促血管生成素1和血管内皮生长因子基因合适剂量配比有效促进血管生成

目的在大鼠后肢缺血模型中,探讨联合转染促血管生成素1和血管内皮生长因子基因治疗对新生血管数量和侧支循环形成的影响,并观察不同基因剂量配比对新生血管形态和功能的影响。方法制备大鼠后肢血管闭塞病变模型,采用电脉冲法介导转基因,按不同剂量配比进行肌肉组织转染携带促血管生成素1基因的质粒和/或携带血管内皮生长因子基因的质粒。采用逆转录聚合酶链反应检测外源基因的表达、免疫组织化学染色检测缺血局部毛细血管和小动脉的数量和分布、通过后肢血管造影检测侧支循环建立的情况。结果单独转促血管生成素1基因或血管内皮生长因子基因治疗组血管生成数量略有增加,联合基因治疗组中,血管内皮生长因子质粒剂量为100μg时,随着促血管生成素1质粒剂量的增加,小动脉计数较同期空载质粒对照组分别增加0.90倍、1.40倍和1.45倍;当促血管生成素1质粒剂量为100μg时,随着血管内皮生长因子质粒剂量的增加,小动脉计数较同期空载质粒对照组分别增加1.90倍、1.07倍和1.39倍。血管通透性检测表明,随着血管内皮生长因子质粒剂量的增加,血管通透性渐增加,促血管生成素1质粒能降低新生血管的通透性;其中促血管生成素1质粒200μg联合血管内皮生长因子质粒100μg组在有效促进小动脉形成的同时血管通透性与正常对照组最为接近。结论联合血管生成素1和血管内皮细胞生长因子基因治疗能更有效促进小动脉形成,其中血管内皮生长因子质粒与促血管生成素1质粒比例为1:2时血管通透性最接近生理状态,是比较理想的联合基因治疗剂量配比。     

联合基因治疗小型猪冠状动脉闭塞的实验研究

目的　探讨联合血管内皮生长因子 (VEGF)及促血管生成素 1(Ang 1)基因治疗小型猪冠脉闭塞的疗效。方法　在小型猪冠脉闭塞模型的心肌内注射质粒PCD2 /VEGF和 (或 )PCD2 /Ang 1,检测外源基因在心肌中的表达并观察其生物学作用。结果　逆转录 PCR及免疫组化染色均检测到外源基因的表达。转PCD2 /VEGF和 (或 )PCD2 /Ang 1基因治疗组毛细血管计数及小动脉计数均高于空载质粒对照组 (P <0 0 5 ) ,联合基因治疗组血管计数最高。冠状动脉造影证明联合基因治疗组促进闭塞冠脉侧支循环建立更为有效。结论　心肌内联合注射PCD2 /VEGF及PCD2 /Ang 1基因能够获得外源基因mRNA及蛋白的有效表达 ,与单基因治疗相比联合基因治疗能更有效地促进血管生成及侧支循环的建立。     

肌肉电转促血管形成素-1基因治疗大鼠下肢血管闭塞病

目的观察肌肉电转促血管形成素-1（angiopoietin-1,Ang-1）基因对大鼠下肢血管闭塞病的治疗效果。方法制作大鼠下肢血管闭塞病模型,通过电转Ang-1进行基因治疗,以RT-PCR及免疫组化方法观察Ang-1基因的表达,应用微血管计数、血管造影技术及相关临床指标观察Ang-1基因导入大鼠体内的生物学效应。结果（1）肌肉电转pcD2Ang-1基因可在大鼠局部转基因肌肉组织高效表达Ang-1;（2）转pcD2Ang-1基因组大鼠下肢肌肉组织新生血管及侧支循环数目明显多于同期空载质粒对照组。结论转Ang-1基因可促进大鼠局部组织新生血管形成和侧支循环建立,恢复闭塞部位血供,从而有效治疗血管闭塞病。     

