糖尿病心肌病影像诊断研究进展

糖尿病心肌病(diabetic cardiomyopathy,DCM)可导致糖尿病患者心力衰竭甚至死亡,是糖尿病的一种严重的并发症。早期表现为能量代谢、心肌脂肪变性、微循环灌注异常等,逐步发展为心肌细胞重塑、心功能异常和心肌纤维化等。DCM起病隐匿,致病分子机制尚未完全阐明,诊断标准不明确。目前可通过一些影像学检查间接检测心肌细胞可能存在的病理改变,做出诊断,及时干预,并监测病情发展,逆转或延缓病情,改善预后。本文就DCM病程中不同阶段的影像检测方法进行相关综述。     

Myc/his-TDP-43融合蛋白对tau外显子10可变剪接的影响

目的:构建带myc/his标签的pcDNA3.1/TDP-43及其截断体质粒,探讨myc/his-TDP-43融合蛋白对tau外显子10可变剪接的影响。方法:采用PCR法扩增TDP-43基因及其各截断体,将扩增产物和载体pcDNA3.1双酶切后回收,连接载体和目的片段,获得重组质粒。在HEK-293FT细胞中转染pcDNA3.1/TDP-43·myc·his及其截断体质粒,Western印迹法检测相关蛋白的表达。TDP-43或siTDP-43和tau迷你基因SI9/SI10共转,RT-PCR检测TDP-43对tau外显子10可变剪接的影响。结果:成功构建了pcDNA3.1/TDP-43·myc·his全长及各截断体质粒,并在HEK-293FT细胞中检测到其蛋白表达。Myc/his-TDP-43融合蛋白可呈剂量依赖性促进4R-tau的表达(P0.05或0.01)。结论:Myc/his-TDP-43融合蛋白可促进4R-tau的表达,携带myc/his标签不影响TDP-43对tau外显子10可变剪接的促进作用,为后续研究奠定了基础。     

调整阈值的D-二聚体联合Wells评分、修正Geneva评分诊断下肢深静脉血栓形成患者非高危肺血栓栓塞症风险的价值北大核心CSCD

目的评价根据年龄断层调整的D-二聚体阈值水平联合Wells评分、修正Geneva评分对下肢深静脉血栓形成(DVT)患者非高危肺血栓栓塞症(PTE)风险的诊断价值。方法选取2018年8月1日至2021年5月30日在南通大学附属医院住院的下肢DVT患者为研究对象。根据CT肺动脉造影结果,将患者分为单纯下肢DVT组和伴非高危PTE组。分析两组患者调整阈值的D-二聚体阳性比例、Wells评分、修正Geneva评分,绘制受试者工作特征曲线(ROC),以评价诊断伴非高危PTE风险的价值。结果入组475例患者中单纯下肢DVT组346例(72.8%),伴非高危PTE组129例(27.2%)。调整阈值的D-二聚体、Wells评分、修正Geneva评分、调整阈值的D-二聚体+Wells评分、调整阈值的D-二聚体+修正Geneva评分诊断下肢DVT伴非高危PTE的ROC曲线下面积(AUC)分别为0.553(95%CI=0.497~0.610)、0.724(95%CI=0.670~0.779)、0.698(95%CI=0.645~0.750)、0.751(95%CI=0.701~0.802)、0.727(95%CI=0.678~0.777)。Wells评分、修正Geneva评分AUC均大于调整阈值的D-二聚体,调整阈值的D-二聚体+Wells评分AUC大于调整阈值的D-二聚体+修正Geneva评分。结论调整阈值的D-二聚体联合Wells评分对下肢DVT伴非高危PTE的诊断价值较高。     

转化生长因子β信号传导阻断对鼠肝星状细胞培养激活的影响

目的 研究转化生长因子β(TGF-β)信号传导通路的阻断,对鼠肝星状细胞(HSC)培养激活的影响。 方法 利用腺病毒AdT β-ExR的表达产物阻断HSCs中TGF-β信号传导。用酶联免疫吸附试验,Western blot及免疫组织化学等方法检测HSCs中Ⅰ型胶原蛋白、α-平滑肌肌动蛋白(α-SMA)的表达及细胞增殖。 结果 感染AdT β—ExR与感染AdLacZ的HSCs相比,Ⅰ型胶原蛋白的表达量为对照组的42.99％(q=9.100,,P<0.001),α-SMA的表达明显被抑制,而细胞增殖指标5-溴脱氧尿苷的参入量,后者为前者的49.24％(q=7.835,.P<0.001)。 结论 阻断TGF-β信号传导能显著地抑制HSCs的培养激活,但促进HSCs的分裂。通过腺病毒AdT β-ExR的表达产物阻断TGF-β信号传导作为抑制肝纤维化的方法,还需深入一步的研究。     

