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简介:AbstractBackground:Macrophages play an important role in renal ischemia reperfusion injury, but the functional changes of macrophages under hypoxia/reoxygenation and the related mechanism are unclear and need to be further clarified.Methods:The effects of hypoxia/reoxygenation on functional characteristics of RAW264.7 macrophages were analyzed through the protein expression detection of pro-inflammatory factors TNF-α and CD80, anti-inflammatory factors ARG-1 and CD206. The functional implications of C-X3-C motif chemokine receptor 1(CX3CR1) down-regulation in hypoxic macrophages were explored using small interfering RNA technology. Significance was assessed by the parametric t-test or nonparametric Mann-Whitney test for two group comparisons, and a one-way ANOVA or the Kruskal-Wallis test for multiple group comparisons.Results:Hypoxia/reoxygenation significantly increased the protein expression of M1-related pro-inflammatory factors TNF-α, CD80 and chemokine C-X3-C motif chemokine ligand 1 (CX3CL1)/CX3CR1 and inhibited the protein expression of M2-related anti-inflammatory factors ARG-1 and CD206 in a time-dependent manner in RAW264.7 cells. However, the silencing of CX3CR1 in RAW264.7 cells using specific CX3CR1-siRNA, significantly attenuated the increase in protein expression of TNF-α (P < 0.05) and CD80 (P < 0.01) and the inhibition of ARG-1 (P < 0.01) and CD206 (P < 0.01) induced by hypoxia/reoxygenation. In addition, we also found that hypoxia/reoxygenation could significantly enhance the migration (2.2-fold, P < 0.01) and adhesion capacity (1.5-fold, P < 0.01) of RAW264.7 macrophages compared with the control group, and CX3CR1-siRNA had an inhibitory role (40% and 20% reduction, respectively). For elucidating the mechanism, we showed that the phosphorylation levels of ERK (P < 0.01) and the p65 subunit of NF-κB (P < 0.01) of the RAW264.7 cells in the hypoxic/reoxygenation group were significantly increased, which could be attenuated by down-regulation of CX3CR1 expression (P < 0.01, both). ERK inhibitors also significantly blocked the effects of hypoxic/reoxygenation on the protein expression of M1-related pro-inflammatory factors TNF-α, CD80 and M2-related anti-inflammatory factors ARG-1 and CD206. Moreover, we found that conditioned medium from polarized M1 macrophages induced by hypoxia/reoxygenation, notably increased the degree of apoptosis of hypoxia/reoxygenation-induced TCMK-1 cells, and promoted the protein expression of pro-apoptotic proteins bax (P < 0.01) and cleaved-caspase 3 (P < 0.01) and inhibited the expression of anti-apoptotic protein bcl-2 (P < 0.01), but silencing CX3CR1 in macrophages had a protective role. Finally, we also found that the secretion of soluble CX3CL1 in RAW264.7 macrophages under hypoxia/reoxygenation was significantly increased.Conclusions:The findings suggest that hypoxia/reoxygenation could promote M1 polarization, cell migration, and adhesion of macrophages, and that polarized macrophages induce further apoptosis of hypoxic renal tubular epithelial cells by regulating of CX3CL1/CX3CR1 signaling pathway.
