乳腺癌雌激素受体、孕激素受体和C-erbB-2的表达及其临床意义

目的 :研究乳腺癌中雌、孕激素受体 (ER、PR)与C -erbB -2癌基因的表达情况及临床意义。方法 :应用免疫组化S -P法 ,对 15 8例原发性乳腺癌组织进行了ER、PR和C -erbB -2检测 ,并进行统计学分析。结果 :ER、PR和C -erbB -2的表达率分别为 6 8 4 %、6 2 0 %、5 4 4 %。ER的表达与PR的表达呈显著正相关 ( P <0 0 1) ;C -erbB -2的表达与淋巴结转移呈正相关 (P <0 0 5 ) ,与ER、PR的表达呈显著负相关 (P <0 0 1)。C -erbB -2表达阴性及ER、PR表达阳性 4年无复发生存率及总生存率较高。结论 :ER、PR的测定有利于选择乳腺癌内分泌治疗。C -erbB -2可作为判断乳腺癌预后的重要指标 ,C -erbB -2阴性者预后较好。     

乳腺癌辅助化疗诱发2型糖尿病患者预后因素及生存分析

目的总结乳腺癌辅助化疗诱发2型糖尿病(type 2 diabetes mellitus,T2DM)的临床病理特征,探讨其预后影响因素及生存情况。方法选取2009年1月30日—2014年1月30日沈阳军区总医院肿瘤科收治的女性乳腺癌辅助化疗患者184例,将化疗诱发T2DM者46例作为观察组,未诱发T2DM者138例作为对照组。比较两组临床病理特征,采用Cox单因素和多因素回归分析乳腺癌辅助化疗患者不良预后影响因素,并随访观察两组3年、5年无病生存率。结果两组文化程度、糖尿病家族史、体重指数(BMI)、血脂异常、C-反应蛋白(CRP)、血清胱抑素C(Cys-c)、雌激素受体(ER)阴性、孕激素受体(PR)阴性、化疗方案、辅助放疗、辅助靶向治疗方面比较差异均有统计学意义(P0.05或P0.01)。Cox多因素回归分析显示:ER阴性、CRP多次异常、化疗诱发T2DM是乳腺癌辅助化疗患者不良预后的独立危险因素(95%CI:0.294-0.702,P=0.006;95%CI:1.405-2.931,P=0.039;95%CI:1.592-2.837,P=0.019)。观察组3年、5年无病生存率(65.2%、54.3%)显著低于对照组(81.9%、70.3%),差异有统计学意义(χ~2=4.911,P=0.026;χ~2=4.324,P=0.038)。结论 ER阴性、CRP多次异常、化疗诱发T2DM是乳腺癌辅助化疗患者不良预后的独立危险因素,乳腺癌辅助化疗诱发T2DM者较未诱发T2DM者无病生存期缩短。     

乳腺导管上皮病变中细胞增殖和凋亡作用及其与ER、PR、C-erbB-2表达关系的研究

目的 :探讨乳腺导管上皮病变中细胞增殖和凋亡作用及其与ER、PR、C -erbB - 2表达关系的临床病理意义。方法 :采用免疫组织化学Envision二步法和TUNEL法检测正常乳腺导管上皮 2 0例 ,增生性乳腺导管上皮 4 6例 ,导管原位癌 12例和 6 0例乳腺浸润性导管癌增殖指数 (ProliferativeIndex ,PI)和凋亡指数 (ApoptoticIn dex ,AI)以及ER、PR、C -erbB - 2表达 ,分析乳腺导管上皮病变中细胞增殖指数和凋亡指数及其与ER、PR、C-erbB - 2表达率的关系。结果 :正常乳腺导管上皮、增生性导管上皮、导管原位癌、浸润性导管癌及其随病理组织学分级愈高 ,细胞增殖指数 (PI)和C -erbB - 2阳性表达指数 (CI)逐渐增加 ,细胞凋亡 (AI)逐渐减少 ,且正常乳腺导管上皮与乳腺导管原位癌 (重度非典型增生 )细胞增殖指数 (PI)、细胞凋亡 (AI)、C -erbB - 2阳性表达 (CI)及ER、PR表达 ,差异有显著性 (P <0 0 5 )。乳腺导管上皮病变细胞增殖与ER、C -erbB - 2表达呈正相关 (r=0 5 4 ,r=0 79,P <0 0 5 ) ;乳腺导管上皮病变细胞凋亡与ER、C -erbB - 2表达呈负相关 (r =-0 6 2 ,r=- 0 73,P <0 0 5 )。结论 :细胞增殖和凋亡、ER、PR及C -erbB - 2在乳腺导管上皮病变发生发展和预后判断过程中可能起重要作用 ,且它们皆可能作为乳     

