Effect of Xiaoyu Zhixue Tablet (消瘀止血片) on Expression ofPlatelet Membrane Glycoproteins in Patients withHemorrhagic Thrombopathy

Objective: To observe the effect of Xiaoyu Zhixue tablet (消瘀止血片,XYZXT) on the expression of platelet membrane glycoproteins in patients with hemorrhagic thrombopathy, and to explore its possible mechanism. Methods: The total of 148 patients with hemorrhagic thrombopathy were randomly divided into two groups, the traditional Chinese medicicne (TCM) group (n=98) treated with XYZXT and the Western medicine (WM) group (n=50) treated with adrenosin, vitamins C, K and P, both for 6 months. The therapeutic effect and the recovery rate of platelet aggregation in the two groups were observed. And platelet membrane glycoprotein (GP) Ⅰb/Ⅸ, GPⅡb/Ⅲa complexes, GPⅠb, GPⅡb, GP Ⅲa and P-selectin were analyzed by flow cytometry in both groups before and after treatment and also in 34 normal healthy subjects. Results: The total effective rate of hemostasis was 89. 8% in TCM group and 54. 0% in the WM group (x2=45.83, P<0.01), and the recovery rate of platelet aggregation was 72.4% and 4.0% respectively (x2=62.06, P<0.01). The fluorescence intensity of GP Ⅰ b/Ⅸ, GPⅡb/Ⅲa complexes, GPⅠb, GPⅢa and P-selectin were lower in both groups before treatment than those in the healthy subjects. Expression of above-mentioned marks was elevated in TCM group after 6 months' therapy, which was insignificantly different as compared with the healthy subjects (P>0.05) and higher than those in the WM group (P<0.05). Conclusion: One of the mechanisms in treating hemorrhagic thrombopathy with XYZXT is that it could regulate the expression of GP Ⅰb/Ⅸ, GPⅡ b/Ⅲa complexes, GPⅠb, GPⅢa and P-selectin at the level of receptor protein.

Expression of Platelet Membrane Glycoprotein Ib/Ⅸ/Ⅴ Complex,a Receptor of Thrombin,in Patients with Hemorrhagic Thrombopathy

To investigate the role of platelet membrane glycoprotein （GP） Ib/Ⅸ/Ⅴ complex and its subunit GP Ibα in patients with hemorrhagic thrombopathy （HT）, the expressions of GP Ib/Ⅸ/Ⅴ complex and GP Ibα,defined as mean fluorescence intensity （MFI）, were assessed by flow cytometry. The maximum aggregation of platelet was determined by turbidity method. These indicators were compared among 68 HT patients with the presenting complaint of hemorrhage, 33 well-controlled HT patients and 32 normal healthy subjects. The results showed that the MFI of GP Ib/Ⅸ /Ⅴ complex and GP Ibα was markedly lower in HT patients with current hemorrhage than that in the healthy subjects, with difference being statistically significant （P〈0.05）. There was no significant difference in the expressions of GP Ib/ Ⅸ/ Ⅴ complex and GP Ibα between well-controlled HT patients and normal healthy subjects （P〉0.05）. It was concluded that the expression of GP Ib/Ⅸ /Ⅴ complex, the receptor of thrombin and von Willebrand factor, was down-regulated in HT patients with current hemorrhage, which might result in the dysfunction of platelet aggregation and recurrence of HT.

Association of the Platelet Membrane Glycoprotein Ⅰa C807T Gene Polymorphism with Aspirin Resistance

To explore the correlation between the C807T polymorphism of platelet membrane glycoprotein Ⅰa （GP Ⅰa） gene and aspirin resistance in Chinese people, 200 patients with high-risk of atherosclerosis took aspirin （100 mg/d） for 7 days. Platelet aggregation function was detected using adenosine diphosphate （ADP） and arachidonic acid （AA） before and after the administration of aspirin. Then the subjects were divided into three groups according to the results of platelet aggregation function： an aspirin resistant （AR） group, an aspirin semi-responder （ASR） group and an aspirin-sensitive （AS） group. Platelet GP Ⅰa gene 807CT polymorphism was examined by means of polymerase chain reaction-sequence specific primers （PCR-SSP）. The results showed that T allelic frequency in AR group and ASR group were higher that of AS group （P〈0.005）, and the prevalence of genotypes （TT＋TC） of these two groups was significantly higher than that in AS group （P〈0.05）. Platelet GP Ⅰa T allele was significantly associated with aspirin resistance as revealed by multiple logistic regression （OR=3.76, 95% CI： 2.87-9.58）. The results suggest that inherited platelet GP Ⅰa variations may have an important impact on aspirin resistance and the presence of GP Ⅰa T allele may be a marker of genetic susceptibility to aspirin resistance.

