ABO Blood Groups and Their Relationship with Coagulation Factor VIII in Healthy Adults

Background: ABO blood group distribution defers with racial and geographic variations. They are related to diseases like cardiovascular diseases, cerebral thromboembolism. ABO blood group system may influence coagulation factor VIII which may increase the future risk of thrombosis. Aim: To assess the relation of ABO blood group with coagulation factor VIII in healthy adults. Material and Methods: A prospective type of analytical cross-sectional study was conducted in the Department of Physiology, Dhaka Medical College, Dhaka from July 2019 to June 2020. After obtaining ethical clearance, a total of 190 healthy adults were selected from different areas of Dhaka city based on inclusion and exclusion criteria, with ages ranging from 18 - 45 years. The subjects were interviewed and detailed history regarding personal, family, medical and drug were taken. Prior to sample collection, informed written consent was taken from the participants. Individuals of blood group A were selected as group A, blood group B as group B, blood group AB as group AB and blood group O as group O. Coagulation factor VIII was measured in the Department of Hematology and BMT Unit, Dhaka Medical College Hospital, Dhaka. Blood grouping was done in the Department of Physiology, Dhaka Medical College, Dhaka. Statistical Analysis: For statistical analysis, ONE way ANOVA followed by Bonferroni test were considered using SPSS 25.0 version. Results: In this study, blood group B was most common (33.2%). Coagulation factor VIII was significantly higher (p < 0.001) in blood group A (105.76% ± 11.82%), B (112.00% ± 15.02%), AB (109.80% ± 11.93%) than blood group O (82.00% ± 12.86%). No significant difference was observed among A, B and AB blood groups regarding coagulation factor VIII. Conclusions: It can be concluded that blood group A, B, AB individuals may have more chance of thrombosis due to significantly higher coagulation factor VIII than blood group O individuals.

The expression of vascular endothelial growth factor and Factor VIII related anti-gen in the border zone after intracer ebral hemorrhage in rats

AIM:To observe the expression of vascula r endothelial growth factor(VEGF)and Factor VIII related antigen(FVIII-R Ag )in rats brain tissue in the bor-der zone of the hematoma,and explore the relationship between the expres-sion of VEGF and the brain angionesis after intracerebral hmorrhage(ICH).METHODS:Rat of model of ICH was induced by inje ction of collagenase physiological saline into the right caudate nucleus.The immunohistoch emical methods were performed by using diff erent antisera of VEGF and FVIII-R Ag in serial sections of rats brain at 12h,1d,3d and 7d after ICH respec-tively.RESULTS:T he amount of VEGF positive cells was markedly in-creased,and reached the highest at 7th day in the border zone after ICG(P<0.01),but there were no significant diffe rence among the mean value of A of cells at different time points(P>0.05).Compared with the controls,both the FVIII-R Ag positive endothelial cells and the A of cells were signific antly different in experimental ICH goups after 1day.CONCLUSION:The up-regulated expression of VEGF might i nduce proliferation of endothelial cell and angiogenesis in the border zone a fter ICH in rats.

Pharmacokinetic Studies of Factor VIII in Chinese Boys with Severe Hemophilia A： A Single-Center Study

