A comparative study of angiogenic and cytokine responses after laparoscopic cholecystectomy performed with standard- and low-pressure pneumoperitoneum

Background  Surgical procedures enhance production of pro- and anti-inflammatory cytokines and angiogenic factors that play a pivotal role in the immunological response to surgical trauma and take part in the pathogenesis of tumor growth and adhesions formation. The purpose of the study was to access the influence of low-pressure CO₂ pneumoperitoneum on the inflammatory and angiogenic responses during the postoperative period after laparoscopy. Methods  The study group consisted of 40 patients, operated on due to cholelithiasis using standard-pressure (n = 20) and low-pressure (n = 20) CO₂ pneumoperitoneum. Serum concentration of interleukin (IL)-6, IL-8, IL-10, vascular endothelial growth factor (VEGF)-A, and endostatin were measured before and at 6, 24, and 48 h after surgery with commercially available enzyme-linked immunosorbent assay (ELISA). Results  Concentrations of IL-6 increased significantly after the operations in both groups. No differences were observed between the groups in regards to IL-6, IL-8, and IL-10 levels. Concentrations of VEGF-A measured at 6 and 48 h were significantly lower in patients who underwent laparoscopies performed with low-pressure pneumoperitoneum. No significant variations were observed in endostatin serum concentration. Concentrations of the studied parameters were not influenced by duration of surgery or by age, gender, or body mass index (BMI) of the patients. Conclusions  The results obtained in our study do not show any significant differences between studied operative procedures with regards to systemic inflammatory response. Changes in the concentrations of VEGF-A and endostatin observed in the studied population may suggest this technique is more favorable with regards to angiogenesis process intensity, along with all its consequences and implications.     

hIL-2分泌肽序列在HepG2细胞中对hEndostatin表达和分泌差异的影响

目的:了解分泌肽序列在HepG2细胞中对人内皮抑素基因(hEndostatin)的cDNA表达和分泌差异的影响。方法:构建不含hIL-2分泌肽序列的真核表达载体pBlast-hEndo质粒, 转染人的HepG2细胞株, 用RT-PCR的方法检测HepG2(pBlast-hIL2-hEndo), HepG2(pBlast-hEndo), HepG2(pBlast-Mcs)和HepG2中hEndostatin的mRNA表达水平, 及制备4种细胞的总蛋白和收集各自的培养上清, 进行Westernblot分析蛋白表达和分泌的差异。结果:发现HepG2(pBlast-hIL2-hEndo)中hEndostatin的mRNA的水平显著高于HepG2(pBlast-hEndo)。而仅在HepG2(pBlast-hIL-2-hEndo)的细胞总蛋白和上清, 及HepG2(pBlast-hEndo)的细胞总蛋白中检测到hEndostatin表达, 在其它各株细胞总蛋白及上清中, 未检测到hEndostatin。结论:hIL-2分泌肽序列能促进hEndostatin基因在HepG2细胞中表达及分泌。     

小鼠S180移植瘤血管生成作用及其调节机制的研究

目的：观察肉瘤180(S180)移植瘤发展过程中血管生成及血管生成调节因子的变化,并对其调节机制进行探讨。 方法： 利用Km小鼠的S180移植瘤模型，采用FⅧ因子免疫组化染色检测肿瘤血管生成，ELISA和EIA法检测荷瘤鼠肿瘤组织和血浆中血管内皮生长因子（VEGF）和内皮抑制素（endostatin）水平，采用多元回归分析肿瘤组织微血管计数、血管形态与瘤重变化的关系。 结果： 随着荷瘤时间延长，肿瘤组织内微血管计数，瘤内血管相对总量增加，血管的相对面积增大（P<0.05）；肿瘤组织匀浆中VEGF水平在荷瘤10 d、15 d均显著高于5 d组（P<0.05）；endostatin在肿瘤匀浆和血浆中均在荷瘤15 d达到最高（P<0.05）；V/E比值无显著变化；微血管计数、血管相对总面积与瘤重变化有相关性（P<0.01）。 结论： S180移植瘤病期发展中微血管数目增加，血管口径增大，且与瘤重变化呈正相关；肿瘤发展过程中肿瘤局部血管生成正调节因子逐渐增加，促进血管生成；肿瘤局部血管生成调节因子处于相对的平衡。     

Time-resolved fluorometric assay for the detection of endostatin in chromatographically separated extracts of natural peptides

