Examining the combinatorial model of motor neuron survival by expression profiling of trophic factors and their receptors in the embryonic Gallus gallus

During embryogenesis, limb‐innervating lateral motor column (LMC) spinal motor neurons (MN) are generated in excess and subsequently nearly half of them die. Many motor neuron survival factors (MnSFs) have been shown to suppress this default programmed cell death (PCD) program through their receptors (MnSFRs), raising the possibility that they are involved in matching specific MNs with their target muscles. Published observations suggest a combinatorial model of MnSF/Rs function, which assumes that during the PCD phase, MNs are expressing combinations of MnSFRs, whereas the limb muscles innervated by these MNs express cognate combinations of MnSFs. We tested this model by expression profiling of MnSFs and their receptors in the avian lumbosacral spinal cord and limb muscles during the peak PCD period. Our findings highlight the complexity of MnSF/Rs function in the control of LMC motor neuron survival. Developmental Dynamics 239:965–979, 2010. © 2010 Wiley‐Liss, Inc.     

Depletion of CD4+CD25+ regulatory T cells enhances natural killer T cell‐mediated anti‐tumour immunity in a murine mammary breast cancer model

Both invariant natural killer T (NK T) cells and CD4⁺CD25⁺ T regulatory cells (T_regs) regulate the immune system to maintain homeostasis. In a tumour setting, NK T cells activated by α‐galactosylceramide (α‐GalCer) execute anti‐tumour activity by secreting cytokines. By contrast, T_regs intrinsically suppress antigen‐specific immune responses and are often found to be elevated in tumour patients. In this study, we have shown that T_regs regulate NK T cell function negatively in vitro, suggesting a direct interaction between these cell types. In a murine mammary tumour model, we demonstrated that administration of either α‐GalCer or anti‐CD25 antibody alone markedly suppressed tumour formation and pulmonary metastasis, and resulted in an increase in the survival rate up to 44% (from a baseline of 0%). When treatments were combined, depletion of T_regs boosted the anti‐tumour effect of α‐GalCer, and the survival rate jumped to 85%. Our results imply a potential application of combining T_reg cell depletion with α‐GalCer to stimulate NK T cells for cancer therapy.     

Cancer-cell microsurgery using nonlinear optical endomicroscopy

Near-infrared laser-based microsurgery is promising for noninvasive cancer treatment. To make it a safe technique, a therapeutic process should be controllable and energy efficient, which requires the cancer cells to be identifiable and observable. In this work, for the first time we use a miniaturized nonlinear optical endomicroscope to achieve microtreatment of cancer cells labeled with gold nanorods. Due to the high two-photon-excited photoluminescence of gold nanorods, HeLa cells inside a tissue phantom up to 250 μm deep can be imaged by the nonlinear optical endomicroscope. This facilitates microsurgery of selected cancer cells by inducing instant damage through the necrosis process, or by stopping cell proliferation through the apoptosis process. The results indicate that a combination of nonlinear endomicroscopy with gold nanoparticles is potentially viable for minimally invasive cancer treatment.     

犬S100β蛋白的体外克隆表达及抗体制备

目的:制备犬S100β蛋白抗血清,为组织工程和神经损伤及修复研究提供实验基础。方法:提取犬坐骨神经总RNA,采用RT-PCR方法扩增S100β基因,并克隆入pGEM-T载体。经测序验证后,将S100β基因亚克隆入原核表达载体pGEX-4T-1中,IPTG诱导GST-S100β融合蛋白在大肠杆菌BL21(DE3)中表达。GST-S100β融合蛋白经Glutathione Sepharose亲和层析纯化后免疫新西兰大白兔,获得抗血清,经酶联免疫吸附试验(ELISA)法、Western Blot和免疫组化法检测抗体的效价和特异性。结果:测序结果表明,扩增的犬S100β基因序列与GenBank中的序列一致;并得到了较高纯度的GST-S100β融合蛋白和兔源性抗S100β血清,经Western Blot、免疫组织化学及ELISA实验证实所得抗体具有较高的特异性及效价。结论:构建了犬S100β基因的重组原核表达质粒,并获得了高效表达。成功制备了兔抗犬S100β抗血清,并具有较高的效价和特异性,为进一步研究神经损伤与修复机制提供实验基础。     