Redistribution of Platelet Membrane Glycoprotein IV and Release of Intracellular α-granule Thrombospondin in Patients with Chronic Myelogenous Leukemia

The redistribution of platelet membrane glycoprotein IV (GPIV) and the release of intracellular Q-granule thrombospondin (TSP) were examined and the inhibition of 5-thromboglobulin (&TG) and platelet factor 4 (PF4) in patients with chronic myelogenous leukemia (CML) was observed and quantitation of β-TG and PF4 in sera was conducted. GPIV in inactive platelet from CML was 36080±17010 molecules/platelet as compared with 13190±4810 from the controls (P<0,01), No abnormality was found in the distribution of platelet membrane GPIb and GPIIb/III.(P>0. 05). The GPIV redistribution on active platelet membrane induced thrombin (1U/ml) from CML and healthy donors was 44320132310 and 228001 12700 molecules/platelet respectively (P<0. 01 ). The difference in the release of intracellular Q-granule TSP between CML and the control group was not found (P>0.05). There was no direct correlation between GPIV expression and TSP binding after platelet activation. The high leveIs of β-TG and PF4 in sera inhibited release of intracellular a-granule TSP in vitro. These results indicate that the abnormality of platelet membrane GPIV is a common marker in CML, therefore the specific increase of platelet GPIV in patients with CML may be a useful tool for the diagnosis and monitoring of the platelet dysfunction. The release of interna1 TSP pools is hindered by either β-TG or PF4 in sera.

Membrane glycoprotein M6A promotes μ-opioid receptor endocytosis and facilitates receptor sorting into the recycling pathway

The interaction of p-opioid receptor （MOPr） with the neuronal membrane glycoprotein M6a is known to facilitate MOPr endocytosis in human embryonic kidney 293 （HEK293） cells. To further study the role of M6a in the post-endocytotic sorting of MOPr, we investigated the agonist-induced co-internalization of MOPr and M6a and protein targeting after internalization in HEK293 cells that co-expressed HA-tagged MOPr and Myc-tagged M6a. ,We found that M6a, MOPr, and Rab 11, a marker for recycling endosomes, co-localized in endocytotic vesicles, indicating that MOPr and M6a are primarily targeted to recycling endosomes after endocytosis. Furthermore, co-expression of M6a augmented the post-endocytotic sorting of 6-opioid receptors into the recycling pathway, indicating that M6a might have a more general role in opioid receptor post-endocytotic sorting. The enhanced post-endocytotic sorting of MOPr into the recycling pathway was accompanied by a decrease in agonist-induced receptor down-regulation of M6a in co-expressing cells. We tested the physiological relevance of these findings in primary cultures of cortical neurons and found that co-expression of M6a markedly increased the translocation of MOPrsfrom the plasma membrane to intracellular vesicles at steady state and significantly enhanced both constitutive and agonist-induced receptor endocytosis. In conclusion, our results strongly indicate that M6a modulates MOPr endocytosis and post-endocytotic sorting and has an important role in receptor regulation.

In situ tumor vaccination with adenovirus vectors encoding measles virus fusogenic membrane proteins and cytokines

