骨髓增生异常综合征骨髓血管新生的研究

目的 探讨骨髓增生异常综合征(MDS)患者骨髓是否存在血管新生,及其在MDS发病机制、病情进展及预后判断中的价值.方法 用CD34单抗免疫组织化学染色方法标记MDS患者和正常人骨髓绀织m管内皮细胞,盲法计数骨髓微血管密度(MVD).结果 MDS患者MVD显著高于正常对照组(P＜0.01),并且随着病程的进展患者的骨髓MVD呈增高趋势(P＜0.01).结论 MIDS患者骨髓MVD增高,可能与白血病进展有关.     

三氧化二砷抗骨髓增生异常综合征骨髓血管新生机制的初步探讨

目的探讨骨髓增生异常综合征(MDS)患者骨髓是否存在血管新生及三氧化二砷(As2O3)的抗血管新生作用。方法①采用CD34单抗免疫组织化学染色法标记骨髓组织血管内皮细胞,盲法计数骨髓微血管密度(MVD)。②采用四甲基偶氮唑蓝方法观察As2O3对A组(HUVEC)及B组(经SKM-1细胞株上清液干预组HUVEC)的增殖抑制效应。结果①MDS患者骨髓微血管密度(MVD)显著高于正常对照组(P<0.01),并且随着病程的进展患者的MVD呈增高趋势(Rs=0.86 133,P<0.05);②As2O3对HUVEC及SKM-1细胞株干预组的作用均表现时间、剂量依赖性的增殖抑制效应(F=10.92,P<0.05);③HUVEC中加入SKM-1上清液共同培养对促进细胞生长有明显作用(P<0.01)。结论①MDS患者存在骨髓微血管密度增高,并与MDS患者的疾病进展正相关,血管新生在MDS的发生发展中起到重要作用;②As2O3以时间-剂量依赖性方式抑制HUVEC细胞的生长;③提示MDS细胞可分泌促进内皮细胞生长的细胞因子,通过旁分泌作用刺激内皮细胞生长。     

Expression of proliferating cell nuclear antigen (PCNA) in multiple myeloma: its relationship to bone marrow microvessel density and other factors of disease activity

The expression of proliferating cell nuclear antigen (PCNA) was studied in plasma cells in bone marrow biopsies from patients with multiple myeloma (MM) using a double immunostaining method. In the same samples, microvessel density (MVD), after staining with anti-CD34 antibodies, was determined before and after chemotherapy. The correlation of PCNA expression and MVD with other myeloma parameters (clinical stage, bone marrow plasma cell infiltration and serum interleukin-6 (IL-6)) was also investigated. The study population included 51 newly diagnosed MM patients, 15 patients in plateau phase after treatment and 15 normal controls. Pretreatment mean +/- SE values of PCNA, MVD, plasma cell infiltration and serum IL-6 were significantly higher than post treatment values and controls. Pretreatment PCNA expression correlated significantly with bone marrow MVD (p<0.05) plasma cell infiltration (p<0.01) and IL-6 (p<0.01). These findings show that the proliferative activity of plasma cells is related to the angiogenic activity in the bone marrow of multiple myeloma patients. Both PCNA and MVD correlate with markers of disease activity thus may provide additional information when included in the initial evaluation of myeloma bone marrow biopsies.     

Angiogenic molecules in Hodgkin's disease: results from sequential serum analysis

Increased angiogenic activity has been demonstrated in lymphoproliferative diseases including Hodgkin's disease. In the current study, the levels of circulating angiogenic molecules in 60 Hodgkin's patients were determined prior to and after treatment and correlated to disease stage and prognostic score. Hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were increased in Hodgkin's patients in comparison to healthy controls (p<0.001). Angiogenin and angiopoietin-2 levels did not differ from controls. HGF, VEGF, TNF-alpha and angiogenin decreased significantly in Hodgkin's patients after standard treatment (p<0.001 for HGF, p<0.05 for VEGF, TNF-alpha and angiogenin). Furthermore, HGF and TNF-alpha increased with advancing stage of disease (p<0.05). HGF and VEGF correlated significantly with IL-6 (r=0.56, p<0.0005 and r=0.57, p<0.001 respectively). In conclusion, Hodgkin's disease displays an angiogenic activity as depicted by the increased serum levels of a number of angiogenic cytokines. HGF seems to be the prominent molecule in Hodgkin's disease, which may be used to monitor the disease status and the response to treatment.     

