Characterization of neuroblastoma bone invasion/metastasis in established bone metastatic model of SY5Y and KCNR cell lines

Objects

To determine the mechanism of neuroblastoma (NB) bone invasion/metastasis, it is necessary to investigate the bone invasion/metastasis-related factors in the bone invasion/metastasis process. Some evidence has suggested that various proteins were involved in bone osteolytic response. The invasion/metastasis property and gene expression of NB, however, are still unknown.

Methods

Single-cell suspensions of SY5Y and KCNR cells were injected directly into the femur of nude mice. Radiological and histological analyses, immunohistochemistry analyses, and western blot assay were performed to characterize bone metastasis mechanism in these bone metastasis models.

Results

SY5Y and KCNR NB cells result in osteolytic responses in bone metastasis model. Osteoprotegerin (OPG), receptor activator of NF-kappaB ligand (RANKL), parathyroid hormone-related peptide (PTHrP), endothelin 1 (ET-1), and CXCR4 were examined and compared among in vitro, in vivo, and normal bone, respectively. PTHrP, OPG, RANKL, and ET-1 except CXCR4 in SY5Y and KCNR NB cells xenografts were strikingly upregulated compared with normal bone and NB cells. However, significantly stronger expression of PTHrP and RANKL was presented than ET-1 and OPG; furthermore, the ratios of expression of PTHrP, RANKL to OPG, and ET-1 were also markedly increased in vivo versus in vitro.

Conclusions

Our study provided evidence that NB cell may enhance bone invasion through PTHrP, OPG, RANKL, and ET-1, especially PTHrP and RANKL which may display stronger effects. CXCR4 appeared not participating in bone invasion, but in tumor growth, and homing to bone. Targeting PTHrP, OPG, ET-1, and RANKL may provide a new insight and method for patient therapy by inhibiting NB bone metastasis and invasiveness.     

Amyloid plaque pathogenesis in 5XFAD mouse spinal cord: retrograde transneuronal modulation after peripheral nerve injury

The spinal cord is composed of distinct neuronal groups with well-defined anatomic connections. In some transgenic (Tg) models of Alzheimer’s disease (AD), amyloid plaques develop in this structure, although the underlying cellular mechanism remains elusive. We attempted to explore the origin, evolution, and modulation of spinal β-amyloid (Aβ) deposition using Tg mice harboring five familiar AD-related mutations (5XFAD) as an experiential model. Dystrophic neuritic elements with enhanced β-secretase-1 (BACE1) immunoreactivity (IR) appeared as early as 2 months of age, and increased with age up to 12 months examined in this study, mostly over the ventral horn (VH). Extracellular Aβ IR emerged and developed during this same period, site-specifically co-existing with BACE1-labeled neurites often in the vicinity of large VH neurons that expressed the mutant human APP. The BACE1-labeled neurites almost invariably colocalized with β-amyloid precursor protein (APP) and synaptophysin, and frequently with the vesicular glutamate transporter-1 (VGLUT). Reduced IR for the neuronal-specific nuclear antigen (NeuN) occurred in the VH by 12 months of age. In 8-month-old animals surviving 6 months after a unilateral sciatic nerve transection, there were significant increases of Aβ, BACE1, and VGLUT IR in the VN of the ipsilateral relative to contralateral lumbar spinal segments. These results suggest that extracellular Aβ deposition in 5XFAD mouse spinal cord relates to a progressive and amyloidogenic synaptic pathology largely involving presynaptic axon terminals from projection neurons in the brain. Spinal neuritic plaque formation is enhanced after peripheral axotomy, suggesting a retrograde transneuronal modulation on pathogenesis.     

