木通所致肾小管间质肾病及其临床病理特点分析

目的探讨木通及相关中成药物所致肾小管间质肾病(MT-TIN)的临床病理特点,初步分析影响肾功能的相关因素及预后.方法回顾性分析51例MT-TIN病人的临床及病理资料,根据病理分为急性、慢性轻型和慢性重型组,对临床和病理指标进行半定量及相关分析,探讨其服药与发病、临床病理、治疗与转归特点,并与抗菌药物所致急性肾小管坏死(A-ATN)或急性间质性肾炎(A-AIN)加以比较.结果MT-TIN以乏力、消化道症状、多尿及夜尿增多为主要临床症状,常有明显肾小管功能障碍并可伴肾小球功能异常,特征为无明显尿镜检异常,以及病变早期无贫血.主要病理特点为急性期肾小管上皮细胞严重变性、坏死、脱落,形成裸基底膜,细胞再生差,肾间质少有细胞浸润,纤维化病变出现早;慢性期肾小管逐渐萎缩,肾间质弥漫纤维化.病人血肌酐(Scr)增高,并与贫血及肾间质纤维化程度密切相关.经停药,多数病人在2个月内病情可稳定.MT-TIN病人的临床和病理特点均与A-ATN或A-AIN有明显不同;其多数与过量或长期间断应用木通煎剂及含关木通的中成药有关,少数病人常规用药也可发病.结论MT-TIN是一类特殊类型的药物性肾小管间质疾病,主要与应用木通类药物不当有关,应尽可能避免应用含马兜铃酸的木通类药物,并对用药病人加强监测.     

肾间质纤维化中的微血管病变

肾间质纤维化是各种进展性肾脏疾病的共同通路。以往针对肾间质纤维化的研究 ,多数集中于肾小管上皮细胞、间质细胞活化 ,趋化因子、粘附分子的表达 ,炎症细胞的浸润 ,生长因子的作用 ,细胞外基质 (ＥＣＭ)产生和降解的失衡。有关肾间质微血管病变 (主要指肾小管周围毛细血管内皮细胞的损伤 )在肾间质纤维化中的作用 ,近年来才开始得到重视 ,其研究主要集中在以下两方面 :①微血管内皮细胞的损伤和修复 ;②微血管病变和肾小管间质慢性缺氧性损伤及纤维化的关系。1　微血管病变在间质纤维化中的作用肾间质微血管主要是指源于出球小动脉并分布…     

术通所致肾小管间质肾病及其临床病理特点分析

目的 探讨木通及相关中成药物所致肾小管间质肾病（MT-TIN）的临床病理特点，初步分析影响肾功能的相关因素及预后。方法 回顾性分析51例MT-TIN病人的临床及病理资料，根据病理分为急性、慢性轻型和慢性重型组，对临床和病理指标进行半定量及相关分析，探讨其服药与发病、临床病理、治疗与转归特点，并与抗菌药物所致急性肾小管坏死（A-ATN）或急性间质性肾炎（A-AIN）加以比较。结果 MT-TIN以乏力、消化道症状、多尿及夜尿增多为主要临床症状，常有明显肾小管功能障碍并可伴肾小球功能异常，特征为无明显尿镜检异常，以及病变早期无贫血。主要病理特点为急性期肾小管上皮细胞严重变性、坏死、脱落，形成裸基底膜，细胞再生差，肾间质少有细胞浸润，纤维化病变出现早；慢性期肾小管逐渐萎缩，肾间质弥漫纤维化，病人血肌酐（Scr)增高，并与贫血及肾间质纤维程度密切相关。经停药，多数病人在2个月内病情可稳定。MT-TIN病人的临床和病理特点均与A-ATN或A-AIN有明显不同；其多数与过量或长期间断应用木通煎剂及含关木通的中成药有关，少数病人常规用药也可发病。结论 MT-TIN是一类特殊类型的药物性肾小管间质疾病，主要与应用木通类药物不当有关，应尽可能避免应用含马兜铃酸的木通类药物，并对药病人加强监测。     