AIM： To evaluate whether intratumoral expression of measles virus fusogenic membrane glycoproteins H and F （MV-FMG）, encoded by an adenovirus vector Ad.MV-HI F, alone or in combination with local coexpression of cytokines （IL-2, IL-12, IL-18, IL-21 or GM-CSF）, can serve as a platform for inducing tumor-specific immune responses in colon cancer.METHODS： We used confocal laser scanning microscopy and flow cytometry to analyze cell-cell fusion after expression of MV-FMG by dye colocalization. In a syngeneic bilateral subcutaneous MC38 and Colon26 colon cancer model in C57BL/6 and BALB/c mice, we assessed the effect on both the directly vector-treated tumor as well as the contralateral, not directly vector- treated tumor. We assessed the induction of a tumorspecific cytotoxic T lymphocyte （CTL） response with a lactate dehydrogenase （LDH） release assay.RESULTS： We demonstrated in vitro that transduction of MC38 and Colon26 cells with Ad.MV-H/F resulted in dye colocalization, indicative of cell-cell fusion, in addition, in the syngeneic bilateral tumor model we demonstrated a significant regression of the directly vector-inoculated tumor upon intratumoral expression of MV-FMG alone or in combination with the tested cytokines. We observed the highest anti-neoplastic efficacy with MV-FMG and lL-21 coexpression. The degree of tumor regression of the not directly vector-treated tumor correlated with the anti-neoplastic response of the directly vector-treated tumor. This regression was mediated by a tumor-specific CTL response.CONCLUSION： Our data indicate that intratumoral expression of measles virus fusogenic membrane glycoproteins is a promising tool both for direct tumor treatment as well as for tumor vaccination approaches that can be further enhanced by cytokine coexpression.

原发与继发免疫性血小板减少症患者血小板膜糖蛋白特异性抗体及其与出血评分关系的研究

目的探讨原发与继发免疫性血小板减少症(ITP)患者抗GPⅡb/Ⅲa及GPΙb/Ⅸ抗体的表达、抗体阳性表达与出血评分的关系,以及不同抗体类型出血评分的差异,为原发与继发ITP患者的诊治和出血严重程度提供临床依据。方法选取2013年10月—2020年8月在新疆医科大学第一附属医院诊断为原发ITP患者(42例)及继发ITP患者(42例)为研究对象,所有患者入院时采用ITP出血评分量表行出血评分。应用改良MAIPA法检测患者抗GPⅡb/Ⅲa和抗GPΙb/Ⅸ抗体。结果原发与继发组患者抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体阳性率比较,差异无统计学意义(P>0.05),原发组患者抗体表达以抗GPⅡb/Ⅲa抗体为主,继发组抗体表达以抗GPΙb/Ⅸ抗体为主,两者抗体阳性构成比较,差异有统计学意义(P<0.05)。继发组患者整体出血程度较原发组重(P<0.05)。抗体阳性表达与出血程度的关系:(1)抗GPⅡb/Ⅲa抗体阳性患者出血程度较抗体阴性患者重(P<0.05)。(2)抗GPΙb/Ⅸ抗体阳性患者出血程度较抗体阴性患者重(P<0.05),双抗体阳性患者出血程度较抗体阴性患者重(P<0.05)。结论抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体阳性表达对原发与继发ITP无鉴别意义,但原发ITP患者抗体表达以抗GPⅡb/Ⅲa抗体为主,继发ITP患者抗体表达以抗GPΙb/Ⅸ抗体为主。继发ITP患者临床出血程度较原发ITP患者重。抗GPⅡb/Ⅲa抗体、抗GPΙb/Ⅸ抗体及抗体双阳性患者出血程度较抗体阴性患者重。

龙柴降血方对原发性血小板增多症患者凝血功能及血小板活化蛋白的影响

目的观察龙柴降血方对原发性血小板增多症(ET)患者凝血功能及血小板活化蛋白的影响。方法选取2020年1月—2021年10月中国中医科学院西苑医院收治的ET患者22例,给予龙柴降血方治疗4个月,观察治疗前后血栓弹力图指标[凝血时间(R)、凝血速率(K)、纤维蛋白功能(Angle)、血栓最大振幅(MA)、血栓最大振幅时间(TMA)、即时振幅(A)、30 min振幅(A30)、从血样开始检测到曲线分叉所需时间(SP)、血凝块溶解时间(CLT)]、高凝状态发生率和积分变化。选取其中15例ET患者,采用平行反应监测(PRM)技术检测患者治疗前后血小板活化蛋白[血小板膜糖蛋白Ibα(GPIbα)、糖蛋白VI(GPVI)、选择素-P(SELP)]表达水平,分析比较血小板聚集改善患者和血小板聚集增强患者上述各蛋白表达水平的差异;取5例健康者和10例ET患者治疗前后血清,采用Western blot法检测血清GPIbα表达情况;分析15例ET患者MA与血清GPIbα表达水平的相关性。结果与治疗前比较,龙柴降血方治疗后MA、CLT、高凝状态发生率、高凝状态积分和血清GPIbα、GPVI表达水平均明显降低(P均<0.05),血清SELP表达水平无明显变化(P>0.05)。治疗后血小板聚集改善患者血清GPIbα表达水平明显低于血小板聚集增强患者(P<0.05),GPVI表达水平无明显变化(P>0.05)。与健康人相比,10例ET患者治疗前血清GPIbα相对表达量明显增高(P<0.05);10例ET患者治疗后血清GPIbα相对表达量较治疗前明显降低(P<0.05)。龙柴降血方治疗后MA下降与GPIbα表达下调呈正相关(r=0.6986,P<0.05)。结论龙柴降血方能够抑制ET患者血小板聚集,改善高凝状态,其机制可能与抑制GPIbα蛋白介导的血小板活化信号通路有关。

