孕妇血浆中sCD40及其配体与子痫前期发病及肾功能损害的关系

目的 探讨孕妇血浆中可溶性白细胞分化抗原(sCD40)和sCD40配体(sCD40L)水平变化与子痫前期发病及肾功能损害的关系.方法 选择2008年8月-2010年6月在青岛大学医学院附属医院产科分娩的轻度子痫前期孕妇28例(轻度子痫前期组),重度子痫前期孕妇35例(重度子痫前期组);另选同期妊娠结局良好的健康孕妇30例为对照组.比较3组孕妇分娩孕周及血压变化、血小板计数并检测其血常规、C反应蛋白(CRP)、尿常规、24h尿蛋白定量,以及血清尿酸(UA)、肌酐(Cr)、尿素氮(BUN)等生化指标.采用ELISA法检测3组孕妇血浆中sCD40和sCD40L的水平,并对血浆sCD40和sCD40L的水平与各临床指标的相关性进行分析.结果 (1)血常规及l临床指标:重度子痫前期组和轻度子痫前期组孕妇血浆CRP水平(分别为10.8及7.1 mg/L)均明显高于对照组(3.3 mg/L),前后两者分别比较,差异均有统计学意义(P＜0.05);重度子痫前期组高于轻度子痫前期组,两组比较,差异也有统计学意义(P＜0.05).重度子痫前期组孕妇分娩孕周(32.5周)明显低于轻度子痫前期组(37.2周)和对照组(38.6周),分别与对照组比较,差异均有统计学意义(P＜0.01);轻度子痫前期组与对照组比较,差异无统计学意义(P＞0.05).重度子痫前期组孕妇血小板计数(132×109/L)明显低于轻度子痫前期组(212×109/L)和对照组(216×109/L),分别比较,差异有统计学意义(P＜0.01);轻度子痫前期组与对照组比较,差异无统计学意义(P＞0.05).3组孕妇之间血红蛋白水平及白细胞数分别比较,差异均无统计学意义(P＞0.05).(2)血浆sCD40及sCD40L水平:重度子痫前期组、轻度子痫前期组和对照组孕妇血浆sCD40水平分别为133.6、126.5和90.7 ng/L,sCD40L水平分别为12.5、10.4和4.4 ng/L,24h尿蛋白定量分别为4.5、0.8 g和0,UA水平分别为486、289和162 μmol/L,重度子痫前期组以上各指标均明显高于轻度子痫前期组和对照组,差异均有统计学意义(P＜0.01);轻度子痫前期组也明显高于对照组,差异也有统计学意义(P＜0.01).重度子痫前期组孕妇血浆Cr(89 μmol/L)、BUN(5.32 mmol/L)水平高于轻度子痫前期组(分别为66μmol/L及4.49mmol/L)和对照组(分别为57 μmol/L及3.32 mmol/L),分别比较,差异均有统计学意义(P＜0.05);轻度子痫前期组与对照组比较,差异无统计学意义(P＞0.05).(3)相关性分析:轻、重度子痫前期组孕妇血浆sCD40水平与24 h尿蛋白定量呈正相关(r=0.434,P＜0.05),与UA、CRP呈明显正相关(r=0.536、0.528,P＜0.01),与收缩压、舒张压、分娩孕周、Cr、BUN、血小板计数无明显相关(r分别为0.135、0.183、-0.133、0.190、0.167、-0.221,P均＞0.05).轻、重度子痫前期组孕妇血浆sCD40L水平与24 h尿蛋白定量、UA、CRP均呈明显正相关(r分别为0.591、0.445、0.539,P均＜0.01),与收缩压、舒张压、分娩孕周、Cr、BUN、血小板计数无明显相关(r分别为0.178、0.212、-0.292、0.144、0.135、-0.273,P均＞0.05).轻、重度子痫前期组孕妇血浆sCD40L水平与sCD40呈明显正相关(r均为0.707,P＜0.01).对照组孕妇血浆sCD40、sCD40L水平与各项临床指标均无相关性(P＞0.05).结论 子痫前期孕妇血浆中sCD40和sCD40L水平明显升高,可能参与了子痫前期的发病,并导致肾功能损害.sCD40和sCD40L水平变化也与子痫前期的严重程度相关.

