Inhibition of melanoma growth by hemocyanin occurs via early apoptotic pathways

BACKGROUND: We hypothesized that keyhole limpet hemocyanin (KLH) would reduce cellular proliferation and effect apoptosis of melanoma cell lines in vitro. METHODS: Two human melanoma cell lines (HTB68 and HTB72) were subjected to a dose-response treatment regimen of KLH (0.4 microg to 100 microg/well). Cell viability was tested by MTT assay (SIGMA, St Louis, MO) at 72 hours. Apoptosis and necrosis were measured by the Annexin V FITC assay (Biovision Inc, Mountain View, CA). RESULTS: Melanoma cell proliferation was significantly reduced in the HTB68 cell line treated with 6.3 microg or higher doses of KLH. A significant reduction in cell growth was also observed in the HTB72 cells at 50 and 100 microg of KLH. KLH increased early apoptotic activity, whereas both late apoptosis and necrosis were decreased by the addition of KLH. CONCLUSIONS: KLH significantly reduces cellular proliferation in vitro in melanoma, via early apoptotic pathways. The results warrant in vivo studies into the effects of KLH in melanoma.     

Proteomic analysis of SEG-1 Human Barrett’s-associated esophageal adenocarcinoma cells treated with keyhole limpet hemocyanin

Keyhole limpet hemocyanin (KLH) is an immune stimulant derived from a circulating glycoprotein of the marine mollusk Megathura crenulata. We previously reported that KLH inhibited the growth of human Barrett’s-associated esophageal adenocarcinoma in vitro via apoptotic and nonapoptotic mechanisms. We hypothesize that KLH reduces the growth of Barrett’s cancer cells by altering protein expression profiles. A cell line (SEG-1) derived from Barrett’s-associated adenocarcinomas of the distal esophagus was selected. Cells were administered KLH (500 μg/ml) or vehicle. After 24 hours, cytosolic fractions were separated through two-dimensional gel electrophoresis. Statistical analysis was performed with Evolution Pro software to identify spots that were differentially expressed between the KLH and control groups. Proteins displaying a twofold or greater change in expression levels were selected for identification. In a total of 420 spots, 31 were differentially expressed between the KLH and control groups. In all, 12 were upregulated and 19 were downregulated. Of the 31, 17 were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Proteomic evaluation shows downregulation of proteins associated with metabolic processes (glycolysis, protein synthesis). KLH also induced proteins indicative of oxidative stress (heat shock 70 family and UDP-glucose 6-dydrogenase). Our results indicate that growth arrest by KLH is accompanied by a cellular stress response and attenuation of metabolic processes. The use of KLH as adjuvant or topical therapy for Barrett’s adenocarcinoma provides a promising development in the treatment of this disease. Presented in part at the Forty-Fifth Annual Meeting of The Society for Surgery of the Alimentary Tract, New Orleans, Louisiana, May 15–19, 2004 (poster presentation). Supported by the Bernard Zimmerman Foundation (to the Department of Surgery) and the WVU Robert C. Byrd Health Science Center Proteomic Core Laboratory (COBRE in Signal Transduction RR16440 as a grant of time).     

利用锁孔技术行幕下硬膜外血肿清除术的意义

目的探讨锁孔技术在幕下硬膜外血肿清除术中的作用。方法将60例创伤性幕下硬膜外血肿患者随机分成两组各30例（分别称为观察组及对照组，简称A组及B组），A组利用锁孔技术清除硬膜外血肿（必要时使用神经内镜辅助），B组使用传统后颅窝开颅术清除硬膜外血肿，比较两组在手术时间、手术出血量及疗效方面的差别。结果平均手术时间A组为46min，B组为113min（P〈0．01）；平均手术出血量A组为81ml，B组为425ml（P〈0．01）；两组硬膜外血肿均完全清除，无复发。结论在幕下硬膜外血肿清除术中，利用锁孔技术比传统后颅窝开颅术具有手术时间短、出血量少、疗效确切等优点，值得推广应用。     

经额外侧锁孔入路切除鞍区颅咽管瘤

目的探讨经额外侧锁孔入路切除鞍区颅咽管瘤的应用价值。方法对我院17例鞍上直径在2.5-4.8cm的颅咽管瘤患者采用额外侧锁孔入路发迹内做6.5cm左右皮切口,铣出面积约（3×2.5）cm^2的椭圆形小骨瓣,术中根据肿瘤位置利用不同解剖间隙切除肿瘤。结果术中15例病人肿瘤全部切除,2例次全切除。术后7例病人出现不同程度的尿崩症及电解质紊乱,经药物治疗后好转,9例病人视力改善。结论与传统翼点入路及眶上锁孔入路相比,额外侧锁孔入路切口小、创伤小,能提供足够的鞍上区手术空间,显露鞍区病变及其邻近结构充分,且减少了脑牵拉和手术创伤,有利于颅咽管瘤的全切除,是一种安全有效的处理鞍上颅咽管瘤的手术入路途径。     