Reduced high serum hepatocyte growth factor levels after successful cardioversion in patients with atrial fibrillation

1. Serum hepatocyte growth factor (HGF) is considered to be a potent marker of vascular endothelial injury. The present study was designed to examine serum HGF levels in atrial fibrillation and after successful direct current (DC) cardioversion. 2. We measured serum HGF levels before and 7 days and 1 month after DC cardioversion in 39 patients with atrial fibrillation in whom sinus rhythm was maintained for at least 7 days after DC cardioversion and in 30 age- and sex-matched normal control subjects with sinus rhythm. We also measured acetylcholine-induced changes in forearm blood flow (FBF) using venous occlusive plethysmography in 10 patients. 3. Serum HGF levels were significantly higher in the atrial fibrillation patients (both lone atrial fibrillation and with underlying heart disease) than in the controls (0.16 +/- 0.07 vs 0.10 +/- 0.04 ng/mL; P < 0.001). Seven days after successful DC cardioversion, the patients' serum HGF levels had decreased significantly (0.16 +/- 0.07 vs 0.12 +/- 0.06 ng/mL; P < 0.05) and in the 24 patients maintaining sinus rhythm 1 month after DC cardioversion, serum HGF levels decreased to control values (0.10 +/- 0.08 ng/mL at 1 month). Serum HGF levels of the 15 patients who had relapsed into atrial fibrillation 1 month after DC cardioversion tended to decrease 7 days after DC cardioversion, but increased again 1 month after DC cardioversion. Percentage changes in FBF between baseline and the highest dose of acetylcholine before and after DC cardioversion were 180 +/- 98 and 323 +/- 196%, respectively (P = 0.0051). The rate of increase in FBF at the highest dose of acetylcholine between before and after DC cardioversion correlated negatively with the rate of decrease in serum HGF levels between before and after DC cardioversion (r = -0.837; P = 0.0025). 4. This study is the first to demonstrate that serum HGF levels increase in atrial fibrillation and decrease after successful DC cardioversion. This may reflect the fact that atrial fibrillation induces vascular endothelial injury.     

Increased level of serum hepatocyte growth factor/scatter factor in liver cancer is associated with tumor metastasis.

Hepatocyte growth factor/scatter factor (HGF/SF) has been shown to play an important role in tumor migration and metastasis. We therefore investigated the relationship between HGF/SF expression and metastasis of human liver tumors. The serum HGF/SF levels in 41 patients with primary hepatocellular carcinoma (HCC; 22 patients with metastasis, 19 patients without metastasis); 4 patients with benign hepatic tumor, 4 patients with secondary hepatic carcinoma, and 12 healthy blood donors, were measured by sandwich enzyme-linked immunosorbent assay (ELISA) in this study. Our results show that liver tumor patients have significantly higher serum levels of HGF/SF compared to healthy blood donors. However, there is no difference between the serum HGF/SF levels in patients with primary HCC and patients with secondary HCC or benign hepatic tumors. In addition, we measured significantly higher levels of HGF/SF in serum from HCC patients with metastasis compared to HCC patients without metastasis, indicating that the elevations in serum HGF/SF level correlated positively with the tumor metastasis in human HCC. These findings appear to suggest that HGF/SF may be a useful serological biomarker for clinical diagnosis and follow-up of HCC metastasis, and suggest that larger scale studies in patients with hepatomas are warranted.     

