Effect of Xiaoyu Zhixue Tablet (消瘀止血片) on the expression of platelet membrane glycoprotein I b/IX/V complex in patients with chronic renal failure

ABSTRACT Objective： To investigate the effect of Xiaoyu Zhixue Tablet （消瘀止血片, XYZXT） on the expression of platelet membrane glycoprotein （GP） Ⅰ b/Ⅸ/Ⅴ complex and GP Ⅰ b α in patients with chronic renal failure （CRF） in early metaphase. Methods： Fifty-one patients with CRF in early metaphase （treated group） were treated with XYZXT, 3 months as the course of treatment for 2 courses. The previous therapies remained unchanged. Flow cytometry was used to assess the expression of platelet GP Ⅰb/Ⅸ/Ⅴ complex and GP Ⅰb α in patients with CRF, and turbidity method was used to determine the platelet maximum aggregation rate （MAR）, meanwhile the renal function was measured. The final data were compared with those before the treatment, and with those in the normal control group （31 healthy subjects）. Results： Compared with the normal control group, expressions of GP Ⅰ b/Ⅸ/Ⅴ complex and GPⅠb α, and platelet MAR in CRF patients were significantly lower （P=0.007,P=0.001,P=0.009） before the treatment; after the treatment with XYZXT, the above indexes in CRF patients were remarkably increased （P=0.033,P=0.026, P=0.045）, but still lower than those in the normal control group, however, it was not statistically significant. Conclusion： （1） The expression of GP Ⅰ b/Ⅸ/Ⅴ complex in CRF patients of early metaphase was decreased, which lead to platelet aggregation dysfunction. This might be one of the reasons for the hemorrhagic trend in CRF. （2） XYZXT was able to upgrade expressions of GP Ⅰb/Ⅸ/Ⅴ complex and GPⅠb α in CRF patients, improve platelet function and clown-regulate platelet activation in patients with CRF.     

出血性血小板病患者凝血酶受体GPIb／IX／V复合物表达研究

目的观察出血性血小板病患者的血小板膜糖蛋白（GP）Ib／IX／V复合物及其组分GP Ibα的表达。方法采用流式细胞术检测68例出血性血小板病患者在出血期的血小板GPIb／IX／V复合物和GP Ibα的表达，采用比浊法检测其血小板最大聚集率（MA），并与33例出血控制者及32例正常健康者比较。结果出血性血小板病患者在出血期间GPIb／IX／V复合物、GP Ibα表达明显低于正常组（P均〈0．01），出血控制者的GPIb／IX／V复合物、GP Ibα表达与正常组比较无显著性差异（P均〉0．05）。结论血小板GPIb／Ⅸ／V复合物作为凝血酶和血管性血友病因子的受体，在出血性血小板病患者体内的表达下降，导致血小板聚集功能障碍，可能是本病反复发作出血倾向的重要原因之一。     

益气活血方治疗出血性血小板病临床疗效观察

目的：观察益气活血方治疗出血性血小板病的临床疗效。方法将212例出血性血小板病患者随机分为两组，中药组142例给予益气活血方治疗；西药组70例给予安络血治疗，两组均治疗8周。观察两组治疗后止血疗效及血小板聚集缺陷恢复率。结果中药组止血总有效率为83.82％，西药组为46.15％，两组比较差异有统计学意义（ P＜0.01）；中药组血小板聚集功能恢复率为67.94％，西药组仅为5.17％，中药组明显优于西药组（ P＜0.01）。结论益气活血方治疗出血性血小板病疗效优于常规西药。     

