Cloning of Human α-defensin-1 (HNP-1) Gene and Construction of Its Eukaryotic Expression Vector

1 Introduction Defensins are small cationic antimicrobial peptides that function in the host's innate immune system. The human defensin family includes three small peptides from the azurophil granules of polymorphonuclear cells named human neutrophil peptide (HNP)-1, HNP-2, HNP-3,which consist 5%-7% of the protein of human neutrophil. HNP-4 is approximately one hundred times less abundant. They demonstrate antibacterial, antifungal and antiviral properties in vitro. HNPs are important component of nonoxidative mechanism in macrophages, and can direct inactivate the enveloped viruses. Because only special cells express defensins. And it is hard to extract them naturally and the production is few. So researcher expect to obtain defensins highly through heterogenous expression by gene engineering technology. In order to express HNP-1, we cloned the cDNA of HNP-1 from human polymorphonuclear cells in peripheral blood, and constructed its eukaryotic expression vector, which provided a base for the further study on its mechanism of antimicrobial effect.     

A Study of Early Pregnancy Factor Activity in Preimplantation

PROBLEM and METHOD: Early pregnancy factor (EPF), an Immunosuppressive substance, which appears in pregnant women's sera 48 h after fertilization, is a kind of pregnancy-specific protein. To determine whether the EPF activity could be a super early indicator of pregnancy, we used rosette inhibition assay to detect EPF activity in the sera, collected from 70 women 2–7 days after ovulation intending to conceive monitored by ultrasonography. Simultaneously we selected 40 non-pregnant sera and 12 early-pregnant sera as negative control and positive control, respectively. RESULTS: The results of this study demonstrated that EPF activity is detected in 35 women's sera out of 70 women within 2–7 days after ovulation, and 28 women out of the 35 were pregnant, which was known by follow-up, and 7 were not pregnant, possibly due to either false positive results or embryo loss because of preimplantation failure, thus causing no pregnancy. The other 35 out of 70 had no EPF activity and 34 of them were not pregnant, which was known by follow-up, but one case became pregnant, which was false negative result. Our study showed that diagnosis of the super early pregnancy could be made by detecting EPF activity in maternal serum within the time of preimplantation. The accuracy of pregnancy diagnosis by this method is 88.6%, with a false negative rate of 3.4% and a false positive rate of 17.1%. The β-HCG level was measured from the above 70 women's sera in order to contrast EPF activity. All of the sera collected 2–6 days following ovulation indicated that there were lower β-HCG values in very early pregnancy (≥a5 mIU/ml). On the seventh day after ovulation, EPF activity was detected in 11 out of 15 sera with only 2 of them with a b-HCG level that reached or slightly surpassed that of the early pregnancy diagnosis (5 mIU/ml and 5.4 mIU/ml, respectively). This demonstrated that β-HCG is not the earliest signal of pregnancy; otherwise the EPF activity is one that appears 2–6 days earlier than β-HCG appears. We measured the progesterone level of the 48 sera from the 70 collected above within 2–7 days postovulation and found most of them reached the level of progesterone in the luteal phase (7.5–98.3 nmol/L). This indicated that ovulation had taken place in these women, which was in accordance with observations by ultrasonography. CONCLUSIONS: Our study showed that diagnosis (of 88.6%) of super early pregnancy could be made with an accuracy of 88.6% by detecting EPF activity in maternal serum within 2-days after ovulation. This offers a basis for pregnancy diagnosis for the women who attempt to terminate their pregnancy safely or who conceive unexpectedly, and it contributes to family-planning.     

