Oncogenic BRAF and KRAS mutations in endosalpingiosis

Endosalpingiosis, a microscopic lesion composed of ectopic Fallopian tube epithelium, frequently involves the peritoneum and lymph nodes in patients with ovarian serous borderline tumour or low-grade serous carcinoma, but its pathogenic significance remains unclear. Using laser-capture microdissection and droplet digital PCR, we investigated whether endosalpingiosis harbours the driver mutations in BRAF and KRAS that characterise ovarian low-grade serous neoplasms. Somatic mutations were detected in 14 (33%) of 43 endosalpingiotic lesions analysed. Of 21 women with endosalpingiosis associated with a synchronous or metachronous ovarian low-grade serous tumour, mutations were identified in endosalpingiotic lesions from 11 (52%) women, with most cases (10/11, 91%) demonstrating identical mutations in both tumour and endosalpingiosis. In contrast, of 13 cases of endosalpingiosis not associated with an ovarian tumour, only one harboured a KRAS mutation. The proliferative activity as assessed by Ki-67 immunohistochemistry was lower in endosalpingiosis than in low-grade serous tumours, and endosalpingiosis with either a BRAF or KRAS mutation had a significantly lower Ki-67 index than those without. Ectopic expression of KRAS^G12V in Fallopian tube epithelial cells led to ERK phosphorylation, p21 induction, growth arrest and cellular senescence. In conclusion, we demonstrate that endosalpingiosis represents an interesting example of cancer driver mutations in deceptively normal-appearing cells, which may be prone to neoplastic transformation upon bypass of endogenous oncosuppressive mechanisms. © 2019 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

第四跖背动脉逆行岛状皮瓣在第二足趾再造手指供区趾蹼皮肤缺损中的临床应用

目的探讨第四跖背动脉逆行岛状皮瓣修复第二足趾再造手指后供区趾蹼皮肤缺损的临床应用效果。方法2007年1月~2009年12月,均为切取第二足趾再造手指时携带面积不等的跖背皮瓣13例患者的临床资料进行分析。结果术后13例转移皮瓣全部存活,术后随访时间为3~16个月,皮瓣质地、感觉良好,趾蹼外形满意,供足行走功能满意。结论最大限度地保护了供足的功能,减少了植皮后并发症,是较为理想的手术方式。     

Common occurrence of an antiidiotypic antibody that recognizes T14+ anti-DNA antibodies in patients with systemic lupus erythematosus

Objective. To investigate whether antibodies to a T14 anti-DNA antibody can be found in patients with systemic lupus erythematosus (SLE). Methods. Seventy-six serum samples (37 from patients with SLE) were randomly selected from among sera submitted for routine antinuclear antibody testing. Short, overlapping peptides based on the partial V_H (variable region of the heavy chain) sequence of the T14 antibody were synthesized on multipins and screened for reactivity with SLE sera. In addition, selected peptides from T14 and related proteins were synthesized in bulk and screened for reactivity with both SLE and control sera. A monoclonal antibody was generated to determine the prevalence of the T14 idiotype (T14+ Id) in the different study populations. Results. Antibodies were detected by a peptide based on the third complementarity-determining region (CDR3) of the T14 protein in 15 (41%) of 37 patients with SLE or 15 (54%) of 28 who had anti-DNA antibodies, in 3 (9%) of 34 patients without anti-DNA antibodies (9 of whom had SLE), and in 6 (10%) of 57 healthy controls. In SLE sera, the antiidiotypic (anti-Id) responses (IgM and IgG) correlated well with the anti-DNA responses (IgG), and both responses correlated well with the T14+ Id activity in SLE sera. Control peptides based on the 18/2 (16/6+ Id) and S107 proteins detected low antibody activities in SLE sera, attributable to cross-reactivity with the T14 peptide. A peptide based on an unrelated human antibody was not reactive with these sera. Conclusion. Anti-Id antibodies directed to T14 V_HCDR3 were found commonly in the sera of patients with SLE, and they appeared to be induced by the anti-DNA antibodies present in the sera. Based on these findings, these secondary antibodies may be pathogenic in SLE.     

bcl-xL纳米微粒的制备及其对大鼠心肌细胞的高效转染

目的:制备包载含人类抗凋亡基因bcl-xL的纳米微粒,评价其转染大鼠心肌细胞的能力. 方法:用超声双乳化蒸发法制备包载bcl-xL的纳米微粒,扫描电镜观察纳米的形态和大小,DNase I消化酶试验检测其抗核酸酶能力;转染体外培养的大鼠心肌细胞后RT-PCR和Western-blot法检测基因表达. 结果:制备的纳米微粒直径约200～260 nm,包封完好,性能稳定并对基因有缓释作用;转染1 wk后bcl-xL表达高于对照组(P<0.05). 结论:成功制备包载真核表达质粒的纳米微粒,并能够高效转染大鼠心肌细胞. 纳米微粒是心肌细胞转染较理想的基因载体之一.     