内镜辅助锁孔手术切除颅内蛛网膜囊肿

目的探讨内镜辅助锁孔手术切除颅内蛛网膜囊肿的临床意义。方法应用内镜辅助锁孔手术切除颅内蛛网膜囊肿19例(囊肿大部分切除加脑池交通)。结果术后3个月至2年随访,19例患者均有不同程度的改善,囊肿闭合消失15例,明显缩小4例,5例癫痫患者在小剂量抗癫痫药物辅助下均较好得到控制,头晕头痛者均获明显改善。结论内镜辅助锁孔手术切除颅内蛛网膜囊肿是一种行之有效的方法 ,疗效满意     

Epitope mapping of a PrP(Sc)-specific monoclonal antibody: identification of a novel C-terminally truncated prion fragment

Monoclonal antibodies (mAbs) against prion proteins (PrPs) are indispensable in research and diagnosis of prion diseases, however the majority of these bind both the cellular (PrP^C) and the disease-associated (PrP^Sc) isoforms. According to the widely accepted protein-only hypothesis the two isoforms share the same sequence, but differ in their conformation. In the present study we set to determine the critical binding residues of our PrP^Sc-specific mAbs with the view of discerning which residues play a key role in the conformational transition between PrP^C and PrP^Sc. Focussing on the V5B2 mAb that provided differential labelling of prion-affected tissue from individuals positive for transmissible spongiform encephalopathies, we performed alanine scanning and phage-display epitope mapping to elucidate the antigenic determinants of this mAb and gain insight into its specificity on a molecular level. We observed that instead of discriminating between the two prion protein isoforms based on conformational differences, V5B2 binds a previously uncharacterized C-terminally truncated form of PrP^Sc that ends with the residue Y226, which we named PrP226*. The addition of a single C-terminal amino-acid residue completely abolished V5B2 binding, while Western blots using recombinant full-length PrPs and PrPs terminating at Y226 confirmed that the V5B2 mAb discriminates between the two based on their difference in length.     

超早期翼点小切口经侧裂-岛叶入路显微手术治疗高血压基底节区脑出血的疗效观察

目的探讨高血压基底节区脑出血（HBGH）新的手术治疗方法和疗效。方法60例HBGB患者中,30例经超早期翼点小切口经侧裂-岛叶入路显微手术清除血肿（侧裂组）,另30例经常规骨瓣开颅、皮质造瘘口直视下手术清除血肿（皮质组）。结果侧裂组近期和远期疗效均明显优于皮质组（P〈0．05）。结论超早期翼点小切口经侧裂一岛叶入路显微手术治疗HBGH创伤小、疗效佳、预后好。     

Keyhole limpet hemocyanin: an effective adjunct against melanoma in vivo

BACKGROUND: We have previously demonstrated the potent in vitro antiproliferative effects of keyhole limpet hemocyanin (KLH) against melanoma. Our prior studies directed us to hypothesize that KLH would be effective in vivo against melanoma, alone and in combination with conventional immunotherapy. METHODS: Mice were inoculated with 2 x 10(7) HTB68 cells and randomized to 6 groups. Treatment groups consisted of control, KLH 200 microg, alpha interferon (AIFN) 1000 IU, interleukin-2 (IL-2) 5000 IU, KLH + AIFN, and KLH + IL-2. RESULTS: KLH + IL-2 exhibited the greatest reduction in tumor volume (30%) as compared to control (P = .014), followed by KLH + AIFN (28%, P = .031). Singly treated animals had less tumor inhibition: IL-2 (30%, P = .022), KLH (18%, not significant), and AIFN (16%, not significant). CONCLUSIONS: KLH augments the effects of AIFN, one of the standard immunotherapeutic agents against melanoma in vivo. Further in vivo and early clinical studies into the effects of KLH as both a single and combined agent are warranted.     

经穹隆间第三脑室底锁孔入路的显微解剖

目的 探索经穹隆间第三脑室底锁孔入路的可行性和手术方法。 方法 设计经穹隆间第三脑室底锁孔入路(第三脑室底切口起自灰结节向后,经乳头体间,止于后穿质)。运用解剖学方法在导航辅助下在16例尸头标本上模拟经穹隆间第三脑室底锁孔入路手术,在手术显微镜下对手术显露进行观察,利用导航作解剖学测量。 结果 导航辅助下能顺利完成16例尸头标本的经穹隆间第三脑室底锁孔入路手术。冠状缝与矢状缝交点到室间孔上缘、丘脑间黏合、乳头体和中脑导水管上缘的距离分别为（68.4±4.6）mm、(66.3±6.0)mm、(86.3±5.3)mm、(82.0±7.6)mm,冠状缝与矢状缝交点到基底动脉末端分叉的操作距离为(91.8±5.0)mm。灰结节向后经乳头体间止于后穿质切开第三脑室底可获得长（9.5±2.6）mm的手术通道。术中经第三脑室底切口能清晰显露脚间池内的基底动脉末段、大脑后动脉P1段、P2段、小脑上动脉、后交通动脉以及它们的穿通支血管。向前解剖Liliequist膜可显露斜坡和鞍背,侧方可显露出动眼神经,向后显露出脚间窝。基底动脉末端分叉多偏于左侧（68.8%）,两侧大脑后动脉多向前外侧斜行（68.8%）。大部分大脑后动脉夹角上有1～4支小穿支血管自基底动脉末端分出。 结论 经穹隆间第三脑室底锁孔入路在技术上可行,深入研究可望应用于基底动脉末端动脉瘤的直接手术。