马桑内酯所致癫痫发作大鼠红细胞免疫粘附功能的研究

作者用红细胞Ｃ＿３ｂ受体花环试验和红细胞免疫复合物花环试验对马桑内酯所致癫痫发作大鼠红细胞免疫粘附功能的变化进行了观察，结果表明，癫痫组动物红细胞Ｃ＿３ｂ受体花环率明显低于对照组，而红细胞免疫复合物花环率相差不显著．提示癫痫发作可导致大鼠红细胞免疫粘附功能降低，因此在癫痫治疗中注意调整和增强患者的红细胞免疫功能具有重要意义。     

Organ distribution of severe acute respiratory syndrome (SARS) associated coronavirus (SARS-CoV) in SARS patients: implications for pathogenesis and virus transmission pathways

We previously identified the major pathological changes in the respiratory and immune systems of patients who died of severe acute respiratory syndrome (SARS) but gained little information on the organ distribution of SARS-associated coronavirus (SARS-CoV). In the present study, we used a murine monoclonal antibody specific for SARS-CoV nucleoprotein, and probes specific for a SARS-CoV RNA polymerase gene fragment, for immunohistochemistry and in situ hybridization, respectively, to detect SARS-CoV systematically in tissues from patients who died of SARS. SARS-CoV was found in lung, trachea/bronchus, stomach, small intestine, distal convoluted renal tubule, sweat gland, parathyroid, pituitary, pancreas, adrenal gland, liver and cerebrum, but was not detected in oesophagus, spleen, lymph node, bone marrow, heart, aorta, cerebellum, thyroid, testis, ovary, uterus or muscle. These results suggest that, in addition to the respiratory system, the gastrointestinal tract and other organs with detectable SARS-CoV may also be targets of SARS-CoV infection. The pathological changes in these organs may be caused directly by the cytopathic effect mediated by local replication of the SARS-CoV; or indirectly as a result of systemic responses to respiratory failure or the harmful immune response induced by viral infection. In addition to viral spread through a respiratory route, SARS-CoV in the intestinal tract, kidney and sweat glands may be excreted via faeces, urine and sweat, thereby leading to virus transmission. This study provides important information for understanding the pathogenesis of SARS-CoV infection and sheds light on possible virus transmission pathways. This data will be useful for designing new strategies for prevention and treatment of SARS.     

Activation of the Notch signaling pathway disturbs the CD4+/CD8+, Th17/Treg balance in rats with experimental autoimmune uveitis

Objective and design

The present study aimed to investigate the relationship between the disturbed balance of CD4⁺/CD8⁺, Th17/Treg and the activation of the Notch signaling pathway in experimental autoimmune uveitis (EAU).

Methods

An EAU rat model was induced in Lewis rats, and pathology analysis was performed by hematoxylin and eosin (H&E) staining. CD4⁺, CD8⁺, Th17, and Treg levels in spleen, lymph nodes and eye tissues were determined by flow cytometry. Meanwhile, the expression of Notch1, DLL4, IL-10, and IL-17 was determined by quantitative polymerase chain reaction (Q-PCR) and enzyme-linked immunosorbent assay (ELISA). In addition, the inhibitory effect of N-(N-(3,5-difluorophenacetyl-l-alanyl))-S-phenylglycine t-butyl ester (DAPT) on Th17 differentiation by Notch signaling in vitro was further investigated using T lymphocytes from EAU rats on day 12 post-immunization by flow cytometry.

Results

The pathological results showed that inflammatory cell infiltration occurred in ocular tissues in EAU rats. The CD4⁺/CD8⁺ and Th17/Treg ratios in EAU rats were apparently higher than those in normal control individuals. Q-PCR and ELISA analyses indicated the expression of Notch1, DLL4, IL-10, and IL-17 in EAU rats gradually increased on day 6 after immunization, peaked on day 12, and then gradually decreased. The dynamic trends in Notch1 and DLL4 expression in EAU rats were identical to those of CD4⁺/CD8⁺ and Th17/Treg levels. DAPT can significantly inhibit the activation of Notch signaling, decrease Th17 cell differentiation, and attenuate the level of the Th17 cell lineage, contributing to the balance of the Th17/Treg ratio.