We present a heterogeneous non-competitive immunological detection assay for peptide and protein antigens from crude extracts of biological sources. This time-resolved fluoroimmunoassay (TR-FIA) has been designed in a solid-phase mode using 96-well microtiter plates. Using the rare-earth metal europium as a fluorescent marker, a highly sensitive, selective and efficient procedure was developed. This technique prevents from interferences of intrinsic protein fluorescence which is highly important for antigen measurement in complex matrices. The TR-FIA has been applied for the detection of circulating forms of the potential anti-tumor agent endostatin, a C-terminal fragment of collagen XVIII, and its close homolog collagen XV (restin) from hemofiltrate. Endostatin was detected with a limit of detection of 3 ng (150 fmol/well) and a broad dynamic range from 10-1000 ng/well.     

内抑素(endostatin)对移植大肠癌新生血管密度影响的动物实验研究

目的：探讨内皮细胞生长抑素（内抑素endostatin）对裸鼠大肠血管生成的影响及其作用机理，为endostatin的临床应用提供理论依据。方法：采用免疫组化方法检测肿瘤新生血管密度，对用药过程中瘤 体积进行测量，对endostatin抑制肿瘤的效果进行分析。结果：endostatin能够抑制裸鼠大肠癌肿瘤血管生成，减轻肿瘤的血管密度。结论：endostatin通过抑制血管形成抑制肿瘤的生长。     

人内皮抑素在毕赤酵母中的组成型表达及其活性检测

为获得大量具有生物学活性的重组人内皮抑素(Endostatin),在毕赤酵母中进行组成型表达的研究。将由毕赤酵母偏爱密码子组成的Endostatin cDNA插入组成型载体pGAPZαA中,构建表达载体pGAPZαA-ENDO,并转化到毕赤酵母X-33中。重组菌株在甘油醛-3-磷酸脱氢酶(GAP)启动子的调控作用下,进行组成型表达Endostatin蛋白。重组人Endostatin产量达到102mg/L。Western blot显示:表达蛋白能与兔抗Endostatin的多克隆抗体特异性结合。纯化后的重组Endostatin具有抑制人血管内皮细胞系ECV-304细胞增殖的活性。利用毕赤酵母组成型表达系统能获得大量具有生物活性的Endostatin蛋白。     

Elevated serum endostatin levels is associated with favorable outcome in acute myeloid leukemia

Introduction

Endostatin is the C-terminal antiangiogenic fragment of the extracellular matrix protein collagen XVIII, and is generated by tumor-derived proteases. The levels and the prognostic relevance of serum endostatin in AML patient is not fully clear.

Aim

To evaluate serum levels of endostatin in acute myeloid leukemia patients before chemotherapy and after achieving complete remission and to correlate endostatin levels with patients outcome.

Materials and methods

Serum samples from 30 adult patients (22 males and 8 females, median age 37, range 19–66 years) with AML had been taken before chemotherapy was administered. In addition 20 out of 30 patients were reinvestigated again at complete remission (CR). Ten samples from healthy normal persons of matched age and sex were evaluated as a reference control group. Serum endostatin levels were determined using enzyme linked immune sorbent assay (ELISA).

Results

Endostatin serum levels were not significantly different in the pretreatment AML patients as compared to that in normal controls (P>0.05). In AML patients the baseline endostatin levels were significantly lower than at CR (P=0.001). No significant correlation were detected between pretreatment serum endostatin levels and age, peripheral blood white cell counts, platelet counts, bone marrow blast cell counts, blast cell distribution ratio. The prognostic value of sE was also evaluated by dividing AML patients into high and low sE groups using the 75 percentile sE levels of the patients group as cutoff. The authors found that patients group in the high sE group survived for significantly longer time than those patients in the low sE group.

Conclusions

Elevated endostatin levels at AML diagnosis is a good prognostic marker for patients’ outcome. Wide scale study is recommended in order to establish the clinical value of this study.