益气活血解毒法对溃疡性结肠炎血浆血小板活化的临床及实验研究

[目的]探讨益气活血解毒法对溃疡性结肠炎(UC)血浆血小板活化的影响。[方法]分临床研究及动物实验两部分;临床研究以90例溃疡性结肠炎复发患者按随机数字表法随机分为2组,中药治疗组45例,西药对照组45例。治疗3个月后对完全缓解及有效病例(其中治疗组39例,对照组32例)进行为期6个月的随访。治疗组给予益气活血解毒立法的中药汤剂,对照组给予柳氮磺胺吡啶治疗,并各与20例作正常对照。动物实验用三硝基苯磺酸(TNBS)复制实验性大鼠UC模型,将其随机分为模型组、西药对照组、中药治疗组,并设正常对照组。进行反映血小板活化程度的特异性标志物P-选择素的检测。[结果]临床研究中,治疗前UC患者血浆P-选择素水平明显高于正常健康人(P<0.01);治疗组治疗后及随访时含量明显低于西药对照组(P<0.05或P<0.01)。动物实验中,在治疗10 d时,两治疗组皆可降低血浆P-选择素水平,西药组水平明显低于中药组(P<0.05);治疗30 d时中药组与西药组相比无显著差异(P>0.05);停药10 d时,中药组明显低于西药组(P<0.05)。[结论]以益气活血解毒立法的中药溃结复发方与西药比较,可有效降低血浆P-选择素水平,从而阻抑血小板活化,可能是抗溃结复发作用机制之一。     

腺苷二磷酸在凝血酶信号传递过程中的作用

目的比较腺苷二磷酸（ADP）在两类凝血酶受体活化过程中对血小板聚集率以及血小板膜表面活化标记物表达的影响,探讨ADP在凝血酶信号传递机制中的作用。方法分别以凝血酶受体（PAR）活化肽PAR1-AP与PAR4-AP诱导血小板活化,观测腺苷三磷酸双磷酸酶（Apyrase,ADP抑制剂）ⅤⅡ作用过程中血小板聚集以及血小板膜表面糖蛋白（GP）Ib与P-选择素的改变。结果两类凝血酶受体都能诱导血小板活化,产生完全的聚集波,血小板膜GPIb则呈现先进行性减少后逐渐回升的可逆性变化,P-选择素水平持续升高。ApyraseⅤⅡ作用后,PAR1-AP诱导的血小板聚集反应受到部分抑制,而PAR4-AP诱导的血小板聚集反应未有明显改变。ApyraseVⅡ加速PAR1途径GPIb回复细胞表面,对PAR4途径中GPIb的改变则没有显著影响。两种活化途径中P-选择素的表达都不受ApyraseVⅡ的影响。结论ADP在凝血酶信号传递过程中发挥重要作用,其中PAR1途径与ADP参与密切相关．     

老年慢性肺心病患者GPⅡb／Ⅲa和P—selectin的变化及意义

摘要：目的探讨老年慢性肺心病患者血小板活化标志物GPⅡb／Ⅲa、p-selectin的变化及其与Pa02、PaC02的关系。方法用三色全血流式细胞术测定36例老年慢性肺心痛患者外周血中血小板GPⅡb／Ⅲa、P—selectin的表达水平,检测患者动脉血氧分压、二氧化碳分压,并与28例老年健康对照者及28例非老年健康对照者比较。结果老年慢性肺心病GPⅡb／Ⅲa、P-selectin均明显高于老年健康对照组及非老年健康对照组（均P〈0．001）。且与动脉血氧分压呈负相关、与动脉血二氧化碳分压呈正相关。老年健康对照组GPⅡb／Ⅲa、P—selectin阳性表达率亦高于非老年健康对照组（均P〈0．05）。结论老年慢性肺心病患者血小板明显活化,其活化程度与PaO2、PaCO2关系密切。     

Two-year observation of the clinical efficacy in treating chronic hepatitis B patients with Ganxian recipe and lamivudine