Increased dopamine receptor signaling and dopamine receptor-G protein coupling in denervated striatum

Chronic interruption of the nigrostriatal dopaminergic pathway leads to sensitized dopaminergic responses in striatum. We attempted to explore the mechanism(s) underlying this dopaminergic supersensitivity by assessing dopamine receptor signaling and receptor-G protein coupling in unilateral 6-hydroxydopamine-lesioned rats. Dopamine-stimulated adenylyl cyclase activity as well as dopamine-activated guanosine 5'-O-(3-[(35)S]thiotriphosphate) ([(35)S]GTPgammaS) binding and [(3)H]palmitate incorporation by Galpha proteins were enhanced in tissues obtained from denervated striata without apparent changes in Galpha protein levels. Moreover, high-affinity binding sites of the D(1) dopamine receptor increased in lesioned compared with control striata without altering the expression level of the receptor. These denervation-mediated changes appear to correlate with the increase in D(1) dopamine receptor binding sites that co-immunoprecipitated with Galphas(olf)/q(11) proteins. In contrast, the total number of D(2) receptor binding sites was increased, yielding an increase in absolute number of high-affinity sites without significant changes in the proportion of high-affinity sites. Stimulation of the D(2) dopamine receptor enhanced coupling to Galphai protein; this was increased in the striata lesioned. The results provide an important molecular mechanism by which dopamine receptor-regulated signaling is enhanced following denervation of dopaminergic input to striatum. Although D(1) dopamine receptor supersensitivity appears to be mediated by enhanced coupling of the receptor to its G proteins, sensitization in the D(2) dopamine receptor system is mediated by increased D(2) receptor density and enhanced D(2) receptor-Gi protein coupling.     

FOLFOX与TCF方案治疗晚期胃癌的临床观察

目的观察比较FOLFOX方案与TCF方案治疗晚期胃癌的疗效、毒副反应。方法85例晚期胃癌随机分为FOLFOX组与TCF组。两组均以28d为1周期,重复3个周期。近期疗效及毒副反应按WHO标准进行评价。结果FOLFOX组患者总有效率、初治有效率、Karnofsky评分改善率、中位肿瘤进展时间、中位生存期分别是48．84％、56．67％、72．09％、6．5个月、11．5个月,TCF组则分别是30．95％、35．48％、38．09％,5．5个月、9．7个月,两组有效率与中位生存期比较差异有统计学意义（P〈0．05）。两组主要毒副反应为骨髓抑制、胃肠道反应、神经毒性。结论FOLFOX方案治疗晚期胃癌安全、有效,优于TCF方案。     

Long non-coding RNA ncRuPAR regulates gastric cancer cell proliferation and apoptosis via phosphoinositide 3-kinase/protein kinase B signaling

Objective: To determine the effect and mechanism of the long non-coding RNA (lncRNA) ncRuPAR (non-protein coding RNA, upstream of coagulation factor II thrombin receptor [F2R]/protease-activated receptor-1 [PAR-1]) in human gastric cancer.Methods: HGC-27-ncRuPAR overexpression and MGC-803-ncRuPAR-RNAi knockdown gastric cancer cell lines were established. We assessed the effect of ncRuPAR on cell proliferation, apoptosis, migration, and invasion using Cell Counting Kit 8, flow cytometry, scratch and transwell assays, respectively. Differentially expressed genes in HGC-27-ncRuPAR overexpression and HGC-27-empty vector cell lines were identified using Affymetrix GeneChip microarray analysis. Ingenuity Pathway Analysis (IPA) of the microarray results was subsequently conducted to identify ncRuPAR-enriched pathways, followed by validation using real time-quantitative PCR (RT-qPCR). As one of the top enriched pathways, phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway was further examined by western blotting to determine its role in ncRuPAR-mediated regulation of gastric cancer pathogenesis.Results: ncRuPAR inhibited human gastric cancer cell proliferation and induced G1/S phase arrest and apoptosis, but did not affect migration or invasion in vitro. Overexpression of ncRuPAR in vitro was found to inhibit its known target PAR-1, as well as PI3K/Akt signaling. The downstream targets of PI3K/Akt, cyclin D1 was downregulated, but there was no change in expression level of B-cell lymphoma 2 (Bcl-2).Conclusions: We showed that lncRNA-ncRuPAR could inhibit tumor cell proliferation and promote apoptosis of human gastric cancer cells, potentially by inhibiting PAR-1, PI3K/Akt signaling, and cyclin D1. The results suggest a potential role for lncRNAs as key regulatory hubs in GC progression.     