Background： Although much attention has been paid to the pharmacokinetics （PKs） of different factor VIII （FVIlI） concentrates in persons with hemophilia A （HA）, limited information is available in young boys with severe HA. In this study, we aimed to assess the PK parameters of FVIII products in boys with severe HA in China. Methods： A total of 36 boys （plasma-derived [pd]-FVIII, n= 15; recombinant [r] FVIII, n = 21） were enrolled between January 2015 and May 2016 in Beijing Children＇s Hospital. PK characteristics of FVIII products were studied according to a reduced 4-sampling time point design （1 h, 9 h, 24 h, and 48 h postinfusion）. Results： The mean FVIII half-life （t1/2） was 10.99 ± 3.45 h （range 5.52-20.02 h）, the mean in vivo recovery （IVR） was 2.01 ± 0.42 IU/dl per IU/kg （range 1.24-3.02 IU/dl per IU/kg） and mean clearance （CL） of FVIII is 4.34 ± 1.58 ml·kg^-1·h ^-1 （range 2.29-7.90 ml·kg^-1·h·^-1）. We also analyzed the influence of several parameters that potentially modulate FVIII PK. The age was closely associated with FVIII half-life （R^2 = 0.32, P 〈 0.01 ）. The t1/2 of FVIII increased by 0.59 h per year. Besides age, yon Willebrand factor antigen （VWF：Ag） also was associated with FVIII half-life （R^2 = 0.52, P 〈 0.01）. Patients with blood Group O had a shorter FVIII halt-life than patients with non-O blood group （9.40 ± 0.68 h vs. 12.3 ± 0.79 h, t = 2.70, P = 0.01）. The FVIII IVR correlated with age （R^2 =0.21, P 〈 0.01） and VWF：Ag level （R^2 = 0.28, P 〈 0.01 ）. CL rates were taster in young patients and in those with low-VWF：Ag levels. CL rates of FVIII are higher in blood Group O versus non-blood Group O persons （5.02 ± 0.38 vs. 4.00 ± 0.32 ml·kg^-1·h^-1 , t = 2.53, P = 0.02）. Conclusions： Chinese boys with severe HA have similar PK values to other ethnic groups and large differences in FVIII PK between individual patients. Age, blood group, and VWF：Ag levels are important determining factors for FVlll CL.

中国人动脉血栓性疾病与活化蛋白C裂解FV和FVIII基因位点突变的相关性研究

目的 :检测脑血栓 (CT)和急性心肌梗死 (AMI)患者活化蛋白C(APC)抵抗 (APCR)及APC裂解凝血因子V (FV )和凝血因子VIII(FVIII)肽链的基因位点突变。方法 :用单链构象多态性分析 (SSCP)方法 ,检测 10 2例CT、46例AMI和 10 5例健康人的APC裂解FV肽链 (Arg3 0 6、Arg 5 0 6、Arg 679和结合部位 1865～ 1875 )、裂解FVIII肽链 (Arg3 3 6、Arg 5 62和结合部位 2 0 0 5～ 2 0 18)的基因位点。结果 :对照组和患者组均未检出FV的突变基因。结论 :中国人动脉血栓性疾病患者无APC裂解FV和FVIII肽链的基因突变 。

早孕先兆流产患者血FVⅢ活性、PAI-1水平及其对保胎结局的影响

目的:探究凝血因子Ⅲ(FVⅢ)活性、纤溶酶原激活物抑制剂-1(PAI-1)对早孕先兆流产保胎结局的预测价值。方法:回顾性分析本院于2020年1月-2022年12月收治的345例早孕先兆流产患者临床资料,根据保胎结局分为保胎失败组124例、成功组221例,检测两组血FVⅢ活性、PAI-1水平,超声检测子宫内膜厚度(EST),血清β-人绒毛膜促性腺激素(β-hCG)、雌二醇(E_(2))、孕酮(P),采用多因素logistic回归分析早孕先兆流产保胎成功影响因素。结果:失败组FVⅢ活性(168.31±25.84)%、PAI-1(54.39±6.05μg/l)均高于成功组(138.64±24.61)%、(49.31±5.87μg/l),EST(10.06±3.26 mm)、β-hCG(1763.86±706.77 U/L)、E_(2)(1051.16±369.57 poml/L)、P(32.96±15.72 noml/L)水平均低于成功组(12.55±1.91 mm、3299.46±1831.59 U/L、3571.36±1126.29 poml/L、64.38±17.24 noml/L)(均P<0.05)。多因素logistic回归分析显示,FVⅢ活性升高,β-hCG、E_(2)、P水平降低为影响早孕先兆流产保胎成功的危险因素(P<0.05),而PAI-1为保胎成功非危险因素。结论:FVⅢ活性升高影响保胎成功,临床可予以参考,减少盲目保胎带来的风险。

Milk fat globule epithelial growth factorⅧ(MFG-E8)sustains survival of cancer cells by prompting tumor angiogenesis and suppressing host immunities