     

肝动脉化疗栓塞术中注入重组人血管内皮抑制素治疗中晚期肝癌的疗效及安全性分析

目的：观察重组人血管内皮抑制素经肝动脉灌注联合TACE治疗中晚期肝癌的近期及远期疗效。方法将64例中晚期肝癌患者分为对照组（n=30例),仅予以TACE治疗,和观察组（n=34例）,在行TACE治疗时,同时经肝动脉注入重组人血管内皮抑制素,比较两组近、远期疗效,观察治疗前后肿瘤直径、KPS评分及AFP变化,分析两组毒副反应。结果观察组治疗总有效率（64.7%）明显高于对照组（43.3%,P<0.05）;两组患者治疗后肿瘤直径[（4.0±1.2） cm和（4.9±1.3） cm]较治疗前[（6.7±2.0） cm和（6.6±2.2） cm]明显缩小（P<0.05）,KPS评分明显升高[（92.5±9.8）对（76.1±4.5）和（84.8±8.6）对（75.2±3.4）,P<0.05],AFP明显降低[（547.2±160.7）μg/L对（1275.3±316.8）μg/L和（558.5±176.4）μg/L 对（1218.8±337.9）μg/L,P<0.05],且观察组肿瘤直径[（4.0±1.2） cm、KPS评分（92.5±9.8）较对照组缩小（4.9±1.3） cm或升高（84.8±8.6）更为显著,P<0.05];两组毒副反应比较差异无显著性（P>0.05）;随访1年,观察组患者病死率（5.9%）和肝外转移率（14.7%）明显低于对照组患者（16.7%和26.7%, P<0.05）。结论在行TACE治疗时加用重组人血管内皮抑制素治疗中晚期肝癌患者可明显提高近期及远期疗效,缩小肿瘤直径和改善KPS评分,且不会加重毒副反应。     

Circulating angiogenic biomolecules at rest and in response to upper-limb exercise in individuals with spinal cord injury

Objective

Individuals with spinal cord injury (SCI) show structural and functional vascular maladaptations and muscle loss in their lower limbs. Angiogenic biomolecules play important roles in physiological and pathological angiogenesis, and are implicated in the maintenance of muscle mass. This study examined the responses of angiogenic molecules during upper-limb aerobic exercise in patients with SCI and in able-bodied (AB) individuals.

Methods

Eight SCI patients with thoracic lesions (T₆–T₁₂, ASIA A) and eight AB individuals performed an arm-cranking exercise for 30 minutes at 60% of their VO_2max. Plasma concentrations of vascular endothelial growth factor (VEGF-A₁₆₅), VEGF receptor 1 (sVEGFr-1), VEGF receptor 2 (sVEGFr-2), metalloproteinase 2 (MMP-2), and endostatin were measured at rest, after exercise, and at 1.5 and 3.0 hours during recovery.

Results

The two-way analysis of variance showed non-significant main effects of “group” and significant main effects of “time/exercise” for all angiogenic biomolecules examined (P < 0.01–0.001). The arm-cranking exercise significantly increased plasma concentrations of VEGF, sVEGFr-1, sVEGFr-2, MMP-2, and endostatin in both groups (P < 0.001–0.01). The magnitude of the increase was similar in both patients with SCI and AB individuals, as shown by the non-significant group × time interaction for all angiogenic parameters.

Conclusions

Upper-limb exercise (arm-cranking for 30 minutes at 60% of VO_2max) is a sufficient stimulus to trigger a coordinated circulating angiogenic response in patients with SCI. The response of angiogenic molecules to upper-limb aerobic exercise in SCI appears relatively similar to that observed in AB individuals.     

骨肉瘤患者肿瘤伴随抑制现象的临床研究

目的探讨骨肉瘤患者的肿瘤伴随抑制现象（CTR）、预测肺转移的高危患者，并对其进行抗肿瘤血管生成因子的治疗提供依据。方法采用ELISA法检测骨肉瘤患者手术前后1周血清中内皮抑素（ES）、血管内皮生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）和转化生长因子-β1（TGF-β1）的水平。术后每月摄胸部X线片和每3个月行胸部CT扫描，以了解是否有早期肺转移。比较出现肺转移组患者与未出现肺转移组患者手术前后ES、VEGF、bFGF和TGF-β1变化值的临床意义。结果在肺转移组8例与无肺转移组9例手术前后血清中ES变化值和VEGF升高值的差异有显著性（P均〈0．05）。两组手术前后血清TGF-β1、bFGF变化值的差异无显著性（P均〈0．05）。结论本组骨肉瘤患者可能存在肿瘤伴随抑制现象。血清VEGF、ES手术前后的变化值有望成为骨肉瘤患者术后早期筛查肺转移的预测因子及判断预后的指标之一。