Voxel Based Analysis of Surgical Revascularization for Moyamoya Disease: Pre- and Postoperative SPECT Studies

Moyamoya disease (MMD) is a chronic, progressive, cerebrovascular occlusive disease that causes abnormal enlargement of collateral pathways (moyamoya vessels) in the region of the basal ganglia and thalamus. Cerebral revascularization procedures remain the preferred treatment for patients with MMD, improving the compromised cerebral blood flow (CBF). However, voxel based analysis (VBA) of revascularization surgery for MMD based on data from pre- and postoperative data has not been established. The latest algorithm called as Diffeomorphic Anatomical Registration Through Exponentiated Lie Algebra (DARTEL) has been introduced for VBA as the function of statistical parametric mapping (SPM8), and improved registration has been achieved by SPM8 with DARTEL. In this study, VBA was conducted to evaluate pre- and postoperative single photon emission computed tomography (SPECT) images for MMD by SPM8 with DARTEL algorithm, and the results were compared with those from SPM8 without DARTEL (a conventional method). Thirty-two patients with MMD who underwent superficial temporal artery-middle cerebral artery (STA-MCA) bypass surgery as the first surgery were included and all patients underwent pre- and postoperative 3D T1-weighted imaging and SPECT. Pre- and postoperative SPECT images were registered to 3D T1-weighted images, then VBA was conducted. Postoperative SPECT showed more statistically increased CBF areas in the bypassed side cerebral hemisphere by using SPM8 with DARTEL (58,989 voxels; P<0.001), and increased ratio of CBF after operation was less than 15%. Meanwhile, postoperative SPECT showed less CBF increased areas by SPM8 without DARTEL. In conclusion, VBA was conducted for patients with MMD, and SPM8 with DARTEL revealed that postoperative SPECT showed statistically significant CBF increases over a relatively large area and with at most 15% increase ratio.     

VEGF受体KDR胞外区基因的克隆及其在昆虫细胞中的表达

VEGF(血管内皮细胞生长因子,Vascular Endothelial Growth Factor)是刺激内皮细胞增殖和新生血管形成的最重要因子,与多种实体瘤的生长和转移密切相关。应用其可溶性受体阻断它的病理作用是一个非常有前景的课题。将VEGF受体KDR胞外区前三个Loop 969碱基对的cDNA片段克隆到杆状病毒表达载体pFastBacI,与杆状病毒表达载体Bacmid同源重组后,转染昆虫细胞SF9,获得重组杆状病毒并证明了目的基因的高效表达。经Western blot证实表达产物的特异性。经ELISA和体外生物学活性检测表明表达产物可阻断VEGF的生物学活性,抑制鸡胚CAM血管的生长。     

VEGF 家族及其在肿瘤生长中作用的研究

血管内皮生长因子(Vascular Endothelial Growth Factor,VEGF)家族是一类多功能的细胞因子,在血管生成和淋巴管生成中具有直接和间接的调控作用,可促进内皮细胞增殖、促进血管生成以及增加血管的通透性。VEGF/VEGFR轴由多重配基和受体质量叠加交错组成,并且受体与配基结合具有专一性,在不同的细胞中具有不同的细胞类型表达和功能.启动VEGF信号通路,触发了一个网状的信号过程,从而促进血管内皮细胞生长、转移和存活。进来研究发现,VEGF的一个重要作用表现为可动员内皮祖细胞从骨髓向远处转移从而形成新生血管,因而有必要设计和发展针对这一途径的抑制因子。随着研究的深入,VEGF促进肿瘤血管生成的作用和与人类癌症的发病机制的关系是确定的,因此,抑制VEGF途径被确认为是一种重要的有效的抗癌模式     

用腺相关病毒载体介导人血管内皮生长因子受体KDR胞外区基因的有达

新生血管大量形成在实体瘤的生长和转移中起着关键的作用。血管内皮生长因子 (VEGF)是介导肿瘤血管生成的最主要因素。从原代培养的人脐静脉血管内皮细胞 (HUVEC)提取细胞总RNA ,采用逆转录PCR(RT PCR)方法得到VEGF受体KDR胞外区cDNA片段。将获得的受体基因克隆到AAV基因治疗载体pSNAV中 ,得到重组质粒pSNAV KDR。重组质粒转染BHK细胞 ,加入辅助病毒后 ,获得了表达目的蛋白的重组AAV。重组病毒表达的KDR在体外实验中具有与VEGF结合的活性。在体内实验中 ,重组AAV感染的黑色素瘤细胞在小鼠中形成肿瘤的血管化程度明显低于对照组。     