幽门螺杆菌感染与胃癌P53、C-erbB-2表达的关系

目的　探讨幽门螺杆菌感染与胃癌P53 、C erbB - 2的关系。方法　应用Warthin -Starry染色法检测幽门螺杆菌 ,免疫酶组化SP法检测P53 、C -erbB - 2基因蛋白。结果　Hp感染率在胃癌和癌前病变组中均高于对照组 ,而前两组差异无显著性 ,Hp感染与C erbB 2过度表达有关。P53 仅在胃癌组织中表达 ,Hp感染者与P53 过度表达有关 (P <0 0 5 )。结论　Hp感染可能作为促进剂使P53 失活及C -erbB - 2基因激活 ,说明Hp感染在胃癌发生中起一定作用     

环氧合酶-2、微卫星稳定表达与结直肠癌术后患者含奥沙利铂化疗方案预后的关系

目的 探讨结直肠癌组织环氧合酶-2(COX-2)、微卫星稳定(MSS)表达与结直肠癌患者术后含奥沙利铂化疗方案预后的关系.方法 选取70例结直肠癌患者作为研究对象,患者均在本院行结直肠癌根治术,且术后采用含奥沙利铂辅助化疗方案治疗,观察结直肠癌组织COX-2、MSS表达和患者无病生存时间.结果 70例患者中,COX-2和MSS阳性表达者34例(双阳组),COX-2阳性表达者12例和MSS阳性表达者11例(单阳组),COX-2和MSS阴性表达者13例(双阴组);双阳组、单阳组患者TNM分期Ⅲ期者占比分别为58.82％、43.40％,均高于双阴组,差异有统计学意义(P＜0.05);双阳组患者中位无病生存期为14个月(95％ CI为13.22～14.78),短于单阳组的24个月(95％ CI为21.53 ～26.47)和双阴组的34个月(95％ CI为27.35 ～ 40.65),差异有统计学意义(P＜0.05);Cox比例风险回归分析结果显示,TNM分期、COX-2、MSS表达是结直肠癌患者术后含奥沙利铂化疗方案预后的影响因素(HR=2.354、1.103、1.155,P＜0.05).结论 COX-2、MSS阳性表达可影响结直肠癌患者术后应用含奥沙利铂化疗方案的效果,缩短无病生存时间.     

即刻早期反应基因-1在乳腺癌新辅助化疗后表达的变化及对预后的意义

目的探讨即刻早期反应基因-1(IEX-1)在乳腺癌新辅助化疗(NCT)后表达的变化及在预后判断中的意义。方法选取2012年1月至2017年1月首诊首治并接受NCT的110例乳腺癌女性患者作为研究对象进行回顾性研究,采用二步法免疫组化方法检测NCT前后乳腺癌组织中IEX-1表达的变化,采用Kaplan-Meier和Log-rank法分析其对无病生存期(DFS)及总生存期(OS)的影响。结果 NCT前乳腺癌组织中IEX-1蛋白均阴性;NCT后,IEX-1蛋白阳性表达45例(40. 9%),阴性65例。生存分析显示,IEX-1蛋白阳性表达组患者的DFS、OS均低于IEX-1蛋白阴性表达组(P均<0. 01)。多因素Cox比例风险回归模型分析发现,IEX-1蛋白阳性表达为乳腺癌预后的独立风险因素[HR=4. 921,95%CI=(1. 273~19. 018),P=0. 021]。结论乳腺癌NCT前后检测IEX-1有助于预后判断,IEX-1可能成为乳腺癌NCT疗效的预后指标。     