UC-MSCs对UUO大鼠肾间质ET-1、VEGF、肾小管周Ⅷ因子表达水平的影响

目的 观察人脐血间充质干细胞(UC-MSCs)对单侧输尿管梗阻(UUO)大鼠肾间质中内皮素-1(ET-1)、血管内皮生长因子(VEGF)、肾小管周Ⅷ因子表达的影响.方法 24只大鼠随机分为3组,各8只.模型组与治疗组均行右侧输尿管结扎术,假手术组手术入路方式同模型组,进腹腔后分离右输尿管但不结扎,术后注射生理盐水0.5mL;模型组术后注射生理盐水0.5 mL;治疗组术后注射UC-MSCs 1×106个(0.5 mL).术后14 d处死各组动物.术后14 d取梗阻侧肾组织,观察肾小管间质损伤程度,免疫组化法检测肾组织中ET-1、VEGF及肾小管周Ⅷ因子的水平.结果 假手术组肾间质中ET-1、VEGF及肾小管周Ⅷ因子相对表达量分别为1.178 8±0.884 2、0.456 3 ±0.079 3、13.16±8.01,模型组分别为9.623 8±3.323 4、1.346 3±0.612 9、7.36±1.07,治疗组分别为5.243 7±2.657 4、4.290 0±0.594 6、10.29±0.06,三组相比,P均＜0.05.肾小管周Ⅷ因子与ET-1表达呈负相关(r=-0.881,P＜0.01),与VEGF表达呈正相关(r=0.861,P＜0.01).结论 UC-MSCs可以抑制ET-1的表达,上调VEGF水平,保护肾间质微血管,延缓肾间质纤维化,从而发挥肾保护作用.     

单克隆免疫球蛋白相关肾小管间质疾病诊治进展

单克隆免疫球蛋白相关肾小管间质疾病主要指单克隆轻链(κ,λ) 所导致的肾小管间质病变, 包括轻链相关 肾小管损伤及肾小管坏死、轻链相关肾小管间质性肾炎、轻链近端肾小管病及轻链管型肾病等四类疾病。临床上以 肾功能不全为主要特征,可伴有小分子为主的蛋白尿;病理以单克隆轻链在肾小管上皮胞浆、肾小管基底膜及管型 沉积为共同特征;电镜与免疫电镜证实单克隆轻链在上皮胞质溶酶体及其结晶等包涵体内的蓄积,具有重要的诊断 价值。轻链导致不同的病理损伤类型,可能与轻链的负荷量、轻链的特定基因亚型及其不同的结构变异有关,详细 的发病机制有待进一步研究。     

缬沙坦和雷米普利对5/6肾切除大鼠肾间质微血管的作用

目的 :观察大鼠 5 /6肾切除模型中间质微血管病变的特点及药物缬沙坦和雷米普利对其的干预作用。　 方法 :以SD大鼠 5 /6肾切除建立肾间质纤维化 (RIF)动物模型 ,设正常组 (n =6 ) ,假手术组 (n =6 ) ,手术组(n =7) ,氨氯地平组 (n =6 ) ,缬沙坦组 (n =10 ) ,雷米普利组 (n =10 )。术后定时测定 2 4h尿蛋白排泄量及血压。 12周后取材 ,留取血样 ,2 4h尿样及肾组织。常规病理检查判断RIF及肾小管间质损伤程度 ;应用免疫组化方法显示肾脏毛细血管。　 结果 :与正常组和假手术组相比 ,手术组RIF明显 ,间质微血管病变严重 ,肾小管周围毛细血管(PTC)密度明显减少 (36 3 2 9± 70 .99vs 798.17± 6 6 .5 3,776 .10± 5 1.2 9,P <0 0 1)。而缬沙坦和雷米普利能延缓RIF及间质微血管病变 ,并改善肾功能 ,与手术组相比 ,两组PTC密度均明显改善 (45 5 5 0± 6 2 .98,P <0 .0 5和 4 6 5 6 0±5 7 38,P <0 0 1)。而氨氯地平血压控制组与手术组PTC密度无差异。　 结论 :间质微血管病变不仅是RIF的特征性病理改变 ,而且在RIF及肾功能减退中可能起重要作用。缬沙坦和雷米普利延缓RIF和对肾功能的保护作用与间质微血管病变的改善有关 ,其机制可能与血管紧张素Ⅱ相关联。     

