Protective effect of scutellaria baicalensis stem-leaf total flavonoid on the injury of microvascular architecture and lipid peroxidation induced by cerebral ischemia reperfusion in rats

目的:探讨黄芩茎叶总黄酮(SSTF)预处理对脑局灶性缺血再灌注大脑皮质的保护作用及其机制.方法:线栓法制备局灶性脑缺血再灌注模型,造模成功后24 h测脑梗死体积,单宁酸-氯化铁媒染法显示大脑皮质微血管,并用Mivnt图像分析系统对皮质微血管密度(MVD)及微血管面积密度(MVA)进行定量分析;检测脑皮质组织中超氧化物歧化酶(SOD)活力、丙二醛(MDA)含量.结果:SSTF预处理可改善术后大鼠的MVD和MVA;降低因缺血再灌注引起的脑皮质组织MDA含量增加,提高SOD活力.结论:SSTF预处理可改善脑皮质微血管形态结构,抵制脂质化过氧化反应,减轻自由基损害,对缺血再灌注损伤脑组织具有保护作用.     

丁苯酞预处理对大鼠脑缺血再灌注损伤的保护作用及可能机制

目的丁苯酞预处理对大鼠脑缺血再灌注损伤后脑水肿、血脑屏障和基质金属蛋白酶-9(MMP-9)表达的影响。方法选取48只健康雄性大鼠,随机分为假手术组(Sham,16只),缺血再灌注组(I/R,16只)和丁苯酞干预组(TanⅡA,16只),用线栓法阻塞大鼠右大脑中动脉制作局灶性脑缺血再灌注模型。干湿重法测定脑含水量反映脑水肿程度,比色法测定脑组织伊文思蓝(EB)含量反映血脑屏障的损伤程度,免疫组织化学方法法和Western blot方法检测各组大鼠脑组织MMP-9蛋白表达,Real-time PCR检测MMP-9m RNA表达。结果相比于假手术组,缺血再灌注组大鼠脑含水量及EB含量显著升高,而NBP预处理组脑含水量及EB含量明显降低(0.01)。相比于假手术组,缺血再灌注组大鼠脑组织MMP-9蛋白及及m RNA的表达水平均显著升高,NBP预处理组则显著降低(0.01)。结论 NBP预处理对大鼠脑缺血再灌注损伤的保护作用,可能与调节MMP-9的表达、降低血脑屏障通透性和减轻脑水肿相关。     

葛根素预处理对脑缺血再灌注大脑皮质微血管和血脑屏障的预防性保护作用

目的:探讨葛根素(Pue)对缺血再灌注(IR)大鼠大脑皮质微血管和血脑屏障预防性保护作用。方法:SD大鼠随机分为假手术组(Sham组)、IR组、葛根素组和尼莫地平组(NIM组);葛根素组、NIM组分别于术前给予葛根素50 mg·kg~(-1)·d~(-1)、NIM 1mg·kg~(-1)·d~(-1)预处理,制备大脑中动脉阻塞脑缺血再灌注模型;单宁酸-氯化铁法(TA-Fe法)显示大脑皮质微血管,图像定量分析微血管密度(MVD)和微血管面积比(MVA);伊文思蓝(EB)和干湿重法检测脑组织含水量和血脑屏障(BBB)通透性,透射电镜观察大脑皮质BBB的超微结构变化。结果:与IR组相比,葛根素组脑组织含水量和微血管通透性明显降低,MVD、MVA显著增高,葛根素组与尼莫地平组差异无统计学意义。葛根素组、尼莫地平组的大脑皮质缺血区BBB超微结构变化接近,都表现为内皮细胞肿胀减轻,紧密连接趋于正常,基膜松散但连续,胶质膜排列较致密。结论:葛根素预处理可通过减轻微血管损伤、增加再通数量,降低血脑屏障的通透性,减轻脑水肿,预防性保护缺血再灌注损伤脑组织。     