慢性非缺血性心力衰竭患者血小板PAC-1和CD62p的表达及其与心功能的关系

目的探讨慢性非缺血性心力衰竭患者血小板膜糖蛋白纤维蛋白原受体(PAC-1)和P选择素(CD62p)的表达及其与心功能的关系。方法选取2019年1月至2022年1月天津市南开医院收治的185例慢性非缺血性心力衰竭患者作为观察组,根据纽约心脏病学会(NYHA)心功能分级将观察组患者分为心功能Ⅱ级组58例、心功能Ⅲ级组88例和心功能Ⅳ级组39例,根据左室射血分数(LVEF)将观察组患者分为射血分数保留心衰组(HFpEF)123例、射血分数临界值心衰组(HFmrEF)25例和射血分数下降心衰组(HFrEF)37例,再以B型钠尿肽(BNP)中位数为界将观察组患者分为高BNP组109例和低BNP组76例;另选取同期心功能正常的就诊者55例作为对照组。比较各组患者的基本临床信息、生化、BNP指标和PAC-1、CD62p的表达水平,采用Pearson相关分析法分析PAC-1、CD62p水平与BNP、LVEF的关系。结果观察组患者的PAC-1、CD62p水平分别为(23.67±16.54)%、(10.43±9.19)%,明显高于对照组的(9.33±9.63)%、(4.24±5.49)%,差异均有统计学意义(P<0.05);不同心功能分级患者对比发现,PAC-1、BNP水平为Ⅳ级>Ⅲ级>Ⅱ级>对照组,CD62p水平为Ⅳ级>Ⅲ级、Ⅱ级>对照组,差异均有统计学意义(P<0.05);不同LVEF分级患者对比发现,PAC-1水平为HFrEF组>HFmrEF组、HFpEF组>对照组,BNP水平为HFrEF组>HFmrEF组>HFpEF组>对照组,HFpEF组、HFmrEF组、HFrEF组CD62p水平分别为(10.72±9.17)%、(9.87±11.19)%、(9.84±7.88)%,明显高于对照组的(4.24±5.49)%,差异均有统计学意义(P<0.05);高BNP组患者的PAC-1、CD62p分别为(26.97±16.94)%、(11.05±9.62)%,明显高于低BNP组的(13.80±12.82)%、(6.97±7.54)%,差异均有统计学意义(P<0.05);经Pearson相关分析结果显示,PAC-1、CD62p与BNP呈正相关(r=0.363、0.182,P<0.05),PAC-1与LVEF呈负相关(r=-0.297,P<0.05)。结论慢性非缺血性心力衰竭患者的PAC-1、CD62p水平均明显增高,且与心功能有关,检测其水平有助于为患者的病情评估及临床治疗提供参考依据。

Deletion of the first glycosylation site promotes Lassa virus glycoprotein-mediated membrane fusion

The Lassa virus(LASV)is endemic in West Africa and causes severe hemorrhagic Lassa fever in humans.The glycoprotein complex(GPC)of LASV is highly glycosylation-modified,with 11 N-glycosylation sites.All 11 N-linked glycan chains play critical roles in GPC cleavage,folding,receptor binding,membrane fusion,and immune evasion.In this study,we focused on the first glycosylation site because its deletion mutant(N79Q)results in an unexpected enhanced membrane fusion,whereas it exerts little effect on GPC expression,cleavage,and receptor binding.Meanwhile,the pseudotype virus bearing GPC_(N79Q)was more sensitive to the neutralizing antibody 37.7H and was attenuated in virulence.Exploring the biological functions of the key glycosylation site on LASV GPC will help elucidate the mechanism of LASV infection and provide strategies for the development of attenuated vaccines against LASV infection.