急性马兜铃酸肾病大鼠肾脏微血管损伤的研究

目的探讨急性马兜铃酸肾病（AAAN）肾脏微血管损伤的机制。方法将48只雌性SD大鼠随机分为观察组及对照组各24只。观察组采用关木通水煎剂灌胃5d制作AAAN模型;对照组灌相同体积自来水。灌胃后第3、5、7天两组分别处死大鼠各8只,留取血、尿、肾组织标本分别行肾功能（24h尿蛋白量及血BUN和SCr水平）、肾脏病理（电镜）、肾小球毛细血管密度及HIF-lα、VEGF表达（免疫组化法）等检测。结果①肾功能：观察组第3、5、7天各检测指标逐渐升高,且明显高于对照组,P均〈0.05;②病理：观察组示肾小球基底膜断裂,内皮细胞破裂;对照组无明显异常;③毛细血管密度：观察组第3、5、7天血管密度逐渐减少,且明显低于对照组,P均〈0.05;④HIF-1α及VEGF表达：对照组HIF-1α几乎没有表达;观察组第3天仅微量表达,至第7天表达明显增强,与对照组比较,P均〈0.05。对照组VEGF大量表达;观察组表达逐步减少,且明显低于对照组,P均〈0.05。结论 AAAN大鼠存在明显的肾脏微血管损伤;VEGF表达降低可能是其原因。     

肾组织粘附分子及树突状细胞与IgA肾病小管间质病变的关系

目的:观察粘附分子P-选择素与树突状细胞(DC)在IgA肾病(IgAN)患者肾组织中的表达与分布,探讨P-选择素及DC与肾小管间质病变以及肾功能损害的关系.方法:选择经肾活检和临床资料确诊的IgAN患者45例,根据小管间质病变程度分为3组:轻度组29例,中度组10例,重度组6例;10例正常人肾组织作为对照.采用免疫组化法观察IgAN患者肾组织中P-选择素表达;免疫双标记染色与荧光图像分析法,观察CD1a CD80 DC在肾组织中的分布,并探讨DC与患者肾小管间质病变及肾功能损害的关系.结果:①P-选择素在正常肾组织中基本不表达,在IgAN肾组织中以肾小管上皮细胞为主广泛高表达,重度组肾小管间质表达明显强于轻、中度组,且与肾小管间质病变程度轻重显著相关.②正常人肾组织中基本未见CD1a CD80 DC分布,IgAN肾组织中以肾小管间质为主分布明显增多;其在患者肾小管间质的分布于重度组明显高于轻、中度组,亦与肾小管间质病变程度及血肌酐显著相关.此外,CD1a CD80 DC在患者肾小管间质分布与P-选择素表达明显相关.结论:P-选择素尤其DC可能参与了IgAN肾小管间质病变的免疫病理损伤机制,且推测DC肾炎组织迁移聚集可能与P-选择素介导有关.     

移植肾缺血-再灌注损伤与炎症和微血管病变

移植肾早期不可避免地存在缺血再灌注损伤(IRI),对移植肾的早期恢复和远期预后均有重要影响.缺血-再灌注除损伤肾小管上皮细胞外,还导致间质炎症和徽血管病变;而后者是IRI引起肾功能障碍的关键.缺血后微血管病变以内皮细胞肿胀和随之而来的微血管闭塞为特征,导致再灌注不能顺利进行(无复流现象).阐明内皮细胞功能障碍机制,为移植肾IRI的治疗提供新思路.     