血管内皮生长因子基因治疗小型猪冠状动脉闭塞的实验研究

目的探讨应用血管内皮生长因子(VEGF)基因治疗动物实验性冠状动脉闭塞的疗效.方法中国实验用小型猪19只,结扎左冠状动脉前降支中远段,心肌内多点注射自行构建的pcD2/hVEGF121真核表达质粒.应用逆转录聚合酶链反应、VEGF免疫组化染色、Ⅷ因子相关抗原免疫组化染色等方法检测VEGF基因在心肌中的表达及其生物学作用,用常规冠状动脉造影观察VEGF基因治疗对闭塞冠脉侧支循环建立的作用.结果转移VEGF基因后,小型猪心肌内VEGF mRNA高表达,VEGF免疫组化染色提示VEGF蛋白表达水平升高;Ⅷ因子相关抗原免疫组化染色显示心肌毛细血管增加;冠状动脉造影证明VEGF基因治疗能够促进闭塞冠脉侧支循环的建立.结论心肌内注射pcD2/hVEGF121真核表达质粒能够获得VEGF mRNA和蛋白的有效表达,促进心肌毛细血管增生,促进侧支循环建立.     

人血管内皮细胞生长因子-165和人血管生成素-1促进大鼠缺血下肢血管新生

目的　观察肌肉转染腺病毒(adenovirus ,Ad )携带的人血管内皮细胞生长因子- 1 6 5(ves cularendothelialgrowfactor ,Ad5 VEGF1 65)和人血管形成素- 1 (angiopoietin- 1 ,Ad5 Ang -1 )基因对大鼠缺血下肢的生血管效应。方法　制作大鼠下肢血管闭塞模型,肌肉内注射Ad5 VEGF1 65 或 和Ad5 Ang -1 ,以Westernblot法检测生长因子蛋白表达,免疫组化检测基因导入后在缺血肌肉引起的效应。结果　肌肉经注射Ad5 VEGF1 65和Ad5 Ang- 1后高表达人VEGF1 65 蛋白和人Ang- 1蛋白。术后7d处理组与对照组新生毛细血管数目无明显差别。但术后1 4d和2 1dAng- 1和VEGF1 65 组的新生毛细血管 肌肉数目比明显高于对照组(P <0 . 0 1 ) ,VEGF1 65 Ang 1两因子合用组明显高于单用组(P <0. 0 1 )。Ad -VEGF1 65组及Ad VEGF1 65 Ad Ang 1合用组出现大量包围着一厚层αSMA阳性平滑肌细胞的血管,其数量与肌肉数比值明显高于对照组(P <0 . 0 1 )。因子单用组和合用组肌肉内出现大量溴化脱氧尿嘧啶阳性细胞浸润,部分细胞表面呈现C- Kit阳性,新生血管内皮细胞中也有C Kit阳性标记。结论　Ad5- VEGF1 65和Ad5 -Ang -1能促进缺血肢体血管新生,二者合用的生血管效应更为显著;VEGF1 65能促进包被平滑肌细胞的形似微小动脉的血管数量增多     

人血管生成素-1和血管内皮生长因子165基因克隆及复制缺陷型腺病毒载体构建

本研究克隆人血管生成素 1(Angiopoietin 1,Ang1)和血管内皮生长因子 (VEGF) 165的全长编码基因 ,构建高转染效率和表达水平的复制缺陷型腺病毒重组体。研究结果显示 ,Ang1和VEGF联合应用具有抗细胞凋亡的作用。1.材料与方法 :( 1)通过反转录聚合酶链反应从人类流产胎儿组织中以特异性引物扩增人VEGF165和Ang1全长基因并测序。将目的基因分别亚克隆接入腺病毒穿梭载体 ,将携带目的基因的腺病毒穿梭载体与复制缺陷型腺病毒基因组载体进行同源重组获得重组复制缺陷型腺病毒载体Ad VEGF165和Ad Ang1。将Ad VEGF165和Ad Ang1分别转…     