缺血性股骨头坏死患者血清透明质酸及层粘连蛋白的研究

为探讨细胞外间质主要成分透明质酸（ＨＡ）与层粘连蛋白（ＬＮ）在缺血性股骨头坏死（ＩＮＦＨ）中的生理病理过程及临床价值，采用放射免疫分析法对４５例不同病因的ＩＮＦＨ患者进行了血清ＨＡ与ＬＮ的定量分析，并与３０例正常人对照。结果显示，ＩＮＦＨ患者血清ＨＡ含量极显著地高于对照组（ｔ＝３－２９；Ｐ＜０－０１）。尤以激素性ＩＮＦＮ增高最为显著（ｔ＝３－６２；Ｐ＜０－０１）。ＩＮＦＨ患者ＬＮ含量亦明显高于对照组（ｔ＝２－８４；Ｐ＜０－０１）。同时，ＨＡ与ＬＮ含量增高与病程发展密切相关。故定量检测ＨＡ与ＬＮ可作为ＩＮＦＨ早期诊断及判断预后的良好指标     

外源性凝血途径和内皮细胞损伤与动脉粥样硬化之间的关系研究

目的探讨外源性凝血途径在动脉粥样硬化(AS)发病中的作用。方法均采用ELISA法测定心肌梗死、脑梗死、脑出血病人及正常对照组血浆TF、FⅦa的含量及不同时期这些物质浓度的变化、同时通过测定内皮细胞损伤的分子标记物vWF、内皮素 -1(ET -1)、总组织因子途径抑制物(TFPI)、组织因子(TF)探讨外源性凝血途径的活化与水平状况及内皮细胞损伤之间的关系。结果心肌梗死、脑梗死、脑出血病人TF、TFPT、ET -1、vWF、FⅦa,血中的浓度较正常对照组高 (P<0.01) ,有非常显著意义。结论外源性凝血途径与内皮细胞损伤和AS有密切相关。     

大学生个体印象管理对其社会网络质量的影响

目的:研究个体印象管理对其社会网络质量的影响。方法:选用社会网络调查法、大学生印象管理量表,对46名大学生进行调查。结果:印象管理建构的提高不利于朋友网和沟通网的强联系人数量的增加(β分别为-1.04和-1.33,P分别小于0.05和0.01;F值分别为4.19和6.45,P分别小于0.05和0.01),而印象管理动机对强联系人的数量并没有影响,并且印象管理动机和印象管理结构都不影响个体为中心网络的紧密性。结论:只有个体印象管理的建构部分不利于其社会网络中的强联系人数量的增加。其他部分对社会网络无明显影响。     

Cariporide对兔心房电重构的影响

目的：探讨钠氢交换体Ⅰ型(NHE-1)特异性抑制剂cariporide对快速起搏所致兔心房电重构的影响。 方法： 30只兔随机等分为3组：对照组、起搏组和cariporide组。起搏组和cariporide组给予6 h 600 beats/min的快速心房起搏。测定各组不同时点的心房有效不应期（AERP200，AERP150，AERP130），连续刺激6 h后取左右心耳组织，用Western blotting测定NHE-1的含量。 结果： 在快速起搏后1 h后，起搏组的AERP200较起搏前明显缩短，2 h时达高峰，相对缩短量为（15.63±9.04）ms，而对照组和cariporide组AERP未发生明显变化，起搏2h时相对缩短量为(1.43±2.44)ms和(1.43±6.90)ms（P<0.05，与起搏组相比），这些变化一直保持至快速起搏后6 h；起搏组的AERP频率适应性下降, 起搏前AERP130较AERP200缩短(11.88±15.57)ms，起搏后6 h只缩短(4.38±5.63)ms。而cariporide组的AERP频率适应性则未发生显著变化。起搏组右心耳组织的NHE-1含量显著少于对照组（P<0.05），cariporide组的右心耳组织NHE-1含量与对照组差异不显著。 结论： Cariporide可有效阻止快速心房起搏引起的AERP缩短，但不影响起搏引起的NHE-1含量的下降。     