用腺相关病毒载体介导人血管内皮生长因子受体KDR胞外区基因的表达

新生血管大量形成在实体瘤的生长和转移中起着关键的作用。血管内皮生长因子(VEGF)是介导肿瘤血管生成的最主要因素。从原代培养的人脐静脉血管内皮细胞(HUVEC)提取细胞总RNA,采用逆转录PCR(RT-PCR)方法得到VEGF受体KDR胞外区cDNA片段。将获得的受体基因克隆到AAV基因治疗载体pSNAV中,得到重组质粒pSNAV/KDR。重组质粒转染BHK细胞,加入辅助病毒后,获得了表达目的蛋白的重组AAV。重组病毒表达的KDR在体外实验中具有与VEGF结合的活性。在体内实验中,重组AAV感染的黑色素瘤细胞在小鼠中形成肿瘤的血管化程度明显低于对照组。     

VEGF、KDR和细菌L型感染在卵巢肿瘤中的表达及临床相关性研究

目的探讨血管内皮生长因子（VEGF）及其受体VEGFR-2（KDR）和细菌L型感染在卵巢肿瘤中的表达及临床相关性研究。方法应用免疫组化、原位杂交和革兰染色等方法检测了120例卵巢肿瘤中的VEGF、KDR蛋白及mRNA的表达,以及细菌L型的检出率。并对97例卵巢乳头状癌和23例卵巢乳头状瘤主要临床资料和病理分级参数进行比较,用χ^2检验进行统计学处理。结果VEGF、KDR蛋白及mRNA阳性表达率恶性肿瘤明显高于良性肿瘤（P〈0．005）。细菌L型检出阳性率与卵巢良、恶性肿瘤差异无显著性（P〉0．5）。VEGF、KDR蛋白及mRNA阳性表达以及细菌L型检出阳性率与卵巢乳头状癌的临床分期、病理分级和腹腔淋巴结有转移、腹水差异有显著性（P〈0．001—0．05）。细菌L型阳性患者中VEGF、KDR阳性明显高于L型阴性患者中阳性表达,2组差异具有非常显著性（P〈0．0001）。结论VEGF、KDR蛋白及mRNA在卵巢肿瘤中有不同程度的异常表达,与卵巢癌的临床分期、病理分级和浸润、转移呈正相关,L型感染极有可能成为诱发肿瘤因素之一,它们可能有协同致瘤作用。研究L型感染与卵巢肿瘤的关系,具有重要的临床应用价值。     

Role of HLA-DP Polymorphisms on Chronicity and Disease Activity of Hepatitis B Infection in Southern Chinese

Background and Aims

The association between HLA-DP single nucleotide polymorphisms (SNPs) and chronic hepatitis B virus (HBV) infection varies between different populations. We aimed to study the association between HLA-DP SNPs and HBV infection and disease activity in the Chinese population of Hong Kong.

Methods

We genotyped SNPs rs3077 (near HLA-DPA1) and rs9277378 and rs3128917 (both near HLA-DPB1) in 500 HBV carriers (hepatitis B surface antigen [HBsAg]-positive), 245 non-HBV infected controls (HBsAg- and antibody to hepatitis B core protein [anti-HBc]-negative), and 259 subjects with natural HBV clearance (HBsAg-negative, anti-HBc-positive). Inactive HBV carriers state was defined by HBV DNA levels <2,000 IU/ml and persistently normal alanine aminotransferase level for least 12 months.

Results

Compared to the non-HBV infected subjects, the HBV carriers had a significantly lower frequency of the rs3077 T allele (p = 0.0040), rs9277378 A allele (p = 0.0068) and a trend for lower frequency of rs3128917 T allele (p = 0.054). These alleles were associated with an increased chance of HBV clearance (rs3077: OR = 1.41, p = 0.0083; rs9277378: OR = 1.61, p = 0.00011; rs3128917: OR = 1.54, p = 0.00017). Significant associations between HLA-DP genotypes and HBV clearance were also found under different genetic models. Haplotype TAT was associated with an increased chance of HBV clearance (OR = 1.64, p = 0.0013). No association was found between these SNPs and HBV disease activity.