GPIa C807T和G873A基因多态性对血小板聚集抑制率的影响

目的:探讨GPIa C807T和G873A基因多态性对血小板聚集抑制率的影响,为临床个体化抗血小板聚集治疗提供实验数据支持。方法:收集正在使用阿司匹林抗血小板聚集治疗的住院患者80例,采用血栓弹力图仪检测血小板聚集抑制率,根据聚集抑制率检测结果分为阿司匹林抵抗(aspirin resistance,AR)组(共10例)和阿司匹林敏感(aspirin sensitivity,AS)组(共70例),分析GPIa C807T、G873A基因型及其等位基因在两组分布状况以及两者患者临床资料与实验检测指标关系。结果:AR组807T、873G等位基因高于AS组(χ^(2)=4.039、4.354,P=0.044、0.037);AR组高血压病史患病率、卒中史高于AS组(χ^(2)=4.612、4.737,P=0.032、0.030);AR组血浆D-二聚体(D-D)、血清甘油三酯(TG)高于AS组(t=2.499、2.276,P=0.016、0.033),而AR组血小板聚集抑制率、血清葡萄糖低于AS组(t=15.352、3.076,P<0.001、0.005)。结论:GPIa C807T和G873A基因多态性与AR存在相关性,建议联合检测两基因位点的多态性并结合血小板聚集抑制率,共同指导临床个体化抗血小板聚集治疗。

血清上皮细胞膜结合黏液素1、α1-酸性糖蛋白水平与急性胆囊炎胆道炎症及预后的关系

目的分析血清上皮细胞膜结合黏液素1(MUC1)、α1-酸性糖蛋白(AAG)水平与急性胆囊炎病人炎症以及预后的关系。方法收集2020年7月至2022年4月内蒙古北方医院收治的急性胆囊炎病人136例,病人根据严重程度分为轻度组、中度组与重度组,另选取同期体检健康者130例作为对照组,制备研究对象血清标本,ELISA法检测血清MUC1、AAG水平,比较对照组与急性胆囊炎组、不同严重程度急性胆囊炎病人白细胞计数、中性粒细胞、血清总胆红素、血清MUC1、AAG、C反应蛋白(CRP)水平;根据急性胆囊炎病人治疗后预后情况将病人分为预后良好组与预后不良组,比较两组血清MUC1、AAG水平;lo⁃gistic回归分析血清MUC1、AAG水平与急性胆囊炎病人预后情况的关系;受试者操作特征(ROC)曲线分析血清MUC1、AAG水平对急性胆囊炎病人预后不良的预测价值。结果与对照组相比,急性胆囊炎组病人血清MUC1[(134.29±22.35)ng/L比(85.17±14.11)ng/L]、AAG水平[(0.84±0.14)g/L比(0.45±0.07)g/L]升高(P<0.05);与轻度组相比,中度组、重度组病人白细胞计数、中性粒细胞、血清总胆红素、血清MUC1、AAG、CRP水平升高(P<0.05);与中度组相比,重度组病人白细胞计数、中性粒细胞、血清总胆红素、血清MUC1、AAG、CRP水平升高(P<0.05);与预后良好组相比,预后不良组急性胆囊炎病人血清MUC1、AAG升高(P<0.05);血清MUC1、AAG是急性胆囊炎病人预后不良的危险因素(P<0.05);血清MUC1、AAG、二者联合检测对急性胆囊炎病人预后不良的预测AUC分别为0.80(灵敏度为61.4%,特异度为86.4%)、0.81(灵敏度为95.5%,特异度为53.4%)、0.88(灵敏度为88.6%,特异度为79.5%)。结论血清MUC1、AAG在急性胆囊炎中均升高,且与病人的炎症程度有关,对病人术后预后不良具有较高预测作用。