脑缺血再灌注损伤后血脑屏障结构与功能的变化及丁苯酞预处理的干预作用

 目的:探讨脑缺血再灌注损伤（CIRI）大鼠血脑屏障结构与功能的变化,以及丁苯酞（NBP）预处理对其的干预作用。 方法: 120只雄性SD大鼠随机分为假手术组（sham组）、模型组（IR组）、NBP预处理组低剂量组（NBPⅠ组）、NBP预处理组中剂量组（NBPⅡ组）和NBP预处理组高剂量组（NBP Ⅲ组）,每组24只。采用线栓法制作CIRI模型,缺血2 h再灌注24 h后测定脑含水量和伊文思蓝（EB）含量,透射电镜下观察血脑屏障病理形态学变化,分别采用免疫组化法和real-time PCR法检测基质金属蛋白酶9（MMP-9） 蛋白和mRNA的表达。 结果: 脑缺血再灌注损伤后脑水肿加重,EB含量明显增加,血脑屏障损伤严重,MMP-9表达上调（均P<0.01）;NBP预处理后能明显减轻脑水肿,减少EB含量,减轻血脑屏障损伤,降低MMP-9的表达（均P<0.01）;NBPⅡ、Ⅲ组较NBPⅠ组改变更为明显（P<0.05）,Ⅱ、Ⅲ组之间差异无统计学意义（P＞0.05）。 结论: 丁苯酞预处理脑缺血再灌注损伤大鼠后,可降低MMP-9的表达,减轻血脑屏障损伤程度,发挥预防性保护作用。     

丁苯酞预处理对脑缺血再灌注损伤大鼠基质金属蛋白酶-9、基质金属蛋白酶组织抑制因子-1和血脑屏障的影响

目的:探讨丁苯酞预处理对脑缺血再灌注损伤大鼠基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶组织抑制因子-1(TIMP-1)和血脑屏障的影响.方法:90只雄性SD大鼠随机分为假手术组,模型组,NBP预处理低剂量组、中剂量组、高剂量组.采用线栓法制作大脑中动脉栓塞(MCAO)模型,缺血2h再灌注24 h后以干湿重法测定脑组织含水量,伊文思蓝(EB)评估血脑屏障的破坏程度,实时PCR检测MMP-9及TIMP-1 mRNA的表达水平.结果:脑缺血再灌注后,脑组织含水量及EB含量明显增加,MMP-9和TIMP-1表达均增强,与假手术组比较差异有统计学意义.丁苯酞预处理各组脑组织含水量及EB含量较模型组明显下降,MMP-9表达显著减少,TIMP-1表达明显增加,丁苯酞预处理中、高剂量组差异无统计学意义.结论:丁苯酞预处理对脑缺血再灌注损伤大鼠可通过调节MMP-9/TIMP-1的表达,降低血脑屏障通透性,减轻脑水肿,发挥预防性保护作用.     

雌激素对慢性脑缺血大鼠海马微血管构筑的影响

目的:探讨雌激素对慢性脑缺血大鼠海马微血管构筑的影响.方法:采用永久性结扎去势大鼠双侧颈总动脉的方法建立慢性脑缺血模型,项背部皮下注射雌激素.单宁酸-氯化铁媒染法观察大鼠海马微血管构筑的动态变化,采用MiVnt图像分析系统对大鼠海马水平部的微血管密度(MVD)和微血管面积密度(MVA)进行定量分析.结果:模型组大鼠各时间点海马的MVA、MVD值均较正常对照组大鼠明显降低,且随着时间的延长,MVA、MVD值逐渐下降.雌激素治疗组大鼠各时间点海马的MVA、MVD值高于模型组同时间点,且随着用药时间的延长MVA、MVD逐渐上升.结论:雌激素可改善慢性脑缺血大鼠海马的微血管构筑,降低缺血、缺氧对于海马神经元的损伤.     

黄芩茎叶总黄酮对大鼠大脑海马区微血管和血脑屏障缺血再灌注损伤的预防作用

目的 探讨黄芩茎叶总黄酮（SSTF）对脑缺血再灌注海马区微血管、血脑屏障（BBB）损伤的预防性保护作用。 方法 SD大鼠90只,随机分为假手术组（sham组）、模型组（IR组）和SSTF预处理组,造模前1周SSTF各组灌胃给药,低、中、高剂量组分别给50、100、200 mg/(kg·d),共7d。IR组和SSTF各组制作脑缺血再灌注模型,术后评价神经功能缺损变化,干湿重法和伊文思蓝法检测脑组织含水量和微血管通透性,单宁酸 氯化铁媒染（TA-Fe）法显示并观测大鼠微血管密度(MVD)和微血管面积比(MVA),Real-time PCR法检测水通道蛋白4（AQP4） mRNA的表达水平,电镜观察血脑屏障完整性。 结果 SSTF各组与IR组比,神经功能缺损评分、脑组织含水量、微血管通透性明显降低（P＜0.01,P＜0.05）,MVD、MVA增加（P＜0.01）, AQP4 mRNA表达增强(P＜0.01),BBB损伤不同程度减轻,内皮细胞肿胀渐消退,紧密连接松解好转,基膜渐连续、清晰,胶质细胞足板胞质溶解减轻,渐接近正常;SSTF 中、高组上述表现较SSTF 低组好转的更明显（P<0.01）,SSTF 中组与SSTF 高组比较差异无统计学意义（P＞0.05）。 结论 SSTF干预对海马区微血管、血脑屏障和神经损伤有预防性保护作用,其作用机制可能是通过增加有效微血管再通数量,维持血脑屏障完整和功能,减轻脑水肿实现的。SSTF的有效预防剂量为100mg/(kg·d)。     

