猪纤维蛋白原免疫原性研究

目的 研究猪纤维蛋白原的免疫原性和免疫效果,为该药物安全评价和进一步优化提供实验依据.方法 建立生物素-亲和素系统改良的ABC-ELISA检测抗体;新西兰白兔创伤试验考察血清抗体效价及其动态变化;免疫器官质量及脏器指数分析、噻唑蓝(MTT)比色法检测脾淋巴细胞增殖、氯化硝基四氮唑蓝(NTB)还原法检测中性粒细胞吞噬能力,揭示猪纤维蛋白原对小鼠免疫系统的影响.结果 1周后新西兰白兔血清即可检测出抗体,且第2周抗体含量达到峰值后下降至较稳定水平,抗体效价也呈现相同的变化趋势;小鼠免疫器官质量及脏器指数均未发生明显改变;与对照组相比,各实验组小鼠淋巴细胞增殖指数无明显差异;实验组小鼠中性粒细胞吞噬能力显著性增强.结论 猪纤维蛋白原刺激新西兰白兔机体后产生体液免疫应答,但在小鼠体内未表现出特异性免疫反应.     

海南汉族人群九个纤维蛋白原基因多态性及其与血浆纤维蛋白原水平的关系

目的 调查海南汉族健康人群αTaqⅠ和βBclⅠ、HinfⅠA／C、448G／A、βBsmA ⅠG／C、＋1689T／G、-148C／T、-249C／T、-455G／A多态性的等位基因频率及其与血浆纤维蛋白原（fibrinogen，Fg）水平的关系。方法 用比浊法测定238名健康个体的血浆Fg浓度，用PCR-限制性片段长度多态性方法及测序法分析多态性基因型，并用协方差分析比较9个位点的基因型与血浆Fg水平的关系。结果 海南汉族人群αTaqⅠ、βBclⅠ、HinfⅠA／C、C448、βBsmAⅠG／C、＋1689T／G、-148C／T、-249C／T、-455G／A稀有位点的等位基因频率分别为0．445、0．239、0．134、0．235、0．273、0．241、0．265、0．441、0．254，9个位点之间存在连锁不平衡；在总人群中，-455GA和AA基因型、-148CT和TT基因型、αTaqⅠT1T1基因型组的血浆Fg水平比野生型组高（P值均〈0．01）；在男性人群中，-455GA和AA基因型、-148CT和TT基因型、αTaqⅠT1T1、αTaqⅠT1T2基因型组的血浆Fg水平比野生型组高（P值均〈0．01）。在女性人群中，携带稀有位点A^-455、T^-148、αTaⅠT1的基因型组与野生型组之间的血浆赡水平差异无统计学意义。结论 在9个多态性位点之间存在连锁不平衡；A^-455、T^-148、αTaqⅠT1位点与血浆Fg水平增高关联，β-Fg-455G／A、-148C／T及α-TaqⅠ多态性与男性血浆Fg水平关联。     

EBV-associated diffuse large B-cell lymphoma arising in the chest wall with surgical mesh implant

In Japan, most cases of malignant lymphoma arising in the thorax are pyothorax-associated lymphoma, which develops in patients who have undergone artificial pneumothorax, used in the past as surgical therapy for pulmonary tuberculosis. Pyothorax-associated lymphoma consist mostly of diffuse large B-cell lymphoma and have a strong association with EBV. Herein is reported the case of a diffuse large B-cell lymphoma arising in the left thoracic wall after left lung resection for squamous cell carcinoma and chest wall reconstruction with polyethylene terephthalate (PET) surgical mesh. The tumor was found 20 years after the operation and was confined to the chest wall adjacent to the PET mesh. The patient did not have a clinical history of pyothorax after surgery. The lymphoma cells were of the large cell type and were positive for CD20, EBV-encoded small RNA--in situ hybridization, LMP-1 and EBNA-2. The present case demonstrates that EBV-related B-cell lymphoma can occur after surgery other than artificial pneumothorax. In the present case, long-standing chronic inflammation induced by PET mesh may have been associated with the development of lymphoma.     