Objective:To evaluate the clinical efficacy of Ganxian recipe (GXR) and lamivudine (LVD) in a two-year treatment of chronic hepatitis B (CHB).Methods: One hundred and twenty patients with CHB were randomly divided into the combinedly treated group (combined group) of 40 CHB patients who were treated with GXR combined with LVD. Another 40 CHB patients were treated with LVD alone (WM group), and still another 40 CHB patients were treated with GXR alone (TCM group). All these cases were randomly controlled and observed for two years.Results: Comprehensive efficacy: Total effective rate of the combined group (complete response and partial response) was 92.5%, while that of the WM group was 67.5% and TCM group 57.5%, respectively, with the difference between them was significant (P<0.01); after treatment, the hepatic functions (AST, ALT, SB) of the three groups were all reduced, and the reduction in the combined group was particularly significant in comparison with the WM group or TCM group,P< 0.05 orP<0.01 respectively, suggesting that the effect in the combined group was better than that in the other two groups; the rate of tyrosine-methionine-aspartate-aspartate (YMDD) virus mutation: it was 7.5% in the combined group, 40.0% in the WM group, and 5.0% in the TCM group; liver fibrosis improvement parameter: after treatment, the results in the combined group got better than those in the other two groups.Conclusion: GXR could inhibit the appearance of YMDD after long-term application of LVD, and combined use has marked synergism.     

血小板膜糖蛋白Ⅱb／Ⅲa受体激活在再灌注心肌无复流形成中的作用

OBJECTIVE: To observe the changes in the activity of platelet glycoprotein (GP) IIb/IIIa receptor following the occurrence of no-reflow after myocardial reperfusion, to explore the measures for clinical management of this condition. METHODS: Nine canine models of no-reflow following myocardial ischemia verified by reperfusion myocardial contrast echocardiography were used in this investigation. The peripheral venous blood (PVB) and sinus venous blood (SVB) were sampled before myocardial reperfusion (after a 3-hour myocardial ischemia) and after a 3-hour reperfusion for determinating GP IIb/IIIa receptor activities by means of enzyme-linked immunosorbent assay (ELISA). RESULTS: The activity of platelet GP IIb/IIIa of PVB, either in 3-hour ischemic group or in 3-hour reperfusion group, was elevated significantly (P < 0.001), suggesting platelet activation. Elevated platelet GP IIb/IIIa receptor activity in SVB occurred only in 3-hour ischemic group, and the 3-hour reperfusion group (P < 0.001) and the no-reflow group had significantly decreased activities, which was more obvious in the latter group (P < 0.01). The findings signified the consumption of the platelet GP IIb/IIIa receptors in the myocardial microcirculation and the presence of platelet aggregation. CONCLUSIONS: Myocardial ischemia may activate the platelets to express GP IIb/IIIa receptors through stress response mechanism and damage the endothelia of the microvasculature to incur adhesion and aggregation of the platelets within the microcirculation, which may be one of the pathogeneses for myocardial no-reflow phenomenon.     

剪切力作用对血小板膜糖蛋白分子表达的影响

目的　探讨剪切力活化血小板的作用机制。方法　用改制的锥板黏度计对健康人的全血标本分别施加 10 0、15 0、10 0 0、30 0 0s-1的剪切力作用后 ,以血小板膜糖蛋白 (GP)的荧光标记抗体、单色或三色荧光法染色血小板 ,用流式细胞术检测血小板膜PAC 1(活化的GPIIb/IIIa)、CD6 2P(P 选择素 )、GPIb/IX和GPIIb/IIIa的水平。结果　PAC 1、CD6 2P在静止的血小板表面表达很低 ,阳性率分别为 1 7%± 1 1% (n =5 )和 0 9%± 0 5 % (n =5 ) ;受到高剪切力作用后表达水平增加 ,以PAC 1增加更为迅速、明显 ,于 30 0 0s-1剪切力作用 0 5min时达峰值 (73 6 %± 7 4 % ) ,之后很快开始下降 ,CD6 2P的表达相对较缓慢 ,但在观察时间内呈持续上升趋势 ,30 0 0s-1剪切力作用 7min时 ,阳性率为2 6 4 %± 3 5 %。GPIb/IX、GPIIb/IIIa存在于 96 5 %～ 99 4 %的静止血小板表面。血小板膜GPIb/IX水平在低剪切力作用下无明显变化 ,高剪切力作用 1min内有不同程度增加 ,但随后开始下降 ,30 0 0s-1作用 7min时 ,平均荧光强度由静止时的 72 4± 6 7降低到 4 4 9± 4 9(n =5 )。GPIIb/IIIa水平在高剪切力作用下迅速增加 ,30 0 0s-1作用 7min时 ,平均荧光强度由静止时的 98 5± 12 1增高到 15 9 5±2 3 6 (n =5 )。结论　高     

