Multifocal metachronous giant cell tumour in bilateral upper limb: a rare case presentation

Giant cell tumour (GCT) of bone is a benign but locally aggressive tumour and accounts for 20 % of all benign bone tumours and 5 % of all bone tumours. Multicentric GCT of bone is a rare entity and has increased prevalence of involvement of the small bones of hands and feet in multicentric GCT. The clinical behaviour in multicentric GCTs tends to be aggressive as in recurrent GCTs. En-bloc resection remains the most successful surgical technique for treating both multicentric and solitary lesions. We report a 14-year-old female patient presenting with metachronous benign GCT located at the right proximal humerus and subsequent lesions in left hand and left proximal humerus. The case was treated with multimodality therapy including en-bloc resection along with bisphosphonate therapy over a period of 5 years.     

Designing the epitope flanking regions for optimal generation of CTL epitopes

The flanking amino acids that surround epitopes are critical for effective antigen processing and maintenance of epitope integrity. In the present study, the frequency and characteristics of each amino acid that flanked the peptides generated from the proteasomal degradation of three different subtypes of HIV-1 Gag-p24 were determined. Synthetic flanking regions were designed based on the highest and the lowest frequencies of amino acid with the ideal characteristics at positions upstream and downstream of the proteasomal cleavage site. Peptides were synthesized that contained known CD8+ CTL-epitopes from HIV-1 Gag, CMV pp65, and vaccinia proteins HRP-2, and C16, flanked by amino acid sequences specifically designed to either generate or inhibit the known CD8+ CTL-epitopes. As predicted, the known CD8+ CTL-epitopes were effectively generated from the peptides with synthetic flanking regions specifically designed to promote epitope generation in a proteasome-dependent manner. The majority of the proteasome-generated epitopes were cleaved immediately after the C-terminal amino acid of the specific CTL-epitope. The synthetic peptide sequences containing known CD8+ CTL-epitopes with the flanking regions that promote epitope generation were effectively processed and presented to epitope specific CD8+ T-cells resulting in the production of IFN-γ. These results highlight the importance of flanking regions in promoting efficient antigen processing and presentation. This concept can have important implications in vaccine design and development strategies.     

Development and preclinical testing of HNVAC,a cell culture-based H1N1 pandemic influenza vaccine from India

Several limitations of the use of embryonated eggs and the threat of pandemics have highlighted the need for other platforms for the production of influenza vaccines. We report the indigenous development and pre-clinical testing of an MDCK-based H1N1 pandemic influenza vaccine HNVAC from India. The cell bank and virus seed were characterized extensively. The cells were characterized by PCR, electron microscopy, and karyotyping, and found to be of female canine epithelial origin. The virus was confirmed by neutralization, haemagglutination inhibition, neuraminidase inhibition, and PCR and nucleotide sequencing. Adventitious agent testing was performed by both in vitro and in vivo studies. The in vitro studies included culturing, haemadsorption, haemagglutination, PCR and RT-PCR, whereas in vivo studies included passage in embryonated eggs and in laboratory animals. Both cell bank and virus seed were free of adventitious agents. MDCK cell lysates as well as cellular DNA did not produce tumours in newborn or adult laboratory animals. The bioprocess parameters were standardized to recover antigen with minimal levels of process-related impurities. The vaccine bulk was tested for the presence of specific antigen, and quantified by single radial immunodiffusion. Finally, non-adjuvanted and aluminium hydroxide adjuvanted vaccine formulations were found to be safe in preclinical toxicity studies in mice, rats, guinea pigs and rabbits, and immunogenic in mice and rabbits. This is the first and only cell culture-based influenza vaccine platform developed in any developing country.     

Involvement of Protease-Activated Receptor 2 in Nociceptive Behavior in a Rat Model of Bone Cancer

Treatment for bone cancer pain remains a clinical challenge due to a poor understanding of the underlying mechanisms. Protease-activated receptor 2 (PAR2), a receptor for inflammatory proteases, has been implicated in nociceptive signaling under both normal and pathologic pain states. However, little is known of the role of PAR2 in cancer-induced bone pain. Here we investigated the potential role of PAR2 in a rat model of bone cancer pain. The model of bone cancer pain was induced by inoculating Walker 256 into the tibia bone cavity of rats and verified by X-ray imaging, pathology, and behavior assessments. The rats with bone cancer exhibited marked mechanical allodynia, thermal hyperalgesia, and signs of spontaneous nocifensive behavior. Subcutaneous administration of the PAR2 antagonist FSLLRY-NH2 almost completely abolished mechanical allodynia and thermal hyperalgesia but had no effects on spontaneous pain behavior in the rats with bone cancer. Immunohistochemical study revealed that the expression of PAR2 was significantly increased in large- and medium-sized dorsal root ganglia (DRG) neurons but not in small-sized neurons after Walker 256 inoculation. These results suggest that the increased expression of PAR2 in the DRG may contribute to the development of mechanical allodynia and thermal hyperalgesia associated with bone cancer rats. PAR2 might become a novel target for the treatment of pain in patients with bone cancer.     