Epigenetic regulation of B cells and its role in autoimmune pathogenesis

B cells play a pivotal role in the pathogenesis of autoimmune diseases. Although previous studies have shown many genetic polymorphisms associated with B-cell activation in patients with various autoimmune disorders, progress in epigenetic research has revealed new mechanisms leading to B-cell hyperactivation. Epigenetic mechanisms, including those involving histone modifications, DNA methylation, and noncoding RNAs, regulate B-cell responses, and their dysregulation can contribute to the pathogenesis of autoimmune diseases. Patients with autoimmune diseases show epigenetic alterations that lead to the initiation and perpetuation of autoimmune inflammation. Moreover, many clinical and animal model studies have shown the promising potential of epigenetic therapies for patients. In this review, we present an up-to-date overview of epigenetic mechanisms with a focus on their roles in regulating functional B-cell subsets. Furthermore, we discuss epigenetic dysregulation in B cells and highlight its contribution to the development of autoimmune diseases. Based on clinical and preclinical evidence, we discuss novel epigenetic biomarkers and therapies for patients with autoimmune disorders.     

A Prognostic Model Generated from an Apparent Diffusion Coefficient Ratio Reliably Predicts the Outcomes of Oral Tongue Squamous Cell Carcinoma

This study aimed to develop an apparent diffusion coefficient (ADC) ratio-based prognostic model to predict the recurrence and disease-free survival (DFS) of oral tongue squamous cell carcinoma (OTSCC). A total of 188 patients with cT1-2 oral tongue squamous cell carcinoma were enrolled retrospectively. Clinical and laboratory data were extracted from medical records. The ADC values were measured at the regions of interest of the tumor and non-tumor tissues of the MRI images, and the ADC ratio was used for comparison between the patient with recurrence (n = 83 case, 44%) and patients without recurrence (n = 105 cases, 56%). Cox proportional hazards models were generated to analyze the risk factors of cancer recurrence. A nomogram was developed based on significant risk factors to predict 1-, 5- and 10-year DFS. The receiver operator characteristic (ROC) curves of predictors in the multivariable Cox proportional hazards prognostic model were generated to predict the recurrence and DFS. The integrated areas under the ROC curve were calculated to evaluate discrimination of the models. The ADC ratio, tumor thickness and lymph node ratio were reliable predictors in the final prognostic model. The final model had a 71.1% sensitivity and an 81.0% specificity. ADC ratio was the strongest predictor of cancer recurrence in prognostic performance. Discrimination and calibration statistics were satisfactory with C-index above 0.7 for both model development and internal validation. The calibration curve showed that the 5- and 10-year DFS predicted by the nomogram agreed with actual observations.     

紫外线消毒运行成本及灼伤事件原因分析

目的 了解紫外线消毒的运行成本和紫外线灼伤事件的原因,为进一步高效安全运行紫外线消毒提供参考依据。方法 收集2014年1-12月温州医科大学附属第一医院所有紫外线消毒区域(包括治疗室、处置室、输液室、血透室、分娩室、细胞分离室等)紫外线消毒的运行情况和紫外线灼伤事件发生情况,分析紫外线消毒的流动成本和固定成本以及紫外线灼伤事件发生的主要原因。结果 温州医科大学附属第一医院2014年紫外线消毒运行成本中的流动成本约为1 653 715元,固定成本约为88 872元,流动成本远大于固定成本;流动成本中,紫外线强度检测人工成本约为106 378元,紫外线每日消毒操作人工成本约为1 534 910元,紫外线灯管额外安装与维修费约为10 000元,电费约为2 427元;固定成本中,紫外线设备成本约为78 292元,紫外线设备安装成本约为10 580元;2014年共发生紫外线灼伤事件2起,主要原因为紫外线灯误开与照明用灯同时使用未发现和新护工的意外操作。结论 紫外线消毒运行中人力成本支出较高,应进行量化精准监控,优化管理流程,控制成本,并杜绝人为暴露灼伤,确保紫外线消毒的有效性和安全性。