。方法以密度梯度离心分离骨髓基质细胞,在神经干细胞培养液中培养,采用四唑盐(MTT)法观察在培养液中添加碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)对BMSCs增殖的影响;观察添加脑源性神经生长因子(BDNF)、神经生长因子(NGF)和维甲酸(RA)对rBMSCs的诱导分化情况;采用免疫组织化学法(ABC)检测诱导后的细胞表达神经元特异性烯醇化酶(NSE)、神经元核蛋白(NeuN)和胶质原性纤维酸性蛋白抗体(GFAP)等特异性标志物的情况;以流式细胞分选确定神经元的比例。结果bFGF和EGF能在体外促进rBMSCs增殖,BDNF、NGF和RA能诱导rBMSCs来源的神经干细胞(NSCs)表达NSE、GFAP等特异性标志物。结论EGF、bFGF、BDNF、NGF、RA及适宜的培养液可使rBMSCs定向转化为NSCs,获得足够的目的细胞,进而分化为神经元样和神经胶质样细胞。     

140例系统性红斑狼疮患者的脑电图分析

目的了解不同表现类型的SLE患者在病程不同时期表现于脑电图上的变化,并利用脑电图检查来探讨SLE患者脑部受累的情况.方法140例不同表现类型的SLE患者按脑部不同的表现分为四组,进行发作期、治疗期、稳定期以及复发期的常规脑电图检查,又对伴有头痛症状的SLE患者与血管性头痛症的患者进行脑电图比较.结果在发作期,A组和B组的脑电图异常率为31%和37%;C组和D组为26%和100%.经过二周的治疗,脑电图的高度异常率自40%下降至零,中度异常率自60%下降至26%,但轻度异常的脑电图却难以转入正常.对于血管性头痛症患者和SLE伴头痛的患者在头痛期和非头痛期中前者的脑电图异常率自100%下降至9%,而后者几乎没有下降.结论不同表现类型的SLE患者中都可能有机会出现脑电图的异常者,反映了脑部受累的存在.但经治疗和症状的稳定,对于轻度异常的脑电图仍然难以转入正常,提示了脑部的基础损害可能是不可逆的.复发期患者的脑电图异常程度往往和其原先的脑电图异常程度基本相同.     

外周血CD34+细胞形貌在系统性红斑狼疮患者与健康人的差异比较

背景：系统性红斑狼疮患者CD34+细胞是否存在异常目前仍有争议。 目的：以健康人为对照，应用原子力显微镜观察比较二者外周血CD34+细胞形貌特征。 设计：病例-对照分析。 对象：6例系统性红斑狼疮患者，男3例，女3例，平均年龄26.3岁，平均病程19.0个月，均符合1982年美国风湿病学会制定的诊断标准。3例健康志愿者为对照。两组对象基线资料具有可比性。 方法：系统性红斑狼疮患者均静脉注射环磷酰胺4 g/m2，白细胞<1.0×109 L-1时给予粒细胞集落刺激因子5 μg/kg，待白细胞> 5.58×109 L-1及CD34+>2%时采集外周血干细胞。3例健康志愿者采取单用粒细胞集落刺激因子为动员方案。向外周血干细胞中加入CD34 MicroBeads FcR和FcR Blocking Reagent制备细胞悬液，离心弃上清，重悬后过滤，应用免疫磁珠细胞分选技术纯化CD34+细胞。 主要观察指标：原子力显微镜观察细胞表面形貌，每例患者随机测20个细胞，每个细胞不同部位做2 μm×2 μm图像5幅，将5幅图像的直径、平均粗糙度、均方根粗糙度、平均高度的均值作为该细胞的参数。 结果：成像范围在12 μm×12 μm时，可将细胞整体模拟成像。成像范围在2 μm×2 μm时，两组细胞表面均可见大小、深浅不一的凹陷及椭圆形球状突起，粗糙不平。与健康志愿者比较，系统性红斑狼疮患者外周血CD34+细胞的直径、平均粗糙度、均方根粗糙度、平均高度等参数经单向方差分析差异无显著性意义（F=0.203~4.553，P均> 0.05）。 结论：应用原子力显微镜选择成像范围在2 μm×2 μm时可清晰观测外周血CD34+细胞的表面结构，且系统性红斑狼疮患者和健康人的外周血CD34+细胞形貌基本相似。     

大鼠骨髓基质细胞生物学特性研究

目的 观察大鼠骨髓基质细胞的生物学特性。方法 取SD大鼠骨髓，分离培养骨髓基质细胞，相差显微镜下观察其形态，传代后MTT法绘制其生长曲线。结果 原代培养20天后可分离得到骨髓基质细胞，典型的骨髓基质细胞可分为两个类型，传代后2小时贴壁率达70％以上。结论 骨髓基质细胞具有多态性和贴壁生长特性,通过贴壁培养方法能够较容易地对其进行分离扩增，可作为多种疾病细胞治疗和基因治疗的载体。     

