联合应用SSH和cDNA表达谱芯片筛选肝星状细胞早期活化的相关基因

目的利用抑制消减杂交(suppressionsubtractivehybridization,SSH)和cDNA表达谱芯片筛选早期活化的肝星状细胞(hepaticstellatecell,HSC)和大鼠正常肝脏组织、轻度肝纤维化组织、重度肝纤维化组织中差异表达的基因。方法从SSH构建的大鼠轻度肝纤维化、重度肝纤维化2个差异cDNA文库中挑选1000条上调显著的基因,与正常大鼠4136条基因克隆制作成一张芯片,筛选早期活化的肝星状细胞相关基因。结果获得与HSC早期活化相关的上调基因有202条,下调基因有80条,其中血清和糖皮质激素调节蛋白激酶(serumandglucocorticoidsensitivekinase,SGK)在正常大鼠基因克隆和2周及8周SSH上调差异基因中均呈上调信号。结论联合应用SSH和cDNA表达谱芯片是筛选和鉴定不同样本中差异表达基因的快速、经济和有效的方法;SGK可能作为多种细胞信号传导通路和细胞磷酸化级联反应的一个功能性交汇点,参与了肝星状细胞的早期活化和信号传导。     

Liver regeneration in acute severe liver impairment: a clinicopathological correlation study.

BACKGROUND: Although normally quiescent, the adult mammalian liver possesses a great capacity to regenerate after different types of injury. Major players in the regeneration process are mature residual cells, including hepatocytes, cholangiocytes and stromal cells. However, if the regenerative capacity of mature cells is impaired, hepatic progenitor cells (HPCs) are activated and expand into the liver parenchyma. Upon transit amplification, the progenitor cells generate new hepatocytes and biliary cells to restore liver homeostasis. AIMS/METHODS: To study the relationship between different histopathological parameters as well as their correlations with clinical parameters and outcome, we examined liver specimens from 74 patients with acute or subacute severe liver impairment by immunohistochemistry for CK7/CK19 (evaluation of HPCs activation/differentiation), Mib1(Ki 67)/P21 (evaluation of proliferative activity/proliferation arrest of hepatocytes) and hematoxylin and eosin (evaluation of hepatocyte loss). RESULTS: Of the 74 patients, 32% survived without transplantation, 14% died without transplantation and 54% were transplanted. Our results show that a threshold of 50% loss of hepatocytes, associated with significant decrease in the proliferative activity of remaining mature hepatocytes, is needed for extensive hepatic progenitor cell activation. Such activation is a sign of disease severity and occurs early (within 1 week) in the disease course. However, development of intermediate hepatocytes, suggesting HPCs differentiation towards mature hepatocytes, takes at least 1 week's time. We found a positive correlation between histopathological parameters (percentage hepatocyte loss, number of proliferating hepatocytes and number of HPCs) and clinical parameters of liver impairment such as model for end stage liver diseases (MELD). Surviving patients compared with those who either died or were transplanted had significantly less hepatocyte loss, less HPCs activation and more mature hepatocyte proliferative activity. Hepatocyte proliferative activity and degree of hepatocyte loss were the most important independent histopathological parameters in predicting outcome. CONCLUSION: Liver biopsy can provide important additional information in a patient with severe acute liver impairment.     

Fetal cardiac screening by three- and four-dimensional ultrasound

Objective. The objective of our paper is to show that the spatio-temporal image correlation (STIC) and tomographic ultrasound imaging (TUI) is very convenient and helpful for the fetal screening of complex congenital heart defects (CHD).

Methods. Ultrasound examinations were performed using a Voluson 730 EXPERT or PRO system (GE Medical system, Kretztechnik, Zipf, Austria), and the transabdominal probe (RAB 4-8 MHz or 1-5 MHz) was used to acquire the STIC volumes. Various complex CHD including heterotaxia, ventricular septal defect (VSD), atrio-ventricular septal defect (AVSD), tetralogy of fallot (TOF), transposition of great artery (TGA), hypoplastic left heart syndrome (HLHS) were examined between 20 weeks and 35 weeks. After routine cardiac screening and examination by two-dimensional ultrasound, three- and four-dimensional ultrasound were performed by gray-scaled and color flow mapping. After the examination detailed analysis of CHD were performed by STIC and TUI.

Results. In the case of heterotaxia, STIC and TUI was useful for the detection of stomach and cardiac apex in the different slices. In the case of VSD and AVSD, they were useful for the exact determination of septal defect location. In the cases of TOF, TGA and HLHS, they were helpful for observation of outflow tract and exact diagnosis.

Conclusion. In the screening of complex CHD including heterotaxia, VSD, AVSD, TOF, TGA and HLHS, STIC is very useful and powerful tool.     

Can Electrocardiographic Criteria Predict Adverse Cardiac Events and Positive Cardiac Markers?