Milk fat globule epithelial growth factor VIII(MFG-E8) is a novel adhesion protein mainly produced by macrophages and dendritic cells; it is expressed in most of the human tissues and functions to prompt cancer progression and survival. MFG-E8 contains a signal sequence for secretion, two epidermal growth factor(EGF)-like domains at the NH2 terminus and two discoidin domains with blood-clotting factor V/factor Ⅷ(C1 and C2) at the COOH terminus. The second EGF domain contains an arginine-glycine-aspartic(RGD) integrin-binding motif that engages α_vβ_5 integrins to facilitate cell adhesion and induce integrinmediated signal transduction. Integrin α_vβ_3 associates with VEGF receptor 2, engagement of integrins can promote angiogenesis, which plays key roles in growth, proliferation, and survival of cancer cells. VEGF stimulates the expression of α_vβ_3 and α_vβ_5 integrins on angiogenic vasculature, thereby potentiating effects of VEGF receptor engagement. Mice expressing a mutant form of α_vβ_3 integrin are unable to undergo tyrosine phosphorylation, confirming the important role that this integrin plays in pathological angiogenesis and providing important mechanistic insights. The C-terminus discoidin-like domains promote binding to membrane phospholipids, functioning close to VEGF like angiogenesis. MFG-E8 is an opsonin for apoptotic cells, and it acts as a bridging protein between apoptotic cells and phagocytes. It also influences cell immunities by altering CD4^+ and/or CD8^+ cells. Antibody or small peptide works with MFG-E8 at different functional sites or interacts with EGF-like domains and/or discoidin-like domains may play an important role in anti-angiogenesis or immune restoration. Altering the structures and/or functions of MFG-E8 and/or its domains is promising for development of novel anti-cancer strategies.

一种分泌和活性改善的新型人凝血因子VIII变构体

甲型血友病基因治疗疗效受凝血因子VIII(FVIII)表达水平低下的制约。FVIII分泌低效,且活化产物的A2区易于自发脱落使其容易失活。犬属FVIII(cFVIII)的胞内Furin水解酶裂解位点变异,使其主要以单链多肽形式分泌,而人源FVIII(hFVIII)受Furin水解,主要以重链(HC)和轻链(LC)非共价二聚体形式分泌,这种差异使cFVIII分泌性和比活性均高于hFVIII。我们前期的工作在人源B结构域缺失型FVIII(hBDD-FVIII)HC的A2与轻链LC的A3区引入链间二硫键表明,HC和LC二聚体的分泌性得到改善,活化产物稳定性和血浆凝血活性提高。本文构建一种新型hBDD-FVIII变构体F8C-RH,包含链间二硫键突变M226C/D1828C和Furin水解位点突变R1645H。培养的CHO和BHK细胞转基因表明,F8C-RH的分泌性和活性有明显改善。小鼠门静脉注射肝靶向转基因,血浆F8C-RH浓度和凝血活性均明显升高。为进一步腺相关病毒(AAV)载体转F8C-RH的甲型血友病基因治疗研究提供了依据。

Successful reconstruction of an ankle defect with free tissue transfer in a hemophilia A patient with repetitive hemoarthrosis:A case report

BACKGROUND Hemophilia, an uncommon yet consequential hereditary bleeding disorder, manifests as two clinically indistinguishable forms that hinder the normal functioning of the coagulation cascade. This impairment renders individuals more susceptible to excessive bleeding during significant surgical interventions. Moreover, individuals with severe hemophilia frequently encounter recurring hemarthrosis, resulting in progressive joint destruction and, subsequently, the need for hip and knee replacement surgeries.CASE SUMMARY The patient was a 53-year-old man with hemophilia A as the underlying disease and had self-injected factor Ⅷ twice weekly for several decades. He had undergone ankle fusion surgery for recurrent hemarthrosis at the Department of Orthopedic Surgery 1 mo prior and was referred to our department because of skin necrosis after a hematoma at the surgical site. An anterolateral thigh perforator free flap was created after three cycles of factor Ⅷ administration in addition to the concomitant administration of tranexamic acid(TXA)(Transamin 250 mg cap, 1 cap tid, q8h). After the operation, from postoperative days(PODs) 1-5, the factor Ⅷ dose and interval were maintained, and q12h administration was tapered to q24h administration after POD 6. Because the patient’s flap was stable 12 d after the operation, factor Ⅷ administration was tapered to twice a week. At 6 mo follow-up, the patient recovered well without any complications.CONCLUSION To the best of our knowledge, there are very few reports of successful free flaps in patients with hemophilia, and none have been reported in patients with hemophilia A. Moreover, there are several reports on the efficacy of TXA in free flaps in general patients;however, there are no case reports of combining factor Ⅷ and TXA in patients with hemophilia. Therefore, we report this case to contribute to future academic research.