急性胰腺炎患者血清肿瘤坏死因子α与肝细胞生长因子水平及临床意义的研究

目的探讨急性胰腺炎(AP)患者血清肿瘤坏死因子α(TNF-α)、肝细胞生长因子(HGF)的水平变化,以分析其变化规律及对病情判断的价值。方法应用ELISA测定39例急性胰腺炎(AP)及20例对照组血清TNF-α和HGF水平。结果AP患者血清TNFα和HGF水平明显高于对照组(P<0.01),在发病第1天,重症AP患者TNF-α水平明显高于轻症AP患者,而HGF则轻症AP患者较高,差异有统计学意义(P<0.01)。结论TNF-α和HGF参与了急性胰腺炎的发生发展过程,可作为AP早期诊断的参考指标。     

Bcl-2 与急性白血病患者骨髓组织血管新生的关系

目的通过检测急性白血病（AL）患者骨髓病理切片中血管内皮生长因子（Vascular Endothelial Growth Factor,VEGF）及Bcl-2蛋白的表达和微血管的生成情况及VEGF及Bcl-2蛋白的表达和微血管生成之间的相关性。方法免疫组织化学染色方法检测35例AL患者和35例对照组骨髓病理切片中的,Bcl-2表达;ELISA检测患者血清中VEGF-C的表达。结果对照组患者血清中VEGF-C为（126.34±21.22）pg/ml,实验组患者血清中VEGF-C为（324.45±34.12）pg/ml,35例AL患者,微血管密度（microvessel density,MVD）的平均光密度值为（0.3573±0.030）。35例对照组骨髓病理免疫组织化学片中,MVD的平均光密度值为（0.2192±0.037）。AL患者的MVD平均光密度值明显高于对照组,有显著性差异。结论 AL患者的骨髓组织中由于VEGF的表达,诱导产生更多的Bcl-2蛋白来降低AL细胞凋亡,从而增加AL的恶性程度。     

食管鳞癌患者前血青HGF、VEGF-C的表达及临床意义

目的:探讨食管鳞癌患者术前血清HGF、VEGF-C的表达及临床意义.方法:以手术组57例食管鳞癌患者和对照组20例健康体验者为研究对象,所有患者术前抽取血清标本,酶联免疫吸附反应方法(ELISA法)测定HGF、VEGF-C的表达水平.结果:食管癌患者组治疗前的血清HGF和VEGF-C表达水平均明显高于正常对照组(P<0...     

胃癌患者血清肝细胞生长因子的检测及意义

目的探讨血清肝细胞生长因子（HGF）在胃癌患者血清水平变化的临床意义。方法选取手术和临床病理证实为胃癌的60例患者,采用酶联免疫吸附法（ELISA）检测其血清HGF水平,其中40例行胃癌根治术,20例患者因已有远处转移而丧失根治手术机会。分别检测胃癌患者术前、术后7d和3个月者血清HGF的水平;同时选取加例慢性胃炎患者以及40例门诊健康体检者作为对照。结果胃癌患者血清HGF水平明显高于慢性胃炎组和正常对照者（P〈0．01）。肿瘤血管侵犯者血清HGF明显升高（P〈0．05）;有淋巴结转移者,血清HGF水平明显高于无淋巴结转移者（P〈0．05）;肝脏转移者血清HGF水平明显高于无肝脏转移者（P〈0．05）;肿瘤最大直径t〉3cm的胃癌患者血清HGF水平明显高于最大直径〈3cm的胃癌患者（P〈0．05）;按胃癌TNM分期标准,Ⅰ期和Ⅱ期患者血清HGF水平明显低于Ⅲ期和Ⅳ期患者（P〈0．05）;胃癌患者血清HGF水平与性别、年龄及组织学类型无明显关系（P〉0．05）。40例手术治疗患者术后1周血清HGF水平明显降低,术后3个月降低更为明显（P〈0．05和P〈0．001）。结论HGF在胃癌的发生发展过程中可能起重要作用,血清HGF水平的检测可成为胃癌病情程度监测和预后判断的重要指标。     