癌胚抗原相关细胞黏附分子源性多肽KM17对血小板活化作用的研究

目的 探讨癌胚抗原相关细胞黏附分子（CEACAM1）源性多肽KM17对血小板聚集、释放、黏附功能的影响。方法 采用血小板聚集仪观察多肽KM17对凝血酶、胶原、花生四烯酸（AA）、二磷酸腺苷（ADP）等激动剂诱导的血小板聚集的影响;流式细胞术观察多肽KM17对ADP激活血小板后P-选择素释放的影响;显微镜下观察多肽KM17对血小板静态黏附于胶原基质的影响。结果 多肽KM17能显著促进ADP诱导的血小板聚集且呈剂量依赖（P <0.05）,而对AA、胶原及凝血酶诱导的血小板聚集差异均无统计学意义（P>0.05）;此外,多肽KM17对ADP激活血小板后P-选择素释放及血小板静态黏附于胶原基质的差异也无统计学意义（P>0.05）。结论 多肽KM17可明显促进ADP诱导的血小板聚集,但对ADP活化血小板后P-选择素释放及血小板静态黏附于胶原基质未见明显作用。     

中西医结合治疗成人原发性肾病综合征50例

[目的]观察采用中西医结合方法治疗原发性肾病综合征的临床疗效。[方法]全部96例病人随机分为中西医组(n=50)和西医组 (n=46)。西医组给予常规治疗,中西组在此基础上应用中药治疗,均用活血化瘀治疗 (血府逐瘀汤加减或血府逐瘀胶囊加减)。[结果]中西医组总有效率为86 9%,西药组总有效率为53 3%,两组比较有显著性差异(P<0.01)。两组均有不同程度的糖皮质激素或/和细胞毒性药物副作用,中西医组副作用发生明显轻于西医组,两组比较有显著性差异(P<0.01)。[结论]采用中西医结合方法治疗成人原发性肾病综合征可以提高疗效,降低副作用发生。     

Use of Traditional Chinese Medicine in Chinese Patients with Coronary Heart Disease

Objective To study the use of traditional Chinese medicine(TCM)or both TCM and guideline-recommended Western medicine(WM)in Chinese patients with coronary heart disease(CHD).Methods A cross-sectional nationwide survey of 2803 CHD outpatients was completed by collecting information,including general demographic data,disease history,and use of drugs(including TCM and WM).Results Of the 2712 CHD outpatients with complete drug treatment data,only 3.1% received TCM without any WM for CHD,30.0% received both TCM and WM recommended by current CHD guidelines,and 66.9% received only WM.Patients with a longer history of CHD or with a history of stroke,were more likely to use TCM.However,90.6% of CHD patients who used TCM also used certain WM.Furthermore,patients who used more types of TCM tended to use much less WM recommended by current guidelines.Conclusion A substantial proportion of Chinese CHD outpatients use both TCM and WM for secondary prevention of CHD.It is important to assess the effect of combined TCM and WM on major clinical outcomes in Chinese CHD patients.     

人血小板功能抗体及其片段的识别

目的研究特发性血小板减少性紫癜（ITP）患者血小板膜糖蛋白（GP）特异性IgG抗体及其片段的免疫活性及对血小板聚集功能的影响.方法用改良单克隆抗体特异性俘获血小板抗原技术（MAIPA）检测84例慢性ITP患者血浆中抗GPⅡb/Ⅲa、GPⅠb/Ⅸ及GPⅥ自身抗体,比浊法血小板聚集试验筛选出自身抗体阳性并能抑制血小板聚集的患者,用蛋白A柱纯化其血浆IgG抗体并用胃蛋白酶制备F（ab＇）2片段.检测纯化的IgG抗体及其酶切片段与血小板GP的结合活性及对正常人血小板聚集功能的影响.结果 （1）84例ITP患者血浆中,48例（57.1%）抗GPⅡb/Ⅲa和/或GPⅠb/Ⅸ和/或GPⅥ自身抗体阳性,其中7例（14.6%）明显抑制了二磷酸腺苷、瑞斯托霉素或胶原诱导的血小板聚集;（2）纯化的IgG及F（ab＇）2片段具有与相应血小板GP的结合活性;（3）4例患者纯化IgG及F（ab＇）2片段具有抑制血小板聚集的功能.结论 F（ab＇）2片段是IgG自身抗体的功能片段,它不但保留了良好的抗原结合活性且可部分抑制血小板聚集功能,为人源化血小板糖蛋白特异性抗体的制备奠定了基础.     