Bricker术后TU-Pouch尿流复道(附1例报告并文献复习)

目的:探讨根治性全膀胱切除术术后尿流复道关键技术的设计以及适应证的选择。方法:我们对1例行根治性全膀胱切除+回肠膀胱术后患者施行回肠+乙状结肠原位新膀胱术以实现尿流复道,术前准备主要包括患者心理准备、肿瘤预后的相关检查和尿道括约肌功能判断。金属尿道探子支撑下分离尿道残端以备用。选取乙状结肠20cm纵行剖开形成肠袋,作为新膀胱的"U"部分。切除Bricker腹壁造瘘口,并以此段回肠形成半套叠状抗反流结构及新膀胱的"T"部分。"T"肠袋与"U"肠袋侧-侧吻合,共同形成"TU-Pouch"新膀胱。新膀胱最低点与尿道残端吻合。结果:术后随访1年,患者自行排尿通畅,尿量350~400ml/次,日间排尿3~5次,控尿良好;夜间排尿2~3次,轻度尿失禁。逆行膀胱造影未见造影剂向输尿管反流,静脉肾盂造影见双侧肾盂输尿管显影良好,无积水。结论:全膀胱切除术后的尿流复道手术是复杂的、高难度和高风险手术,需严格掌握适应证。有效的抗反流机制和良好的控尿功能是决定手术是否成功、影响患者术后生活质量的关键。     

SiRNA联合介导Livin和Survivin基因沉默诱导人肾癌细胞786-O化疗敏感性的实验研究

目的:观察siRNA联合介导Livin和Survivin基因的沉默诱导人肾癌细胞786-O化疗敏感性的作用。方法:构建Livin和Suvivin联合靶向的小干扰RNA(siRNA)重组表达载体shRNA,然后将目的载体转染于人肾癌细胞株786-O。应用实时荧光定量聚合酶链反应(Real-time PCR)及Western blot方法分别检测转染前细胞经顺铂、5-FU和丝裂霉素处理后的Livin和Survivin基因的mRNA与蛋白水平的变化。应用CCK-8试验检测转染前后细胞对顺铂、5-氟尿嘧啶(5-FU)和丝裂霉素的半数致死量(IC50)、细胞增殖的变化。结果:CCK-8结果显示,联合介导Livin和Survivin基因沉默组细胞较分别单独介导细胞组化疗的敏感性明显增强(P0.05),且转染前后细胞对顺铂、5-FU和丝裂霉素的IC50发生显著变化(P0.05)。结论:SiRNA联合介导Livin和Survivin基因表达下调,发挥协同增强的siRNA作用。     

经后外侧入路椎间盘摘除椎间融合术治疗胸椎间盘突出症

【摘要】 目的：评价经后外侧入路椎间盘摘除椎间融合术治疗胸椎间盘突出症的安全性和有效性。方法：2006年1月~2012年8月采用经后外侧入路椎间盘摘除椎间融合术治疗胸椎间盘突出症（TDH）患者24例,男15例,女9例,年龄37~64岁,平均45.3岁。病程4个月~2年,平均14个月。均为单节段椎间盘突出,中央型突出16例,旁中央型8例;突出为硬性18例,软性6例。突出部位：T7/8 2例,T8/9 4例,T9/10 4例,T10/11 5例,T11/12 9例。其中3例合并胸椎黄韧带骨化,4例合并胸椎后纵韧带骨化。手术前后进行JOA评分和Otani分级,并通过Bridwell标准评价骨融合情况。结果：患者均顺利完成手术,未出现神经损伤。1例切口脂肪液化延迟愈合,其余均Ⅰ期愈合。3例术中硬脊膜损伤,术中修补后2例术后未发生脑脊漏,1例发生脑脊液漏,经置管引流后治愈。24例均获随访,随访时间0.5~3.5年,平均2.1年。术后定期门诊复查。患者症状均得到不同程度改善,随访期间未见神经症状加重及脊柱不稳等情况。术前改良JOA评分为4.4±2.4分,术后3个月为8.7±2.3分,末次随访时为9.0±2.3分,术后3个月和末次随访时与术前比较差异均有统计学意义（P＜0.05）。末次随访时临床改善率为（74.6±16.6）%。根据Otani分级,优9例,良11例,可2例,差2例,优良率为83.3%。2例截瘫患者中,1例无明显恢复,1例术后半年由Frankel A级恢复至C级。术后3个月随访植骨块均达到Ⅱ级或Ⅲ级愈合,术后6个月时Ⅰ级愈合6例,其余均达Ⅱ级愈合,19例随访9个月以上的患者9个月时均达到Ⅰ级愈合。随访期内无内固定松动、断裂和节段间塌陷等。结论：采用经后外侧入路椎间盘摘除椎间融合术治疗胸椎间盘突出症可获得较好的临床效果。     

盐制对杜仲治疗去卵巢大鼠骨质疏松症影响的研究

目的：研究杜仲及其盐制品对去卵巢大鼠骨质疏松症的治疗作用,并探讨盐制对其治疗作用的影响。方法40只SD雌性大鼠分为伪手术组、模型对照组、杜仲生品治疗组和杜仲盐制品治疗组,除假手术组施行假手术外,其余均行手术彻底摘除卵巢,术后4周开始杜仲生品及盐制品水提液灌胃给药（4．0 g／kg?d）,实验过程称体质量,连续给药12周后,颈动脉取血,测定血钙（S－Ca）、血磷（S－P）含量,ELISA试剂盒双抗体夹心法测定血清碱性磷酸酶（ALP）、骨钙素（OCN）、雌二醇（E2）含量;动物处死后剥取完整子宫称重,采用HE染色法进行子宫病理学检查,剥离双侧股骨,双能X射线衍射法测定大鼠股骨骨密度。 HE染色法观察股骨骨小梁形态和成骨细胞数量,micro－CT扫描观察股骨骨小梁微体系结构。结果杜仲生品及盐制品能有效抑制去卵巢所致的大鼠体重增加,升高血清钙、雌二醇的含量,降低血磷、碱性磷酸酶和骨钙素的含量。增加大鼠股骨骨密度,改善股骨骨小梁微体系结构,增加股骨骨小梁成骨细胞数量,且长期用药对子宫无明显刺激作用。结论杜仲生品及盐制品对于大鼠去卵巢所引起的骨质疏松症有良好治疗作用,且盐制品效果优于生品。