Conclusion

HLA-DP SNPs rs3077, rs9277378 and rs3128917 were associated with chronicity of HBV disease in the Chinese. Further studies are required to determine whether these SNPs influence the disease endemicity in different ethnic populations.     

烟雾病5 例临床分析

目的:研究烟雾病(moyamoya disease,MMD)的临床及影像学特征。方法:回顾性分析5例烟雾病患者,分析其临床及影像学特点。结果:本组病例既往均无阳性病史,中、青年起病,男性居多,均以缺血性脑血管病起病,肢体瘫痪不重,经颅多普勒(TCD)及头颅核磁改变明显,数字减影血管造影(DSA)检查均存在血管闭塞及侧支开放,烟雾血管网形成,1例MRA证实烟雾血管网形成。结论:对于无既往史的中、青年脑卒中患者,要考虑MMD的可能,需完善TCD、头核磁检寻找证据,最后完善数字减影血管造影确诊。     

烟雾病5例临床分析

目的：研究烟雾病（moyamoya disease,MMD）的临床及影像学特征。方法：回顾性分析5例烟雾病患者,分析其临床及影像学特点。结果：本组病例既往均无阳性病史,中、青年起病,男性居多,均以缺血性脑血管病起病,肢体瘫痪不重,经颅多普勒（TCD）及头颅核磁改变明显,数字减影血管造影（DSA）检查均存在血管闭塞及侧支开放,烟雾血管网形成,1例MRA证实烟雾血管网形成。结论：对于无既往史的中、青年脑卒中患者,要考虑MMD的可能,需完善TCD、头核磁检寻找证据,最后完善数字减影血管造影确诊。     

Role of VEGF in organogenesis

The cardiovascular system, consisting of the heart, blood vessels and hematopoietic cells, is the first organ system to develop in vertebrates and is essential for providing oxygen and nutrients to the embryo and adult organs. Work done predominantly using the mouse and zebrafish as model systems has demonstrated that Vascular Endothelial Growth Factor (VEGF, also known as VEGFA) and its receptors KDR (FLK1/VEGFR2), FLT1 (VEGFR1), NRP1 and NRP2 play essential roles in many different aspects of cardiovascular development, including endothelial cell differentiation, migration and survival as well as heart formation and hematopoiesis. This review will summarize the approaches taken and conclusions reached in dissecting the role of VEGF signalling in vivo during the development of the early cardiovasculature and other organ systems. The VEGF?mediated assembly of a functional vasculature is also a prerequisite for the proper formation of other organs and for tissue homeostasis, because blood vessels deliver oxygen and nutrients and vascular endothelium provides inductive signals to other tissues. Particular emphasis will therefore be placed in this review on the cellular interactions between vascular endothelium and developing organ systems, in addition to a discussion of the role of VEGF in modulating the behavior of nonendothelial cell populations.     

Role of VEGF in organogenesis

The cardiovascular system, consisting of the heart, blood vessels and hematopoietic cells, is the first organ system to develop in vertebrates and is essential for providing oxygen and nutrients to the embryo and adult organs. Work done predominantly using the mouse and zebrafish as model systems has demonstrated that Vascular Endothelial Growth Factor (VEGF, also known as VEGFA) and its receptors KDR (FLK1/VEGFR2), FLT1 (VEGFR1), NRP1 and NRP2 play essential roles in many different aspects of cardiovascular development, including endothelial cell differentiation, migration and survival as well as heart formation and hematopoiesis. This review will summarize the approaches taken and conclusions reached in dissecting the role of VEGF signalling in vivo during the development of the early cardiovasculature and other organ systems. The VEGF-mediated assembly of a functional vasculature is also a prerequisite for the proper formation of other organs and for tissue homeostasis, because blood vessels deliver oxygen and nutrients and vascular endothelium provides inductive signals to other tissues. Particular emphasis will therefore be placed in this review on the cellular interactions between vascular endothelium and developing organ systems, in addition to a discussion of the role of VEGF in modulating the behavior of nonendothelial cell populations.Key words: VEGF, VEGF receptors, organogenesis, mouse, angiogenesis, cardiovascular, conditional mutagenesis, Cre-loxP system     