脂质体介导血管内皮生长因子基因在内皮细胞的稳定表达

建立稳定表达血管内皮生长因子的人脐静脉内皮细胞系 ,为其在组织化工程血管的应用奠定基础。将真核表达载体PCD2 VEGF1 2 1 用阳离子脂质体介导 ,转染人脐静脉内皮细胞系细胞 ,G4 18筛选 ,获得G4 18抗性单克隆细胞 ,扩增后分别用逆转录聚合酶链反应、免疫组织化学及血管通透性实验检测血管内皮生长因子的转录、蛋白质的表达及其生物学活性。结果发现 ,逆转录聚合酶链反应检测出了转录血管内皮生长因子的稳定转染细胞克隆 ,该单克隆细胞的免疫组织化学检测血管内皮生长因子蛋白质表达呈阳性 ,血管通透性实验和细胞生长实验发现其表达产物具有生物学活性 ,而作为对照的转空白质粒细胞和未转染细胞上述实验结果皆为阴性。结果表明 ,该方法成功建立了稳定表达血管内皮生长因子的人脐静脉内皮细胞系单克隆细胞     

血管紧张素Ⅱ及其受体1在食管鳞癌血管生成中的作用

目的 探讨血管紧张素Ⅱ（AngⅡ）及其受体1（AT1R）在食管鳞癌组织中的表达及其在血管生成中的作用。方法 应用免疫组化SP法检测30例食管鳞癌患者癌组织及其食管断端正常鳞状上皮中AngⅡ、AT1R和血管内皮生长因子（VEGF）蛋白的表达。结果 AngⅡ、AT1R和VEGF蛋白表达主要定位于鳞癌细胞iAngⅡ、AT1R在鳞癌组织中的表达均显著高于正常鳞状上皮（P均〈0．01）;癌组织中AngⅡ的表达程度与VEGF的表达程度呈正相关（ra′=0．4249,P〈0．05）,AT1R的表达程度与VEGF无相关性（ra′=0．1298,P〉0．05）。结论 食管鳞癌癌组织中高水平表达AngⅡ及AT1R,二者可能通过诱导VEGF的产生促进肿瘤新生血管的形成。     

血管内皮细胞生长因子对肾小球内皮细胞通透性的影响

目的 :观察血管内皮细胞生长因子 (VEGF)对肾小球内皮细胞通透性的影响 ,并与脐静脉内皮细胞进行比较分析。　 方法 :采用二室弥散系统检测内皮细胞通透性。肾小球内皮细胞 (MGEC)和人脐静脉内皮细胞(HUVEC)接种于细胞培养嵌套的微孔滤膜 (孔径 0 4 5 μm)上 ,待细胞生长融合后 ,加入不同浓度的VEGF作为处理因素 ,以生物素标记的牛血清白蛋白 (biotin BSA)作为通透性指示剂 ,采用ELISA方法检测不同时间段biotin BSA通过细胞单层的百分数。　 结果 :正常培养条件下 ,生长在滤膜上的MGEC和HUVEC单层的通透性没有明显差异。VEGF可显著增加MGEC和HUVEC的通透性 ,呈剂量和时间依赖性。 1μg/LVEGF作用 12h可使MGEC对白蛋白的通透率增加近 2 5 % (0 31± 0 0 3vs 0 2 5± 0 0 3) ;VEGF浓度达 10 μg/L始显著增加HUVEC通透性 (0 2 8± 0 0 7vs0 2 1± 0 0 4 ) ,且远小于MGEC通透性增加的幅度 (P <0 0 1) ;VEGF浓度达 5 0 μg/L时 ,其增加内皮细胞通透性的作用基本达到饱和。 5 0 μg/LVEGF作用 0 5h即可使MGEC和HUVEC的蛋白通透率分别增加 10 0 % (0 12± 0 0 2vs0 0 6± 0 0 1)和 6 0 % (0 0 8± 0 0 3vs0 0 5± 0 0 1) ,并至少持续到 12h。　 结论 :首次在体外直接证实了VEGF具有增加MGEC     

糖尿病鼠肾小管周围新生血管与促血管生长因子关系研究

目的探讨促血管生成素-2(Ang-2)和血管内皮生长因子(VEGF)在糖尿病肾脏血管新生中的作用及其意义。方法2004年3月至2005年3月在四川大学华西医院用SD大鼠建立链唑霉素(STZ)诱导的糖尿病肾病模型,定量分析肾脏VEGF和Ang-2的表达变化,用逆转录聚合酶链式反应技术(RT-PCR)检测肾脏VEGF、Ang-2的mRNA的表达。结果糖尿病组肾脏VEGFmRNA表达从2周开始持续上调,16～20周达高峰;免疫组化显示糖尿病组各时点肾小管的VEGF着染均强于对照组;糖尿病组仅于16周和20周时探及肾组织Ang-2mRNA表达,12～24周免疫组化显示皮质区管周Ang-2着染管周微血管,整个实验期间对照组未探及Ang-2mRNA和蛋白表达;VEGF与Ang-2的改变呈正相关。结论糖尿病肾脏中后期皮质区存在Ang-2着染的新生管周微血管,VEGF与Ang-2参与糖尿病肾脏新生血管生成。     

