Infantile-onset spinocerebellar ataxia type 5 associated with a novel SPTBN2 mutation: A case report

BackgroundSpinocerebellar ataxia type 5 (SCA5), a dominant spinocerebellar ataxia is caused by spectrin beta nonerythrocytic 2 gene (SPTBN2) mutation. It typically consists of a slow progressive cerebellar ataxia with an onset principally in adulthood. Here, we report on the first Japanese patient with infantile-onset SCA5 associated with a novel heterozygous SPTBN2 mutation.Case reportThe patient, a 6-year-old girl, developed delayed motor development and unsteady arm movement during infancy. She also showed gaze-evoked nystagmus, saccadic eye pursuit, dysarthria, dysmetria, intention tremor and mild intellectual disability. Brain MRI revealed moderate cerebellar atrophy and mild pontine atrophy. Comprehensive target capture sequencing to identify the causative gene identified a novel missense mutation in SPTBN2 (c.1309C<G, p.R437G), which was thought to be pathogenic.DiscussionTwo patients with infantile-onset SCA5 associated with another novel heterozygous SPTBN2 mutation have recently been reported; these SPTBN2 mutations, which may have a significant impact on protein function, were located in the second spectrin. Our findings indicate that SPTBN2 mutations may be associated with infantile-onset cerebellar ataxia accompanied with global developmental delay.     

β位淀粉样前体蛋白裂解酶2基因敲除斑马鱼动物模型的构建及其初步应用

目的 构建β位淀粉样前体蛋白裂解酶2(β-site APP-cleaving enzyme 2,BACE2)基因敲除斑马鱼动物模型并及其初步探索其应用。方法 运用CRISPR/Cas9技术,在斑马鱼中设计针对靶点的gRNA,使之诱导靶点突变,经测序鉴定得到突变体。结果 BACE2纯合突变体在早期部分无法存活,RT-PCR结果显示BACE2基因在斑马鱼的早期发育中持续表达。结论 BACE2基因敲除的斑马鱼动物模型将为进一步研究BACE2基因在斑马鱼胰腺发育中的作用和针对糖尿病的药物靶点筛选提供有效工具。     

Overview of Transforming Growth Factor β Superfamily Involvement in Glioblastoma Initiation and Progression

Glioblastoma, also known as glioblastoma multiforme (GBM), is the most aggressive of human braintumors and has a stunning progression with a mean survival of one year from the date of diagnosis. High cellproliferation, angiogenesis and/or necrosis are histopathological features of this cancer, which has no efficientcurative therapy. This aggressiveness is associated with particular heterogeneity of the tumor featuring multiplegenetic and epigenetic alterations, but also with implications of aberrant signaling driven by growth factors. Thetransforming growth factor β (TGFβ) superfamily is a large group of structurally related proteins including TGFβsubfamily members Nodal, Activin, Lefty, bone morphogenetic proteins (BMPs) and growth and differentiationfactor (GDF). It is involved in important biological functions including morphogenesis, embryonic development,adult stem cell differentiation, immune regulation, wound healing and inflammation. This superfamily is alsoconsidered to impact on cancer biology including that of GBM, with various effects depending on the member. TheTGFβ subfamily, in particular, is overexpressed in some GBM types which exhibit aggressive phenotypes. Thissubfamily impairs anti-cancer immune responses in several ways, including immune cells inhibition and majorhistocompatibility (MHC) class I and II abolishment. It promotes GBM angiogenesis by inducing angiogenicfactors such as vascular endothelial growth factor (VEGF), plasminogen activator inhibitor (PAI-I) and insulinlikegrowth factor-binding protein 7 (IGFBP7), contributes to GBM progression by inducing metalloproteinases(MMPs), “pro-neoplastic” integrins (αvβ3, α5β1) and GBM initiating cells (GICs) as well as inducing a GBMmesenchymal phenotype. Equally, Nodal promotes GICs, induces cancer metabolic switch and supports GBMcell proliferation, but is negatively regulated by Lefty. Activin promotes GBM cell proliferation while GDFyields immune-escape function. On the other hand, BMPs target GICS and induce differentiation and sensitivityto chemotherapy. This multifaceted involvement of this superfamily in GBM necessitates different strategiesin anti-cancer therapy. While suppressing the TGFβ subfamily yields advantageous results, enhancing BMPsproduction is also beneficial.     