丁苯酞预处理对脑缺血再灌注大鼠皮质神经元Fas/FasL和超微结构的影响

目的:探讨丁苯酞对大鼠脑缺血再灌注(IR)皮质神经元的预防性保护作用及其可能机制。方法:采用改良线栓法制备大鼠右侧脑局灶性缺血再灌注模型;免疫组织化学法和电子显微技术观察缺血再灌注皮质神经元Fas/Fas L的表达和超微结构改变。结果:假手术组偶见Fas、Fas L阳性细胞表达,神经元形态正常;模型组Fas、Fas L阳性表达明显增多,神经元坏死多见,胞膜断续或崩解,线粒体肿胀、嵴断裂,高尔基复合体肿胀扩张,核膜曲折不光滑;与模型组比较,丁苯酞预处理组Fas、Fas L阳性表达面积率降低(P0.01),多数神经元较正常,核模较完整,染色质分布较均匀,线粒体损伤减轻,内质网趋于正常,凋亡细胞少见。结论:丁苯酞预处理可下调皮质神经元Fas、Fas L阳性表达,减轻缺血再灌注神经元超微结构的损伤,对脑IR损伤有一定的预防性保护作用。     

大鼠海马微血管构筑的定量分析及衰老变化

目的:观察并定量分析大鼠海马的微血管构筑及衰老变化。方法:采用单宁酸-氯化铁媒染微血管的方法分别观察青龄、老龄大鼠海马与额叶皮质的微血管构筑,并采用MiVnt图像分析系统对海马与额叶皮质的微血管密度(MVD)及微血管面积密度(MVA)进行定量分析。结果:老龄大鼠海马、皮质的微血管数量显著减少,分布杂乱、扭曲缠结。定量分析显示老龄大鼠海马、皮质的MVD值和MVA值均明显低于青龄大鼠,差异具有统计学意义。结论:老龄大鼠海马及额叶皮质的MVD值、MVA值均明显低于青龄大鼠,这是老年血管性痴呆发生机制的主要形态学依据。     

TA-Fe法显示微血管及大脑不同区域微血管的定量研究

目的　定量分析研究大脑不同区域微血管的直径、密度及分布。方法　应用TA Fe法媒染大鼠大脑微血管 ,光镜下观察其形态结构 ;用目镜测微尺按体视学方法测定大脑相关区域微血管直径、密度 (MVD) ;用CMIAS图像分析系统测定微血管面积比 (MVA )。结果　血管直径 :顶、枕、颞叶皮质差别不明显 (P >0 .0 5 ) ,皮质与髓质、内囊均有显著性差异 (P<0 .0 5 )。MVD、MVA值 :顶区皮质 >枕区皮质 >颞区皮质 >髓质 >内囊 ,皮质间MVD无明显差别 ,皮质各区与髓质、内囊均有显著性差异。结论　TA Fe法是一种研究微血管构筑的理想方法 ,大脑各区皮质内MVD、MVA基本相同 ,皮质明显高于髓质和内囊。     

Increased microvessel density in malignant and borderline mammary phyllodes tumours

AIMS: Tumour vascularity is considered a prognostic indicator in breast carcinoma, but its utility in mammary phyllodes tumour has not been explored. The authors report the correlation between intratumoral microvessel density and the histological grade of phyllodes tumour. METHODS AND RESULTS: Forty cases of phyllodes tumour were reviewed for stromal cellularity, overgrowth, cytological pleomorphism, mitotic count and margin pattern. Using established criteria, these were diagnosed as benign (n=28), borderline (n=10) and malignant (n=2). Microvessel density was counted on CD31-stained slides as the number of vessels per high power field. For benign phyllodes tumour, the range was 7-26.2 (mean 13.1); for borderline phyllodes tumour the range was 17.2-32.5 (mean 22.4); for malignant phyllodes tumour the range was 25.9-33.3 (mean 29.6). The difference between the benign and borderline groups was significant (P < 0.0001) but that between the borderline and malignant groups was not, due to the small number of malignant cases. CONCLUSIONS: There is a significant difference in stromal microvessel density between benign and borderline phyllodes tumour. Although the small number of cases of malignant phyllodes tumour limits further interpretation, we believe that microvessel density can be used as an additional objective histological parameter in the evaluation of phyllodes tumour.     