全麻下扁桃体切除术对OSAHS患儿发音特征及血液中PCT、GMP-140、T淋巴细胞亚群的影响

目的探讨全麻下扁桃体切除术对阻塞性睡眠呼吸暂停低通气综合征(OSAHS)患儿发音特征及血液中降钙素原(PCT)、血小板膜蛋白-140(GMP-140)、T淋巴细胞亚群等的影响。方法随机选取2020年6月—2022年3月在西安市儿童医院耳鼻咽喉头颈外科行全麻下扁桃体切除术的OSAHS患儿43例。分别于术前及术后第1、3个月时检测患儿发音特征及血清中PCT、GMP-140、T淋巴细胞亚群的水平,同时检测患儿的睡眠情况。结果与术前比较,术后第1、3个月患儿发/a/时最长发音时间明显延长,第二共振峰明显增强(F=43.07、39.65,P<0.05);与术前比较,术后第1、3个月患儿发/i/时最长发音时间明显延长,第四共振峰明显增强(F=21.43、44.25,P<0.05)。与术前比较,术后第1、3个月患儿血液中PCT、GMP-140、CD8^(+)水平及呼吸暂停低通气指数明显降低(F=29.67~112.86,P<0.05),血液中CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平及夜间最低血氧饱和度水平、魁北克睡眠问卷评分明显升高(F=12.15~234.11,P<0.05)。结论全麻下扁桃体切除术能够使OSAHS患儿发/a/、/i/时最长发音时间明显延长,发音特征明显改善,同时可降低患儿炎症反应,改善睡眠质量,并对恢复免疫功能有积极作用。

A Thylakoid Membrane Protein Functions Synergistically with GUN5 in Chlorophyll Biosynthesis

Chlorophyll(Chl)is essential for photosynthetic reactions and chloroplast development.While the enzymatic pathway for Chl biosynthesis is well established,the regulatory mechanism underlying the homeostasis of Chl levels remains largely unknown.In this study,we identified CBD1(Chlorophyll Biosynthetic Defect1),which functions in the regulation of chlorophyll biosynthesis.The CBD1 gene was expressed specifically in green tissues and its protein product was embedded in the thylakoid membrane.Furthermore,CBD1 was precisely co-expressed and functionally correlated with GUN5(Genome Uncoupled 5).Analysis of chlorophyll metabolic intermediates indicated that cbd1 and cbd1gun5 mutants over-accumulatedmagnesium protoporphyrin Ⅸ(Mg-Proto Ⅸ).In addition,the cbd1 mutant thylakoid contained less Mg than the wild type not only as a result of lower Chl content,but also implicating CBD1 in Mg transport.This was supported by the finding that CBD1 complemented a Mg^(2+)uptake-deficient Salmonella strain under low Mg conditions.Taken together,these results indicate that CBD1 functions synergistically with CHLH/GUN5 in Mg-Proto IX processing,and may serve as a Mg-transport protein to maintain Mg homeostasis in the chloroplast.

Biosynthesis of Callose and Cellulose by Detergent Extracts of Tobacco Cell Membranes and Quantification of the Polymers Synthesized in vitro

The conditions that favor the in vitro synthesis of cellulose from tobacco BY-2 cell extracts were determined. The procedure leading to the highest yield of cellulose consisted of incubating digitonin extracts of membranes from 11-day-old tobacco BY-2 cells in the presence of 1 mM UDP-glucose, 8 mM Ca^2＋ and 8 mM Mg^2＋. Under these conditions, up to nearly 40% of the polysaccharides synthesized in vitro corresponded to cellulose, the other polymer synthesized being callose. Transmission electron microscopy analysis revealed the occurrence of two types of structures in the synthetic reactions. The first type consisted of small aggregates with a diameter between 3 and 5 nm that associated to form fibrillar strings of a maximum length of 400 nm. These structures were sensitive to the acetic/nitric acid treatment of Updegraff and corresponded to callose. The second type of structures was resistant to the Updegraff reagent and corresponded to straight cellulose microfibrils of 2-3 nm in diameter and 200 nm to up to 5 μm in length. In vitro reactions performed on electron microscopy grids indicated that the minimal rate of microfibril elongation in vitro is 120 nm/min. Measurements of retardance by liquid crystal polarization microscopy as a function of time showed that small groups of microfibrils increased in retardance by up to 0.047 nm/min per pixel, confirming the formation of organized structures.