Abstract：

Objective To investigate the variance levels of plasma soluble leukocyte differentiation antigens CD40 (sCD40) and soluble CD40 ligand (sCD40L) in preeclamptic patients with renal damage and its relationship. Methods A total of 63 pregnant women attended the Department of Obstetrics, Affiliated Hospital of Qingdao University Medical College between August 2008 and June 2010. In the present study included 28 pregnant women with mild preeclampsia and 35 patients with severe preeclampsia. Thirty matched normotensive pregnant women were enrolled in the study as the control group. Expression of sCD40 and sCD40L were determined by ELISA. At the same time, the blood routine, C reaction protein ( CRP),urine routine, 24 hours urine protein excretion, and serum uric acid (UA), creatinine (Cr), blood urea nitrogen (BUN) were measured. The correlation analysis was performed between the sCD40/sCD40L and the blood biochemical indexes in 3 groups. Results ( 1 ) The median levels of CRP in severe preeclampsia (10. 8 mg/L)and mild preeclampsia group(7. I mg/L)are significantly higher than that of control group (3. 3 mg/L,P ＜ 0. 05 ); The level of CRP in severe preeclampsia group was also higher than that of mild preeclampsia group ( P ＜ 0. 05 ). The median gestational age at delivery in severe preeclampsia ( 32. 5 weeks)was significantly less than that of mild preeclampsia group ( 37. 2 weeks) and normal group ( 38. 6 weeks,P ＜ 0. 05). However no significant differences were observed between mild preeclampsia group and normal group ( P ＞0. 05 ). The platelet count in severe preeclampsia ( 132 × 109/L) was significantly less than those of mild preeclampsia group (212 × 109/L) and normal group ( 216 × 109/L, P ＜ 0. 01 ), but no significant differences were observed in blood platelet amount between mild preeclampsia group and normal group ( P ＞0. 05 ). There was no significant difference in hemoglobin level and white blood cell in three groups ( P ＞0. 05). (2) The sCD40 plasma concentration in severe, mild preeclampsia and normal group was 133.6,126. 5 and 90. 7 ng/L, respectively. The sCD40 L plasma concentrations were 12. 5, 10. 4 and 4. 4 ng/L respectively in the 3 groups. 24 hours urinary protein quantitative was 4. 5 g/d,0. 8 g/d and 0 in the 3 groups respectively. And the UA level was 486 μ mol/L,289 μmol/L and 162 μmol/L. In the above three groups,the monitoring indicators were significantly higher in women with severe preeclampsia group compared with mild preeclampsia and control groups (P ＜ 0. 01 ), and there were also higher in mild preeclampsia group than that in control groups ( P ＜ 0. 01 ). The level of plasma Cr ( 89 μmol/L) and BUN ( 5. 32 mmol/L) in severe preeclampsia group were higher than those of mild preeclampsia group (66 μmol/L and 4. 49mmol/L) and control group ( 57 μmol/L and 3.32 mmol/L, P ＜ 0. 05 ). There was no significant difference between mild preeclampsia group and normal group (P ＞ 0. 05 ). (3) The correlation analysis indicated that the level of sCD40 has a positive correlation with 24 hours urinary protein quantitative( r = 0. 434, P ＜ 0. 05 ),also significant positive correlation( r =0. 536,0. 528 ,P ＜ 0. 01 ) between the level of sCD40 and UA or CRP in women with preeclampsia. There was no significant correlation between the level of sCD40 and systolic blood pressure, diastolic blood pressure, delivery gestational age, Cr, BUN, and platelet count(r =0. 135,0. 183, -0. 133,0. 190,0. 167, -0. 221 ,all P ＞0. 05 ). There were positive correlation between the level of sCD40L and 24 hours urine protein excretion, either UA or CRP( r =0. 591,0. 445,0. 539 ,all P ＜0. 01 ). No significant correlation was found between sCD40 L and systolic blood pressure, diastolic blood pressure,delivery gestational age, Cr, BUN, and platelet count( r =0. 178,0. 212, -0. 292,0. 144,0. 135, -0. 273,all P ＞0. 05). There was significant positive correlation between plasma sCD40 and sCD40L ( r =0. 707 ,P ＜0. 01 ). There was no relationship between the level of sCD40, sCD40L and the blood biochemical indexes in normotensive pregnant women ( P ＞ 0. 05 ). Conclusions The plasma concentrations of sCD40 and sCD40 L are significantly higher in pregnant women with preeclampsia compared with the control, which may be involved in the development of preeclampsia and contribute to the kidney damage. The variance levels of sCD40 and sCD40L may be also related to the severity of preeclampsia.     