Haemopoietic growth factors in the treatment of myelodysplastic syndrome

The myelodysplastic syndromes (MDS) are clonal stem cell disorders characterised by ineffective haematopoiesis, various degrees of cytopenia and risk of transformation to acute myelogenous leukaemia. The allogeneic haematopoietic stem cell transplantation (AHSCT) is the only potentially curative treatment for MDS; unfortunately, it can only be prescribed to a small percentage of patients. Although the high-risk group of patients are candidates for AHSCT, low-risk MDS patients are generally assigned to low-intensity therapies (e.g., supportive care, haematopoietic growth factors (HGF), biological response modifiers or immunosuppression) with the therapeutic goals to improve the cytopenias and to enhance the quality of life. Clinically available HGFs enable the stimulation of erythropoiesis, granulocytopoiesis and, with lesser efficacy, the megakaryocytopoiesis; however, they are not devoid of imperfections. This patent review focuses on the latest developments of new promising HGFs that could be significantly utilised in MDS.     

The role of basic-fibroblast growth factor (b-FGF) in cyclosporine-induced nephrotoxicity

BACKGROUND: The effect of the b-fibroblast growth factor (b-FGF) on cyclosporine A (CsA)-induced nephrotoxicity in the rat kidney was investigated. MATERIALS AND METHODS: The rats were divided into six groups: A (control), B (b-FGF-treated), C, D: (CsA-treated and sacrificed on days 14 or 21), E, F (Cs A- and b-FGF- treated and sacrificed on days 14 or 21). The antibody mouse anti-rat CD31 was used to evaluate the kidney vessels present in histological preparations. RESULTS: The kidney vessels in group B were increased in comparison with the control group (p<0.05). Reduction of kidney vessels in groups C and D (p<0.05) in comparison with the controls was observed, while in groups E and F they were increased when compared to group C (p<0.05) and D (p<0.05), respectively. CONCLUSION: The angiogenic role of b-FGF was confirmed in normal rats and a possible "protective" role of b-FGF was shown in rat kidney with CsA-induced nephrotoxicity.     

肝细胞生长因子基因联合骨髓间充质干细胞治疗糖尿病周围动脉硬化闭塞症及糖尿病足近期观察

目的 探讨利用肝细胞生长因子(HGF)基因联合骨髓间充质干细胞治疗糖尿病周围动脉硬化闭塞症(PAD)和糖尿病足的临床疗效.方法 将Fontaine分级Ⅲ～Ⅳ级的糖尿病PAD及糖尿病足患者30例完全随机分为2组.研究组(16例)采用携带人HGF基因的质粒DNA( HGF-PVAX1 )4 mg联合骨髓动员后的于细胞,局部注射于患肢腓肠肌;对照组(14例)于患肢腓肠肌局部注射单纯骨髓动员后的于细胞.分别于移植术后1、3、6个月比较2组在干细胞局部注射治疗前后的临床疗效.结果 2组患者间歇性跛行有好转.研究组术前与术后6个月的下肢踝/肱指数(ABI)分别为(0.346±0.036)、(0.596±0.036),对照组是(0.379±0.031)、(0.468±0.032).2组术后与术前相比,差异均有统计学意义(均P＜0.05);术后研究组ABI水平高于对照组,差异亦有统计学意义(P＜0.05).研究组术后皮温平均为(35.9±1.8)℃,术前为(33.4±2.1)℃;对照组术后皮温平均为(33.1±1.3)℃,术前为(33.7±1.5)℃.2组皮温变化程度比较,差异有统计学意义(P＜0.05).2组移植后3个月复查下肢动脉彩色超声,股浅动脉、腘动脉未见明显变化.研究组移植前足背动脉血流量≥20 ml/min者8例,15 ～ 19 ml/min者4例,1～ 14 ml/min者2例,0 ml/min者2例;移植后≥20 ml/min者10例,15 ～ 19 ml/min者2例,1～14 ml/min者2例,0ml/min者2例.对照组移植前足背动脉血流量≥20 ml/min者6例,15～19 ml/min者3例,1～14 ml/min者3例,0ml/min者2例;移植后≥20 ml/min者7例,15～19 ml/min者2例,l～ 14 ml/min者3例,0 ml/min者2例.研究组移植后足背动脉血流量改善情况好于对照组,差异有统计学意义(P＜0.05).2组间不良反应发生率差异无统计学意义.结论 注射HGF-PVAXl联合骨髓干细胞移植可在一定程度上改善肢体缺血,可能成为治疗PAD的一种新手段.但其远期疗效和安全性还有待观察.     