OBJECTIVES: To determine electrocardiogram (ECG) predictors of positive cardiac markers and short-term adverse cardiac events in an undifferentiated chest pain population presenting to emergency departments (EDs). The authors hypothesized that specific ECG findings, other than those previously identified in higher-risk populations, would be predictive of cardiac outcomes and positive cardiac markers. METHODS: This study used data from a prospectively collected, retrospectively analyzed Internet-based data registry of undifferentiated chest pain patients (i*trACS). Logistic regression modeling was performed to determine the ECG findings that were predictive of 1) positive cardiac markers and 2) short-term adverse cardiac events. RESULTS: ST-segment elevation (STE), ST-segment depression (STD), pathological Q-waves (PQW), and T-wave inversion were associated with increased odds of percutaneous coronary intervention or catheterization, myocardial infarction, or coronary artery bypass grafting. The odds of creatine kinase-MB (CK-MB) measuring positive were increased if STE, STD, or PQW were present [odds ratio (OR) 2.495, 2.582, and 1.295, respectively]. A right bundle branch block tended to decrease the odds of CK-MB measuring positive (OR 0.658). A similar pattern of results was observed for troponin I (OR 3.608 for STE, 3.72 for STD, 1.538 for PQW). Troponin T showed an increased odds of measuring positive if any of STE, STD, left bundle branch block, or T-wave inversion were evident (OR 2.313, 2.816, 1.80, and 1.449, respectively). CONCLUSIONS: Initial ECG criteria can be used to predict short-term cardiac outcomes and positive cardiac markers. These findings can be important aids in the risk-stratification and aggressive treatment regimens of chest pain patients presenting to EDs.     

猪喉软骨硫酸软骨素的制备和自由基清除活性

直接用木瓜蛋白酶水解软骨,三氯醋酸除蛋白质,乙醇沉淀干燥得硫酸软骨素粗多糖,经DEAE-Sepharose fast flow离子交换层析分离纯化,高效凝胶渗透色谱和琼脂糖凝胶电泳测定其相对分子质量和纯度,比较硫酸软骨素粗品和纯品的体外自由基清除活性。结果显示硫酸软骨素粗品和纯品提取率分别为31.56±0.46%和19.79±0.78%,后者相对相对分子质量为75 174,粗品的DPPH.和.OH清除活性最强,随着产品纯度的提高,活性降低,而对于O2-.清除活性结果则相反。     

A study exploring the role of intercostal nerve damage in chronic pain after thoracic surgery

Objective: Our aim was to investigate the prevalence of intra-operative nerve damage and its association with chronic pain. Methods: Our prospective study of 33 patients used nerve conduction studies to assess intercostal nerve function during elective thoracic surgical procedures. We used two methods to study nerve conduction: pre-operative magnetic stimulation (in 10 patients) and intra-operative nerve conduction studies (in all patients) We correlated these findings with specific intra-operative parameters, pain and psychological questionnaires pre-op and 3 month post-op and altered cutaneous sensation. Results: Magstim (magnetic stimulation) assessments were not reliable and were therefore abandoned. Intraoperative intercostal nerve studies revealed two distinct patterns of nerve injury and also that nerve injury was less in those cases where a rib was not resected. However, intercostal nerve damage detected at the time of operation is not associated with chronic pain or altered cutaneous sensation at 3 months post-op. Conclusions: The study findings suggest that either the amount of intra-operative intercostal nerve damage is not indicative of long-term nerve damage or that there is a more significant cause for chronic pain other than intercostal nerve injury.     

类风湿性关节炎药物治疗的进展

近年来,随着类风湿性关节炎发病机制及某些致炎因子的发现,出现了一系列新药。环氧合酶-Ⅱ特异性抑制剂与传统的非甾体类抗炎药相比具有疗效好、副作用小的优点;早期联合使用改变病情性抗风湿药在近期内有较好的临床疗效。此外重组可溶性肿瘤坏死因子受体融合蛋白 (etanercept)、人体抗肿瘤坏死因子-α单克隆抗体(adalimumab)和阿那白滞素(anakinra)等生物学治疗及中药治疗均显示了新的治疗前景。     

胰岛素强化治疗对严重创伤患者炎性反应及预后的影响

Objective To investigate the effects of intensive insulin therapy on inflammatory re-sponse and prognosis of patients with severe trauma. Methods Eighty severely injured patients were di-vided into intensive insulin therapy group (n = 40, IT) and routine therapy group (n = 40, RT) in random pair. At the time of admission, a continuous infusion of insulin (2 -4 U/h) was pumped into the patients of IT group to maintain blood glucose level at 6 -8 mmol/L. Patients in RT group were given routine treatment without administration of insulin. Fever, organ injury, and mortality of patients in 2 groups were recorded. Venous blood was drawn from patients of 2 groups on the morning of post treatment day (PTD) 1, 3, 5, and 7. Values of TNF-α, C-reactive protein (CRP), IL-2, and IL-10 in plasma were assayed. Results High fever appeared in 9 patients in IT group, and WBC exceeded 10.0×109 for more than 3 days in 17 patients in this group, versus 20 and 29 patients respectively in RT group. Dysfunction of 1 organ appeared in 31 pa-tients in IT group and 30 patients in RT group. Dysfunction of 3 organs appeared in 10 patients in IT group and 19 patients in RT group. Dysfunction of 4 organs appeared in 7 patients in IT group and 12 patients in RT group. In IT group, 4 patients died within 3 post-injury day (PID), and 1 patient died after PID 3 (total case fatality: 12.5% ). In RT group, 5 patients died within 3 PID, and 4 patient died after PID 3 (total case fatality: 22.5%). Plasma levels of TNF-α and CRP of patients in IT group were significantly lower than those of patients in RT group on PID 3 - 7 ( P<0.05 or P<0.01 ), while levels of IL-2 and IL-10 of patients in IT group were significantly higher than those of patients in RT group ( P<0.05 or P<0.01 ). Plasma levels of TNF-α ( 1.3±0.6 μg/L) and CRP (55±16 mg/L) of patients in IT group on PTD 7 were lowered to the trough level, and they were significantly lower than those of patients in RT group (3.0±0.8μg/L, 89±20 mg/L, respectively, P <0.01 ). Conclusions Intensive insulin therapy can mitigate systemic inflammatory response and improve prognosis of patients with severe trauma.