帕金森病患者脑脊液中miR-9a的表达及其临床意义

目的研究miR-9a在帕金森病(PD)患者脑脊液中的表达,为帕金森病的早期诊断提供临床检测指标。方法采用实时荧光定量PCR检测42例PD患者、24例普通头痛患者和24例健康人脑脊液中miR-9a表达水平情况。分析脑脊液中miR-9a表达水平与PD相关临床指标的关系。结果 PD组患者脑脊液中miR-9a表达水平明显高于头痛组和健康对照组(P<0.05),且PD患者脑脊液中miR-9a表达与帕金森综合评分量表(UPDRS)分级呈正相关(r=0.70,P<0.05)。ROC曲线确诊PD患者的脑脊液miR-9a临界值为0.667,其诊断敏感度为85.7%,特异度为77.9%。结论脑脊液miR-9a的检测不仅有助于PD的诊断,也可能作为判断PD病情的指标之一。     

血清和脑脊液中载脂蛋白E水平与脑卒中预后的相关性研究

目的 :通过测定脑卒中患者血清和脑脊液中载脂蛋白E(ApoE)浓度 ,对疾病的诊断及预后作初步判断。方法 :测定100例健康人和67例脑卒中患者 (其中脑梗死38例、脑出血29例 )治疗前后血清和脑脊液中ApoE浓度。结果 :脑卒中患者血清ApoE浓度(43.8±7.2)mg/L ,比健康人(37.6±6.1)mg/L显著升高 (P<0.01) ,脑卒中患者治疗前后血清ApoE浓度无显著性变化 ,治疗后显效组(7.4±1.1)mg/L和有效组(7.2±1.0)mg/L脑脊液ApoE浓度均比无效组(5.9±0.9)mg/L有显著性升高 (P<0.01)。结论 :血清和脑脊液中ApoE浓度可作为诊断脑卒中及判断预后的一项有效指标。     

结核性脑膜炎患者脑脊液中左氧氟沙星浓度监测分析

目的评价结核性脑膜炎(结脑)患者脑脊液中左氧氟沙星浓度监测的临床意义。方法运用高效液相色谱法测定8例结脑患者脑脊液中左氧氟沙星峰浓度时间(tmax)左右3个时间点(2、4、6 h)的浓度。结果结脑患者脑脊液tmax左右3个时间点的平均左氧氟沙星浓度分别是(0.94±0.39)(2 h)、(2.36±0.42)(4 h)、(2.14±0.32)(6 h)μg·mL-1,所有测定值均高于最小抑菌浓度(MIC,0.5μg·mL-1)。每位结核性脑膜炎患者tmax左右3个时间点的最大浓度均高于最小杀菌浓度(MBC,1.0μg·mL-1)。结论结核性脑膜炎患者口服左氧氟沙星400 mg,1次·d-1,脑脊液中浓度能达到治疗水平。     

帕金森氏病患者血小板5-羟色胺浓度研究

目的 探讨帕金森氏病(parkinson disease,PD)患者血小板5-羟色胺(5-hy-droxytryptamine,5-HT)浓度的价值.方法 采用高效液相色谱法检测35例PD患者血小板5-HT浓度,并与30例正常对照组的测定结果相比较.结果 PD患者血小板5-HT浓度明显低于正常对照组(P＜0.05),并且在高龄、病情重和抑郁患者的血小板5-HT浓度降低更为显著(P＜0.05).结论 血小板5-HT浓度的降低与PD的发病以及病情的进展密切相关,早期纠正PD患者血小板5-HT浓度异常具有重要意义.     

神经胶质瘤中组织型谷氨酰胺转氨酶的表达及其意义

目的研究神经胶质细胞瘤(Neuroglioma,Ng)中组织型谷氨酰胺转氨酶(tTG)的表达情况,探讨其与神经胶质细胞瘤恶性程度的关系。方法采用免疫组织化学(Immunohistochemistry)方法检测tTG蛋白在星形细胞瘤、少突胶质细胞瘤及正常脑组织中的表达。并用图像分析系统测定tTG蛋白的表达面积;统计分析肿瘤恶性程度与tTG蛋白表达的变化的相互关系。结果随着神经胶质细胞瘤分化程度的降低,肿瘤细胞内tTG表达明显增加,tTG蛋白表达与肿瘤的恶性程度有关。结论神经胶质细胞瘤细胞内tTG表达的检测可用来评价星形细胞瘤恶性进展程度,有助于对胶质细胞瘤防治。     