骨髓间充质干细胞对系统性红斑狼疮患者T细胞活化的影响

背景：骨髓间充质干细胞具有免疫调节作用,可治疗自身免疫性疾病。 目的：探讨在体外人骨髓间充质干细胞对系统性红斑狼疮患者T细胞活化的影响。 方法：分离、培养人骨髓间充质干细胞,培养至第3代以后的细胞胰蛋白酶消化后用流式细胞仪确定其浓度,按照不同浓度将骨髓间充质干细胞设为1×108,1×107L-1两组。取肝素抗凝新鲜27例系统性红斑狼疮患者患者外周血10 mL,分离培养T淋巴细胞,以1×109 L-1的细胞浓度,100 µL/孔,接种两组骨髓间充质干细胞上,以单纯统性红斑狼疮患者的T淋巴细胞作为对照组。通过流式细胞术计算CD3+ T细胞CD25(IL-2R)和CD38细胞的表达率。 结果与结论：与对照组比较,1×108 L-1浓度组的骨髓间充质干细胞对系统性红斑狼疮患者CD25及CD38的表达呈明显抑制作用(P < 0.01),1×107 L-1浓度组,无明显变化。结果表明：骨髓间充质干细胞抑制系统性红斑狼疮患者T细胞的活化,且对这种抑制作用具有数量依赖性。     

系统性红斑狼疮患者肺部感染的分析与护理

目的对186例住院的系统性红班狼疮(SLE)患者并发肺部感染的情况进行回顾研究,分析护理干预在SLE并发肺部感染时的意义。方法对所有患者活动性指数进行评分,分析狼疮活动评分及治疗措施与肺部感染的关系,并发肺部感染患者的病原学特点及其与患者预后的关系。结果中、高度狼疮活动组肺部感染的患病率明显高于轻度活动组,使用较大剂量糖皮质激素的患者发生肺部感染的几率升高,发生肺部感染的78例患者中40例细菌培养阳性,住院天数明显延长,病死率增高。结论SLE患者有易发生感染的倾向。加强原发病的治疗,激素及抗生素的合理运用,都是预防和控制肺部感染重要条件。加强患者的健康教育及护理干预,增加患者对自身疾病和治疗的认识,尽可能的减少肺部感染,对改善患者的预后具有重要意义。     

骨髓基质细胞联合细胞因子对脐血单个核细胞表面抗原表达的影响**

背景：课题组已建立胎儿骨髓基质细胞联合细胞因子的造血细胞体外培养体系,该培养体系能否有效扩增各个发育阶段的造血细胞有待验证。 目的：观察骨髓基质细胞联合细胞因子培养体系对脐血单个核细胞表面抗原CD133、CD34表达的影响。 方法：将从脐血标本中分离出来的单个核细胞接种于无血清培养体系,实验分为3组：①F组：干细胞因子+Flt3配体+促血小板生成素+单个核细胞。②S组：基质细胞+单个核细胞。③SF组：基质细胞+干细胞因子+Flt3配体+促血小板生成素+单个核细胞。在第0,6,10,14天检测有核细胞总数、CD133+、CD34+、CD133+CD34+细胞数以及集落形成单位数。 结果与结论：SF组有核细胞总数在各个检测时间点均比其他两组高;除了第14天外,第6、10天两个时间点SF组中CD133+、CD34+、CD133+CD34+细胞及集落形成单位数均高于其他组;含骨髓基质细胞的S组和SF组中CD133+细胞/有核细胞、CD34+细胞/有核细胞、CD133+CD34+细胞/有核细胞的比例保持在较高的水平。结果说明骨髓基质细胞联合细胞因子能有效的扩增脐血单个核细胞及其中的CD133+、CD34+、CD133+CD34+细胞,基质细胞对维持造血干细胞的原始性具有重要的作用。     

系统性红斑狼疮患者血清免疫抑制物的初步观察

目的　初步探讨系统性红斑狼疮 (SLE)患者血清中的免疫抑制物质。方法　选用 SLE患者、狼疮肾患者、慢性肾小球肾炎患者血清。以正常志愿者血清为对照。检测血清及其超滤后不同组分对由刀豆蛋白 (ConA)诱导的淋巴细胞转化的影响。结果　与正常对照组相比 ,狼疮性肾炎患者的血清可十分显著地抑制正常淋巴细胞转化 (抑制率在 90 %以上 ) ,超滤后大于相对分子质量为 3 0 0 0 0的组分具有抑制作用 ,而且其抑制强度与不作超滤处理的血清相同。相对分子质量 <3 0 0 0 0的组分未表现出任何抑制作用。临床诊断未明显累及肾脏的 SLE疮患者 ,其血清也具有明显抑制作用 (抑制率在 5 0 %左右 )。慢性肾小球肾炎患者的血清则对正常淋巴细胞转化几乎无抑制作用 (抑制率小于 2 0 % )。结论　 SLE患者血清中可能存在一种能抑制 T淋巴细胞转化的大分子物质 (相对分子质量 >3 0 0 0 0 ) ,它与由神经系统介导生成的应激免疫抑制蛋白的关系值得进一步探讨。     