肝纤维化大鼠血小板衍生生长因子受体β亚单位的表达及其与细胞外基质成分的相关性

目的 研究血小板衍生生长因子(PDGF)受体β亚单位在肝纤维化组织中的动态表达及其与细胞外基质成分的相关性。 方法 将动物分为正常对照组和模型组,用C Cl4复制肝纤维化模型,用免疫组化方法动态检测PDGF受体β亚单位、α～平滑肌抗体(α-SMA)、Ⅰ、Ⅲ型胶原在肝纤维化组织中的表达,将免疫组化结果行图像扫描半定量后进行统计学分析并计算其相关性。 结果 随着肝纤维化程度的加重,PDGF受体β亚单位、α—SMA表达逐渐增加,Ⅰ、Ⅲ型胶原的表达也逐步增加。在2周时PDGF受体β亚单位与Ⅰ、Ⅲ型胶原的相关性不明显,但与α-SMA呈显著相关,其相关系数为0.6 2(P<0.05);在4周时其与Ⅰ、Ⅲ型胶原和α—SMA的相关系数分别为0.74、0.60和0.69(P<0.05);在6周时其与Ⅰ、Ⅲ型胶原和α-SMA的相关系数分别为0.83、0.67和0.81(P<0.05)。 结论 PDGF受体β亚单位在肝纤维化的发生发展中起重要作用,抑制PDGF受体β亚单位的表达可望能减少细胞外基质的合成。     

SYT16 is a prognostic biomarker and correlated with immune infiltrates in glioma: A study based on TCGA data

BackgroundGlioma is the most lethal primary brain tumor. Lower-grade glioma (LGG) is the crucial pathological type of Glioma. Immune-infiltration of the tumor microenvironment positively associated with overall survival in LGG. SYT16 is a gene has not been reported in cancer. We assess the role of SYT16 in LGG, via the publicly available TCGA database.MethodsGene Expression Profiling Interactive Analysis (GEPIA) was used to analyze the expression of SYT16 in LGG. We evaluated the influence of SYT16 on survival of LGG patients by survival module. Then, datasets of LGG were downloaded from TCGA. The correlations between the clinical information and SYT16 expression were analyzed using logistic regression. Univariable survival and Multivariate Cox analysis was used to compare several clinical characteristics with survival. we also explore the correlation between SYT16 and cancer immune infiltrates using CIBERSORT and correlation module of GEPIA. Gene set enrichment analysis (GSEA) was performed using the TCGA dataset. In addition, we use TIMER to explore the collection of SYT16 Expression and Immune Infiltration Level in LGG and to explore cumulative survival in LGG.ResultsThe univariate analysis using logistic regression, indicated that increased SYT16 expression significantly correlated with the tumor grade. Moreover, multivariate analysis revealed that the up-regulated SYT16 expression is an independent prognostic factor for good prognosis. Specifically, SYT16 expression level has significant negative correlations with infiltrating levels of B cell, CD4+ T cells, Macrophages, Neutrophils and DCs in LGG. In addition, GSEA identified ingle organism behavior, gated channel activity, cognition, transporter complex and ligand gated channel activity  in Gene Ontology (GO) were differentially enriched in the high SYT16 expression phenotype pathway. Neuroactive ligand receptor interaction, calcium signaling pathway, long term potentiation, type II diabetes mellitus and long term depression were identified as differentially enriched  pathway in Kyoto Encyclopedia of Genes and Genomes (KEGG).ConclusionSYT16 is a Prognostic Biomarker and Correlated with Immune Infiltrates in LGG.     