利奈唑胺在脑膜炎患者脑脊液中的药动学及药效分析

目的分析利奈唑胺在脑膜炎患者脑脊液中的药动学及药效。方法选取2012年4月至2015年6月我院神经外科收治的15例脑外室引流(EVD)导致的革兰阳性球菌感染引起的脑膜炎重症患者作为研究对象,所有患者均采用利奈唑胺治疗,在药物达到稳定状态后对脑脊液中的药动力学参数进行测定,结合微生物学对其药效学指标进行评价。结果患者在用药2.5 h之后达到Cmax(峰浓度)(5.6±2.4)μg/m L,在用药0及12 h时到达的Cmm(谷浓度)分别为(3.2±1.2)μg/m L和(3.4±1.7)μg/m L;药物对脑脊液平均穿透率达到65%左右。结论利奈唑胺针对脑膜炎患者的脑脊液具有良好的穿透率,常规剂量即可达到高效的治疗浓度,值得临床选择和积极推广。     

ICU颅内感染患者血清及脑脊液中万古霉素浓度影响因素分析

目的 探讨影响ICU颅内感染患者血清和脑脊液中万古霉素浓度的相关因素.方法 收集48例ICU颅内感染患者的临床资料,采用简单线性回归和多重线性回归分析影响患者血清和脑脊液中万古霉素谷浓度的相关因素.结果 48例患者血清万古霉素谷浓度为(19.26±8.96) mg/L,血药浓度达标率为31.25％;脑脊液万古霉素谷浓度...     

反相高效液相色谱法测定隐球菌性脑膜炎患者脑脊液中两性霉素B的含量

目的:建立反相高效液相色谱法测定人脑脊液中两性霉素B,并用于隐球菌性脑膜炎患者脑脊液样品的测定。方法:采用Agilent C18(5μm,4.6 mm×250 mm)色谱柱,流动相为0.005 mol.L-1乙二胺四乙酸二钠-甲醇(21∶79),流速1.0 mL.min-1,检测波长380 nm,室温下进样20μL。结果:由于患者脑脊液中药物的含量测定范围较大,故采用2个浓度线性范围:1.25～30μg.mL-1(r=0.9999),高、中、低3种浓度的平均回收率为94.9%,日内RSD≤1.38%,日间RSD≤2.67%;0.11～5μg.mL-1(r=0.9999),高、中、低3种浓度的平均回收率为93.1%,日内RSD≤1.73%,日间RSD≤4.09%。结论:本法灵敏度高,重现性好,专属性强,可用于椎管内持续给予两性霉素B的临床药代动力学研究及脑脊液中该药物浓度的临床检测。     

血清和脑脊液B细胞趋化因子-1的水平对临床孤立综合征转归的意义

目的探讨临床孤立综合征(CIS)患者血清及脑脊液中B细胞趋化因子-1(BLC-1/CX-CL13)浓度与病情变化的关系,与临床表现、磁共振成像(MRI)检查、诱发电位的相关性。方法入选CIS患者73例,包括脊髓型CIS患者20例,脑干型CIS患者18例和视神经型CIS患者19例,神经系统非炎性疾病(NND)16例。在患者发病期进行扩展残障状态量表(EDSS)评分、MRI检查、诱发电位检查,用酶联免疫吸附法(ELISA)检测患者血清及脑脊液中CXCL13浓度。结果脊髓型CIS组、视神经型CIS组、脑干型CIS组血清及脑脊液CXCL13浓度与NND患者比较,差异有统计学意义(P<0.01);脑干型CIS组脑脊液CXCL13浓度高于脊髓和视神经型CIS组,差异有统计学意义(P<0.01);诱发电位异常组与正常组血清和脑脊液CXCL13浓度差异无统计学意义。血清与脑脊液CXCL13浓度呈正相关(r=0.750,P<0.01),血清及脑脊液CXCL13浓度与EDSS评分呈正相关(r=0.837,P<0.01;r=0.689,P<0.01)。结论 CXCL13浓度增高可导致CIS转化为多发性硬化(MS),对预测CIS的转归具有重要的临床意义。     

脑脊液中利奈唑胺质量浓度测定方法的建立及临床应用

目的建立人脑脊液中利奈唑胺质量浓度测定方法并开展临床检测。方法使用高效液相色谱仪(HPLC),采用外标法。色谱条件为:色谱柱:Agilent 5TC-C18(2),250mm×4.6mm;流动相:甲醇-水(40∶60);流速:1.0mL·min~(-1);柱温:25℃;紫外检测波长:250nm。结果利奈唑胺质量浓度在0.31~40 mg·L~(-1)范围内线性关系良好(r=0.999 6),定量下限为0.31mg·L~(-1),方法回收率、提取回收率均大于85%,高、中、低3个质量浓度日内、日间精密度的RSD值均小于5%。药物干扰试验中美罗培南、头孢曲松及头孢他啶对利奈唑胺的测定无影响。对8例患者开展27例次脑脊液利奈唑胺质量浓度测定,平均质量浓度为2.67±1.11mg·L~(-1)。结论建立的利奈唑胺脑脊液质量浓度测定方法简单、方便、灵敏、准确,可用于临床检测;利奈唑胺在脑脊液中的质量浓度个体差异较大。     