晚期乳腺癌应用多西他赛联合卡培他滨行新辅助化疗的临床观察

目的探讨晚期乳腺癌应用TX（多西他赛联合卡培他滨）方案行新辅助化疗的效果及安全性。方法回顾性分析55例Ⅲ-Ⅳ期用TX方案行新辅助化疗的晚期乳腺癌的临床资料。结果新辅助化疗的总有效率为89.09%（49/55）,有67.27%（37/55）的患者分期降低,患者的无病生存期平均为59.3个月,5年生存率为38.18%。新辅助化疗主要不良反应为胃肠道反应和骨髓抑制,多为Ⅰ级或Ⅱ级,Ⅲ/Ⅳ级不良反应主要为中性粒细胞减少（12.72%,7/55）和手足综合征（9.09%,5/55）,所有患者均无化疗相关性死亡。结论应用TX方案行新辅助化疗能降低晚期乳腺癌患者的分期,为手术创造最大机会,减少或延缓肿瘤复发、转移,并可延长晚期乳腺癌患者的无病生存期,不良反应可以耐受。     

非蒽环类辅助化疗方案治疗老年乳腺癌患者的安全性和耐受性观察

探讨非蒽环类辅助化疗方案治疗老年乳腺癌患者的安全性和耐受性。选取我院收治的老年乳腺癌患者50例,按随机分组原则分为AC化疗组和PC化疗组,比较两组患者化疗毒性反应、复发率、无病生存期和总生存期。与AC化疗组比较,PC化疗组各项毒性反应发生率明显降低(P0.05);两组患者复发率、2年无病生存率和2年总生存率比较差异无显著性(P0.05)。PC化疗方案在耐受性和安全性上优于AC化疗方案,适合应用于老年乳腺癌患者的化疗中。     

P-糖蛋白在乳腺癌组织中表达与预后的关系

目的 研究P-糖蛋白（P-glycoprotein，PGP）在乳腺癌组织中的表达。评估其在乳腺癌预后中的作用。方法 采用免疫组织化学方法（Immunohistochemistry，IHC）检测60例手术切除的乳腺癌组织中PGP的表达，并分析其与临床、病理特征的关系及对预后的影响。结果 PGP在乳腺癌组织中的阳性表达率为81．7％（49／60例），其中高表达者26例（43．3％）；PGP的表达与月经状况、肿瘤大小、淋巴结转移、组织分级和激素受体状况差异均无显著性意义（P〉0．05）；Kaplan-Meier生存分析结果表明PGP表达与无病生存期和总生存期差异均有显著性意义（P〈0．01）；Cox多因素分析显示肿瘤大小、腋淋巴结转移和雌激素受体状况是影响无病生存期和总生存期的重要因素（P〈0．05），同时也显示PGP表达和总生存期明显相关，而和无病生存期无关。结论 PGP在乳腺癌组织中具有较高的表达，PGP有可能成为判断乳腺癌预后的有用指标。     

乳腺癌雌激素受体在新辅助化疗中的预期意义

目的探讨雌激素受体 (ER)在新辅助化疗中乳腺癌组织表达的预期意义。方法采用免疫组织化学法检测 14 2例乳腺癌组织中ER的表达状况 ,并分析其在新辅助化疗中的预期意义。结果ER的表达与肿瘤的临床化疗反应显著相关 (P <0 .0 1) ,新辅助化疗前后ER的表达无明显变化 (P >0 .0 5 )。结论ER在新辅助化疗乳腺癌患者中有重要的预期意义 ,在决定化疗后患者是否行内分泌治疗时 ,应考虑化疗对ER功能的影响。     