Characterization of Bleeding and Laboratory Phenotype in Hemophilia A Carriers: A Cross-Sectional Study in Benin

Background: In Africa, hemophilia is underdiagnosed and carriers have long been considered free from bleeding symptoms. However, recent research has begun to reveal hemostatic abnormalities and bleeding manifestations in carriers of hemophilia A, particularly due to excessive inactivation of normal X chromosomes. Objective: To describe the bleeding symptoms and hemostatic abnormalities in carriers of hemophilia A (HA) in Benin. Methods: This study was conducted as a prospective cross-sectional investigation between April 2021 to March 2022. The study population consisted of identified through pedigrees of persons with hemophilia A being treated in various hospitals in Benin. Data were collected through interviews conducted by trained physician and each carrier underwent a biological workup. Results: A total of 71 hemophilia A carriers were included and 38 of whom were obligatory carriers. Thirty-one carriers (43.7%) reported abnormal bleeding symptoms. Menorrhagia has (71%) being the most important manifestation, followed by bleeding during or after childbirth (45.2%). Among the 71 carriers, 45 were of reproductive age. Of whom 22 (48.8%) had a Higham score exceeding 100. Activated partial thromboplastin time was prolonged in 7 carriers (9.9%). The mean activity factor VIII:C (FVIII:C) levels were 68.8 ± 34.9 IU/dL. The average FVIII:C level in obligatory carriers was 56.9% and among potential carriers, the average FVIII:C level was higher at 80.4%. However twelve female carriers (16.9%) had FVIII:C levels < 40%. The FVIII:C/FvWAg ratio was below 0.7 in 73.2% of female drivers. Obligatory carriers (p = 0.00003) and FVIII;C/FvWAg ratio = 0.003) were statistically associated with abnormal bleeding symptoms, while blood group O (p = 0.0002) and FVIII/FvWAg ratio = 0.0016) were associated with a higher risk of menorrhagia. Conclusion: In Benin, carriers of haemophilia A present bleeding symptoms and haemostatic abnormalities. Further studies on a larger number of carriers are needed to better characterize and manage these patients.

大鼠原代脊髓微血管内皮细胞的体外分离培养及鉴定

目的:建立一种简单高效的大鼠原代脊髓微血管内皮细胞培养方法。方法:分离大鼠脊髓组织,经剪碎、细胞筛网过滤、Ⅱ型胶原酶消化后接种培养。通过细胞形态学观察、FⅧRag免疫细胞化学染色鉴定所培养的目的细胞。结果:接种的脊髓微血管段呈“短棍样”,培养24 h后有少量短梭形细胞从血管段周围迁移爬出,48 h后逐渐形成“岛屿状”的细胞集落,72 h后细胞铺满皿底,呈典型的单层、“鹅卵石样”、镶嵌式排列生长;细胞FⅧRag免疫细胞化学染色显示细胞质呈棕红色,表达为阳性。结论:该方法可成功分离培养出活性好、纯度高的大鼠原代脊髓微血管内皮细胞。

单筛过滤法体外分离培养原代大鼠脑微血管内皮细胞及其鉴定

目的 采用单筛过滤法获取脑微血管以培养纯度高、活性好的原代大鼠脑微血管内皮细胞。方法 取4周龄SD大鼠脑皮质,经剪碎、单层细胞筛网过滤、收集网上截留组织、Ⅱ型胶原酶消化后,置于CO_(2)培养箱中进行原代培养。通过细胞形态学观察、第Ⅷ因子相关抗原免疫细胞化学染色检测鉴定所培养的目的细胞。结果 体外培养24 h后,短梭形细胞从脑微血管段周围爬出;48 h后“岛屿状”的细胞团簇形成;96 h后细胞融合,呈典型的单层、“铺路石样”、镶嵌式贴壁生长。第Ⅷ因子相关抗原免疫细胞化学染色检测显示细胞胞质呈现棕红色,表达为阳性。结论 单筛过滤法能够成功分离培养出原代大鼠脑微血管内皮细胞。