非那雄胺对前列腺血管内皮生长因子与微血管的影响

目的　探讨非那雄胺对前列腺组织中血管内皮生长因子 (VEGF)与微血管密度的影响。方法　应用免疫组织化学方法检测小剂量用药组 (A组 )、大剂量用药组 (B组 )、对照组 (C组 )及正常组 (D组 )前列腺组织中 VEGF和 CD34 的表达。结果　 A组、B组与 D组前列腺微血管密度、微血管腔内面积及 VEGF染色阳性细胞面积均明显小于 C组 ,差别有显著性意义 (P<0 .0 1 ) ;B组前列腺微血管密度、微血管腔内面积及 VEGF染色阳性细胞面积小于 A组 ,差别有意义 (P<0 .0 5)。结论　非那雄胺可以抑制前列腺组织中血管内皮生长因子的表达 ,减少腺体内微血管的密度     

Effects of mifepristone on invasive and metastatic potential of human gastric adenocarcinoma cell line MKN-45 in vitro and in vivo

AIM： To investigate the effects of mifepristone on the invasive and metastatic potential of human gastric adenocarcinoma cell line MKN-45 and its mechanisms. METHODS: After incubation with various concentrations of mifepristone (5, 10, 20 umol/L), the adhesion to artificial basement membrane, Matrigel, and the migration of MKN-45 cells were assayed using MTT assay and Transwell cell culture chambers, respectively. Enzyme- linked immunoabsorbent assay (ELISA) and flow cytometry were used to determine the expression of vascular endothelial growth factor (VEGF) and integrin 133 in the cells. After subcutaneous transplantation of MKN-45 cells in nude mice, mifepristone (50 mg/kg.d) was administrated subcutaneously for 8 wk to assess its effects on tumor metastasis. Immunohistochemical analysis was used to detect the expression of VEGF and microvascular density (MVD) in xenografted tumors. RESULTS: Mifepristone dose-dependently inhibited the heterotypic adhesion to Matrigel of MKN-45 cells. The inhibition was accompanied by a significant down-regulation of integrin 133 expression in the cells. After incubation with 5, 10, 20 umol/L mifepristone, the number of migrated MKN-45 cells was 72+8, 50+6, 41+5 in experiment group, and 94+16 in control group (P&lt;0.01). Meanwhile, secreted VEGF protein of MKN-45 cells in mifepristone-treated group (14.2+2.9, 8.9+3.1, 5.4+2.1 ng/g per liter) was significantly lower than that in control group (22.7+4.3 ng/g per liter, P&lt;0.01). In vivo, mifepristone decreased the number of metastatic foci in lungs of nude mice and down-regulated the expression of VEGF and MVD in the xenograted tumors. CONCLUSION: Mifepristone can effectively inhibit the invasive and metastatic potential of human gastric adenocarcinoma cell line MKN-45 in vitro and in vivo through inhibition of heterotypic adhesion to basement membrane, cell migration and angiogenesis.     

The expression of vascular endothelial growth factor and the type 1 vascular endothelial growth factor receptor correlate with the size of papillary thyroid carcinoma in children and young adults.

Vascular endothelial growth factor (VEGF) is essential for the growth of many solid tumors, but there are little data regarding VEGH in childhood thyroid cancers. We examined the relationships between VEGF, the type 1 VEGF receptor (FLT-1) and clinical outcome for a group of thyroid cancers in children and young adults. The expression of VEGF and FLT-1 were determined by immunohistochemistry using archival, paraffin-embedded thyroid tissue blocks and compared with the retrospective clinical outcome for each patient. The study included 67 children and young adults with papillary thyroid carcinoma (PTC, n = 42), follicular thyroid carcinoma (FTC, n = 8), benign lesions (n = 15), or controls (n = 2). VEGF expression was greater in PTC (mean intensity 2.23 +/- 0.25, p = 0.002) and FTC (2.8 +/- 0.73, p = 0.01) than benign lesions (1.0 +/- 0.27), and correlated with PTC size (r = 0.42, p = 0.008). FLT-1 expression was greater in PTC (mean intensity 2.8 +/- 0.17) than FTC (1.9 +/- 0.25, p = 0.015) and benign lesions (1.7 +/- 0.32, p = 0.002); and correlated with PTC size (r = 0.41, p = 0.01) as well as VEGF expression (r = 0.52, p = 0.002). Recurrent disease developed exclusively in patients with PTC which expressed VEGF (7/28, 95% CI 10.6%-44.2%). PTC that did not express VEGF (0/8, 95% CI = 0%-31.2%) did not recur; however, the difference was not statistically significant (p = 0.15). We conclude that the expression of VEGF and FLT-1 are directly correlated with the size of PTC in children and young adults.     