胰岛素瘤的解剖分布特点分析

目的胰岛素瘤是最常见的胰腺神经内分泌肿瘤，因其临床表现多样，导致诊断困难。影像学诊断尤其是超声内镜(EUS)在胰岛素瘤的诊断中起着重要作用，拥有较高的敏感性和特异性。本研究拟通过明确胰岛素瘤的解剖分布特点，以期有助于提高影像学的诊断准确率和降低漏诊率，尤其是在教育和培训实践中对于EUS的学习者更具有指导价值。 方法回顾性分析解放军总医院第一医学中心病案资料数据库1993年1月至2019年11月经外科手术、病理确诊为胰岛素瘤的患者的临床资料，检索方法采取搜索术后病理诊断为"胰岛素瘤"的病例，通过查阅病例的方法，提取出胰岛素瘤的大小和解剖分布等数据，进一步分析其特点。 结果共检索到确诊为胰岛素瘤的患者116例，其中，男45例、女71例，年龄13～76岁，平均年龄(44.4±14.85)岁。胰岛素瘤单发110例(94.8%)、多发6例(5.2%)。位置分布：头颈部46例(39.7%)，单发45例、多发1例；体尾部68例(58.6%)，单发65例、多发3例；全胰腺多发2例(1.7%)。病变大小特点：最大径0.4～3.4 cm，平均大小(1.53±0.58)cm。≤1 cm 29例、>1 cm而≤1.5 cm41例、>1.5 cm而≤2.0 cm28例，≤3 cm 15例，>3 cm 3例。年龄与肿瘤的大小相关，≤44岁患者肿瘤平均大小为(1.36±0.51)cm、>44岁患者肿瘤平均大小为(1.70±0.60)cm，P<0.05。头颈部的肿瘤大于体尾部的肿瘤，头颈部肿瘤平均大小(1.66±0.63)cm，体尾部(1.42±0.52)cm，P<0.05。 结论胰岛素瘤在胰腺体尾部较头颈部更好发；绝大多数单发，但可以全胰腺多发；多数小于1.5 cm，肿瘤的大小与患者年龄和肿瘤的解剖分布相关。     

Pathogenesis of carcinoma of the papilla of Vater

Most adenomas and carcinomas of the small intestine and extrahepatic bile ducts arise in the region of the papilla of Vater. In familial adenomatous polyposis (FAP) it is the main location for carcinomas after proctocolectomy. In many cases symptoms due to stenosis lead to diagnosis at an early tumor stage. In about 80%, curative intended resection is possible. Operability is the most relevant prognostic factor. Most ampullary carcinomas resp. carcinomas of the papilla of Vater develop from adenomatous or flat dysplastic precursor lesions. They can be sited in the ampulloduodenal part of the papilla of Vater, which is lined by intestinal mucosa. They also can develop in deeper parts of the ampulla, which are lined by pancreaticobiliary duct mucosa. Intestinal-type adenocarcinoma and pancreaticobiliary-type adenocarcinoma represent the main histological types of ampullary carcinoma. Furthermore, there exist unusual types and undifferentiated carcinomas. Many carcinomas of intestinal type express the immunohistochemical marker profile of intestinal mucosa (keratin 7?, keratin 20+, MUC2+). Carcinomas of pancreaticobiliary type usually show the immunohistochemical profile of pancreaticobiliary duct mucosa (keratin 7+, keratin 20?, MUC2?). Even poorly differentiated carcinomas, as well as unusual histological types, may conserve the marker profile of the mucosa they developed from. These findings underline the concept of histogenetically different carcinomas of the papilla of Vater which develop either from intestinal- or from pancreaticobiliary-type mucosa of the papilla of Vater. Molecular alterations in ampullary carcinomas are similar to those of colorectal as well as pancreatic carcinomas, although they appear at different frequencies. In future studies, molecular alterations in ampullary carcinomas should be correlated closely with the different histologic tumor types. Consequently, the histologic classification should reflect the histogenesis of ampullary tumors from the two different types of papillary mucosa.     