薄层液面培养法提取大鼠原代脑微血管内皮细胞及对其的鉴定

目的:采用薄层液面培养法提取大鼠原代脑微血管内皮细胞(BMECs)。方法:取4周龄SD大鼠脑皮质,经剪碎、过筛、Ⅱ型胶原酶消化后获得脑微血管段,并严格精准控制培养液的用量,将其接种于培养皿中进行原代培养。通过倒置相差显微镜对目的细胞的形态学观察、免疫细胞化学染色法对细胞第Ⅷ因子相关抗原(FⅧRag)的检测进行鉴定。结果:镜下观察到所培养的目的细胞呈典型的单层、铺路石样、镶嵌式排列生长;FⅧRag阳性细胞率达99%以上。结论:薄层液面培养法能够成功分离培养出活性强、纯度高的大鼠原代BMECs。

基于热管冷却反应堆堆芯有效增殖系数的敏感性分析

核数据作为反应堆计算的输入参数,其敏感性对堆芯物理计算至关重要。基于美国橡树岭实验室开发的反应堆蒙特卡洛中子输运程序MCNP6.1,以美国爱荷华国家实验室(INL)设计的热管冷却反应堆INL Design A为研究对象,调用美国的ENDF/B-Ⅷ.0评价核数据库,对堆芯处于热态(1000 K)零功率且控制棒均未插入工况时的堆芯有效增殖系数进行敏感性分析。计算结果表明:有效增殖系数对235U的平均裂变中子数产额、裂变反应截面敏感性系数分别高达0.88702、0.52898;高能区的235U的平均裂变中子数产额和裂变反应截面,238U平均裂变中子数和裂变截面,^(16)O、^(27)Al和^(56)Fe弹性散射截面,中能区的^(238)U(n,γ)反应截面,^(16)O、^(27)Al和^(56)Fe弹性散射截面具有较大的敏感性,在反应堆物理计算过程中应重点关注。

难治性肾病综合征患者血凝学的研究

目的 探讨难治性肾病综合征（RNS）患者血凝学指标的变化。方法 31例RNS为实验组，另设31例正常对照。两组均检测第Ⅷ因子相关抗原（ⅧR:Ag）、纤维蛋白原（Fbg）、抗凝血酶Ⅲ（AT-Ⅲ）、纤溶酶原（Plg）、白陶土部分凝血活酶时间（APTT）、凝血酶原时间（PT）及血小板聚集试验（PAgT）的变化。结果：ⅧR:Ag、Fbg、PAgT与正常组比较显著增高，AT-Ⅲ、Plg与正常组比较显著降低，APTT时间显著缩短，PT与正常对照组比较无显著性差异。ⅧR:Ag、AT-Ⅲ与尿素氮（BUN）、血浆白蛋白（A1b）呈正相关，Fbg、PAgT与A1b呈负相关。结论 RNS存在严重的高凝状态。

重型血友病甲基因诊断新技术的研究及其应用

运用长距离ＤＮＡ扩增技术（ＬＤ－ＰＣＲ）成功地检测出凝血八因子ＦⅧ基因倒位缺陷患者，并检出该倒位的女性携带者。在建立并完善上述技术之后，对５３份ＨＡ患者及家系成员的ＤＮＡ标本进行了双盲试验，检出ＦⅧ倒位者２３份，野生型２７份，携带者３份。结果与用Ｓｏｕｔｈｅｒｎ杂交技术检测的结果完全一致，说明运用本ＬＤ－ＰＣＲ技术检测ＦⅧ基因倒位的灵敏度与准确率均达到百分之百。运用这一新技术，完成了５个重型血友病甲家系的基因诊断。共检出３例基因倒位者，４位女性倒位基因的携带者。该技术较Ｓｏｕｔｈ－ｅｒｎ印迹杂交法简便、快速、高效直观、灵敏准确，便于推广应用，达到国际先进水平。