血管内皮生长因子反义寡核苷酸抑制甲状腺癌血管生成的效应

目的探讨反义寡核苷酸(ASODN)抑制甲状腺癌细胞血管内皮生长因子(VEGF)表达及内皮细胞生长的效应。方法设计合成靶向VEGF的ASODN转染人髓状甲状腺癌细胞系(TT)细胞,并制备相应条件培养基作用内皮细胞ECV304,设正义寡核苷酸(SODN)和空白对照组进行比较。观察细胞生长状态,RT PCR、免疫细胞化学法检测TT细胞VEGFmRNA和蛋白表达,四氮唑蓝法检测TT和ECV304细胞生长抑制率(IR),流式细胞仪、吖啶橙/溴化乙锭染色法检测ECV304细胞凋亡状态。结果ASODN组TT细胞VEGFmRNA和蛋白表达显著低于SODN和对照组(P<0.01),但IR差异无统计学意义(P>0.05);各转染组ECV304细胞IR差异亦无统计学意义(P>0.05);而经各ASODN组TT细胞条件培养基作用的ECV304细胞生长明显受抑,IR(分别为0.21±0.03、0.31±0.01、0.42±0.22)显著高于SODN组(0.05±0.03,P<0.01),并伴明显细胞凋亡,上述效应呈浓度依赖性。结论ASODN可通过特异性封闭甲状腺癌细胞VEGF表达,抑制内皮细胞生长,干扰肿瘤血管生成。     

转染正义血管内皮细胞生长因子cDNA对大鼠肺泡巨噬细胞趋化蛋白1表达的影响

目的探讨博莱霉素(BLM)致大鼠肺微血管内皮细胞紧密连接变化特征以及血管内皮细胞生长因子(VEGF)对肺微血管内细胞中巨噬细胞趋化蛋白1(MCP-1)mRNA表达的影响。方法健康SD大鼠按随机数字表法随机分为对照组和实验组。实验组大鼠制作BLM损伤肺纤维化模型,分别在第3、7、14以及28天行硝酸镧灌注后,制作肺组织病理标本,透射电镜观察血管内皮细胞(EC)紧密连接及镧颗粒分布。组织块培养法行大鼠肺微血管内皮细胞体外培养。体外培养的大鼠肺微血管内细胞中转染正义VEGF cDNA,采用逆转录-聚合酶链反应(RT-PCR)法检测MCP-1 mRNA的表达。结果对照组EC结构完整,基底膜呈连续线状。EC间连接紧密,镧盐颗粒未见通过EC间的紧密连接。实验组不同时间处理的大鼠均可见EC连接间隙增宽,并有高密度的镧颗粒在细胞连接间隙中线状沉淀,其中以第3天为著,分布于血管周围的内皮下区域,大部分通过基底膜到达细胞外间隙。与空白对照组比较,转染正义VEGF cDNA组肺微血管内皮细胞MCP-1 mRNA表达显著增加(P<0.05),空白对照组为0.36±0.06,转染组为1.21±0.22。结论BLM损伤肺微血管内皮细胞间紧密连接呈持续开放状态和MCP-1持续高表达,成为特异性定向诱导炎性细胞渗出和游走到组织间的病理基础,而炎性细胞持续分泌的细胞因子成为启动成纤维细胞过度增殖、诱发肺纤维化的重要因素之一。     

血管内皮生长因子及其受体在肺气肿患者肺组织中的表达

目的探讨血管内皮生长因子(VEGF)及其受体2(VEGF受体2/KDR)在肺气肿患者肺组织中的表达及其与肺气肿的相关性。方法取35例行肺叶切除术患者[A组(吸烟伴肺气肿组)16例,B组(不吸烟肺功能正常组)14例,C组(吸烟但肺功能正常组)5例]的外周肺组织标本,ELISA法检测肺组织匀浆中VEGF的含量,免疫组化法检测KDR蛋白表达,RT PCR检测VEGF和KDRmRNA水平,TUNEL法检测肺泡隔细胞的凋亡。结果A组患者肺组织VEGF、KDR表达均低于B组(P<0.01),肺泡隔细胞凋亡率高于B组(P<0.01)。C组与B组相比,VEGF及KDR表达差异无统计学意义(P>0.05)。结论VEGF及KDR水平减少与肺泡隔细胞凋亡的增加可能与肺气肿的发生相关。     