活性氧在血小板储存损伤引起的GPIbα酶切中的作用研究

目的 活性氧(ROS)在血小板活化和凋亡过程中起重要作用,血小板在发生活化和凋亡时伴随有糖蛋白(GP) Ibα酶切,血小板储存损伤(PSL)主要由血小板活化和凋亡引起。本研究主要探讨ROS在PSL引起的GPIbα酶切中的作用和分子机制。 方法 取健康志愿者单采血小板,(22±2)℃震荡,分别在保存的1、3、5 d用无菌接管机取一定量血小板,离心得到沉淀和上清,上清用酶标仪检测糖萼蛋白(GC);沉淀用缓冲液重悬,经洗涤2次后得到洗涤血小板;洗涤血小板与不同的检测探针孵育后,用流式细胞仪检测细胞内ROS浓度、线粒体跨膜电位去极化、磷脂酰丝氨酸(PS)外翻和P-选择素表达;Western blot检测p38丝裂原活化蛋白激酶(MAPK)的磷酸化。 结果 随着保存时间的延长,血小板P-选择素表达、PS外翻和线粒体跨膜电位去极化逐渐增多,血小板胞内ROS浓度逐渐增多,p38 MAPK磷酸化逐渐增多、血浆中GC含量逐渐增多;ROS拮抗剂抑制血小板胞内ROS增多、抑制p38 MAPK磷酸化、减少血浆GC含量;p38 MAPK抑制剂抑制p38 MAPK磷酸化和减少血浆GC含量,不抑制ROS浓度增加。 结论 ROS在PSL引起的GPIbα酶切过程中起重要作用,ROS可能通过活化p38MAPK进而引起GPIbα酶切。      

阿司匹林抵抗与GPⅢa基因多态性的相关性研究

目的研究阿司匹林抵抗患者的临床特征及其与血小板糖蛋白GPⅢa Leu33Pro基因位点多态性的关系。方法收集规律服用阿司匹林预防失败的脑梗死患者60例及服用相同剂量阿司匹林的社区体检者24例,以对照组95%尿11-脱氢-血栓素B2（11-DH-TXB2）水平的上界为标准判定阿司匹林抵抗,将病例组分为阿司匹林抵抗组（AR）和阿司匹林敏感组（AS）。比较AR组和AS组的临床特征及血小板糖蛋白GPⅢaLeu33Pro基因位点多态性。结果根据尿11-DH-TXB2的水平,发现31例患者存在AR,AR的发生率为51.67%。两组间年龄、性别,既往脑梗死病史、高血压及糖尿病患病率、TC、TG、HDL、LDL、FBG、PLT、WBC、RBC及Hb水平差异均无统计学意义（均〉0.05）。AR组和AS组GPⅢa Leu33Pro基因型及等位基因的分布无差别。结论温州地区阿司匹林预防失败脑梗死人群中AR的发生率是51.67%,与AS组相比,未发现明显临床特征。GPⅢa Leu33Pro基因位点多态性可能与AR的发生无关。     