骨髓增生异常综合征76例的细胞形态学特点分析

目的探讨骨髓增生异常综合征(MDS)患者的骨髓细胞形态学特点。方法用吉姆萨染色法对骨髓增生异常综合征(MDS)患者进行骨髓细胞形态染色,并在光学显微镜下观察分析。结果通过对76例MDS患者骨髓细胞形态的观察分析发现,68例MDS患者骨髓增生程度在活跃和明显活跃之间,MDS粒系异常以P-H细胞(21例,28%)和中粒以下部分细胞质内颗粒减少(20例,26%)最常见;红系异常以老核幼质(34例,45%)、嗜多染(34例,45%)和中空区偏大(29例,38%)最常见;巨核系异常主要以小巨核(14例,18%)改变为主。结论 MDS的诊断主要依靠骨髓的病态造血,但是有病态造血不一定就是MDS,还需结合细胞遗传学、骨髓活检等实验室检查。此外,还要排除其他疾病引起的病态造血,要注意鉴别诊断。     

Immunosuppressive therapy for myelodysplastic syndromes

Myelodysplastic syndromes (MDS) comprise a heterogeneous group of diseases characterized by bone marrow failure, marrow dysplasia, and a tendency to evolve to acute leukemia. Pathophysiologically, low risk MDS are separated from the high risk category by an increased rate of apoptosis of the bone marrow cells which causes the morphological paradoxon of a peripheral cytopenia and hypercellular bone marrow known as ineffective hematopoesis. Laboratory findings and clinical evidence suggest that some patients with myelodysplastic syndrome have immunologically mediated disease. MDS shares some of the features of acquired aplastic anemia and up to 30% of patients with MDS respond to immunosuppressive treatment. In the last decades, significant advances have been made in the diagnostic and prognostic classifications of myelodysplastic syndromes. While allogeneic transplantation still offers the only available option with a probability of cure in a minority of patients, the mainstay of therapy in low-risk patients remains supportive care, stimulation of ineffective hematopoiesis with growth factors, and immunomodulatory therapy. However, the correct selection of patients for the respective therapeutic intervention continues to be an enormous challenge as this will decide about the probability of therapeutic efficacy. This is important not only because of the high costs involved but also because of the possible side effects that can be difficult to manage. Here we review the pathophysiologic basis for the use of immunosuppressive agents in MDS and summarize the trials leading to the establishment of these therapy strategies in a subgroup of low-risk MDS patients.     

肝癌患者经导管动脉化学药物栓塞术前后血清肝细胞生长因子水平变化及其临床意义

目的:研究肝癌患者经导管动脉化学药物栓塞术(TACE)前后血清肝细胞生长因子(HGF)水平的动态变化及其影响因素,探讨其临床意义。方法:以30例慢性乙型肝炎基础上的肝癌患者为研究对象,用酶联免疫法(ELISA)测定TACE术前1天,术后第3、7、28天血清HGF浓度,分析TACE前后血清HGF水平动态变化及其影响因素。以10例健康献血者为正常对照组,10例慢性乙型肝炎患者及10例慢性乙型肝炎肝硬化患者为慢性肝炎组和肝硬化组。结果:肝癌患者TACE术前血清HGF浓度为[(0.63±0.28)ng/mL],显著高于正常对照组[(0.17±0.04)ng/mL]、慢性肝炎组[(0.35±0.08)ng/mL]以及肝硬化组[(0.39±0.09)ng/mL](P<0.05)。TACE术后血清HGF浓度上升高峰出现在术后第3天[(1.37±0.21)ng/mL],术后第7天逐渐下降[(0.79±0.14)ng/mL],术后第28天基本恢复术前水平[(0.66±0.19)ng/mL]。TACE术后有肿瘤转移的肝癌患者,其术前血清HGF水平显著高于无肿瘤转移的患者[(0.94±0.18)ng/mLvs(0.51±0.23)...     