Mechanisms of factor VIIa‐catalyzed activation of factor VIII

Summary. Background: Factor (F)VIIa, complexed with tissue factor (TF), is a primary trigger of blood coagulation, and has extremely restricted substrate specificity. The complex catalyzes limited proteolysis of FVIII, but these mechanisms are poorly understood. Objectives: In the present study, we investigated the precise mechanisms of FVIIa/TF‐catalyzed FVIII activation. Results: FVIII activity increased ～4‐fold within 30 s in the presence of FVIIa/TF, and then decreased to initial levels within 20 min. FVIIa (0.1 nm ), at concentrations present physiologically in plasma, activated FVIII in the presence of TF, and this activation was more rapid than that induced by thrombin. The heavy chain (HCh) of FVIII was proteolyzed at Arg⁷⁴⁰ and Arg³⁷² more rapidly by FVIIa/TF than by thrombin, consistent with the enhanced activation of FVIII. Cleavage at Arg³³⁶ was evident at ～1 min, whilst little cleavage of the light chain (LCh) was observed. Cleavage of the HCh by FVIIa/TF was governed by the presence of the LCh. FVIII bound to Glu‐Gly‐Arg‐active‐site‐modified FVIIa (K_d, ～0.8 nm ) with a higher affinity for the HCh than for the LCh (K_d, 5.9 and 18.9 nm ). Binding to the A2 domain was particularly evident. Von Willebrand factor (VWF) modestly inhibited FVIIa/TF‐catalyzed FVIII activation, in keeping with the concept that VWF could moderate FVIIa/TF‐mediated reactions. Conclusions: The results demonstrated that this activation mechanism was distinct from those mediated by thrombin, and indicated that FVIIa/TF functions through a ‘priming’ mechanism for the activation of FVIII in the initiation phase of coagulation.     

Long-lasting antithrombotic effects of a single dose of human recombinant, active site-blocked factor VII: insights into possible mechanism(s) of action

Summary.  Tissue factor (TF) is important in initiating intravascular thrombosis. We demonstrated that active-site blocked factor VII (FVIIai) inhibits intravascular thrombosis for at least 6 h following a single injection, despite FVIIai plasma half-life was ∼45 min. The aims of the present study were: (a) to determine the duration of the antithrombotic effects of a single injection of FVIIai; and (b) to assess whether FVIIai prolonged effects can be explained by a slow dissociation rate from TF in the arterial wall. Cyclic flow variations (CFVs), obtained in stenotic rabbit carotid arteries with endothelial injury, were abolished by either FVIIai (100 µg kg⁻¹ min⁻¹ for 10 min) or hirudin (1 mg kg⁻¹). After CFVs were abolished, carotid blood flow velocity was recorded continuously for 24 h. CFVs restored in all hirudin-treated animals after 2.1 ± 0.3 h, while they restored in only four of nine FVIIai-treated rabbits in 10.1 ± 2.2 h. Five animals in this group did not show restoration of CFVs up to 24 h. Immunohistochemistry revealed that FVIIai was still bound to the arterial wall 24 h following its administration, despite at this time FVIIai plasma levels were undetectable. Prothrombin time and partial thromboplastin time did not change significantly. FVIIai exerts potent, long-lasting antithrombotic effects without affecting systemic hemostatic parameters; a possible mechanism is a slow dissociation rate of FVIIai from TF. These proprieties make FVIIai particularly attractive as an antithrombotic intervention.     

Binding studies of the enzyme (factor IXa) with the cofactor (factor VIIIa) in the assembly of factor-X activating complex on the activated platelet surface

Summary.  Activated platelet membranes expose binding sites for the enzyme factor (F)IXa, the substrate (FX) and the cofactor (FVIIIa) that colocalize to assemble the FX-activating complex and promote optimal rates of FX activation. To determine the stoichiometry and affinity of binding to activated platelets, coordinate, equilibrium binding studies with enzyme (¹²⁵I-FIXa) and cofactor (¹³¹I-FVIII or ¹³¹I-FVIIIa) were carried out in the presence of saturating concentrations of substrate (FX). Results of these studies indicate that in the presence of FX (1.5 µ m ), the enzyme (active-site-inhibited Glu-Gly-Arg-FIXa, EGR-FIXa) and procofactor (FVIII) bind to an equal number (approximately 700 sites/platelet) of receptors whereas the active cofactor (FVIIIa) binds an additional approximately 500 high-affinity FVIIIa binding sites per platelet (K_d approximately 0.8 n m ). With excess zymogen (FIX) to block shared FIX/FIXa-binding sites, the stoichiometry of ¹²⁵I-FIXa and ¹³¹I-FVIIIa binding was 1 : 4. These FIXa/FVIIIa binding studies together with previously reported evidence of the coordinate binding of FVIIIa and FX to equivalent numbers of binding sites on activated platelets provide strong evidence to support the conclusion that FVIIIa comprises the receptor that presents FX to FIXa for efficient catalysis on the activated platelet membrane.     