von Willebrand因子和凝血因子Ⅷ 水平与急性脑梗死的临床转归

目的探索急性脑梗死患者血浆FVIII（factor VIII,FVIII）和VWF（von Willebrand factor,VWF）水平的升高与疾病的严重性、预后、住院期间发生感染或神经功能损伤加重等事件的关系。方法在2014年12月至2015年3月期间就诊于中国医科大学附属盛京医院神经内科的住院患者中,筛选出急性脑梗死病例组90例,无急性脑梗死的对照组50例,测定血浆FVIII和VWF水平。按两因子水平是否升高将病例组分成4组：FVIII和VWF均不升高组（FVIII-/VWF-）、FVIII升高且VWF不升高组（FVIII↑/VWF-）、FVIII不升高且VWF升高组（FVIII-/VWF↑）和FVIII和VWF均升高组（FVIII↑/VWF↑）。比较各病例组和对照组的临床特点。结果病例组的VWF水平的中位数1521.88 U/L,明显高于对照组的中位数1281.77 U/L（P=0.023）。与FVIII-/VWF-组相比,FVIII↑/VWF↑组急性脑梗死的发病症状较重（入院NIHSS评分〉5分）（OR=3.643,95%CI：1.258-10.549,P=0.017）,疾病预后较差（3个月m RS评分〉2分）（OR=7,95%CI：2.304-21.266,P=0.001）,出院时m RS评分〉2分者所占比例高（OR=3.797,95%CI：1.346-10.713,P=0.012）,住院期间更易发生神经功能损伤加重（OR=5.538,95%CI：1.099-27.908,P=0.038）,且更易并发感染（OR=3.913,95%CI：1.115-13.729,P=0.033）。对各种混杂因素进行校正后,发现FVIII和VWF水平同时升高是急性脑梗死患者预后不良的独立预测因素（OR=4.495,95%CI：1.012-19.957,P=0.048）。结论急性脑梗死患者很可能存在FVIII或VWF水平升高,二者水平同时升高可能对疾病的严重性及临床转归有影响,并且很可能是预后不良的独立预测因素。

血友病A患者血浆抗因子Ⅷ同种抗体的检测及其与因子ⅧC区的关系

本研究应用快速而敏感的ELISA法检测血友病A患者血浆抗因子Ⅷ同种抗体,并探讨其与因子ⅧC区的关系。收集140例血友病A患者样本,其中重型(FⅧ∶C<1%)84例,中型(FⅧ∶C 1-5%)34例,轻型(FⅧ∶C>5%)22例;患者均已经过因子Ⅷ替代治疗。正常对照样本80例。以本室制备的抗因子Ⅷ单克隆抗体包被酶标板,封闭后加入重组人凝血因子Ⅷ,洗涤后加入待测稀释血浆,再次洗涤后加入HRP标记的羊抗人IgG,OPD显色,记录A490,以正常人血浆为对照,样本A490大于正常人对照3 SD定义为因子Ⅷ抗体阳性。以抗His抗体包被酶标板,封闭后加入本室制备的rFⅧ-C1C2区蛋白,洗涤后加入待测稀释血浆,再次洗涤后加入HRP标记的羊抗人IgG,OPD显色,记录A490,以正常人血浆为对照,样本A490大于正常人对照3 SD定义阳性。结果表明,通过ELISA法检测,56例(40%)血友病A患者血浆抗因子Ⅷ同种抗体阳性,其中46例(82.1%)因子Ⅷ同种抗体阳性的血浆样本中存在与因子ⅧC1C2结合的同种抗体。结论:因子ⅧC功能区是中国血友病A患者因子Ⅷ同种抗体的主要结合部位之一。