血管生成素-1对小鼠早期急性肺损伤的保护作用

目的 观察血管生成素-1(Ang-1)对油酸致小鼠早期急性肺损伤的保护作用,探讨Ang-1对小鼠早期急性肺损伤血管内皮细胞生长因子(VEGF)表达的影响.方法 96只BALB/c雌性小鼠按随机数字表法分为4组,每组24只.正常对照组:尾静脉注射生理盐水(0.9 ml/kg);急性肺损伤组:尾静脉注射油酸(0.9 ml/kg)制备急性肺损伤模型;正常对照+Ang-1组:注入生理盐水4 h后腹腔内注射Ang-1(312.5μg/kg);急性肺损伤+Ang-1组:注入油酸4 h后腹腔内注射Ang-1(312.5μg/kg);8 h后各组随机抽12只小鼠处死后行支气管肺泡灌洗(BAL),另外12只小鼠右肺称重后烘干,左肺留取病理标本;分别测定肺湿/干重比、支气管肺泡灌洗液(BALF)中细胞总数、中性粒细胞数及蛋白含量变化;采用酶联免疫吸附(ELISA)法测定血清和BALF中自细胞介素-6(IL-6)、VEGF含量;光镜观察并盲法评分比较肺组织病理学改变;采用免疫组化法检测肺组织中VEGF的表达.采用SPSS 10.0软件.数据以-x±s表示,多组间比较采用单因素方差分析,组间比较采用LSD-t检验,P＜0.05为差异有统计学意义.结果 急性肺损伤+Ang-1组肺湿/干重比、BALF中蛋白含量和细胞总数、中性粒细胞、血清和BALF中IL-6含量、血清VEGF含量分别为4.09±0.14、(176±13)μg/ml、(34.4±4.1)×104/L、(19.85±3.93)×103/L、(1318±62)pg/ml、(652±17)pg/ml、(48±5)pg/ml,与急性肺损伤组[5.32±0.51、(227±12)μg/ml、(42.2±5.2)×104/L、(26.22±2.22)×103/L、(1510±117)pg/ml、(744±13)pg/m1、(74±5)pg/ml]比较差异有统计学意义(t值分别为8.16、9.92、4.02、4.88、5.03、19.18、14.81,P均＜0.01);而急性肺损伤+Ang-1组BALF中VEGF含量[(179±15)pg/ml]与B组[(140±20)pg/ml]比较差异有统计学意义(t=5.31,P＜0.01);急性肺损伤+Ang-1组肺损伤评分为(1.84±0.12)分,急性肺损伤组为(3.44±0.21)分,两组比较差异有统计学意义(t=24.16,P＜0.01).肺组织中VEGF表达吸光度(A)值急性肺损伤+Ang-1组为549±72,与急性肺损伤组(342±85)比较差异有统计学意义(t=5.22,P＜0.01);急性肺损伤+Ang-1组与正常对照组(768±111)比较差异亦有统计学意义(t=5.35,P＜0.01);正常对照+Ang-1组和正常对照组肺组织中VEGF吸光度(A)值表达比较差异无统计学意义(t=0.69,P＞0.05).结论 Ang-1对油酸致小鼠早期急性肺损伤可减轻渗出、抑制炎性细胞浸润和炎症因子动员,同时Ang-1可引起小鼠早期急性肺损伤血清VEGF表达降低,而BALF和肺组织中VEGF表达增加.提示Ang-1对油酸致小鼠早期急性肺损伤有一定的保护作用.     