脑星形细胞瘤微血管密度与临床病理的关系

目的：探讨星形细胞瘤微血管密度与其临床病理的关系。方法：采用免疫组织化学S－P法，对血管内皮细胞行第八因子相关抗原（FⅧRAg）染色，然后测定微血管密度（MVD）。结果：MVD与患者性别、肿瘤生长部位、大小无明显相关；不同病理组织学级别间MVD差异有显著性（P＜0.05)，病理级别越高，MVD越大；瘤周水肿程度不同的星形细胞瘤之间，MVD差异有显著性（P＜0.01)；复发患者MVD高于未复发患者（P＜0.01)。结论：星形细胞瘤MVD与病理级别、术后复发及瘤周组织水肿关系密切，可作为一项有意义的预后指标。     

信号素4D在结直肠癌组织中的表达及其与微血管密度的关系

目的:探讨结直肠癌组织中信号素4D(semaphorin4D,Sema4D)的表达及其与微血管密度(microvessel density,MVD)的相关性.方法:回顾性分析2014至2015年间在云南省肿瘤医院结直肠外科接受结直肠癌根治性手术的61例患者的临床病理资料;采用免疫组织化学SP法检测该组患者的结直肠癌组织标本及其56例癌旁组织中Sema4D蛋白的表达水平;采用微血管密度计数法计数用CD34标记的微血管.结果:Sema4D主要表达于结直肠癌细胞膜及细胞质,癌旁组织细胞质内呈阴性表达.癌组织Sema4D表达阳性率(61/61)明显高于癌旁组织(0/56),差异有统计学意义(x2=113.027,P＜0.001).Sema4D与MVD两者作相关性分析,两者有显著的正相关(r=0.333,P=0.009).MVD表达量在Sema4D表达高低组间存在显著差异(P=0.003).结论:Sema4D在结直肠癌组织高水平表达,且与MVD存在明显的正相关性,提示其有望成为潜在的结直肠癌血管治疗靶点.     

Correlation of adrenomedullin with the erythropoietin receptor and microvessel density in hepatocellular carcinoma

Introduction

Uncontrolled angiogenesis plays an essential role in the occurrence, metastasis and malignant progression of hepatocellular carcinoma (HCC). This study aimed to investigate the expression of adrenomedullin (ADM) in human HCC and its correlation with the expression of erythropoietin receptor (EPOR), microvessel density (MVD) and the tumor pathological characteristics.

Material and methods

Fresh tumor tissues were obtained from 30 HCC patients after hepatectomy. Ten cirrhotic and 10 normal liver tissues were included as controls. Expression of ADM and EPOR was determined by real-time PCR. The MVD was determined by counting the number of microvessels.

Results

The MVD and the mRNA levels of ADM and EPOR in cancer tissues were significantly higher than those in the non-cancer tissues (p < 0.05). Expression of ADM was significantly correlated with the MVD and EPOR (r = 0.68 and 0.74, p < 0.01). Adrenomedullin and EPOR mRNA levels in HCC tissues were correlated with capsule invasion, pathological differentiation and tumor metastasis (p < 0.05).

Conclusions

Our findings suggest that ADM and EPOR may serve as new regulatory factors involved in angiogenesis of HCC and represent novel targets for the treatment of HCC.     

非霍奇金淋巴瘤中的微血管密度及其临床病理意义

目的：探讨非霍奇金淋巴瘤(NHL)中的微血管密度(MVD)及其与NHL恶性程度、免疫学类型、预后的关系；方法：采用生物素标记的荆豆凝集素I(Bio-UEA-I)免疫组化ABC法对102例NHL的MVD进行原位观察和数量分析；结果：MVD随NHL恶性程度的增高而增高，高度恶性组和中度恶性组MVD显著高于低度恶性组MVD；T细胞性NHL中的MVD显著高于B细胞性NHL的MVD；短生存期组的MVD显著高于长生存期组的MVD；结论：NHL中的MVD可作为判断肿瘤恶性程度、免疫学类型及预后估计的有意义的指标，在临床病理中有实际应用价值。     

VEGF in 105 pheochromocytomas: enhanced expression correlates with malignant outcome