Conclusion

The activation of the Notch signaling pathway can regulate Th17 and Treg cell differentiation, disrupt the CD4⁺/CD8⁺ and Th17/Treg balance, and aggravate the severity of EAU; inactivation of the Notch signaling pathway contributes to the CD4⁺/CD8⁺ and Th17/Treg balance in EAU rats. Our findings highlighted that the dynamic change in the CD4⁺/CD8⁺ and Th17/Treg ratio was consistent with the expression trend of Notch signaling in EAU rats, suggesting that Notch signaling may be a potentially important therapeutic target in clinical practice.

     

Neuroprotective effects of Longxue Tongluo Capsule on ischemic stroke rats revealed by LC-MS/MS-based metabolomics approach

Objective: The present study aimed to evaluate the therapeutic effect and explore the underlying mechanisms of Longxue Tongluo Capsule (LTC) on ischemic stroke rats. Methods: Twenty-six rats were randomly divided into four groups, including sham group, sham + LTC group, MCAO group, and MCAO + LTC group. Ischemic stroke rats were simulated by middle cerebral artery occlusion (MCAO), and LTC treatment group were orally administrated with 300 mg/kg of LTC once daily for seven consecutive days. LTC therapy was validated in terms of neurobehavioral abnormality evaluation, cerebral infarct area, and histological assessments. The plasma metabolome comparisons amongst different groups were conducted by UHPLC-Q Exactive MS in combination with subsequent multivariate statistical analysis, aiming to finding the molecules in respond to the surgery or LTC treatment. Results: Intragastric administration of LTC significantly decreased not only the neurobehavioral abnormality scores but also the cerebral infarct area of MCAO rats. The interstitial edema, atrophy, and pyknosis of glial and neuronal cells occurred in the infarcted area, core area, and marginal area of cerebral cortex were improved after LTC treatment. A total of 13 potential biomarkers were observed, and Youden index of 11 biomarkers such as LysoPC, SM, and PE were more than 0.7, which were involved in neuroprotective process. The correlation and pathway analysis showed that LTC was beneficial to ischemic stroke rats via regulating glycerophospholipid and sphingolipid metabolism, together with nicotinate and nicotinamide metabolism. Heatmap and ternary analysis indicated the synergistic effect of carbohydrates and lipids may be induced by flavonoid intake from LTC. Conclusion: The present study could provide evidence that metabolomics, as systematic approach, revealed its capacity to evaluate the holistic efficacy of TCM, and investigate the molecular mechanism underlying the clinical treatment of LTC on ischemic stroke.     

Preliminary evaluation of pelvic vascular bed isolation chemotherapy in the treatment of advanced cervical carcinoma

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糖尿病合并妊娠伴视网膜病变孕妇的临床观察

目的：研究糖尿病合并妊娠伴视网膜病变孕妇的妊娠结局及孕期视网膜病变。方法：回顾性分析１９８１～１９９５年间４９例糖尿病孕妇妊娠情况。伴视网膜病变的１６例分为第Ⅰ组；无视网膜病变的３３例分为第Ⅱ组。第Ⅰ组糖尿病病程平均为８．９２±４．３５年；第Ⅱ组为３．１１±１．７４年。结果：第Ⅰ组孕妇并发症及围产儿发病率较高，但两组比较，差异无显著性（Ｐ＞０．０５）。３１．１％孕妇视网膜病变在孕期加重，但仍属单纯型（背景期）。结论：糖尿病合并妊娠伴眼底病变时，仍可以继续妊娠，但孕期应密切观察眼底变化和严格控制血糖。     

妊娠期外阴阴道念珠菌病的发病分析

目的:了解妊娠期外阴阴道念珠菌病(VVC)的发病情况以及影响因素。方法:回顾性调查2005年1月1日至2005年6月20日在北京大学第一医院分娩的1119例孕妇孕前及孕期VVC的发病情况。结果:1119例孕妇中,孕前曾患VVC共212例(18·95%),孕前9个月内曾患VVC的孕妇较未感染者孕期VVC发生率明显升高(P<0·01)。1119例孕妇中孕期VVC感染102例(9·12%),明显高于孕前9个月内发生率(P<0.01),其中,妊娠早期22例,中期39例,晚期41例;血糖正常孕妇VVC发生率与血糖异常孕妇发生率比较,差异无显著性(P>0.05)。结论:妊娠中、晚期VVC发病率高于早期。妊娠前9个月内曾患有VVC是孕期VVC的高危因素,妊娠期糖代谢异常者孕期VVC的发生率未见明显增加。     