Influence of thalidomide on Bcl2 expression and proangiogenic cytokine levels in short-term culture of peripheral blood and bone marrow mononuclear cells of multiple myeloma patients

Supernatants from short-term culture of peripheral blood and bone marrow mononuclear cells obtained from 22 multiple myeloma patients were used to measure the concentration of TNF-alpha, HGF, IL-6 and its soluble receptor (sIL-6R), VEGF and bFGF. Cells were cultured with or without thalidomide (THAL). We observed statistically significant decrease in TNF-alpha, HGF, IL-6, sIL-6R in supernatants from THAL cultures compared to cells cultured without THAL. Flow cytometry technique was applied to study the Bcl2 expression on CD 4, CD 8 and CD 138 positive cells. The statistically significant decrease in Bcl2 expression on myeloma cells (CD 138+) was observed both in PB and BM cultures. THAL could inhibit the plasma cell growth both by diminishing proangiogenic cytokines production and enhancing myeloma cell apoptosis.     

全反式维甲酸联合细胞因子诱导骨髓间充质干细胞分化为神经元样细胞的作用

目的 探索全反式维甲酸(ATRA)、碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)在诱导骨髓间充质干细胞(BMSCs)分化为神经元样细胞过程中的作用.方法 应用Ficoll分离骨髓中单核细胞,贴壁培养,观测形态并用流式细胞仪检测其细胞表面标志;用含ATRA、bFGF、EGF的条件培养基对培养的细胞向神经细胞诱导分化;采用免疫荧光染色法对分化的细胞进行鉴定.结果 分离培养的贴壁细胞具有典型的BMSCs形态和细胞表面标志,诱导后细胞表达神经元和星形胶质细胞标志神经元特异性烯醇化酶(NSE)和胶质纤维酸性蛋白(GFAP) .结论 ATRA联合细胞因子具有诱导BMSCs在体外分化为神经元样细胞的能力.     

宫颈癌微血管计数和血管内皮生长因子的表达

目的探讨微血管密度(MVD)和血管内皮生长因子(VEGF)表达与宫颈癌的临床病理联系及其相互关系。方法应用免疫组织化学方法检测和分析72例宫颈癌、24例正常宫颈组织中MVD和VEGF的表达情况。结果宫颈癌患者的微血管密度明显高于正常组织(P<0.01);宫颈浸润癌明显高于原位癌(P<0.01),浸润癌Ⅰ期与Ⅱ期差异无显著性(P>0.05);MVD随着病理分级增高呈递增趋势,组织学分级Ⅰ级与Ⅱ级、Ⅲ级差异有显著性(P<0.05);MVD增高与患者的生存期有关,生存期<5年者MVD明显高于生存期>5年者(P<0.05);MVD与临床分型、淋巴结转移无明显相关性(P>0.05)。VEGF在宫颈原位癌呈低表达,原位癌强阳性率明显低于浸润癌(P<0.05);VEGF表达强弱与病理组织学分级有关,分化程度低,恶性程度高,VEGF表达增强。VEGF表达阳性组的宫颈癌标本中MVD明显高于VEGF表达阴性组(P<0.05),且随着VEGF表达的增强,微血管密度随之增加,两者呈正相关。结论VEGF与宫颈癌的血管生成密切相关;MVD和VEGF表达与宫颈癌的恶性程度密切相关。