溃疡性结肠炎患者血浆P-选择素、血小板膜糖蛋白Ⅰb/Ⅸ/Ⅴ复合物与疾病活动性的关系

目的探索溃疡性结肠炎(ulcerative colitis,UC)患者血浆P-选择素(P-selectin)、血小板膜糖蛋白(GP)Ⅰb/Ⅸ/Ⅴ复合物与疾病活动性的关系。方法选择28例UC活动期、16例UC缓解期患者和26例健康志愿者,采用ELISA法检测血浆P-selectin,比浊法检测血小板最大聚集率(MAR),并采用流式细胞仪检测GPⅠb/Ⅸ/Ⅴ复合物。结果活动期UC患者血浆P-selectin、血小板MAR较缓解期患者及健康者明显升高(P0.01或P0.05),而GPⅠb/Ⅸ/Ⅴ复合物的表达显著降低(P0.01或P0.05)。结论溃疡性结肠炎患者存在血小板激活,血浆P-selectin和GPⅠb/Ⅸ/Ⅴ复合物作为血小板激活的标志物,与疾病的活动程度相关,可以作为UC活动期的指标。     

中药自拟方联合低分子肝素钙治疗晚期恶性肿瘤高凝状态的临床研究

[目的] 观察中药自拟方联合低分子肝素钙对晚期恶性肿瘤患者血液高凝状态的临床疗效及安全性。[方法] 选取晚期恶性肿瘤高凝状态患者70例,随机分为对照组（低分子肝素钙）和试验组（中药自拟方+低分子肝素钙）,每组35例。观察两组患者治疗前和治疗14 d后的血小板计数（PLT）、纤维蛋白原（FIB）、D-二聚体（D-D）、中医血瘀证候评分、卡氏评分（KPS）及安全性指标情况。[结果] 两组治疗14 d后的PLT、FIB、D-D、血瘀证候积分、KPS评分均较治疗前改善,且试验组优于对照组,差异有统计学意义（P<0.05）。两组治疗前后安全性方面,对照组有1例患者出现少量牙龈出血,试验组无不良反应发生。[结论] 中药自拟方联合低分子肝素钙可以更好的改善晚期恶性肿瘤患者血液高凝状态,优于单纯低分子肝素钙治疗。     

Effect of Qingre Buyi Decoction on nitric oxide and histomorphology of intestinal mucosa in rats with radiation enteritis

Objective: To explore the mechanism of Qingre Buyi Decoction (QBD) in prevention and treatment of radiation enteritis in rats.Methods: Forty-eight Wistar rats were randomly divided into four groups, the TCM group, the WM group, the model group and the control group, 12 in each group. Rats in the former three groups were given orally with QBD, norfloxacin and normal saline once a day for 7 successive days, after being irradiated with X-ray at single dose of 10 Gy for modeling of radiation enteritis, while rats in the control group were untreated. Animals were sacrificed at the end of the medication. NO concentration, mean height and number of villi per centimeter in their small intestinal mucosa were measured.Results: The intestinal NO concentration was significantly lower in the TCM and WM groups than that in the model group(P<0.05), while the number of villi was significantly more and the height higher in the former two groups than those in the model group (P<C0.01 for both), but no significant difference was shown between the TCM group and the WM group.Conclusion: QBD could inhibit the production of NO, increase the number and height of intestinal villi in rats with radiation enteritis, suggesting that it could reduce the inflammatory reaction of intestinal mucosa to irradiation, protect mucosa from radiation damage, and promote the regeneration of mucosa.     

受体酪氨酸激酶Axl调控糖蛋白Ⅵ介导的血小板活化

目的研究生长静止特异性基因6的受体Axl对胶原蛋白受体糖蛋白Ⅵ介导的血小板活化的调控作用。方法利用Axl基因敲除小鼠和糖蛋白Ⅵ特异性刺激剂,在体外分别研究血小板的聚集和铺展功能,并应用流式细胞术检测血小板的颗粒释放和整合素αⅡbβ3的活化。结果与野生型小鼠相比,Axl基因敲除小鼠的血小板聚集和铺展功能受到明显抑制;流式细胞术检测结果表明,Axl敲除的血小板的颗粒释放标记物CD62P的表达明显降低（P〈0.05）,整合素αⅡbβ3的活化同样受到明显抑制（P〈0.05）。结论生长静止特异性基因6的受体Axl在糖蛋白Ⅵ介导的血小板活化中发挥重要的调控作用。