Pheochromocytomas are rare sympathoadrenal tumors that are highly vascular. Their malignancy is extremely difficult to estimate on the basis of histopathological features. Vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors involved in both tumor growth and metastasis. In our search for new prognostic markers, we investigated the expression of VEGF in normal adrenal gland, in 105 primary pheochromocytomas, and in 6 metastases by using immunohistochemistry and Northern blot analysis. We also calculated the microvessel density of these tumors by staining the endothelial cells with monoclonal CD34 antibody. VEGF messenger ribonucleic acid was found in all pheochromocytomas studied. Immunohistochemically, VEGF was not found in normal adrenal medullary cells. Interestingly, all malignant pheochromocytomas (n=8), regardless of their primary location, had strong or moderate VEGF immunoreactivity, while most benign adrenal pheochromocytomas (26 of 37, 70.3%) were either negative or only weakly positive. The staining was heterogenous in extraadrenal pheochromocytomas as well as in a group of tumors that had histologically suspicious features but had not metastasized, here called borderline tumors (n=29). The microvessel density varied greatly in all of the tumor groups, and no statistical difference was found between these groups. Here we report moderate to strong VEGF expression in malignant pheochromocytomas, and negative or weak expression in benign adrenal pheochromocytomas. Normal medullary cells are immunohistochemically negative. Thus, low VEGF expression in pheochromocytomas favors a benign diagnosis.     

Intratumoral microvascular density in malignant lymphomas of B-cell origin

Sections of surgical lymph-node biopsies of four types of malignant non-Hodgkin's lymphoma of B-cell origin (B-NHL) classified according to the R.E.A.L. terminology or lymphadenitis were immunostained in order to demonstrate endothelial CD34 (QBEnd 10) and to determine the microvascular density and vessel-size distribution using an interactive image-analysis technique. Only microvessels displaying a cross-sectional area corresponding to a diameter of between 3.2 and 34.6 microm were included. The intratumoral microvascular density (iMVD) was found to be significantly higher in chronic lymphatic leukaemia (CLL, n = 13) compared with the clinically more aggressive mantle cell lymphoma (MCL, n = 9) and diffuse large B-cell lymphoma (DLBCL, n = 14). iMVD in CLL was also higher than in the follicular neoplastic parts (FL FOLL) of follicular lymphoma (FL, n = 16). In FL FOLL the microvessel density was, moreover, significantly lower than in the surrounding non-neoplastic FL tissue. In lymphadenitis (LA, n = 10) the iMVD was higher than in DLBCL, FL FOLL and MCL. The data suggest that future studies focusing on the relationship between iMVD and the clinical outcome within each particular NHL group should be carried out in order to verify whether iMVD is a prognostic factor in NHL, as it is in carcinomas.     

乳腺癌中CD105表达及相关因素分析

目的 多项免疫组化（multiple immunohistoehemistry,MIHC）检测内皮因子CD105作为判断患者乳腺癌生物学行为及预后指标的可能性。方法 选取2003年11月-2004年1月经河北医科大学病理科确诊为原发性乳腺癌患者石蜡包埋组织标本87例,癌旁组织标本20例,同期乳腺良性病变标本24例。以CD105单克隆抗体为标记物进行免疫组化染色,根据CD105的表达水平计测微血管密度（mierovessel density,MVD）。结果CD105所标记的MVD在乳腺癌组织和乳腺良性病变以及乳腺良性病变与癌旁组织之间的表达差异均有显著性（P＜0．05）,癌旁组织中CD105几乎表达缺失;淋巴结转移阳性、组织分化差（Ⅲ级）、临床TNM分期晚以及VEGF表达阳性、ER、PR表达阴性的患者中,CD105的表达明显升高,差异均有显著性（P＜0．05）。结论 CD105主要在处于增殖状态的血管内皮细胞上表达,在标记肿瘤组织活性微血管方面特异性极高,是检测乳腺癌患者预后的重要指标。     

胸苷磷酸化酶在结直肠癌中的表达及与血管生成的关系

目的：研究结直肠癌组织中胸苷磷酸化酶（thymidine phosphorylase,TP)表达与癌组织分化程度，淋巴结转移及肿瘤内微血管密度（MVD）的关系。方法：对50例结直肠癌标本及21例正常结直肠黏膜组织进行免疫组化S－P法测定癌组织中TP的表达情况；通过CD34免疫组化染色检测癌组织中的MVD。结果：结直肠癌细胞的TP表达阳性率高于正常结直肠黏膜上皮（P＜0．01）；TP阳性组中局部淋巴结转移率高TP阴性组；TP阳性组的MVD高于TP阴性组；它们的差别均具有统计学意义，结直肠癌中高、中、低分化癌组之间的TP阳性率无明显差异。结论：癌细胞中TP的表达与肿瘤的血管生成及淋巴结转移有关。但与结直肠癌的组织化学级别无关。