血小板膜糖蛋白特异性抗体在ITP患儿中的表达及其与疗效、出血程度的关系分析

目的分析血小板膜糖蛋白(GP)特异性抗体在免疫性血小板减少症(ITP)患儿中的表达情况及其与疗效、出血程度的关系。方法选取2018年2月至2020年12月于唐山市妇幼保健院儿科就诊的102例ITP患儿为研究对象,并将其纳入病例组,另选取同期同院进行体检的110例健康儿童纳入健康组。病例组患儿均接受糖皮质激素冲击疗法,以4周为1个疗程,共治疗5个疗程,并随访3个月。检测并比较两组GP特异性抗体表达情况,统计ITP患儿出血部位并分析不同GP特异性抗体出血程度、疗效及预后。结果病例组抗GPⅠb/Ⅸ抗体、抗GPⅡb/Ⅲa抗体表达水平高于健康组,差异有统计学意义(t值分别为16.521、51.718,P<0.05);102例ITP患儿抗GPⅠb/Ⅸ抗体阳性、抗GPⅡb/Ⅲa抗体阳性、双抗体阳性、双抗体阴性检出率分别为15.69%、4.90%、30.39%、49.02%,0级、1级、2级出血率分别为16.67%、50.00%、33.33%;且0级出血以双抗体阳性占比(0)最低,1级出血以抗GPⅡb/Ⅲa抗体阳性占比(80.00%)最高,2级出血以抗GPⅠb/Ⅸ抗体阳性、双抗体阳性占比(62.50%、48.39%)较高,差异有统计学意义(H=38.237,P<0.05);102例ITP患儿治疗5个疗程后有效率为78.43%,随访3个月后完全缓解率为68.63%;双抗体阴性ITP患儿有效率、完全缓解率均高于抗GPⅠb/Ⅸ抗体阳性、抗GPⅡb/Ⅲa抗体阳性、双抗体阳性ITP患儿,差异有统计学意义(χ^(2)值分别为19.866、27.709,P<0.05)。结论ITP患儿抗GPⅠb/Ⅸ抗体、抗GPⅡb/Ⅲa抗体表达水平较高,且其不同表达水平与患儿出血程度、疗效及预后均有密切关系,临床可通过监测GP特异性抗体来对ITP患儿进行早期诊断,并对其出血程度、疗效、预后等进行预测。

替罗非班联合丁苯酞治疗超时间窗急性脑梗死患者的效果评价

目的:探究与分析替罗非班联合丁苯酞治疗超时间窗急性脑梗死患者的效果。方法:选取2020年1月-2021年9月龙岩市第二医院神经内科收治的108例超时间窗急性脑梗死患者作为研究对象,采取随机数字表法分为对照组与观察组,每组54例。对照组给予丁苯酞治疗,观察组给予丁苯酞联合替罗非班治疗。对比两组临床疗效、治疗前后美国国立卫生研究院卒中量表(NIHSS)评分、凝血功能指标、血清血红素加氧酶-1(HO-1)、血管生成素-1(Ang-1)、P-选择素(P-selectin)及溶酶体膜糖蛋白(CD63),对比治疗期间两组不良反应。结果:观察组总有效率高于对照组,差异有统计学意义(P<0.05)。治疗后14、28 d,观察组NIHSS评分均较对照组低,差异均有统计学意义(P<0.05)。治疗后,观察组血清HO-1、P-selectin、CD63水平均较对照组低,Ang-1水平较对照组高,差异均有统计学意义(P<0.05)。两组治疗前后血小板计数、凝血酶原时间、国际标准化比值比较,差异均无统计学意义(P>0.05)。两组治疗期间不良反应发生率比较,差异均无统计学意义(P>0.05)。结论:超时间窗急性脑梗死患者行替罗非班联合丁苯酞治疗可改善神经功能缺损症状,调节血清HO-1、Ang-1、P-selectin及CD63水平,且不会对凝血功能带来不良影响,未增加明显的不良反应,安全性较高。