Demonstration of the mechanism of action of biguanides

Summary Palmitic acid oxidation in rat diaphragm homogenate is depressed by biguanide concentrations that are still incapable of inhibiting oxidative phosphorylation. Glucose oxidation is not directly effected by the same biguanide concentrations: however, the inhibitory effect of palmitic acid on glucose oxidation is partly removed by biguanides. Inhibition of fatty acid oxidation, which accounts for most of the metabolic effects caused by these drugs, can be regarded as the fundamental mechanism of action of biguanides. There is some evidence suggesting that these drugs might interact with carnitine, thus preventing long-chain fatty acids from being transported across the mitochondrial membrane to the site of oxidation. Traduzione a cura degli AA.     

Lack of correlation of degree of villous atrophy with severity of clinical presentation of coeliac disease

BACKGROUND AND AIM: Both the clinical presentation and the degree of mucosal damage in coeliac disease vary greatly. In view of conflicting information as to whether the mode of presentation correlates with the degree of villous atrophy, we reviewed a large cohort of patients with coeliac disease. PATIENTS AND METHODS: We correlated mode of presentation (classical, diarrhoea predominant or atypical/silent) with histology of duodenal biopsies and examined their trends over time. RESULTS: The cohort consisted of 499 adults, mean age 44.1 years, 68% females. The majority had silent coeliac disease (56%) and total villous atrophy (65%). There was no correlation of mode of presentation with the degree of villous atrophy (p=0.25). Sixty-eight percent of females and 58% of males had a severe villous atrophy (p=0.052). There was a significant trend over time for a greater proportion of patients presenting as atypical/silent coeliac disease and having partial villous atrophy, though the majority still had total villous atrophy. CONCLUSIONS: Among our patients the degree of villous atrophy in duodenal biopsies did not correlate with the mode of presentation, indicating that factors other than the degree of villous atrophy must account for diarrhoea in coeliac disease.     

血吸虫童虫生长发育及其机制的研究进展

血吸虫童虫是宿主免疫系统攻击的重要靶标,包括皮肤型、肺型和肝门型童虫。宿主分子对童虫生长发育具有重要作用。童虫生长发育机制包括免疫调节、信号转导、性别发育及凋亡等。肌动蛋白、组织蛋白酶、烯醇化酶和葡萄糖基转移酶等分子为血吸虫童虫生长发育的重要分子。本文对血吸虫童虫生长发育及其机制的研究进展做一综述。     

124例耐多药结核分枝杆菌基因突变特征分析

目的对临床分离的耐多药结核分枝杆菌相关基因的突变特征进行分析。方法对124例耐多药结核分枝杆菌以及50株敏感株的耐药相关基因(包括异烟肼inh A、kat G、oxyR-ahp C间隔区以及利福平rpo B)进行序列测定,分析其基因突变情况。结果异烟肼耐药inh A基因突变率为14.5%;kat G基因突变率为70.2%(87/124),主要位于315位;oxyR-ahp C间隔区突变率为15.3%;inh A、kat G两种基因同时突变率75.0%,三种基因同时突变率为89.5%。利福平rpo B基因突变的检出率高达95.2%,突变主要发生在531、526、516位点。结论我省耐多药菌异烟肼耐药相关基因最常见突变为kat G 315、inh A C-T(-15)、axyR-ahp C间隔区(-10)C-T,利福平为rpo B531、526、516。结合MDR-TB耐药相关基因的特征分析,可以建立一种快速、准确、特异的适合于我省的检测结核菌耐多药性的新方法。     