Mechanical properties of electrospun fibrinogen structures

Fibrin and fibrinogen have a well-established track record in tissue engineering due to their innate ability to induce improved cellular interaction and subsequent scaffold remodeling compared to synthetic scaffolds. Use of fibrinogen as a primary scaffold component, however, has been limited by traditional processing techniques that render scaffolds with insufficient mechanical properties. The goal of this study was to demonstrate, based on mechanical properties, that electrospun fibrinogen overcomes these limitations and can be successful as a tissue engineering scaffold or wound dressing. Electrospun fibrinogen scaffolds were characterized for fiber diameter and pore area and subsequently tested for uniaxial mechanical properties while dry and hydrated. In addition, uniaxial mechanical testing was conducted on scaffolds treated to regulate scaffold degradation in serum-containing media by supplementing the media with aprotinin or cross-linking the scaffolds with glutaraldehyde vapor. A linear relationship between electrospinning solution concentration and measured fiber diameter was seen; fiber diameters ranged from 120 to 610 nm over electrospinning concentrations of 80 to 140 mg/ml fibrinogen, respectively. Pore areas ranged from 1.3 microm(2) to 13 microm(2) over the same fibrinogen concentrations. Aprotinin in the culture media inhibited scaffold degradation in a predictable fashion, but glutaraldehyde vapor fixation produced less reliable results as evidenced by mechanical property testing. In conclusion, the mechanical characteristics of electrospun fibrinogen strongly support its potential use as a tissue engineering scaffold or wound dressing.     

Effect of fibrinogen on functional activity of blood leukocytes

Fibrinogen intensified luminol-dependent chemiluminescence of blood leukocytes stimulated with opsonized zymosan. It is hypothesized that fibrinogen stimulates and prolongs functional activity of leukocytes during inflammation.     

纤维蛋白原活性在冠心病发展中的作用

目的:探讨纤维蛋白原活性升高在冠心病发展中的可能作用。方法:用血液流变学方法检测冠心病患者稳定期和心绞痛患者的纤维蛋白原活性。结果:冠心病患者血浆纤维蛋白原水平及其血浆粘性显著高于正常对照组(P＜0.01,P＜0.05),并且心绞痛患者纤维蛋白原活性和血小板聚集性(Pt max、Pt H、Pt K)也同时高于正常对照组(P＜0.01),并发现心绞痛组纤维蛋白原活性值与Pt max、PtH呈显著负相关(r＝-0.8379,P＜0.01;r=-0.8784,P＜0.01)。结论:纤维蛋白原活性升高在冠心病发展中起一定作用。     

医用生物蛋白胶及其主体成分纤维蛋白原的免疫原性研究

研究医用生物蛋白胶及其主体成分纤维蛋白原的潜在免疫原性。采用SDS-PAGE电泳、Western blot分离确定主体胶成分中的纤维蛋白原蛋白,建立间接ELISA法检测抗体条件;新西兰白兔创伤实验及BALB/c小鼠实验考察医用生物蛋白胶及纤维蛋白原的免疫原性。结果表明,生物蛋白胶主体成分以0.5μg/ml包被时,可有效检测抗纤维蛋白原抗体。新西兰白兔创伤实验中,实验组兔血清1周后出现抗体,2周后抗体水平明显升高,6周后抗体水平下降,呈先升后降的动态变化趋势;小鼠实验中,NBT试验未发现阳性中性粒细胞,MTT检测到T细胞呈现弱增殖能力。研究表明,医用生物蛋白胶主体胶成分中的纤维蛋白原在兔出现一过性抗体,医用生物蛋白胶对小鼠未表现出明显的免疫原性。     

Dynamics of interrelationships between the content of lipoprotein particles,fibrinogen, and leukocyte count in the plasma from patients with coronary heart disease treated with Kwai

The hypolipidemic effect of Kwai, a preparation based on garlic powder (Allium sativum), depended on the initial content of cholesterol and/or triglycerides. This effect was most pronounced in patients with coronary heart disease with initial cholesterol >7.0 mmol/liter and triglyceride >1.92 mmol/liter. After treatment with Kwai the correlation between triglyceride content and leukocyte count remained unchanged, the correlation between cholesterol content and leukocyte count disappeared, fibrinogen concentration decreased by 11%, and no correlation was found between fibrinogen content and leukocyte count.     