切割海马伞海马中56 kD差异蛋白的质谱分析

目的:探讨切割海马伞海马中具有生物活性的56kD差异蛋白的组成成份及其功能。方法:切割SD大鼠海马伞后14d海马进行非变性聚丙烯酰胺凝胶电泳(NativePAGE),应用电喷雾质谱分析、蛋白质数据库和文献分析等方法对56kD差异蛋白进行检测和功能分析。结果:质谱分析找到了33组肽段,这些蛋白按照其功能可以分为:细胞骨架蛋白、参与代谢的酶、信号传递、蛋白降解、核酸水解、氧化应激、神经递质运输、突触形成、功能未知蛋白。结论:切割海马伞海马中56kD差异蛋白中含有功能涉及神经干细胞迁移、分化的蛋白质。     

CD13抑制剂乌苯美司对A549细胞顺铂敏感性的影响及其机制

  目的   检测CD13抑制剂乌苯美司联合顺铂对A549细胞增殖、细胞凋亡的影响,并探讨其机制。   方法   应用噻唑蓝比色法检测细胞增殖;流式细胞Annexin V-FITC/PI双染色法检测细胞凋亡;Western blot方法检测p53、PARP-1、Bcl-2、Bcl2-xL和Caspase蛋白水平;酶活性底物法检测CD13活性;Hoechst33342染色联合流式细胞术检测侧群细胞。   结果   乌苯美司单药对A549细胞增殖无显著影响（P>0.05）,乌苯美司（100 μg/mL）联合顺铂作用A549细胞24、48、72 h后,乌苯美司可显著增强8、16 μmol/L顺铂对A549细胞的增殖抑制（P < 0.05）,并呈时间依赖性,72h时,8、16μmol/L顺铂组和顺铂联合组的细胞增殖率分别为（62.06± 7.60）% vs.（27.92±5.84）%和（19.22±1.57）% vs.（0.67±0.42）%（P < 0.05）;但对64 μmol/L顺铂的细胞增殖率无明显影响（P>0.05）。16 μmol/L顺铂联合组细胞凋亡的比例明显增加（P < 0.05）。乌苯美司对P53、PARP-1、Bcl-2、Bcl2-xL及Capsize蛋白水平无明显影响,但显著抑制A549细胞CD13活性和降低A549侧群细胞比例（P < 0.05）。   结论   CD13抑制剂乌苯美司体外条件下显著提高低剂量顺铂对A549细胞的抑制作用,可能成为CD13阳性非小细胞肺癌患者顺铂的增敏剂。      

急性髓系白血病Ki-67的表达及其临床意义

目的：探讨增殖相关抗原Ki-67在急性髓系白血病（AML）中的表达及其临床意义。方法选取2012年10月至2016年1月首都医科大学附属复兴医院血液内科住院AML患者45例,其中初治36例,复发9例;以20例健康志愿者作为健康对照。采用流式细胞术（FCM）检测骨髓原始细胞群Ki-67抗原的表达,分析Ki-67水平与患者临床特征的相关性。结果初治、复发AML患者和健康对照者的骨髓原始细胞群Ki-67阳性率分别为（10.38±8.41）%、（20.99±11.49）%和（40.77±11.97）%,初治和复发AML患者的Ki-67阳性率均低于健康对照者（均P＜0.05）,初治AML患者Ki-67阳性率低于复发AML患者（P＝0.006）。初治AML患者的Ki-67水平与患者的年龄、FAB亚型、白细胞计数、有无骨髓增生异常综合征（MDS）病史、乳酸脱氢酶水平、骨髓原始细胞比例、NPM1基因突变、FLT3内部串联重复（ITD）、染色体核型、诱导化疗反应均无关（均P＞0.05）。初治AML患者Ki-67高水平组与低水平组的总生存时间分别为（780±110）d和（788±118）d,两组差异无统计学意义（P＝0.927）。结论 AML患者原始细胞群增殖活性较健康对照低,检测Ki-67水平可能为临床选择细胞周期特异性化疗药物提供依据;对AML患者动态监测Ki-67水平有助于监测疾病进展,预测复发。