不同浓度异丙酚持续输注对家兔脂代谢的影响

目的:观察50 g/L异丙酚的药效及对家兔脂代谢的影响.方法:选择家兔12 只,随机分为10 g/L,50 g/L异丙酚组,每组6 只.10 g/L,50 g/L异丙酚单次首次剂量为6 mg/kg,以0.6 mg/s从家兔耳缘静脉分别匀速注射,记录起效时间.注射完毕后接麻醉注射泵以400 μg/(kg·min)的速度持续输注异丙酚360 min,记录心率、血压和呼吸等生命体征变化及停药后苏醒时间.并于注射前、注射完毕、持续输注后5,30,60,180,360 和365 min分别抽血样,检测血浆药物浓度,及血清甘油三酯(TG),总胆固醇(TC),高密度脂蛋白胆固醇(HDL-c),低密度脂蛋白胆固醇(LDL-c)的浓度.结果:单次静脉注射异丙酚起效时间、各时间点血药浓度两组间无显著差异(P>0.05);持续用药后,10 g/L异丙酚组家兔血清TG 浓度在60,180和360 min与用药前比较均有显著增高(P<0.01),血清TC浓度在用药后360 min较给药前增高(P<0.01);50 g/L异丙酚组的血清TG 浓度只在360 min时与用药前比较有显著增高(P<0.01),但显著低于同时间点10 g/L异丙酚组(P<0.01);两组停药后的苏醒时间无差异(P>0.05).结论:50 g/L异丙酚与10 g/L异丙酚的药效相似,但能延缓脂代谢紊乱的发生时间.     

小鼠IL-12基因转染对人卵巢癌Skov3细胞的生长及细胞因子的表达

目的:观察含小鼠IL-12全长基因的质粒转染到Skov3人卵巢癌细胞中,在脾细胞存在的情况下对肿瘤细胞的影响及相关细胞因子的表达. 方法:脂质体转染技术将基因转染至Skov3人卵巢癌细胞中,ELISA方法检测IL-12及INF-γ的表达,MTT方法检测对Skov3卵巢癌细胞增殖的影响. 结果:转染后48 h检测到IL-12的高表达,约(473±38) ng/L;而未转染组约13～17 ng/L,两者比较差异有统计学意义(P＜0.05). 在脾细胞作用下产生INF-γ,以作用后24 h表达量高,约(173±18) ng/L,较未转染组高,比较有统计学意义(P＜0.05). 转染后癌细胞免疫组化可见IL-12表达,在脾细胞的作用下,IL-12对Skov3卵巢癌细胞有抑制其增殖的作用(P＜0.05). 结论:可以将IL-12基因转染至Skov3人卵巢癌细胞中并表达IL-12,脾细胞检测其活性并有相应的细胞因子产生,对Skov3细胞有抑制其增殖作用,从而杀伤卵巢癌细胞,具有抗肿瘤作用.     

转染试剂及幽门螺杆菌对AGS细胞形态的影响

目的:探讨不同的转染试剂对AGS细胞形态的影响,观察这些转染试剂能否像幽门螺杆菌那样引起AGS细胞蜂鸟状改变．方法:用多聚阳离子转染试剂(多聚乙酰亚胺、梭华-Sofast)和改良的脂类转染试剂 (Effectene Transfection Reagent)分别转染AGS 细胞,观察细胞形态的变化．用幽门螺杆菌 26695菌株攻击AGS细胞,观察细胞形态的变化并与经转染试剂作用的细胞进行形态比较．结果:幽门螺杆菌攻击后4 h能引起AGS细胞发生蜂鸟状改变．两种多聚阳离子转染试剂也均能引起AGS细胞发生蜂鸟状改变,与幽门螺杆菌引起的细胞形态改变相似,大部分细胞拉长,并且不受是否转染DNA的影响．此种变化在转染4 h时便出现,与试剂剂量的增加呈正相关．当PEI的剂量为10 μL时,细胞出现凋亡,15μL时凋亡加重．改良的脂类转染试剂对 AGS细胞的形态未产生影响．结论:在研究引起AGS细胞形态改变的因素时,不宜选择多聚阳离子转染试剂;多聚阳离子引起的AGS细胞形态改变与幽门螺杆菌引起的AGS细胞形态改变之间是否存在关系值得进一步研究．