Glycosylation of tissue factor is not essential for its transport or functions

See also Morrissey JH. Low‐carb tissue factor? This issue, pp 1508–10.DOI: 10.1111/j.1538‐7836.2011.04332.x . Summary. Background: Glycosylation plays an important role in protein function. The importance of glycosylation for tissue factor (TF) function is unclear. Objective: The aim of the present study is to investigate the importance of TF glycosylation in transport to the cell surface and its coagulant and signaling functions. Methods: Endothelial cells and peripheral blood mononuclear cells (PBMC) were treated with tunicamycin to inhibit N‐linked glycosylation. Site‐specific mutagenesis of one or more potential N‐linked glycosylation sites in TF was used to generate TF mutants lacking glycans. TF expression at the cell surface was determined in binding assays using ¹²⁵I‐FVIIa or ¹²⁵I‐TF mAb and confocal microscopy. TF coagulant activity was measured by factor (F) Xa generation assay, and TF signaling function was assessed by measuring cleavage of protease activated receptor 2 (PAR2) and activation of p44/42 MAPK. Results: Tunicamycin treatment reduced TF activity at the endothelial cell surface; however, this reduction was found to be the result of decreased TF protein production in tunicamycin‐treated cells. Tunicamycin treatment had no significant effect on TF activity or antigen levels in PBMC. No significant differences were observed in TF protein expression and procoagulant activity among cells transfected to express either wild‐type TF or TF mutants. A fully non‐glycosylated TF is shown to bind FVIIa and interact with FX with the same efficiency as that of wild‐type TF. Non‐glycosylated TF is also capable of supporting FVIIa cleavage of PAR2 and PAR2‐dependent p44/42 MAPK activation. Conclusions: Glycosylation is not essential for TF transport and coagulant or signaling functions.     

蛋白因子的实验研究

本文系从3～5个月的胎儿肝脏细胞中提取蛋白因子并做理化检查:pH、蛋白含量及分子量、氨基酸、甲胎蛋白、GBsAg 和微量元素、生物学、鸭肝动物模型治疗急性肝衰竭以及对 PLc 细胞分泌HBsAg 的抑制作用试验。结果提示蛋白因子各项指标符合临床肌注要求,具有抑制肝炎病毒的作用,能显著提高肝衰竭动物的存活率,改善肝功能,刺激肝 DNA 合成,加速肝组织修复等功能。     

Molecular mechanisms of mild and moderate hemophilia A

Summary.  Mutations responsible for mild/moderate hemophilia A were extensively characterized over the last 15 years and more than 200 mutations have been identified. However, most of the molecular mechanisms responsible for the reduced factor (F)VIII levels in patients' plasma were determined only recently. Recent progresses in the study of the FVIII molecule three-dimensional structure provided a major insight for understanding molecular events leading to mild/moderate hemophilia A. This allowed prediction of mutations impairing FVIII folding and intracellular processing, which result in reduced FVIII secretion. Mutations potentially slowing down FVIII activation by thrombin were also identified. A number of mutations were also predicted to result in altered stability of activated FVIII. Biochemical analyses allowed identification of mutations reducing FVIII production. Mutations impairing FVIII stability in plasma, by reducing FVIII binding to von Willebrand factor (VWF) were also characterized. Defects in FVIII activity, notably slow activation by thrombin, or abnormal interaction with FIXa, were also recently demonstrated. Biochemical analysis of FVIII variants provided information regarding the structure/function relationship of the FVIII molecule and validated predictions of the three-dimensional structure of the molecule. These observations also contributed to explain the discrepant activities recorded for some FVIII variants using different types of FVIII assays. Altogether, the study of the biochemical properties of FVIII variants and the evaluation of the effects of mutations in three-dimensional models of FVIII identified molecular mechanisms potentially explaining reduced FVIII levels for a majority of patients with mild/moderate hemophilia A. It is expected that these studies will improve diagnosis and treatment of this disease.     