Tissue transglutaminase is not involved in the aggregate formation of stably expressed α-synuclein in sh-sy5y human neuroblastoma cells

Intraneuronal deposition containing alpha-synuclein is implicated in the pathogenesis of synuclein-opathies including Parkinsons disease (PD). Although it has been demonstrated that cytoplasmic inclusions of wild type alpha-synuclein are observed in the brain of PD patients and that alpha-synuclein mutations such as A30P and A53T accelerate aggregate formation, the exact mechanism by which alpha-synuclein forms insoluble aggregates is still controversial. In the present study, to understand the possible involvement of tissue transglutaminase (tTG) in aggregate formation of alpha-synuclein, SH-SY5Y cell lines stably expressing wild type or mutant (A30P or A53T) alpha-synuclein were created and aggregate formation of alpha-synuclein was observed upon activation of tTG. The data demonstrated that alpha-synuclein negligibly interacted with tTG and that activation of tTG did not result in the aggregate formation of alpha-synuclein in SH-SY5Y cells overexpressing either wild type or mutant alpha-synuclein. In addition, alpha-synuclein was not modified by activated tTG in situ. These data suggest that tTG is unlikely to be a contributing factor to the formation of aggregates of alpha-synuclein in a stable cell model.     

Differential expression of tissue transglutaminase protein in mouse ovarian follicle

Tissue transglutaminase (tTG) protein begins to accumulate in apoptotic cells and its mRNA is expressed at the onset of apoptotic change. In the present study, we compared tTG expression with the atretic degree of mouse ovarian follicles. The whole-body gamma-irradiated mouse ovaries were collected and immunohistochemistry for tTG and in situ 3'-end labeling (TUNEL) was performed. Based on the identification of atretic follicles with hematoxylin-eosin and TUNEL immunostaining, tTG expression was evaluated and compared between normal (NF) and atretic follicles (AF). The expression of tTG was different among AF depending on the degree of atretic changes. There was a strong association of tTG expression with the follicular apoptotic changes. Among NF, 24% of follicles expressed tTG protein. This value, however, increased up to 66% in atretic follicles. The present results suggest that the follicular expression of tTG is closely related to the degree of follicle atresia. Therefore, the expression of tTG can be used as a useful marker for the identification of atretic follicles in the ovary.     

Role of tissue transglutaminase in GTP depletion-induced apoptosis of insulin-secreting (HIT-T15) cells

The role of tissue transglutaminase (tTG), a calcium-dependent and GTP-modulated enzyme, in apoptotic death induced by GTP depletion in islet beta-cells was investigated. GTP depletion and apoptosis were induced by mycophenolic acid (MPA) in insulin-secreting HIT-T15 cells. MPA treatment increased in situ tTG activity (but not protein levels) in a dose- and time-dependent manner in parallel with the induction of apoptosis. MPA-induced increases of both tTG activity and apoptosis were entirely blocked by co-provision of guanosine but not adenosine. MPA-enhanced tTG activity could be substantially reduced by co-exposure to monodansylcadaverine or putrescine (tTG inhibitors), and largely blocked by lowering free Ca(2+) concentrations in the culture medium. However, MPA-induced cell death was either not changed or was only slightly reduced under these conditions. By contrast, a pan-caspase inhibitor (Z-VAD-FMK) entirely prevented apoptosis induced by MPA, but did not block the enhanced tTG activity, indicating that GTP depletion can induce apoptosis and activate tTG either independently or as part of a cascade of events involving caspases. Importantly, the morphological changes accompanying apoptosis could be markedly prevented by tTG inhibitors. These findings suggest that the effect of the marked increase in tTG activity in GTP depletion-induced apoptosis of insulin-secreting cells may be restricted to some terminal morphological changes.     

Tissue transglutaminase antibodies in celiac disease: focus on the pediatric population

Celiac disease (CD) is an immune-mediated disorder induced by the ingestion of gluten in genetically susceptible individuals. The enzyme tissue transglutaminase (protein-glutamine gamma-glutamyltransferase 2; tTG) plays an important role in the pathogenesis of the disease and antibodies against tTG are used as serological markers for the diagnosis of CD. Discovery of tTG as the autoantigen for endomysial antibodies changed the diagnostic approach to patients with suspected CD and, importantly, made screening for CD more accurate and easier to perform. Studies in pediatric populations confirmed the high sensitivity and specificity of immunoglobulin A (IgA) tTG antibodies in the diagnosis of CD. The aim of this review is to summarize the data on the role of tTG in the pathogenesis of CD and to present current evidence on the accuracy of IgA tTG, IgG tTG and IgA tTG point-of-care tests in the diagnosis of CD in children. This review shows why IgA tTG antibodies replaced endomysial antibodies as the preferred serological marker due to the ease of their use, their high sensitivity and specificity, and the high correlation between high titers and intestinal mucosal lesions.     