三叶因子2在大鼠下颌下腺的表达及性差

目的:探讨三叶因子2(TFF2)在大鼠下颌下腺的定位及其在不同性别中的表达差异.方法:对大鼠下颌下腺进行TFF2免疫组织化学显色和RT-PCR检测TFF2mRNA表达.结果:TFF2免疫反应阳性物质主要位于闰管、纹状管和小叶间导管上皮细胞胞质,特别是近管腔面更多,纹状管和小叶间导管管腔内亦有阳性物质分布;颗粒区管的少颗粒暗细胞或无颗粒细胞强阳性;多颗粒细胞弱阳性或阴性.雄性大鼠的下颌下腺表达TFF2mRNA明显多于雌性.结论:TFF2主要分布在下颌下腺导管的上皮细胞,以雄性丰富,且主要以腔分泌方式发挥作用.     

Hemifacial spasm patient with ipsilateral total absence of common carotid artery, vertebral artery and aneurysm of the contralateral internal carotid artery

We present a case of a 45-year-old male patient with left hemifacial spasm for 6 years. Magnetic resonance tomographic angiography confirmed that there were small vessels adjacent to the left facial nerve root entry zone, but the left internal carotid artery (ICA) was absent. Magnetic resonance angiography using three-dimensional time of flight technique showed the absence of the left ICA and vertebral artery, But the presence of a 6-mm aneurysm in the bifurcation of the right internal carotid artery. These abnormal arterial structures were further confirmed by angiogram. Aortic arch angiogram revealed a right-sided aortic arch and the Subclavian steal phenomenon. The left common carotid artery and the left Subclavian artery were absent. Right vertebral angiography showed retrograde filling of the left vertebral artery. The left upper limb was fed by this artery. The aneurysm was successfully clipped. Post-operatively, the hemifacial spasm was weakened. However, the operation did not change the neurovascular positional relationship, thus we advised follow-up visits. Cerebral artery anomalies should be taken into consideration when performing imaging and surgical corrections in patients with hemifacial spasm.     

Overexpression of S100A4 is closely related to the aggressiveness of gastric cancer

Elevated levels of the calcium-binding protein S100A4 cause metastasis of benign rat mammary tumor cells. To investigate whether S100A4 plays an important role in the invasion and metastasis of gastric cancers, we examined the gene mutations in the coding regions and expression patterns of the S100A4 in gastric adenocarcinoma in Korea. Moderate to strong expression of S100A4 was found in 53 (68.8%) of the 77 gastric adenocarcinomas, whilst normal gastric epithelium either failed to stain or showed weak staining. Interestingly, S100A4 expression was more frequently observed in gastric cancer patients with advanced gastric cancer (p=0.039), positive lymph node metastasis (p=0.001), and peritoneal dissemination (p=0.022). No gene mutations were found in the analyzed genomic area in 77 gastric adenocarcinomas and 15 gastric cancer cell lines. We found one single nucleotide polymorphism without an amino acid change, A99G, in two cases. These data suggest that the overexpression of S100A4 may be closely related to the aggressiveness of gastric cancer in Korea.     

盐酸吡硫醇静滴致严重静脉炎10例

目的 了解盐酸吡硫醇注射液引起的药物不良反应。方法 收集本院2002年1-3月间急诊留观室和住院10例,患者均因脑外伤而应用盐酸吡硫醇静滴治疗。结果 10例患者分别对盐酸吡硫醇静滴6天后均出现严重静脉炎。结论 盐酸吡硫醇静滴后产生严重静脉炎较少见,故请临床医生加以重视。     

免疫介导脊髓前、后角损伤的对比

目的:探讨实验性自身免疫性灰质病作为肌萎缩侧索硬化(ALS)研究模型的可行性.方法:通过免疫组织化学方法,观察实验性自身免疫性脊髓灰质病过程中脊髓前、后角神经元损伤的特征.结果:神经丝轻链(NF-L)、细胞周期素依赖蛋白激酶5(CDK5)及其激活因子P35/P25抗体阳性的脊髓前角运动神经元数量与对照组相比均明显减少,而脊髓后角阳性的神经元数量与对照组相比差异无统计学意义.结论:在实验性自身免疫性灰质病发病过程中,存在脊髓前角运动神经元相对易感性,实验性自身免疫性灰质病模型在某种程度上可以作为ALS研究模型.