氯硝柳胺悬浮剂的毒性评价

目的评价氯硝柳胺悬浮剂的毒性，为现场大规模应用灭螺提供依据。方法按照中华人民共和国国家标准GB 15670-1995《农药登记毒理学试验方法》和鱼类毒性试验方法进行。结果经口、经皮肤的LDso雌、雄性大鼠均>5 000 mg/kg，经呼吸道的LCso雌、雄性大鼠均>5 000mg/m3，该药经口、经皮肤、经呼吸道毒性均属微毒类药物；兔眼用药后，观察期内无不良反应，对眼无刺激性；皮肤用药后对皮肤无刺激性。与氯硝柳胺原药、氯硝柳胺乙醇胺盐原药和氯硝柳胺乙醇胺盐可湿性粉剂相比，氯硝柳胺悬浮剂对鱼急性毒性最低。结论氯硝柳胺悬浮剂属微毒类药物，对鱼的毒性低于其乙醇胺盐可湿性粉剂，适合于现场应用。     

Assessment of quality of life of parents of children with juvenile chronic arthritis

The aim of the study was to assess the quality of life (QOL) and the psychological status of parents of children with juvenile chronic arthritis (JCA). The QOL, anxiety and depression of the parents of 28 children with JCA were evaluated and compared to those of the parents of 28 healthy children. Mothers of JCA children and mothers of healthy children reported similar QOL. The reported anxiety and depression levels were similar for mothers and fathers in both groups. The parents of children with pauciarticular-type JCA reported lower QOL and higher levels of anxiety and depression than the parents of children with other types, namely polyarticular and systemic JCA. These findings may be explained by the fact that the pauciarticular patients had shorter disease duration and were less frequently seen in the outpatient clinic. The QOL of mothers of children with JCA was found to be slightly impaired in the group of children with pauciarticular JCA. Future larger studies are needed to confirm these results, as the number of subjects in the three groups was rather low. Received: 26 September 2001 / Accepted: 8 February 2002     

Numerical Simulation of the Effect of Rate of Change of Glucose on Measurement Error of Continuous Glucose Monitors

Background

A 5-day in-patient study designed to assess the accuracy of the FreeStyle Navigator® Continuous Glucose Monitoring System revealed that the level of accuracy of the continuous sensor measurements was dependent on the rate of glucose change. When the absolute rate of change was less than 1 mg•dl⁻¹•min⁻¹ (75% of the time), the median absolute relative difference (ARD) was 8.5%, with 85% of all points falling within the A zone of the Clarke error grid. When the absolute rate of change was greater than 2 mg•dl⁻¹•min⁻¹ (8% of the time), the median ARD was 17.5%, with 59% of all points falling within the Clarke A zone.

Method

Numerical simulations were performed to investigate effects of the rate of change of glucose on sensor measurement error. This approach enabled physiologically relevant distributions of glucose values to be reordered to explore the effect of different glucose rate-of-change distributions on apparent sensor accuracy.

Results

The physiological lag between blood and interstitial fluid glucose levels is sufficient to account for the observed difference in sensor accuracy between periods of stable glucose and periods of rapidly changing glucose.

Conclusions

The role of physiological lag on the apparent decrease in sensor accuracy at high glucose rates of change has implications for clinical study design, regulatory review of continuous glucose sensors, and development of performance standards for this new technology. This work demonstrates the difficulty in comparing accuracy measures between different clinical studies and highlights the need for studies to include both relevant glucose distributions and relevant glucose rate-of-change distributions.     

Angiographic diagnosis of the degree of serosal invasion of carcinoma of the colon

Angiography using Prostaglandin El^® was performed on 38 patients with carcinoma of the colon in order to diagnose the degree of serosal cancer invasion. The findings at angiography were classified into four groups:1) AG-S3, abnormal change (irregularity and/or encasement) up to marginal vessels; 2) AG-S2, abnormality up to vasa recta; 3) AG-S1, abnormality of penetrating branches of vasa recta within the wall of the colon; and 4) AG-S0, no distinct findings of abovementioned vessels. These angiographic findings were compared with both macroscopic and microscopic serosal cancer invasion. Angiographic diagnosis is in accord with the macroscopic findings in 84.2 percent of cases. Angiographic diagnosis is in accord with the microscopic findings in 32.4 percent of cases. Macroscopic findings confirm the angiographic diagnosis precisely but the conflict with microscopic findings should not be overlooked. This may be the result of inflammatory change, adhesion, and fibrosis around carcinoma of the colon.