Nestin:A novel angiogenesis marker and possible target for tumor angiogenesis

Abnormal vasculature,termed tumor vessels,is a hallmark of solid tumors.The degree of angiogenesis is associated with tumor aggressiveness and clinical outcome.Therefore,exact quantification of tumor vessels is useful to evaluate prognosis.Furthermore,selective detection of newly formed tumor vessels within cancer tissues using specific markers raises the possibility of molecular targeted therapy via the inhibition of tumor angiogenesis.Nestin,an intermediate filament protein,is reportedly expressed in repair processes,various neoplasms,and proliferating vascular endothelial cells.Nestin expression is detected in endothelial cells of embryonic capillaries,capillaries of the corpus luteum,which replenishes itself by angiogenesis,and proliferating endothelial progenitor cells,but not in mature endothelial cells.Therefore,expression of nestin is relatively limited to proliferating vascular endothelial cells and endothelial progenitor cells.Nestin expression is also reported in blood vessels within glioblastoma,prostate cancer,colorectal cancer,and pancreatic cancer,and its expression is more specific for newly formed blood vessels than other endothelial cell markers.Nestin-positive blood vessels form smaller vessels with high proliferation activity in tumors.Knockdown of nestin in vascular endothelial cells suppresses endothelial cell growth and tumor formation ability of pancreatic cancers in vivo.Using nestin to more accurately evaluate microvessel density in cancer specimens may be a novel prognostic indicator.Furthermore,nestin-targeted therapy may suppress tumor proliferation via inhibition of angiogenesis in numerous malignancies,including pancreatic cancer.In this review article,we focus on nestin as a novel angiogenesis marker and possible therapeutic target via inhibition of tumor angiogenesis.

RNAi介导的Bip表达下调促进基于蛋白质剪接的双链共转基因细胞分泌FⅤⅢ凝血活性

目的探讨用RNA干扰技术下调内质网蛋白伴侣免疫球蛋白重链结合蛋白(Bip)的表达,对基于蛋白质剪接的双链共转凝血因子ⅤⅢ(FⅤⅢ)基因HEK293细胞分泌剪接FⅤⅢ蛋白的量和活性的影响。方法以培养的HEK293细胞,用含蛋白内含子的B区缺失型FⅤⅢ(BDD-FⅤⅢ)的重链和轻链基因表达载体共转染经Bip-siRNA处理的细胞,免疫印迹法检测Bip的表达,四甲基偶氮唑盐(MTT)法检测细胞生长,用酶联免疫吸附(ELISA)和发色分析法分析转基因细胞分泌的剪接BDD-FⅤⅢ蛋白量和活性。结果 RNA干扰下调Bip表达作用明显,细胞生长不受影响;Bip下调的共转基因细胞分泌的剪接BDD-FⅤⅢ和重链量分别为(142±33)μg/L和(197±43)μg/L,明显高于对照细胞[分别为(89±23)μg/L和(120±27)μg/L];Bip下调的共转基因细胞分泌的FⅤⅢ凝血活性为(1 050±160)IU/L,明显高于对照细胞[640±170(IU/L)]。结论 Bip表达下调通过促进剪接BDD-FⅤⅢ的分泌可有效提高基于蛋白质剪接的双载体转BDD-FⅤⅢ基因功效,为进一步动物体内实验提供了依据。

凝血纤溶系统相关因子与多囊卵巢综合征妇女先兆流产预后的关系

目的探讨凝血纤溶系统相关因子与多囊卵巢综合征(polycysticovary syndrome,PCOS)妇女先兆流产后出现的不同妊娠结局的关系以及与胰岛素抵抗(insulin resistance,IR)、血脂、体重指数(BMI)的相关性。方法收集PCOS正常早孕妇女30例,PCOS先兆流产经治疗后继续妊娠28例,保胎失败(难免流产)32例。检测凝血因子VIII(FVIII)活性、凝血因子X(FX)活性、纤溶酶原活化剂抑制物(PAI-1)和组织型纤溶酶原活化剂(t-PA)水平,以及IR、血脂与BMI。结果与正常早孕组相比,继续妊娠组FVIII活性、FX活性升高,差异有统计学意义(P<0.05);难免流产组中FVIII活性、FX活性、PAI-1和t-PA水平升高,差异有统计学意义(P<0.05)。与继续妊娠组相比,PAI-1水平升高,差异有统计学意义(P<0.05)。多元线性回归分析结果显示,难免流产组中PAI-1水平与胰岛素抵抗指数(HOMA-IR)、低密度脂蛋白(LDL)、BMI呈正相关。回归方程的决定系数R^2=0.79。结论难免流产组中,PAI-1水平有明显的升高。PCOS妊娠患者可以通过PAI-1水平,以及HOMA-IR、BMI、血脂测定来预测先兆流产的预后。