人骨髓基质细胞作为核移植供体细胞的相关生物学特性研究

目的研究作为核移植供体细胞的人骨髓基质细胞相关生物学特性。方法应用密度梯度离心法分离培养人骨髓基质细胞，并对其进行形态观察、免疫荧光分析细胞骨架结构、端粒酶活性测定、核型及细胞周期分析。结果培养的细胞具有正常的大小、形态、细胞骨架结构：在融合密度达80%-90%时，G0＋G1期细胞所占的比例较高，约占92．58％；检测传至第7代的人骨髓基质细胞具有正常染色体数目（46，XY），端粒酶活性为0．567。结论形态良好、染色体数目正常的人骨髓基质细胞可以作为核移植的供体细胞。     

新生鼠缺氧缺血性脑病时骨髓基质干细胞的生长特性

背景：骨髓基质干细胞用于缺氧缺血性脑病治疗,其生长特性是否稳定尚需观察。 目的：探讨急性脑缺氧缺血新生大鼠骨髓基质干细胞移植后的生长状态及理化性质的改变。 方法：结扎新生7 d Wistar大鼠双侧颈总动脉制成缺血模型,恢复2 h后,吸入体积分数8%O2、92%N2的混合气体制成缺氧缺血性脑病模型。分为轻度(缺氧1 h)、中度(缺氧1.5 h)、重度(缺氧2.5 h)缺氧缺血性脑病组及假手术组(仅结扎左侧颈总动脉)。观察各组骨髓基质干细胞的生长状态。 结果与结论：假手术组及轻度缺氧缺血性脑病组骨髓基质干细胞在提取培养3 h即可贴壁生长,而中、重度组骨髓基质干细胞贴壁较晚且生长状态较差,假手术组、轻、中度组细胞形态可逐渐恢复。假手术组及轻度组细胞凋亡率基本相同,中、重度组细胞凋亡率明显增高。轻度及中度组培养上清液乳酸脱氢酶有不同程度的增高。提示轻、中度缺氧缺血对骨髓基质干细胞的影响是可逆的,骨髓基质干细胞生长旺盛,对于损伤恢复功能较强。     

系统性红斑狼疮神经系统病变的临床及病理特征

目的探讨系统性红斑狼疮（SLE）神经系统病变的临床及病理特征。方法回顾性分析6例SLE神经系统病变患者的临床及病理资料。结果本组患者累及中枢神经系统4例,其中癫痫1例、脑梗死2例、白质脑病1例;多发性单神经病3例,腓肠神经活检示1例有典型血管炎改变,2例无血管炎改变,腓肠神经内可见有髓纤维丢失、轴索和髓鞘断裂呈块状深染及髓磷脂小球形成。肌活检1例示肌纤维轻度变性坏死。皮肤活检3例示胶原变性,小静脉周围可见炎性细胞浸润。结论SLE可导致神经系统广泛病变,故其临床表现和病理改变复杂多样。     

Neurological and psychiatric symptoms in patients with systemic lupus erythematosus

Sixty patients (51 women/9 men) with diagnosis of SLE were studied for finding out the frequency of nervous system involvement in SLE, the time of of appearance of neurological involvement after diagnosis establishing, the coexistence of the antiphospholipid syndrome, and the character of changes in MR, CT and CSF. Nervous system involvement was found in 40 cases (67%), with 34 cases (56%) had involvement of the CNS, 6 patients (10%) had symptoms of peripheral nervous system dysfunction, and 3 (5%) had involvement of both systems. In 4 cases polineuropathy and transverse spinal cord lesion, and in 3 cases psychiatric symptoms were the first manifestations of SLE. Changes due to involvement of cerebral vessels (TIA, stroke) were observed in 20 patients (33%), and psychiatric symptoms in 16 cases (26.6%). No difference was found in the occurrence of stroke or TIA between SLE patients with and without antiphospholipid syndrome, and no correlations were noted between the presence of neurological or psychiatric symptoms and other SLE symptoms. CT scans demonstrated corticosubcortical atrophy in 28.3% of cases, while in MRI in T2 images small hyperintense lesions were situated mainly in the white matter (33.9%). In 5 cases oligoclonal band was found in the CSF, but without any correlation with specific neurological symptoms.     