Interrelated overexpression of endothelial and inducible nitric oxide synthases, endothelin-1 and angiogenic factors in human papillary thyroid carcinoma

OBJECTIVE: Nitric oxide (NO) and endothelin-1 (ET-1) are involved in carcinogenesis. Overexpression of the ET-1 axis has been demonstrated in papillary thyroid carcinoma (PTC). This study investigated the expression of NO synthases (NOS) and their relationship with expression of ET-1 and angiogenic markers in PTC. DESIGN AND PATIENTS: Expression of NOS, angiogenic markers [vascular endothelial growth factor (VEGF), angiopoietin-1 and angiopoietin-2] and their receptors was studied in surgical thyroid samples obtained from 22 patients aged 15-68 years. Three groups were constituted: normal thyroid (n = 5), Hashimoto's thyroiditis (n = 9) and PTC (n = 8). RESULTS: Immunohistochemistry disclosed NOS2 and NOS3 immunoreactivity in PTC cells, the percentage of positive cells being greater than normal (P < 0.02). Real-time quantitative polymerase chain reaction (RTQ-PCR) showed that NOS2 and NOS3 mRNA levels were, respectively, increased (P < 0.02) by 2.6 +/- 0.6 and 4.2 +/- 1.1 times in PTC. RTQ-PCR demonstrated that VEGF, its receptors VEGFR-1 and VEGFR-2, and angiopoietin-2 and its receptor (Tie2) were also overexpressed (P < 0.05) in PTC. Correlations were found between ET-1 expression and that of NOS2, angiopoietin-1 and -2 (P < 0.05). NOS2 mRNA levels also correlated with those of NOS3 and angiopoietin-2 (P < 0.05). In thyroiditis, NOS2 immunoreactivity was observed in inflammatory cells whereas NOS2 mRNA levels were 12.1 +/- 1.6 times higher than normal (P < 0.005). CONCLUSIONS: This study revealed an activation of the NO pathway in thyroid carcinoma, which is interrelated to the ET-1 axis, both systems being overexpressed in concert with angiogenic factors. This global system might play a role in carcinogenesis and constitutes a potential target for anticancer therapy.     

Influence of the BRAF V600E mutation on expression of vascular endothelial growth factor in papillary thyroid cancer

CONTEXT: The BRAF mutation may influence the expression patterns of molecular markers that are related to the development and progression of thyroid cancer. OBJECTIVE: The objective of the study was to investigate the effects of the BRAF V600E mutation on expression of galectin-3, cyclooxygenase-2, cyclin D1, p53, and vascular endothelial growth factor (VEGF) in papillary thyroid cancer (PTC). DESIGN, SETTING, AND SUBJECTS: One hundred sixty-three PTC and 28 nodular hyperplasia patients were selected retrospectively. The presence of the BRAF V600E mutation and the level of expression of the molecular markers were determined. RESULTS: Of 161 PTC patients, 102 patients (63.4%) were BRAF V600E(+), and these cases had significantly larger tumor sizes (P = 0.01), compared with V600E(-) cases (n = 59, 36.6%). Although PTC tissues had higher expression levels of the selected molecular markers than nodular hyperplasia tissues, expression levels of several molecular markers in BRAF V600E(+) PTC were not significantly different from those of BRAF V600E(-) PTC. But VEGF was significantly up-regulated in BRAF V600E(+) PTC, compared with BRAF V600E(-) PTC. VEGF expression levels were strongly positively correlated to tumor size (P < 0.001), extrathyroidal invasion (P = 0.02), and tumor stage (P = 0.04). Multivariate analysis clearly showed that VEGF expression was up-regulated in BRAF V600E(+) PTC (odds ratio 2.5, confidence interval 1.1-5.6; P = 0.03). CONCLUSIONS: BRAF V600E(+) PTC tended to have larger tumor volumes and higher expression of VEGF. The level of VEGF expression was closely correlated with tumor size, extrathyroidal invasion, and stage. The relatively high levels of VEGF expression may be related to poorer clinical outcomes and recurrences in BRAF V600E(+) PTC.     