β-arrestin1激活JNK信号通路促进慢性髓细胞白血病细胞K562增殖

目的以CML K562细胞为研究对象,探索β-arrestin1促进CML细胞增殖的相关信号通路。方法以β-arrestin1慢病毒载
体感染CML K562细胞,形成稳定的K562-siβ1和K562-β1细胞,及非特异性siRNA对照K562-Ctrl细胞。以此为研究对象,利
用细胞计数与CCK-8实验检测细胞增殖能力;Western blot检测蛋白表达;免疫共沉淀（Co-IP）实验检测蛋白间的相互作用。结
果细胞计数与CCK-8 实验结果显示K562-β1 细胞增殖与细胞存活率能力显著高于K562-Ctrl,而K562-siβ1 显著低于
K562-Ctrl。Western blot结果表明β-arrestin1特异性增强磷酸化JNK表达,JNK抑制剂SP600125能抑制p-JNK表达和K562细
胞增殖;免疫共沉淀实验表明β-arrestin1 能与Src 结合。结论CML K562 细胞中β-arrestin1 与Src 结合,促进JNK信号通路激
活,从而促进细胞增殖。
     

小剂量利多卡因复合氯胺酮对老年胃肠道肿瘤患者术后早期认知功能的影响

目的观察静脉注射小剂量利多卡因和氯胺酮进行麻醉干预对老年胃肠道肿瘤患者术后早期认知功能的影响。方法选
择拟行择期手术的老年胃肠道肿瘤患者60例,年龄63~82岁,ASA分级Ⅰ~Ⅲ级。采用随机数字表法,将患者分为干预组（Ⅰ
组）和对照组（Ⅱ组）,每组30例。静脉诱导气管插管后,I组静注利多卡因和氯胺酮各0.5 mg/kg,继以利多卡因0.5 mg/（kg·h）持
续输注直至术毕;Ⅱ组静注等容量的生理盐水。分别在术前3 d和术后2 d对患者进行神经心理学测试,计算术后成绩与术前基
础值的差值（X值）,记录术后认知功能障碍（POCD）的发生情况。分别于麻醉诱导前（T0）、术毕时（T1）、术后1天（T2）及术后2天
（T3）抽取外周静脉血,ELISA法分别检测S-100β蛋白、神经元特异性烯醇化酶（NSE）及IL-6浓度。结果X值比较,与Ⅱ组比
较,Ⅰ组数字符号测试、累加测试及循迹连线测试A均显著降低（P<0.05）。Ⅰ组和Ⅱ组POCD发生率分别为6.7%和33.3%,Ⅰ
组显著低于Ⅱ组（P<0.05）。T1时点,Ⅰ组S-100β蛋白、NSE及IL-6浓度均显著低于Ⅱ组（P<0.05）;T2时点,Ⅰ组NSE及IL-6浓度
均显著低于Ⅱ组（P<0.05）;T3时点,Ⅰ组IL-6浓度显著低于Ⅱ组（P<0.05）。结论术中静注小剂量利多卡因和氯胺酮进行麻醉
干预可以降低老年胃肠道肿瘤患者术后早期认知功能障碍的发生率,可能与其降低血清S-100β蛋白、NSE及IL-6浓度有关。
     

TGF-β在肿瘤微环境中多重作用的研究进展

转化生长因子β(Transforming growth factor-β,TGF-β)通路在调节细胞生长、分化和迁移中的显著作用决定了该信号通路在癌细胞的进展过程中也具有至关重要的作用。TGF-β通过经典和非经典的这两类信号转导途径来改变细胞的表型,发挥着肿瘤抑制或肿瘤促进的作用。TGF-β通过诱导上皮-间充质细胞转化(Epithelial-mesenchymal transition,EMT)来驱动肿瘤细胞进程,同时在这个过程中上皮细胞获得新的间充质的表型,使肿瘤细胞的转移能力和侵袭能力增强。这证明了TGF-β通路是促进肿瘤发展的因素。然而也有相反的证据表明,癌细胞中TGF-β信号转导途径的缺失与阻断也可以导致肿瘤细胞转移能力的增强。这篇综述回顾TGF-β对肿瘤微环境组成部分的调节和这种调节如何引起肿瘤进展作相关研究。     