兔视网膜中央静脉阻塞模型中小胶质细胞CD40和铁蛋白的表达

目的　分析兔视网膜中央静脉阻塞模型中小胶质细胞ＣＤ４０和铁蛋白表达的变化,探讨其发生机制。方法　将３０只有色兔随机分为空白组、模型组,以激光动力学方法造模,行眼底照相和眼底荧光血管造影法检查,分别于１ｄ、３ｄ、７ｄ、１４ｄ和２８ｄ对两组兔视网膜及视神经标本采用免疫组织化学方法检测ＣＤ４０和铁蛋白表达水平。４０倍光镜照相,测定灰度值和积分光密度（ＩＯＤ）,对测得数据进行统计学分析。结果　小胶质细胞ＣＤ４０的表达结果:３ｄ时空白组和模型组视网膜标本灰度值分别为１９３．４２±９．７２和８６.０３±５．１２,差异有统计学意义（Ｐ＜０．０５）;３ｄ时空白组和模型组视神经标本灰度值分别为１９６．２６±１０．７８和８５．９１±５．０１,差异有统计学意义（Ｐ＜０．０５）;３ｄ时空白组和模型组视网膜标本ＩＯＤ分别为１０５．３６±７．５８和４４７９．３２±３０７．２２,差异有统计学意义（Ｐ＜０.０５）;３ｄ时空白组和模型组视神经标本ＩＯＤ分别为５４．６３±８．７９和１０４３６．４２±１３０６.８３,差异有统计学意义（Ｐ＜０．０５）。小胶质细胞铁蛋白的表达结果:３ｄ时空白组和模型组视网膜标本的ＩＯＤ分别为１６１．９６±３５．３７和１２９５．５９±１２４．７１,差异有统计学意义（Ｐ＜０．０５）。结论　视网膜中央静脉阻塞可以引起视网膜及视神经小胶质细胞的活化,免疫组织化学染色显示小胶质细胞ＣＤ４０和铁蛋白表达均为阳性结果。     

P40家族成员及其在自身免疫性疾病中的作用

作为P40家族中研究最为广泛的细胞因子,白细胞介素-12(Interleukin-12)其最显著的特点是有助于Th1细胞分化,参与由Th1细胞所介导的免疫性疾病。而白细胞介素-23(Interleukin-23)与Th17细胞增殖有关,最新发现Th17介导的免疫反应在实验性变态反应性脑脊髓炎(EAE)和多发性硬化症(MS)疾病方面发挥重要作用,并且其分化途径的提出对传统的Th1/Th2细胞免疫也带来了新的挑战[1]。该家族的另外两个成员P40单体和P40同源二聚体也参与免疫调控,实验证明P40分子不仅在细胞内感染及炎症过程中起着重要的调节作用,而且与银屑病、多发性硬化症、Crohn’s病等多种自身免疫性临床疾病的发病密切相关。     