纤维蛋白原及相关基因多态性与脑梗塞类型的关系

目的 研究纤维蛋白原（fibrinogen，Vg）浓度、分子聚合功能及FgBβ—148、448基因多态性与脑梗塞类型的关系。方法 采用病例对照研究，应用聚合酶链反应-限制性酶切法进行基因多态性分析，并测定血浆Fg浓度及其分子聚合功能参数。结果 FgBβ-148变异基因型人群脑动脉主干支梗塞组血浆Fg浓度高于脑动脉穿通支梗塞组及对照组；448变异基因人群脑动脉主干支梗塞组Fg浓度、纤维蛋白单体聚合速率／最大吸光度、纤维蛋白单体聚合速率高于对照组，Fg浓度高于脑动脉穿通支梗塞组。结论 FgBβ-148、448基因型发生变异后可通过影响Fg浓度而使此人群发生脑动脉主干支梗塞组的危险性升高。     

血浆纤维蛋白原水平在胃癌淋巴结转移诊断中的临床意义

目的观察胃癌患者血浆纤维蛋白原水平与肿瘤临床分期、浸润程度和淋巴结转移的关系。方法结合临床资料,回顾分析我科收治的1064例胃癌患者的血浆中纤维蛋白原含量。结果血浆纤维蛋白原和胃癌分化程度、TNM分期、浸润深度、淋巴结转移及是否有远处转移有密切关系。PN0～PN3血浆纤维蛋白原水平分别为(2.43±0.74)g/L,(3.57±1.16)g/L,(3.98±0.81)g/L,(4.52±0.74)g/L。多因素Logistic回归分析表明高纤维蛋白原血症是胃癌淋巴结转移的一个危险因素。结论术前血浆纤维蛋白原水平可以作为预测胃癌淋巴结转移的一个指标。     

严重急性呼吸综合征患者外周血纤维蛋白原的变化及意义

目的探讨纤维蛋白原(FIB)水平在严重的急性呼吸综合征(Sever acute respiratory syndrome,SARS)中的变化及意义。方法回顾性分析了148例SARS患者外周血纤维蛋白原(Flb)浓度,将148例患者按病程分为初期组(44例)和非初期组(104例)两组;按临床分型分为普通型(87例)和重型(61例)两组;按肺部病变分单叶病变(49例),双叶病变(56例)和弥漫性病变(56例)三组,然后分组进行比较。结果148例SARS患者外周血纤维蛋白原(Flb)水平,其均值(-x±s):522.29±154.87mg/dl,明显高于正常参考值(350mg/dl),差异有统计学意义(P<0.001);初期组血纤维蛋白原水平高于非初期组(P<0.05);随着患者肺部病变范围增大从单叶病变、双叶病变到弥漫性病变,三组患者血纤维蛋白原水平是逐渐升高的(P<0.05)。结论严重急性呼吸综合征(SARS)患者血纤维蛋白原浓度的检测有助于SARS病情的观察和预后的判断,从而提示适时给予纤溶治疗可作为一种提高疗效的尝试。     