GSK3β对骨肉瘤细胞增殖的影响

目的 研究糖原合成激酶3β(GSK3β)在骨肉瘤细胞增殖中的作用及其分子机制。方法 通过Western blot法和NIRKA法,检测正常成骨细胞hFO及骨肉瘤细胞中GSK3β的表达和活性;使用两种GSK3β小分子抑制剂处理骨肉瘤细胞,检测GSK3β活性受抑制下骨肉瘤细胞增殖率和凋亡率;通过GSK3β siRNA降低内源性GSK3β表达,Western blot法检测p27及其下游cyclinD1-CDK-Rb通路因子的蛋白表达及磷酸化;建立骨肉瘤细胞荷瘤小鼠模型,分别给予DMSO作为对照,以及GSK3抑制剂SB216763、AR-A014418,每周3次腹腔给药,观察肿瘤生长和裸鼠的体重变化情况。结果 与正常人成骨细胞相比,骨肉瘤细胞中存在GSK3超表达及活性调节异常;内源性GSK3活性受抑制后抑制骨肉瘤细胞增殖,诱导细胞的凋亡;降低内源性GSK3活性,肿瘤体积与对照组比较,SB216763,AR-A014418组肿瘤增长受到抑制,有统计学意义(P＜0.05);降低骨肉瘤MG-63细胞内源性GSK3蛋白表达,使p27蛋白表达水平明显上调,Rb及其在S780、795部位的磷酸化水平下降,CDK2、4、6蛋白水平降低,cyclinD1的表达上调但其磷酸化水平没有影响。结论 GSK3β通过对p27及cyclinD1-CDK-Rb径路的调节促进骨肉瘤细胞的增殖,可能为骨肉瘤的临床治疗开辟一个潜在的靶点。     

hβ-arrestin2基因重组质粒构建及蛋白表达和定位

目的:构建pEGFP-C1-β-arrestin2融合蛋白表达载体,并证实融合蛋白在细胞内的表达及定位。方法:以pGEX-5X-1-β-arrestin2为模板,PCR法扩增hβ-arrestin2编码序列,亚克隆至pEGFP-C1中,pEGFP-C1-β-arrestin2经双酶切初步鉴定再送测序公司检测正确后瞬时转染人胚肾细胞HEK293,Western blot检测融合蛋白表达。激光扫描共聚焦显微镜观察融合蛋白在HEK293细胞中的分布。结果:hβ-arrestin2编码序列被成功克隆至pEGFP-C1中,Western blot检测到融合蛋白表达,分子量约为77kDa,pEGFP-C1-β-arrestin2定位于HEK293细胞的细胞质中。结论:成功构建hβ-arrestin2基因真核表达载体,证实了融合蛋白定位在HEK293的细胞质中。     

伴ETO或CBFβ阳性的急性髓细胞白血病患者c-kit基因突变的检测及意义

目的:探讨伴AML1-ETO或CBFβ-MYH11阳性的急性髓细胞白血病（acute myeloid leukemia,AML）患者c-kit基因突变情况及其临床意义。方法:采用PCR结合测序的方法,对38例伴AML1-ETO或CBFβ-MYH11阳性的急性髓细胞白血病患者骨髓中c-kit外显子17的突变情况进行检测,并与患者的临床资料、实验室特征及预后的相关性进行综合分析。结果:38例患者中12例检测到c-kit外显子17突变,突变率为31.6％,其中7例为D816V突变,5例为N822K突变,两种突变类型患者的临床资料无显著差异（P>0.05）,突变组骨髓原始细胞比例明显高于野生组（P<0.05）,突变组与野生组患者的年龄、性别、WBC、RBC、PLT和HGB比例无明显差异（P>0.05）。c-kit突变组和野生组化疗后完全缓解率和复发率无统计学差异（P>0.05）,但c-kit突变组的死亡率显著高于野生组（P<0.05）。结论:AML1-ETO或CBFβ阳性的AML患者c-kit D816V和N822K突变常见,突变患者预后差,骨髓原始细胞比例较高,c-kit基因突变检测对患者的治疗和预后有重要意义。     

p28、MMP2、β-catenin在胃癌及淋巴结转移癌中的表达

目的 :探讨胃癌及淋巴结转移癌组织中p28、基质金属蛋白酶2(matrix metalloproteinase 2,MMP2)和β-catenin蛋白表达的差异,并结合肿瘤的病理学特征进行分析。方法:收集123例胃腺癌组织(包括41例不伴淋巴结转移的胃癌组织,82例伴淋巴结转移的胃癌组织及相应的淋巴结转移癌组织)构建组织芯片,采用免疫组织化学方法检测其p28、MMP2和β-catenin蛋白的表达情况。结果:1伴淋巴结转移的胃癌组织中的p28和MMP2阳性表达均明显高于无淋巴结转移的胃癌组织(P<0.05)。2伴淋巴结转移的胃癌病例中,p28与MMP2、p28与β-catenin在原发癌组织及淋巴结转移癌组织中的共表达均呈明显正相关(P0.05)。结论:p28和MMP2高表达的胃癌易发生淋巴结转移;p28与MMP2、p28与β-catenin都具协同性,可能共同参与了胃癌的发生和发展。