Metabolic profiling in human SH-SY5Y neuronal cells exposed to perfluorooctanoic acid (PFOA)

Perfluorooctanoic acid (PFOA) is an abundant per- and polyfluoroalkyl substance (PFAS) detected in both indoor and outdoor environments. While studies suggest exposure concerns for humans, studies investigating PFOA-induced neurotoxicity are lacking. To address this gap, we exposed differentiated human SH-SY5Y cells to PFOA (0.1 μM up to 500 μM) at different time points (4, 24, 48, and 72 h) and measured cell viability, Casp3/7 activity, ATP levels, ATP synthase enzyme activity, mitochondrial membrane potential, reactive oxygen species (ROS), oxygen consumption rates for mitochondrial stress test (XFe24 Flux analyzer), glucose utilization, and global metabolome profiles to assess the potential for PFOA-induced neurotoxicity. Treatment with 10 or 100 μM PFOA did not compromise cell viability nor induce cytotoxicity to SH-SY5Y cells over a 48-hour exposure period. However, >250 μM PFOA compromised cell viability, induced cytotoxicity, and induced caspase 3/7 activity at 48 h. ATP levels were reduced in cells treated with 400 μM PFOA for 24 and 48 h, and with 100 μM PFOA and higher at 72 h. ATP synthase activity was inhibited by 250 μM PFOA but was unchanged by PFOA treatment at 200 μM or less. Conversely, mitochondrial membrane potential was reduced by >10 μM PFOA after 24 h. Total ROS was increased with 100 μM PFOA and higher after 4 h of exposure. Several mitochondria-related endpoints (basal respiration, ATP production, maximum respiration) were negatively affected at 250 μM PFOA at both 24- and 48-hour exposure, but were unaltered at concentrations of 100 μM PFOA or less. One exception was mitochondrial spare capacity, which was reduced by 100 μM PFOA after 24-hour exposure. Similarly, glycolysis, glycolytic capacity, and glycolytic reserve of SH-SY5Y cells were not altered by 10 nor 100 μM PFOA. Nontargeted metabolomics was conducted in cells treated with either 10 or 100 μM PFOA for 48 h, as these two concentrations were not cytotoxic and 28 metabolites differed among treatments. Notable was that 10 μM PFOA had little effect on the SH-SY5Y metabolome, and the metabolic profile was not statistically different from media nor solvent controls. On the other hand, 100 μM PFOA shifted the metabolic signature of the neuronal cells, leading to reduced abundance of ATP-related metabolites (adenine, nicotinamide), neurotransmitter precursors (DL-tryptophan, l-tyrosine), and metabolites that protect mitochondria during oxidative stress (betaine, orotic acid, and l-acetyl carnitine). We hypothesize that this metabolic signature may be associated with the reduced mitochondrial membrane potential observed at lower PFOA concentrations. Metabolic shifts appear to precede compromised cell viability, cytotoxicity, and apoptosis. This study generates mechanistic knowledge regarding PFOA-induced neurotoxicity, focusing on mitochondrial oxidative respiration and the neuronal metabolome.     

穹隆海马伞切割后大鼠海马内RUNX1T1 mRNA和蛋白的表达变化

目的 观察穹隆海马伞切割后不同时间点海马内RUNX1T1 mRNA和蛋白的表达变化及定位。方法 行穹隆海马伞切割术,制备模型大鼠。实验分为正常组、切割3d组、切割7d组、切割14d组、切割21d组、切割28d组。1.提取海马组织总mRNA,用Real-time PCR 检测大鼠海马组织中RUNX1T1mRNA 的表达;2.提取海马组织总蛋白,用Western blotting 检测海马内RUNX1T1蛋白的表达;3.行脑冷冻切片,行RUNX1T1免疫荧光检测和Hoechst标记,观察海马齿状回中RUNX1T1/Hoechst双标阳性细胞。 结果 Real-time PCR检测显示,切割3d组海马组织中RUNX1T1 mRNA的表达明显上调,其余各组差异不明显;Western blotting检测结果显示,正常组海马中有微量RUNX1T1蛋白表达,而切割3d组海马组织中RUNX1T1蛋白表达量明显上升,并达到高峰,7d时仍有较多的表达,此后,14d、21d、28d组中RUNX1T1蛋白表达很快又恢复到正常水平;免疫荧光检测结果表明,正常组海马齿状回中有少量的RUNX1T1/Hoechst阳性细胞,切割3d组海马齿状回中RUNX1T1/Hoechst阳性细胞数量最多,切割7d组逐渐减少,切割14d组、21d组和28d组与正常组差别不明显。结论 穹隆海马伞切割后海马内RUNX1T1 mRNA和蛋白表达在早期呈短暂性上调,并定位于海马齿状回颗粒下层,推测可能与海马神经再生过程中神经干细胞向神经元分化有关。