急性心肌缺血时冠状窦血浆纤维蛋白原的改变及意义

通常人们认为,急性心肌缺血可引起血浆纤维蛋白原(Fg)的增高;但是在缺血区局部Fg的变化并不清楚。本文在17条犬上,以自制微米缩窄器造成冠脉左旋支狭窄与梗塞,观察了冠状窦Fg和血小板数(PC)的改变。结果表明:当冠脉狭窄大于75%后,急性心肌缺血可引起Fg含量的减少,当冠脉大狭窄于90%后,PC也出现减少。病理组织学检查在狭窄部位有内皮细胞的损伤、血小板的粘附及冠状动脉血栓和微血栓的形成。这一结果提示:急性心肌缺血可引起血浆Fg的减少,Fg的减少与血小板的聚集及血栓的形成有关。     

内毒素诱导血小板纤维蛋白原受体显露和磷脂酰肌醇代谢

本文测定了体外纤维蛋白原与人血小板纤维蛋白原受体的结合，研究内毒素对纤维蛋白原受体显露的影响。实验观察到内毒素可以使纤维蛋白原与血小板的结合显著增加，５００μｇ／ｍｌ内毒素使血小板结合的￣（１２５）Ⅰ－纤维蛋白原从对照的５．２±３．５ｎｇ／１０￣８ｃｅｌｌｓ增高为１９．１±６．５ｎｇ／１０￣８ｃｅｌｌｓ（Ｐ＜０．０１，ｎ＝４），表明内毒素可诱导血小板纤维蛋白原受体显露。用Ｓｃａｔｃｈａｒｄ作图法求得Ｋｄ＝２．０５×１０￣－８ｍｏｌ／Ｌ，Ｂｍａｘ＝９６３纤维蛋白原分子／血小板。内毒素能够诱导血小板生成ＰＡ。５００μｇ／ｍｌ内毒素作用９０秒可使ＰＡ增高为对照的１７６±２２％（Ｐ＜０．０５，ｎ＝６）。ＰＡ升高迅速，９０秒时达到高峰，提示ＰＩＰ＿２的分解。上述结果提示内毒素可以直接激活血小板，显露纤维蛋白原受体，可能是通过诱导ＰＩＰ＿２分解，产生三磷酸肌醇（ＩＰ＿３）和甘油二酯（ＤＧ），进而使受体的构型发生改变，显露受体活性。     

一个纤维蛋白原γ链Arg275His突变导致的遗传性异常纤维蛋白原血症家系

目的　对一个遗传性异常纤维蛋白原血症家系进行表型和基因型分析。方法　采集家系3代5人外周血,吸取上层血浆用血凝仪检测活化部分凝血酶原时间、凝血酶原时间、凝血酶时间、蛋白C活性、蛋白S活性和抗凝血酶活性,纤维蛋白原活性和抗原分别用Clauss法和免疫比浊法进行检测。以常规酚-氯仿法抽提家系所有成员外周血基因组DNA,PCR扩增纤维蛋白原基因FGA、FGB和FGG所有外显子及其侧翼序列,PCR产物纯化后直接测序以检测基因突变。结果　先证者活化部分凝血酶原时间、凝血酶原时间正常,凝血酶时间超出正常上限值2倍以上,纤维蛋白原活性明显下降,抗原也低于正常范围,且活性显著低于抗原;其母表型检测结果与之相似。基因分析显示先证者呈纤维蛋白原FGG基因第8外显子g.5 6 78G>A杂合碱基置换,导致Arg2 75 His错义突变,该突变来源于母系。结论　纤维蛋白原γ链Arg2 75 His杂合错义突变是引起该家系异常纤维蛋白原血症的原因。     

Effect of UV-modified fibrinogen on platelet aggregation in platelet-rich plasma

Oxidized UV-modified fibrinogen activates platelets in platelet-rich plasma. Kinetic turbidimetry showed that addition of oxidized fibrinogen to platelet-rich plasma led to platelet aggregation. Reversible aggregation is recorded starting from the 30th second and then constantly grows with the same rate. Nonoxidized fibrinogen produced no such effect. The relationship between aggregation intensity and rate and the degree of fibrinogen oxidation was described by a bell-shaped curve with a peak corresponding to 24% fibrinogen oxidation. The amplitude of aggregation increased with increasing the concentration of irradiated fibrinogen from 0.1 to 1.0 mg/ml and then plateaued. The rate of aggregation little depended on fibrinogen concentration.     