术前sCD_(40)L预测冠状动脉支架后再狭窄的临床价值研究

目的:探讨冠状动脉粥样硬化性心脏病患者支架植入术前sCD40L对再狭窄的诊断预测价值。方法:选择成功接受普通支架置入术的稳定型心绞痛和不稳定型心绞痛患者共92例,分别于支架术前,术后1,5,15d和180d取外周静脉血测定血清sCD40L。所有患者随访6个月。结果:支架内再狭窄率23.9%(22/92)。再狭窄患者支架术前和术后血清sCD40L水平均显著高于无再狭窄患者(P均<0.01);再狭窄患者支架术后高水平sCD40L持续至术后6个月,而无再狭窄患者术后5d则恢复至正常。根据受试者工作特征曲线确定术前血清sCD40L>3.96μg/L为截断值,计算术前sCD40L诊断再狭窄的敏感性、特异性、阳性预测值、阴性预测值、准确度和阳性似然比分别为72.7%、90%、69.6%、91.3%、85.9%和7.27。多变量Logistic回归分析发现,在校正混杂因素后,术前sCD40L是术后再狭窄独立预测因子,OR=1.92(95%CI=1.39～2.64,P=0.013)。结论:再狭窄患者支架术前、术后血清sCD40L水平增加提示sCD40L可能与支架内再狭窄有关。支架术前血清sCD40L是术后再狭窄的独立预测因子,术前sCD40L有可能用于临床支架放置前危险分层的评价。     

氯吡格雷对冠状动脉介入治疗后血清sCD40L及sICAM-1的影响

目的探讨氯吡格雷对冠心病患者冠脉内介入治疗(PCI)疗效的影响。方法选取2012年5月至2014年10月我院收治的冠心病患者45例作为研究组,另取造影正常的45例患者作为对照组,所有患者均接受抗心绞痛治疗,在PCI治疗前给予研究组患者氯吡格雷75 mg/d,使用酶联免疫吸附法(ELISA)检测术前1d、术后1 d和术后5 d患者血清中s CD40L和s ICAM-1的浓度,比较两组之间的差异。结果术前1 d、术后1 d研究组患者血清中s CD40L和s ICAM-1浓度水平明显低于对照组,差异有统计学意义(P<0.05),术后5 d两组s CD40L和s ICAM-1水平比较差异无统计学意义(P>0.05);对照组患者血清中s CD40L和s ICAM-1浓度水平基本维持不变,而研究组患者血清中s CD40L和s ICAM-1浓度水平逐渐降低。结论氯吡格雷可降低冠心病患者PCI术后早期血清s CD40L和s ICAM-1浓度水平,有效预防血栓发生。     

OX40Ig修饰大鼠脂肪间充质干细胞的实验研究

目的构建含OX40融合蛋白(OX40Ig)基因修饰的大鼠脂肪间充质干细胞(ADSCs)即ADSCs^OX40Ig,探讨其在体外对淋巴细胞的免疫调节作用。方法构建pcDNA3.1(+)/OX40Ig融合基因表达载体,采用核转染技术转染大鼠ADSCs,Western blot检测ADSCs中OX40Ig表达水平。ADSCs与大鼠异基因淋巴细胞共培养后分为对照组(反应细胞+刺激细胞)、ADSCs组(反应细胞+刺激细胞+ADSCs)及ADSCs^OX40Ig组(反应细胞+刺激细胞+ADSCs^OX40Ig),MTT法检测淋巴细胞增殖抑制率,逆转录聚合酶链反应(RT-PCR)检测淋巴细胞内干扰素(IFN)-γ、白细胞介素(IL)-10及转化生长因子(TGF)-βmRNA表达。结果经测序鉴定验证,pcDNA3.1(+)/OX40Ig质粒构建成功,转染ADSCs后OX40Ig蛋白呈高表达;与ADSCs组比较,ADSCs^OX40Ig显著增强异基因淋巴细胞增殖抑制率(P<0.05);对照组、ADSCs组及ADSCs<...     