Prevalence of obsessive-compulsive disorder in patients with systemic lupus erythematosus

BACKGROUND: The goal of this pilot study was to investigate the prevalence of obsessive-compulsive disorder (OCD) in a group of patients with systemic lupus erythematosus (SLE). METHOD: Fifty adult patients enrolled in out-patient SLE studies at the National Institute of Arthritis and Musculoskeletal and Skin Diseases (February 1995-October 1996) completed a self-report questionnaire adapted from the Yale-Brown Obsessive Compulsive Scale and an in-person psychiatric clinical interview with a psychiatrist or psychiatric clinical nurse specialist. DSM-IV lifetime diagnosis of OCD was determined by clinical interview. RESULTS: Sixteen subjects (32%) met DSM-IV lifetime diagnostic criteria for OCD and an additional 5 (10%) met criteria for subclinical OCD. Mean +/- SD number of symptoms reported on the self-report questionnaire was significantly higher among subjects diagnosed with OCD on clinical interview (40.7 +/- 23.2) compared with those without OCD (8.9 +/- 11.7; t = 5.8, df = 27, p <.001). CONCLUSION: Obsessive-compulsive disorder was 10 to 15 times more common in this cohort of patients with SLE compared with those in community-based studies of OCD. The use of an OCD self-report rating scale proved helpful in the identification of OCD symptoms among patients with SLE. Results suggest that further studies of OCD in patients with SLE are needed and may provide new insight into the pathophysiology of both disorders.     

Detection of cerebral embolic signals in patients with systemic lupus erythematosus

Background

Involvement of the CNS in systemic lupus erythematosus (SLE) is caused by several pathogenic mechanisms including cerebral embolism.

Aim

To measure the frequency of microembolic signals (MES) by using transcranial Doppler (TCD) ultrasound and to assess their association with cerebral infarction, neuropsychological dysfunction, and biochemical, sonographic and clinical variables in an unselected group of patients with SLE.

Methods

A 1‐h TCD recording from the middle cerebral artery was carried out in 55 patients with SLE having a mean age of 46 (SD 13) years. MRI of the brain, carotid artery ultrasonography with intima–media thickness and atherosclerotic plaque assessments were carried out in addition to a broad biochemical and clinical assessment. All patients underwent a neuropsychological assessment.

Results

Of the 55 patients, MES were detected in 5 (9%) and cerebral infarcts were found in 9 (18%). A significant association was found between MES and cerebral infarcts and considerably more neuropsychological deficits were found in MES‐positive patients compared with the negative group. MES were not associated with other clinical, sonographic and biochemical factors believed to be associated with cerebral embolism.

Conclusions

Cerebral embolism may be one of the important mechanisms responsible for the high prevalence of cerebrovascular events and the neuropsychological deficits observed in patients with SLE. Although the number of MES‐positive patients was small, the lack of a significant association between MES and other known risk factors for MES suggests a complex pathogenesis for the embolisation in these patients.CNS symptoms and signs are common in systemic lupus erythematosus (SLE) and as many as 50% of patients with SLE may have neuropsychiatric involvement.^1,2 Well‐known complications include psychosis, seizures, cerebrovascular accidents and cognitive dysfunction. Women with SLE, aged 18–44 years, are eight times as likely to be admitted to hospital because of stroke as controls.³ The pathogenesis of CNS involvement in SLE has not been clarified, and multiple factors may be associated, such as microvascular damage, small‐vessel vasculopathy, antibodies to nervous tissue and immunologically mediated thromboembolism. Postmortem examinations have shown microinfarcts and microhaemorrhages in cortical and subcortical regions. Several factors are associated with the increased risk of stroke in these patients. These include antiphospholipid antibodies, use of corticosteroids, cardiac involvement and other well‐known risk factors for cerebrovascular disease.^2,4,5Transcranial Doppler (TCD) examination, a non‐invasive technique, can detect cerebral embolisation in the major intracranial arteries.⁶ Microembolic signals (MES) have been detected during cardiac surgery and carotid endarterectomy.^7,8 Long‐term TCD monitoring of the intracranial arteries has shown abnormal signals, indicating clinically silent MES in patients with high‐grade carotid stenosis, with prosthetic heart valves or after recent cerebrovascular events.^9,10,11 Cerebral microemboli may cause cognitive dysfunction if they enter the cerebral circulation in considerably large numbers. This has been studied in detail in patients who have had coronary artery bypass surgery.¹² Some instances showing a positive association between cerebral microemboli detected by TCD and postoperative neuropsychological outcome after cardiac surgery have been reported.^7,10Three studies^13,14,15 that used TCD for embolic detection in patients with SLE have showed conflicting results. It is therefore of interest to carry out further studies on the possible importance of cerebral microembolisation in SLE.We measured the occurrence of MES in a group of patients with SLE and assessed the possible association with cerebral infarcts, neuropsychological function, risk factors for cerebrovascular disease, including carotid atherosclerosis, and biochemical variables associated with cerebrovascular disease.