纤维蛋白原和血常规检测对乙型肝炎的诊断价值

目的 探讨纤维蛋白原和血常规检查对乙型肝炎的诊断意义.方法 选取2012年3月-2015年3月来我院就诊确诊为慢性乙型肝炎的患者80例作为观察组,同时收集健康志愿者40例作为对照组.所有研究对象都检测血常规和纤维蛋白原两类指标,具体血常规检测项目包括红细胞(RBC)、血红蛋白(HGB)、白细胞(WBC)、淋巴细胞(LY),纤维蛋白原(Fbg)检测项目包括纤维蛋白单体聚合反应速率(FMPS)、最大吸光度(A-max)、纤维蛋白原含量(FC)、反应延滞时间(DT)、纤维蛋白原功能指数(FI).结果 观察组的红细胞、血红蛋白、白细胞较对照组明显减少(P＜0.05),而淋巴细胞则显著增多(P＜0.05).观察组的FMPS、A-max、FC、DT、FI各项指标较对照组均明显降低(P＜0.05).结论 血常规检测中红细胞、血红蛋白、白细胞、淋巴细胞可作为乙型病毒性肝炎早期筛查指标.血浆纤维蛋白原在一定程度上可反应肝功能状态和肝细胞坏死程度,可作为早期诊断及判断乙型肝炎疾病严重程度的重要指标.     

Matrix stiffness affects spontaneous contraction of cardiomyocytes cultured within a PEGylated fibrinogen biomaterial

Successful implementation of cardiac cell transplantation for treating damaged myocardium relies on the development of improved injectable biomaterials. A novel biomaterial technology using PEGylated fibrinogen has been developed with controllable physicochemical properties based on the poly(ethylene glycol) (PEG) constituent. In addition, the fibrinogen backbone of the material confers inherent bioactivity to cells. The purpose of this investigation was to explore by in vitro techniques the use of this biomaterial as a scaffold for cardiac tissue regeneration. To this end neonatal rat cardiomyocytes were cultivated in PEGylated fibrinogen constructs. The cell-seeding density and biomaterial composition were optimized to obtain maximum spontaneous contraction of the constructs. Quantitative characterization of the contraction pattern was accomplished by video image analysis. It was possible to demonstrate an inverse correlation between the material stiffness and the amplitude of contraction of the tissue constructs by changing the modulus of the matrix using different compositions of PEG and fibrinogen. The relationship between matrix stiffness, cell density and tissue contraction also provided some insight into the mechanism of cellular remodeling that ultimately leads to synchronized contraction of the constructs. These findings indicate that PEGylated fibrinogen hydrogels can be used as a scaffold for cardiomyocytes, and offer the possibility of controlling cellular remodeling via simple compositional modifications to the matrix.     

Lipoprotein(a): identification of subjects with a superbinding capacity for fibrinogen

We have previously shown that the binding of lipoprotein (a) [Lp(a)] to immobilized fibrinogen involves the domain located in kringles IV-5 to IV-8, but not kringle IV-10. In extending those studies to subjects living in Chicago and in the island of Sardinia, we found that about 6% of them had an Lp(a) with Bmax values of 27.7 + 6.0 fmol, which were about 5–8-fold higher than those of controls (3.4 + 2.8 fmol) and in the range of those observed for free apo(a) derived from the Lp(a) of controls (36.6 + 2.9 fmol). This superbinding phenotype was unaffected by age, sex, type of lipid disorder and hypolipidemic agents, and also had a familial incidence. We are currently exploring the hypothesis that this fibrinogen superbinding phenotype is due to conformational changes of apolipoprotein(a) [apo(a)] resulting from the lipid content and composition of the Lp(a) particle and/or sequence anomalies in the kringle domain IV-5 to IV-8.