抗SARS-CoV抗原的人源Fab段噬菌体抗体库的构建

目的 :利用抗SARS冠状病毒IgG抗体阳性的SARS康复患者外周血淋巴细胞 ,构建人源Fab段抗体文库。方法 :制备外周血淋巴细胞总RNA ,逆转录成cDNA。以其为模板 ,利用针对家族特异性Ig基因的引物扩增重链Fd段和轻链基因 ,并重组到噬菌粒载体pComb3中 ,将重组噬菌粒载体电转化大肠杆菌XL 1Blue,酶切鉴定抗体库的重组率 ,并测定噬菌体抗体库的库容量。结果 :构建了源于SARS康复患者血清中抗Fab段的抗体文库 ,轻链、重链Fd段基因的重组率分别为91%和 75 % ,库容量为 7.2 3× 10 7。结论 :成功地构建了抗SARS CoV抗原的人源Fab段噬菌体抗体库     

氧化应激在溃疡性结肠炎中的作用及中医药防治研究进展

溃疡性结肠炎（UC）是一种临床常见的消化系统疾病,致病机制尚不明确,病情复杂多变,迁延难愈,治疗周期漫长,且无特效药。目前,UC的治疗多采用皮质类固醇、氨基水杨酸及生物制剂等西医手段,短时间起效快,疗效确切。但随着用药时间的延长,部分患者会出现耐药性和加剧病情发展,导致结肠癌的发生。有研究发现,氧化应激是UC重要的致病因素之一,影响着UC的发生、发展。氧化应激是机体内氧化产物与抗氧化系统不平衡的一种应激状态,丙二醛（MDA）、活性氧（ROS）、一氧化氮（NO）等氧化产物的过表达或者超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽（GSH）抗氧化酶的不足都会导致氧化应激的发生。值得注意的是,中医药作为我国独有的医学特色,运用中医药治疗UC疾病已经取得显著的成效。研究表明,中医药一方面通过抑制代谢产物的堆积,有效抑制UC发生,另一方面,通过提高抗氧化系统,达到拮抗UC发展的治疗效果。因此,以中医药调节氧化平衡状态作为诊疗思路,可能是未来治疗UC疾病的新手段、新方向。基于上述研究,该文总结了氧化应激关键致病蛋白与UC发生、发展的作用机制,归纳了中药有效成分、单味中药、中药复方、针灸调控氧化应激上下游靶点蛋白,减轻肠黏膜病理损害,降低结肠损伤指数,以及丰富肠道菌群,增加结肠长度,改善UC临床症状,以期为扩大中医药治疗UC疾病的应用范围,提供可靠的科学理论依据。     

^32P局部照射预防猪冠状动脉再狭窄研究

目的：评价^32P灌注球囊预防再狭窄的有效性、可行性和安全性。方法：对猪冠状动脉左前降支行过度球囊扩张术后,治疗组以^32P作血管内照射,对照组作假照射。术后35d收获目标血管、检测血管形态、细胞增殖百分比等。结果：治疗组血管腔面积较对照组明显增大（P＜0．01）,新生内膜面积、血管狭窄程度和各层PCNA阳性细胞明显减小（P均＜0．01）。结论：^31P灌注球囊照射预防冠状动脉再狭窄有效、安全而且可行。     

慢性乙型肝炎患者肝组织FasL表达与血清可溶性FasL水平的关系

目的：探讨慢性乙型肝炎患者肝组织中FasL表达与血清可溶性FasL水平的关系。方法：用免疫组化方法检测60例慢性乙型肝炎患者肝穿组织FasL的表达,同时用酶联免疫吸附试验检测血清可溶性FasL。结果：重度慢性乙型肝炎患者血清中sFasL水平＞中度＞轻度,各组间差异有显著意义（P＜0．01）;慢性乙型肝炎患者肝组织FasL表达的程度和血清sFasL水平与肝组织病变的活动性一致。结论：（1）肝组织炎症程度加重,肝组织FasL抗原的表达增强,同时血清中sFasL水平升高;（2）Fas介导的肝细胞凋亡在慢性乙型肝炎的发病机制中起重要作用,抑制肝细胞Fas表达有助于减轻肝细胞损伤程度。