丹参饮对高脂血症模型大鼠血小板生理特性和功能的影响机制

目的基于血小板膜糖蛋白4(CD36)/磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)信号通路探讨丹参饮对高脂血症模型大鼠血小板生理特性和功能的影响机制。方法通过喂养高脂饮食复制高脂血症模型大鼠,将其随机分为模型组、辛伐他汀组(0.004 g/kg)和丹参饮高、低剂量组(3.6、0.9 g/kg),另设空白组(喂养基础饲料),每组10只。各给药组大鼠每天灌胃相应药物,其余组灌胃等体积生理盐水,连续4周。末次给药后,检测大鼠血脂生化指标[总胆固醇(TC)、三酰甘油(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)]含量,全血黏度、血浆黏度和血浆中纤维蛋白原(FIB)含量,血小板相关参数[血小板计数(PLT)、血小板分布宽度(PDW)和血小板平均体积(MPV)],血小板生理特性和功能相关因子[血管性血友病因子(vWF)、纤维连接蛋白(Fn)、磷脂酶A2(PLA2)、血栓素B2(TXB2)、6-酮-前列腺素F1α(6-keto-PGF1α)、血栓素A2(TXA2)、前列腺素(PGI2)、环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)、β-血小板球蛋白(β-TG)]水平,血小板聚集率(最大聚集率、60 s聚集率、180 s聚集率)和纤溶系统相关因子[组织型纤溶酶原活化剂(t-PA)、纤溶酶原激活物抑制剂1(PAI-1)]水平,血小板中CD36/PI3K/Akt信号通路相关蛋白[CD36、黏着斑激酶(FAK)、磷脂酰肌醇4-磷酸5-激酶(PIP5K)、PI3K、磷酸化Akt(p-Akt)、p-Akt1/2/3]表达水平。结果与空白组比较,模型组大鼠血浆中TC、TG、LDL-C含量,全血黏度、血浆黏度、血浆中FIB含量,血浆中PLT、MPV、vWF、Fn、PLA2、TXB2、TXA2、cGMP、β-TG水平,血小板最大聚集率、60 s聚集率,血浆中PAI-1水平,CD36、FAK、PIP5K、PI3K、p-Akt、p-Akt1/2/3蛋白表达水平均显著升高(P<0.05);HDL-C含量和6-keto-PGF1α、PGI2、t-PA水平均显著降低(P<0.05)。经丹参饮干预后,上述大部分指标均显著逆转(P<0.05)。结论丹参饮可改善高脂血症模型大鼠血小板生理特性和功能,其作用机制可能与下调CD36/PI3K/Akt信号通路活性、降低血小板活化程度有关。

血清GMP-140、sCD40L表达与重症肺炎预后的相关性

目的 探究重症肺炎患者氧化应激和可溶性CD40配体(sCD40L)、血小板α-颗粒膜糖蛋白140(GMP-140)表达及与预后关系。方法 选取2016年7月至2018年3月汕头市潮阳区大峰医院呼吸科收治的84例肺炎患者为研究对象,依据症状严重程度分为重症肺炎组、非重症肺炎组,选取同期健康体检者50名为对照组。检测受试者sCD40L、GMP-140水平,采用二元Logistic回归分析肺炎严重程度影响因素,采用受试者工作曲线(ROC)分析sCD40L、GMP-140水平对预后的预测价值。结果sCD40L、GMP-140水平:重症肺炎组>非重症肺炎组>健康对照组,存活组患者sCD40L、GMP-140表达水平均低于死亡组,差异有统计学意义(P<0.05)。sCD40L、GMP-140是肺炎病情影响因素,sCD40L、GMP-140预测肺炎患者死亡曲线下面积(AUG)为0.837、0.771。结论sCD40L、GMP-140在重症肺炎患者体内存在异常高表达,是重症肺炎患者的影响因素,对临床预测肺炎患者预后有一定应用价值。

SARS-CoV-2 membrane glycoprotein M antagonizes the MAVS-mediated innate antiviral response

A novel SARS-related coronavirus(SARS-CoV-2)has recently emerged as a serious pathogen that causes high morbidity and substantial mortality.However,the mechanisms by which SARS-CoV-2 evades host immunity remain poorly understood.Here,we identified SARS-CoV-2 membrane glycoprotein M as a negative regulator of the innate immune response.We found that the M protein interacted with the central adaptor protein MAVS in the innate immune response pathways.This interaction impaired MAVS aggregation and its recruitment of downstream TRAF3,TBK1,and IRF3,leading to attenuation of the innate antiviral response.Our findings reveal a mechanism by which SARS-CoV-2 evades the innate immune response and suggest that the M protein of SARSCoV-2 is a potential target for the development of SARS-CoV-2 interventions.