Differential expression of the two VEGF receptors flt and KDR in placenta and vascular endothelial cells

Vascular endothelial growth factor (VEGF) is a newly identified growth and permeability factor with a unique specificity for endothelial cells. Recently the flt-encoded tyrosine kinase was characterized as a receptor for VEGF. A novel tyrosine kinase receptor encoded by the KDR gene was also found to bind VEGF with high affinity when expressed in CMT-3 cells. Screening for flt and KDR expression in a variety of species and tissue-derived endothelial cells demonstrates that flt is predominantly expressed in human placenta and human vascular endothelial cells. Placenta growth factor (PIGF), a growth factor significantly related to VEGF, is coexpressed with flt in placenta and human vascular endothelial cells. KDR is more widely distributed and expressed in all vessel-derived endothelial cells. These data demonstrate that cultured human endothelial cells isolated from different tissues express both VEGF receptors in relative high levels and, additionally, that all investigated nonhuman endothelial cells in culture are also positive for KDR gene expression.     

The Role of Paraoxonase 1 (PON1) Gene Polymorphisms in Coronary Artery Disease: A Systematic Review and Meta-Analysis

Although many studies have investigated the association of paraoxonase 1 (PON1) polymorphisms with coronary artery disease (CAD). However, the outcomes were not consistent and remain uncertain. Therefore, it is the need of the hour to analyze the available literature and evaluate the association of PON1 polymorphisms with the CAD. All the relevant studies published in the English language from January 1, 2000, up to September 20, 2020, were identified by searching through various electronic databases. The two researchers independently extracted the information. The data were analyzed by using the MetaGenyo program. The pooled odds ratio was used to find the associations between CAD and PON1 polymorphisms. In the final analysis, we include 10 studies regarding the association of PON1 polymorphisms (rs662 and rs854560) with CAD. Overall, the Q192R polymorphism increased the risk of CAD in the tested genetic models including the homozygote model: OR 1.35, CI 1.02–1.79; allelic model: OR 1.16, CI 1.00–1.33; dominant model: OR 1.25, CI 1.03–1.52. The L55M polymorphism does not significantly associated with CAD in all the tested genetic models including the homozygote model: OR 1.00 CI, 0.64–1.56; allelic model: OR 1.02, 95% CI 0.84–1.23; dominant model: OR 1.08, CI 0.89–1.31. Further analysis showed no publication bias exists in meta-analysis. Our findings suggested that rs662 in the coding region was significantly associated with the CAD however, rs854560 has no significant association with the disease. Nevertheless, in future, there is a need for more studies with a larger sample size which may provide a more definite conclusion.

Study Registration: PROSPERO registration number CRD42020202278

     

Age-related changes in cardiac expression of VEGF and its angiogenic receptor KDR in stroke-prone spontaneously hypertensive rats

We examined the age-related changes in cardiac expression of angiogenic molecules during the development of cardiac remodeling in stroke-prone spontaneously hypertensive rats (SHRSP) in comparison with those in Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). Vascular endothelial growth factor (VEGF) was highly upregulated in SHRSP aged 20 weeks compared with the same age of WKY, but it was downregulated at 40 weeks. On the other hand, KDR, an angiogenic receptor of VEGF, and endothelial nitric oxide synthase, which is important in the VEGF-mediated angiogenic pathway, were markedly downregulated in SHRSP from 20 weeks of age. Such age-related changes in their expression levels seen in SHRSP were quite different from those in SHR. In both SHR and SHRSP, transforming growth factor-₁ (TGF-₁) expression was increased with age, although SHRSP showed more marked upregulation. Cardiac remodeling in SHRSP was characterized by decreased coronary capillary density, cardiomyocyte hypertrophy, and cardiac fibrosis. We conclude that, in addition to overexpression of TGF-₁, which appears to play a pivotal role in promoting cardiac hypertrophy and fibrosis, a defect of the VEGF-KDR system could result in impaired physiologic coronary angiogenesis in SHRSP, contributing to cardiac deteroration associated with myocardial ischemia in this malignant hypertensive model.     