Potential role for high and low molecular weight tissue transglutaminases in transforming mammalian cell properties

Tissue transglutaminase (tTGase, tTG) is known as being implicated in the intracellular cross-linking of proteins occurring in a growing series of physiological conditions including--just to mention the most relevant ones--programmed cell death (apoptosis), cell adhesion, growth, spreading and differentiation, tumor growth, metastasis, cell adhesion, proliferation, differentiation, extra cellular matrix (ECM) stabilization. In the current work we investigated tTG activity and expression of "normal" and potential transformed cytosolic tTG antigens in mammalian cells. Most cell lines studied showed low tTG activity, which in all cases could be enhanced considerably by treating cell cytosol homogenates with trypsin. The results suggested the existence -in transformed cells- of inactive types of tTGase. We purified cytosolic tTG antigens from these cells utilizing a GTP-agarose resin, and we can therefore conclude that "normal" molecular weight (mw) tTG antigens, but also high molecular weight (hmw) and low molecular weight (lmw) tTG antigens from transformed cells, retain GTP-binding ability. The initial results from our study also allowed us to hypothesize that transformed hmw- and lmw- tTG antigens should not be considered as the result of post-translational modifications of normal mw, cytosolic tTG. The potentially low or absent transamidating functionality of cytosolic tTG species in transformed mammalian cells could be responsible for decreased or even abolished programmed cell death, whereas the unaffected GTP-binding functionality of such proteins in these cells might lead to increased signal transduction and possibly proliferation.     

Antibodies against deamidated gliadin peptides identify adult coeliac disease patients negative for antibodies against endomysium and tissue transglutaminase

Aliment Pharmacol Ther 2010; 32: 254–260

Summary

Background This study was done to evaluate the diagnostic utility of antibodies against deamidated gliadin peptides compared to traditional markers for coeliac disease. Aim To evaluate diagnostic utility of antibodies against deamidated gliadin peptide (DGP). Methods Sera from 176 adults, referred for endoscopy without previous analysis of antibodies against tissue transglutaminase (tTG) or endomysium (EmA), were retrospectively analysed by ELISAs detecting IgA/IgG antibodies against DGP or a mixture of DGP and tTG, and compared with IgA‐tTG and EmA. Seventy‐nine individuals were diagnosed with coeliac disease. Results Receiver operating characteristic analyses verified the manufacturers’ cut‐off limits except for IgA/IgG‐DGP/tTG. In sera without IgA deficiency, the sensitivity was higher for IgA/IgG‐DGP (0.85–0.87) compared with IgA‐tTg (0.76) and EmA (0.61). All tests showed high specificity (0.95–1.00). Eighteen coeliac disease‐sera were negative regarding IgA‐tTG, nine of which were positive for IgA/IgG‐DGP. Sera from coeliac disease‐patients >70 years were more often negative for IgA‐tTG (50%) and IgA/IgG‐DGP (36%) than younger patients (15% and 8% respectively) (P < 0.01). Three of the four IgA‐deficient patients were positive in the IgA/IgG‐DGP assay. Conclusions In this study of patients unselected regarding IgA‐tTg/EmA, thus unbiased in this respect, IgA/IgG‐DGP identified adult coeliac disease patients negative for antibodies against endomysium and tissue transglutaminase. Serology is often negative in elderly patients with coeliac disease; a small bowel biopsy should therefore be performed generously before coeliac disease is excluded.     

302例呼吸内科慢性阻塞性肺疾病住院患者抗菌药物用药调查

目的了解呼吸内科慢性阻塞性肺疾病住院患者抗菌药物使用情况,促进抗菌药物的合理使用。方法回顾性调查某院呼吸内科2011—01—01,12—31出院的慢性阻塞性肺疾病住院患者病历。内容包括患者基本情况、诊断、过敏史、住院时长、用药情况、住院费用、药品费用和用药合理性评价等。结果共302例患者,主要诊断为慢性阻塞性肺病、慢性阻塞性肺病伴急性加重和慢性阻塞性肺病伴肺部感染等;氟喹诺酮类、β-内酰胺类／β-内酰胺酶抑制剂、头孢菌素类和大环内酯类等是最常用的抗菌药物;抗菌药物的使用在给药剂量和频次等方面存在某些不合理现象。结论慢性阻塞性肺疾病的抗菌药物应用基本符合指南的要求,但临床医生应能根据抗菌药物的PK／PD药代动力学特点更好地制订个体化的给药方案。