The age‐related changes in 40 Hz Auditory Steady‐State Response and sustained Event‐Related Fields to the same amplitude‐modulated tones in typically developing children: A magnetoencephalography study

Recent studies have revealed that gamma‐band oscillatory and transient evoked potentials may change with age during childhood. It is hypothesized that these changes can be associated with a maturation of GABAergic neurotransmission and, subsequently, the age‐related changes of excitation–inhibition balance in the neural circuits. One of the reliable paradigms for investigating these effects in the auditory cortex is 40 Hz Auditory Steady‐State Response (ASSR), where participants are presented with the periodic auditory stimuli. It is known that such stimuli evoke two types of responses in magnetoencephalography (MEG)—40 Hz steady‐state gamma response (or 40 Hz ASSR) and auditory evoked response called sustained Event‐Related Field (ERF). Although several studies have been conducted in children, focusing on the changes of 40 Hz ASSR with age, almost nothing is known about the age‐related changes of the sustained ERF to the same periodic stimuli and their relationships with changes in the gamma strength. Using MEG, we investigated the association between 40 Hz steady‐state gamma response and sustained ERF response to the same stimuli and also their age‐related changes in the group of 30 typically developing 7‐to‐12‐year‐old children. The results revealed a tight relationship between 40 Hz ASSR and ERF, indicating that the age‐related increase in strength of 40 Hz ASSR was associated with the age‐related decrease of the amplitude of ERF. These effects were discussed in the light of the maturation of the GABAergic system and excitation–inhibition balance development, which may contribute to the changes in ASSR and ERF.     

二维、三维超声联合应用诊断胎儿唇裂

目的探讨二维超声、三维超声诊断胎儿唇裂的价值。方法对我院门诊孕检的18～40周胎儿,常规二维超声观测胎儿鼻唇情况,然后应用三维超声实时动态面部成像。结果三维超声对18～24周胎儿唇裂成像清晰直观逼真,二维超声对18～24周胎儿唇裂,检查费时,检查者须有经验;三维超声对36～40周胎儿实时成像困难,二维超声则可通过非标准切面面部成像;三维超声及二维超声对24～36周胎儿均能有效成像。结论二维、三维超声均能对胎儿唇裂明确诊断。不同孕期二维超声与三维超声各有优势,三维超声在孕早期诊断唇裂有明显优势,二维超声在孕晚期略显优势。     

联合检测D2-40、calretinin及HBME-1在浆膜腔积液细胞学鉴别诊断中的意义

目的 观察D2-40、calretinin及HBME-1在浆膜腔积液间皮和转移性癌细胞中的表达,评价其用于鉴别诊断的价值.方法 用细胞块技术对87例浆膜腔积液标本进行免疫组化检测,观察D2-40、calretinin和HBME-1在癌细胞和间皮细胞中的表达.结果 D2-40、calretinin和HBME-1标记间皮细胞的敏感度分别为48.1%、81.0%、77.2%,特异度分别为90.2%、69.5%、61.0%.三者联合的敏感度和特异度分别为70.9%和81.7%.三者标记卵巢癌腹水间皮细胞的特异性都不高.结论 D2-40、calretinin和HBME-1合用可辅助鉴别诊断转移性癌与间皮.     

制备淋巴瘤多基因肿瘤疫苗的方法学探索

目的探讨用于淋巴瘤的多基因肿瘤疫苗的制作方式及淋巴瘤的基因治疗方法。方法采用腺病毒转染、脂质体辅助转染、脂质体辅助腺病毒转染的方式,将带GFP的载体转染B系淋巴瘤A20细胞株,并以Hela细胞作为对照,荧光显微镜下观察转染效率。将B7-1与CD40L表达载体联合导入A20荷瘤小鼠肿瘤内,观察肿瘤生长情况,应用肿瘤病理切片和HE染色技术观察肿瘤组织形态及淋巴细胞浸润情况。结果对A20细胞而言,单纯脂质体和单纯腺病毒转染效率均非常低,脂质体辅助腺病毒转染效率有所提高。但均显著低于采用腺病毒转染Hela细胞。瘤内联合注射B7-1和CD40L表达载体可导致肿瘤生长延缓及体积缩小,肿瘤消退效应较明显。肿瘤形态学观察表明,治疗小鼠肿瘤局部有炎性细胞浸润并伴有大面积的坏死,肿瘤局限化。结论B系淋巴瘤细胞株难于通过体外基因导入制作肿瘤疫苗,脂质体辅助腺病毒转染效果较好,质粒载体瘤内注射可作为一种安全有效的肿瘤疫苗作用方式。