血管内皮生长因子受体抑制剂对小鼠支气管哮喘模型气道炎症和气道重塑的影响

目的研究血管内皮生长因子(VEGF)受体抑制剂对变应性气道炎症和气道重塑的影响,阐明VEGF与支气管哮喘(简称哮喘)以及气道重塑的关系。方法BALB/c小鼠按随机数字表法分为正常对照组(A组)、哮喘模型组(B组)、VEGF受体抑制剂治疗组(C组),每组各10只。用酶联免疫吸附测定(ELISA)法对各组小鼠支气管肺泡灌洗液(BALF)和血清中VEGF进行定量分析;用免疫组化法检测VEGF在小鼠肺组织内的表达水平。采用医学图像分析软件测定支气管管壁厚度(WAt/P i)、支气管平滑肌厚度(WAm/P i)、支气管平滑肌细胞计数(N/P i)及肺组织切片中的血管计数、血管壁平滑肌厚度、血管壁平滑肌细胞计数。结果BALF中细胞总数和嗜酸粒细胞比值B组分别为(142±63)×107/L、98.0±46.9,A组分别为(30±14)×107/L、0.7±1.1,C组分别为(41±17)×107/L、4.9±3.5,A组和C组BALF中细胞总数和嗜酸粒细胞比值分别与B组比较差异有统计学意义(P均<0.01)。B组BALF中上清液和血清中VEGF水平分别为(55±26)pg/m l、(72±26)pg/m l,A组分别为(37±9)pg/m l、(49±18)pg/m l,C组分别为(34±3)pg/m l、(43±19)pg/m l,A组与B组、C组与B组间比较差异均有统计学意义(P均<0.05)。免疫组化结果显示,B组大部分支气管平滑肌、黏膜上皮细胞、肺泡上皮细胞和血管周围VEGF均呈阳性表达,而A组和C组几乎没有VEGF的表达。图像分析显示,B组WAt/P i、WAm/P i、N/P i分别为(17±5)μm2/μm、(6.3±2.2)μm2/μm、(0.050±0.020)个/μm,A组分别为(8±3)μm2/μm、(3.2±0.8)μm2/μm、(0.027±0.017)个/μm,A组与B组比较差异有统计学意义(P分别<0.01、0.05)。B组和A组血管计数分别为(19±3)个、(10±5)个,A组与B组比较差异均有统计学意义(P<0.01)。经抑制剂治疗后C组WAm/P i、血管计数分别为(4.5±1.3)μm2/μm、(11±3)个,C组与B组比较差异均有统计学意义(P均<0.05)。结论VEGF在小鼠哮喘模型气道及肺内过度表达,并参与了哮喘的发病和气道重塑过程。VEGF受体抑制剂可明显改善哮喘小鼠的变应性气道炎症和气道重塑的病理生理过程。     

急性白血病患者血管内皮生长因子及其受体表达的动态观察

目的探讨急性白血病(AL)患者治疗前后骨髓中血管内皮生长因子(VEGF)及其受体的表达差异以及这种表达与血管生成的相关性.方法应用EnVision免疫组织化学二步法,检测122例次成人AL患者骨髓中造血细胞VEGF及其两种特异性受体fms-样酪氨酸激酶受体(Flt-1)、激酶插入嵌合受体(KDR)蛋白的表达情况.结果化疗后获得完全缓解(CR)的患者,其VEGF、KDR蛋白的表达在治疗前为6.0(3.3～12.0)和5.3(3.3～8.0),获CR后为5.3(3.3～9.0)和2.0(1.0～4.0)差异有显著性(P<0.05;P<0.01),而在化疗后未获得CR患者中的表达差异无显著性.在缓解后复发患者中的表达又升高到初发时的水平.各组初发患者Flt-1的表达水平与对照组之间差异无显著性,但CR期Flt-1的表达水平在CR组为3.3(1.7～5.3),复发组为3.3(2.0～5.3)与初发及对照组差异有显著性(P<0.01).微血管数处于高水平组的VEGF及KDR表达显著高于微血管处于低水平组者(P<0.01).骨髓原始细胞与急性髓系白血病(AML)初发患者VEGF和KDR的表达之间成正相关(r=0.429,0.359;P=0.005,0.02);与急性淋巴细胞白血病(ALL)初发患者VEGF的表达之间成正相关(r=0.522,P=0.03).结论 VEGF及其两种特异性细胞受体Flt-1, KDR在造血细胞及血管内皮细胞中表达.提示VEGF可能是白血病细胞的一种自分泌因子,同时作为一种旁分泌因子调控患者骨髓中的血管新生反应.VEGF及其细胞受体KDR可能构成抗血管新生和抗白血病治疗的新靶点.