The rediscovery and isolation of TFPI

Summary.  Tissue factor pathway inhibitor (TFPI) is a multivalent Kunitz-type proteinase inhibitor that produces factor (F)Xa-dependent feedback inhibition of the factor VIIa/tissue factor (FVIIa/TF) catalytic complex that is responsible for the initiation of coagulation. Since 1985, when Rapaport and colleagues reported that the lipoprotein fraction of plasma contained a FXa-dependent inhibitor of FVIIa/TF, myriad articles have established its biochemical structure, its mechanism of action, and its physiological importance. This brief personal account reviews historical studies that established the existence of the inhibitor and the events that led to its initial isolation.     

The promise and challenges of bioengineered recombinant clotting factors

The past 10 years of clinical experience have demonstrated the safety and efficacy of recombinant clotting factors. With the adoption of prophylactic strategies, there has been considerable progress in avoiding the complications of hemophilia. Now, insights from our understanding of clotting factor structure and function, mechanisms of hemophilia and inhibitors, gene therapy advances and a worldwide demand for clotting factor concentrates leave us on the brink of embracing targeted bioengineering strategies to further improve hemophilia therapeutics. The ability to bioengineer recombinant clotting factors with improved function holds promise to overcome some of the limitations in current treatment, the high costs of therapy and increase availability to a broader world hemophilia population. Most research has been directed at overcoming the inherent limitations of rFVIII expression and the inhibitor response. This includes techniques to improve rFVIII biosynthesis and secretion, functional activity, half-life and antigenicity/immunogenicity. Some of these proteins have already reached commercialization and have been utilized in gene therapy strategies, while others are being evaluated in pre-clinical studies. These novel proteins partnered with advances in gene transfer vector design and delivery may ultimately achieve persistent expression of FVIII leading to an effective long-term treatment strategy for hemophilia A. In addition, these novel FVIII proteins could be partnered with new advances in alternative recombinant protein production in transgenic animals yielding an affordable, more abundant supply of rFVIII. Novel rFIX proteins are being considered for gene therapy strategies whereas novel rVIIa proteins are being evaluated to improve the potency and extend their plasma half-life. This review will summarize the status of current recombinant clotting factors and the development and challenges of recombinant clotting factors bioengineered for improved function.     

健康成人血小板膜糖蛋白的表达特征

目的　观察健康成人血小板６ 种膜糖蛋白（ Ｇ Ｐ） 在不同性别、不同年龄,静息状态与活化状态时的表达特征。方法　用流式细胞仪测定静息状态下及用凝血酶受体活化肽（ Ｔ Ｒ Ａ Ｐ） 激活的血小板所结合单抗的分子数。结果　男性静息血小板和活化血小板中 Ｇ Ｐ Ⅱｂ／ Ⅲａ 和 Ｇ Ｐ Ⅲａ 显示随年龄减少现象,在静息血小板中 Ｇ Ｐ Ⅱｂ／ Ⅲａ 和 Ｇ ＰⅢａ 值与年龄呈负相关,ｒ 值分别为 － ０ ．４０９ 和－ ０ ．５３４ ,其 Ｐ 值均＜ ０ ．０５ ;而女性中未见到类似结果。 Ｔ Ｒ Ａ Ｐ 激活后 Ｇ Ｐ Ｉｂ 在血小板表面表达下降,而 Ｇ Ｐ Ⅱｂ／Ⅲａ 、Ⅲａ 、 Ｐ选择素均增高,其中 Ｐ选择素在激活后的升高值与激活前的基础值呈负相关。静息血小板的 Ｇ ＰⅠｂ 分子数约为３０ ０００ ,它与 Ｇ Ｐ Ⅴ的比值约３∶１ 。结论　健康人血小板 Ｇ Ｐ表达存在性别和年龄差异。