简介:摘要IgA肾病(IgA nephropathy,IgAN)是儿童及青少年最常见的原发性肾小球肾炎,是慢性肾脏病和终末期肾病的主要原因。目前IgAN发病机制尚未完全清楚,可能与多重免疫打击有关。即半乳糖缺陷型IgA1(galactose-deficient IgA1,Gd-IgA1)形成(第一重打击);抗Gd-IgA1的自身抗体合成(第二重打击); Gd-IgA1与自身抗体结合形成循环免疫复合物(第三重打击);这些含有Gd-IgA1的循环免疫复合物在肾小球系膜中沉积,引发肾脏损害(第四重打击)。Gd-IgA1是IgAN发生发展始发及驱动的关键因素,核心1β1,3-半乳糖基转移酶(core 1,β1,3-galactosyltransferase,C1GALT1)是IgA1 O-糖基化过程中的关键酶,其表达减少和(或)活性下降与Gd-IgA1的产生密切相关。该综述阐述了C1GALT1在IgAN发病、治疗及预后中的作用。
简介:摘要目的探讨肾小球补体C1q及C3c沉积与糖尿病肾病进展的相关性。方法纳入2011年1月至2019年7月在南京医科大学第一附属医院确诊的2型糖尿病肾病患者112例,其中男性83例(74.1%),年龄(51.22±11.12)岁,随访时间19.0(8.5,31.3)个月。根据肾小球C1q和C3c是否沉积分为四组:C1q无沉积C3c无沉积组(n=38)、C1q无沉积C3c沉积组(n=24)、C1q沉积C3c无沉积组(n=14)和C1q沉积C3c沉积组(n=36),检测24 h尿蛋白等临床指标并收集病理资料。采用Cox回归及Kaplan-Meier生存曲线评估肾C1q和C3c沉积对肾脏预后的影响。结果四组间24 h尿蛋白差异有统计学意义[分别为1.84(0.92,3.89),4.19(2.09,6.50)3.30(1.84,6.70),3.64(2.49,7.22)g/24 h,P<0.01],C1q沉积C3c沉积组24 h尿蛋白定量显著高于C1q无沉积C3c无沉积组(P<0.01)。Kaplan-Meier生存曲线结果提示,四组累积生存率差异有统计学意义(Log-rank χ²=8.785,P<0.05),C1q沉积C3c无沉积组累计生存率最低,预后最差。调整后的多因素Cox分析显示,肾小球C1q和C3c共沉积[风险比(HR)2.260,95%可信区间1.329~3.845,P<0.01]、肾小球C1q+C3c+IgM均沉积(HR=4.142,95%可信区间 1.071~16.021,P<0.05)是肾脏预后的独立危险因素。结论肾小球C1q及C3c沉积与糖尿病肾病患者蛋白尿、较差的肾功能和预后不良相关,且肾小球C1q和C3c共沉积是糖尿病肾病进展的独立危险因素。
简介:摘要报道1例抗-c抗体引起的新生儿溶血病患儿。患儿生后1 h出现面部和躯干部皮肤黄染并进行性加重,生后6 h血清总胆红素 168.1 μmol/L,血型O型RhDCcEe,直接抗人球蛋白试验、血清游离抗体试验、红细胞释放抗体试验均阳性;其母血型A型RhDCCee,血清中检出IgG型抗-c抗体,效价1∶4。给予患儿蓝光照射、免疫球蛋白及药物治疗后,痊愈出院。患儿生后1和3个月随访,体格发育及神经行为发育未见异常。
简介:摘要C1q肾病是一种特殊类型的肾小球肾炎,以肾小球系膜补体C1q大量沉积为特征,病理学上常表现为微小病变型肾小球肾炎、局灶节段性肾小球硬化及增生性肾小球肾炎,易发生激素依赖或抵抗,预后相对较差,目前发病机制尚不明确。临床表现差异性较大,糖皮质激素仍是治疗首选药物,免疫抑制剂常用于激素依赖或抵抗的患者,近年来还发现利妥昔单抗对C1q肾病具有较好的疗效。虽然人们对C1q肾病的认识和研究不断深化,但能否将C1q肾病划分为一类独立的疾病及其归类的临床意义还存在较大的争议。该文就C1q肾病的发病机制、病理组织学、临床特点、治疗及预后进行综述,以期对当前C1q肾病的相关研究有更好的理解。
简介:摘要目的探讨德朗热综合征(CdLs)的特点、临床表现及基因突变类型,提高临床医师对该病的认识。方法回顾性分析新乡医学院第一附属医院2019年8月确诊的1例CdLs患儿的临床资料及其基因检测结果。结果患儿,女,2岁8个月,外貌特殊,鼻梁低平,眼距增宽,招风耳,嘴角下斜,腭弓高,小颌畸形,反复癫痫发作,言语及智力发育落后,基因检测结果示SMC1A基因c.2923C>T突变,诊断CdLs 2型。结论CdLs属于较为罕见的遗传代谢病,临床表现常有特殊的面容及体征。SMC1A基因突变在国内仅有1例报道,而SMC1A基因c.2923C>T突变在国内外均未见报道,扩大了SMC1A基因变异谱。
简介:摘要目的探讨胆红素-尿苷二磷酸葡萄糖醛酸酰转移酶(UGT1A1)基因c.1091C>T突变与新生儿高胆红素血症的相关性。方法选取湖南省儿童医院普通足月新生儿病房收治的不明原因高未结合胆红素血症新生儿142例作为观察组,随机选取同期无高胆红素血症新生儿71例作为对照组。收集静脉血,提取基因组DNA,PCR扩增UGT1A1基因启动子、外显子及其附近区域并测序。比较两组患儿及正常人群中c.1091C>T突变的等位基因频率,并根据基因测序结果将观察组进行分组,分析c.1091C>T突变与胆红素水平的关系。结果观察组UGT1A1基因c.1091C>T杂合突变15例,纯合突变2例,等位基因频率为0.0669,对照组未检测到c.1091C>T突变,两组间比较差异有统计学意义(P<0.01)。千人基因组计划数据库中东亚人群该位点的基因携带率为0.0109,与观察组比较差异有统计学意义(P<0.01)。观察组中,c.1091C>T突变组总胆红素峰值与野生型组比较,差异有统计学意义(P<0.05)。结论UGT1A1基因c.1091C>T突变在足月新生儿不明原因高胆红素血症中较为常见,该位点突变与新生儿不明原因高胆红素血症的发生相关。