VEGF、KDR和结核菌L型感染在前列腺癌中的表达及临床意义

目的 探讨血管内皮生长因子(VEGF)及其受体VEGFR-2(KDR)和结核菌L型感染在前列腺肿瘤中的表达及临床相关性研究.方法 应用免疫组化、原位杂交和抗酸染色等方法检测了65例前列腺癌(carcinoma of prostate,PCa)和30例良性前列腺增生(benign prostatic hyperplasia,BPH)中的VEGF、KDR蛋白及mRNA的表达,以及结核菌L型的检出率.并对前列腺肿瘤主要临床资料和病理分级参数进行比较,用χ2检验进行统计学处理.结果 VEGF、KDR蛋白及mRNA阳性表达和结核菌L型检出率前列腺癌明显高于前列腺增生(P<0.001～0.05).VEGF、KDR蛋白及mRNA阳性表达以及结核菌L型检出阳性率与前列腺癌的临床分期、病理分级差异有显著性(P<0.01～0.05).结论 VEGF及KDR蛋白及mRNA在前列腺肿瘤中有不同程度的异常表达,可能与结核菌L型感染有协同致瘤作用.提示2种基因均可作为判断前列腺癌生物学行为及患者预后参考指标.结核菌L型感染可能与前列腺癌的发生发展可能有一定关系,因此研究结核菌L型感染与前列腺癌的关系,具有重要的临床应用价值.     

VEGF signalling controls GnRH neuron survival via NRP1 independently of KDR and blood vessels

Gonadotropin-releasing hormone (GnRH) neurons are neuroendocrine cells that are born in the nasal placode during embryonic development and migrate through the nose and forebrain to the hypothalamus, where they regulate reproduction. Many molecular pathways that guide their migration have been identified, but little is known about the factors that control the survival of the migrating GnRH neurons as they negotiate different environments. We previously reported that the class 3 semaphorin SEMA3A signals through its neuropilin receptors, NRP1 and NRP2, to organise the axons that guide migrating GnRH neurons from their birthplace into the brain. By combining analysis of genetically altered mice with in vitro models, we show here that the alternative neuropilin ligand VEGF164 promotes the survival of migrating GnRH neurons by co-activating the ERK and AKT signalling pathways through NRP1. We also demonstrate that survival signalling relies on neuronal, but not endothelial, NRP1 expression and that it occurs independently of KDR, the main VEGF receptor in blood vessels. Therefore, VEGF164 provides survival signals directly to developing GnRH neurons, independently of its role in blood vessels. Finally, we show that the VEGF164-mediated neuronal survival and SEMA3A-mediated axon guidance cooperate to ensure that migrating GnRH neurons reach the brain. Thus, the loss of both neuropilin ligands leads to an almost complete failure to establish the GnRH neuron system.     

Regulatory molecules for coronary expressions of VEGF and its angiogenic receptor KDR in hypoestrogenic middle-aged female rats

We studied the effects of estrogen deprivation and replacement on the protein and gene expression levels molecules that can be considered to be essential for coronary angiogenesis in middle-aged female rats. The animals were subjected to sham operation, ovariectomy, or ovariectomy with estrogen replacement therapy (ERT). Following ovariectomy, protein and gene expressions of vascular endothelial growth factor (VEGF) and its angiogenic receptor (KDR) showed a marked decline in coronary vessels, as determined by immunohistochemistry and in situ hybridization. ERT resulted in restoration of the ovariectomy-induced changes to intact levels. The coronary expression level of basic fibroblast growth factor was unaffected by estrogen deprivation or treatment. The changes in VEGF and KDR expressions were strongly associated with those in endothelial nitric oxide synthase (eNOS) expression in coronary vessels. Moreover, the age- and gender-dependent accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha) protein appeared to be a determinant molecule of VEGF expression in middle-aged female rats. We reached a conclusion that the VEGF-KDR system plays a key role in coronary angiogenesis in hypoestrogenic elderly women and is critically regulated by estrogen, eNOS and HIF-1alpha.