简介:摘要目的对1个X连锁视网膜劈裂症(XLRS)家系进行基因分析,观察其RS1基因的突变位点。方法回顾性临床研究。一个4代26人的XLRS家系中3例患者及12名家系成员纳入研究。其中,男性8名,女性7名。所有受试者均行眼科常规检查;3例患者中,行光相干断层扫描(OCT)检查2例。抽取所有受试者的外周静脉血,提取全基因组DNA,Panel测序法筛选潜在致病基因。利用软件工具对突变进行保守性分析、致病性分析和蛋白结构预测。并根据美国医学遗传学与基因组学学会(ACMG)指南分析基因突变的致病性。结果先证者,3岁。OCT检查,双眼黄斑区视网膜内核层隆起囊腔样改变,被垂直或斜形桥状组织分割。先证者舅舅,32岁。OCT检查,左眼黄斑区萎缩;右眼黄斑区囊样隆起,被垂直或斜形桥状组织分割。先证者父母及其他家系成员10名眼底检查均未见异常。Panel测序结果显示,先证者(Ⅳ3)及2例患者(Ⅱ1、Ⅲ8)RS1基因第5外显子上存在c.361C>T/p.Q121X半合子突变;其母亲为该基因杂合突变携带者,父亲无变异。该突变基因导致RS1蛋白提前终止,由原来编码224个氨基酸突变为120个氨基酸的截短蛋白。眼底检查正常的10人中,正常者6人;该基因突变携带者4人,均为女性。蛋白序列同源性分析结果显示,该突变位点在12种哺乳动物中均高度保守。RS1蛋白三维结构分析结果显示,突变后的蛋白C端氨基酸序列缺失>50%。ACMG指南分析结果显示,该突变为致病性突变。结论该XLRS家系RS1基因突变位点c.361C>T/p.Q121X为XLRS新的突变位点。
简介:摘要目的总结脊髓性肌萎缩症1c型1例患儿的遗传学特点。方法回顾性分析1例脊髓性肌萎缩症1c型患儿的病例资料,对其进行遗传学分析并文献复习。结果患儿,男,2月龄起病,表现为粗大运动发育落后、肌张力低。多重连接探针扩增技术显示患儿SMN1基因外显子7-8纯合缺失突变,SMN2基因外显子7-8存在重复突变,外显子7/8拷贝数为3/3,其父亲为SMN1基因杂合缺失携带者,SMN2基因8外显子存在纯合突变,外显子7/8拷贝数为2/3,其母亲未发现SMN1基因外显子异常,SMN2基因外显子7/8拷贝数为1/1。结论脊髓性肌萎缩症在早期缺乏特异表现,确诊主要依赖于基因检测,临床医师需要提高警惕,加强对该病的早期认识,改善预后。
简介:摘要目的探究ATP6V1C1在肝细胞癌(HCC)表达情况及功能。方法本研究为基础研究,利用来自癌症基因组图谱数据库基因表达总库(GEO)测序数据,找出HCC差异表达基因(P<0.01,|logFc|≥1),进一步利用来自于癌症基因组图谱(TCGA)在线数据UALCAN、Timer分析ATP6V1C1在HCC不同分期的表达情况及不同表达水平的预后情况,The Human Protein Atlas分析正常肝组织与肿瘤组织免疫组化。TCGA数据库包含女性患者121例,男性患者281例,年龄16~81岁,数据分析后台自行完成。KEGG信号了解ATP6V1C1对相关通路的影响。构建pcDNA3.1(+)-ATP6V1C1载体,购买siRNA干扰片段体外研究ATP6V1C1对肝细胞癌的影响。统计学方法采用独立样本t检验。结果生物信息学分析提示ATP6V1C1在多种癌症中上调(P<0.05),ATP6V1C1的表达水平与HCC的肿瘤分级有关,在肝癌患者中,ATP6V1C1表达上调患者相对下调患者预后更差(P<0.05)。过表达ATP6V1C1后明显促进HCC细胞的侵袭、迁移、增殖及克隆形成能力,同时,干扰ATP6V1C1抑制HCC细胞的侵袭、迁移、增殖及克隆形成能力。结论ATP6V1C1在HCC中表达与预后相关,这个研究为HCC提供了新的可能诊断及治疗靶标。
简介:摘要目的用生物信息学分析筛选骨肉瘤预后相关基因。方法从Gene Expression Omnibus(GEO)下载GSE33382、GSE21257数据集。分析获得差异表达基因(DEGs)。对差异基因进行GO分析,构建蛋白互作网络。Cytoscape进一步筛选关键基因,对关键基因进行生存相关性分析。用临床样本验证关键基因与患者预后情况。两组间差异采用Student’s t检验。结果筛选下调基因43个,上调基因43个,GO分析发现差异基因主要富集在免疫应答、抗原处理和呈递、免疫反应、蛋白质加工和成熟等免疫相关过程。在分子功能中,主要参与细胞内主要组织相容性复合体(MHC)Ⅱ类受体活性、核糖体结构组成、信号和分子转导活性等。在细胞组成成分方面,主要与主要组织相容性复合体蛋白质复合物、裂解空泡、液泡、溶酶体、细胞膜、核糖体等细胞器的功能有关。筛选出10个关键基因:补体C1q A链(CIQA)、补体C1q B链(C1QB)、补体C1q C链(C1QC)、单核细胞分化抗原CD 14(CD14)、单核细胞分化抗原CD 74(CD74)、IgE受体Fc片段(FCER1G)、纤维蛋白原样蛋白2(FGL2)、组织相容性抗原(HLA-DRA)、整合素β2亚基(ITGB2)、酪氨酸激酶结合蛋白(TYROBP)。生存分析发现C1QC与骨肉瘤患者生存呈明显负相关。临床样本证实C1QC与肿瘤Enneking分期、转移显著相关(χ2=4.288和9.809,P<0.05),与患者生存时间呈明显负相关(χ2=5.175,P<0.05)。结论生物信息学分析是有效的筛查方法,C1QC是骨肉瘤预后相关因子。
简介:AbstractBackground:Microglia plays an indispensable role in the pathological process of sleep deprivation (SD). Here, the potential role of microglial CX3C-chemokine receptor 1 (CX3CR1) in modulating the cognition decline during SD was evaluated in terms of microglial neuroinflammation and synaptic pruning. In this study, we aimed to investigat whether the interference in the microglial function by the CX3CR1 knockout affects the CNS’s response to SD.Methods:Middle-aged wild-type (WT) C57BL/6 and CX3CR1-/- mice were either subjected to SD or allowed normal sleep (S) for 8 h to mimic the pathophysiological changes of middle-aged people after staying up all night. After which, behavioral and histological tests were used to explore their different changes.Results:CX3CR1 deficiency prevented SD-induced cognitive impairments, unlike WT groups. Compared with the CX3CR1-/- S group, the CX3CR1-/- SD mice reported a markedly decreased microglia and cellular oncogene fos density in the dentate gyrus (DG), decreased expression of pro-inflammatory cytokines, and decreased microglial phagocytosis-related factors, whereas increased levels of anti-inflammatory cytokines in the hippocampus and a significant increase in the density of spines of the DG were also noted.Conclusions:These findings suggest that CX3CR1 deficiency leads to different cerebral behaviors and responses to SD. The inflammation-attenuating activity and the related modification of synaptic pruning are possible mechanism candidates, which indicate CX3CR1 as a candidate therapeutic target for the prevention of the sleep loss-induced cognitive impairments.
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简介:AbstractBackground:Fourier transform infrared (FTIR) spectroscopy technique has not been used as a diagnostic tool for diabetes in clinical practice. This study was linked to structural changes in hemoglobin (Hb) in type 2 diabetes patients at higher levels of HbA1C using FTIR spectroscopy.Methods:Fifty-three diabetic patients from the Bahawal Victoria Hospital, Bahawalpur, Pakistan were categorized as group A (6% < HbA1C < 7%; n = 25) and group B (HbA1C ≥9%; n = 28). Another group (group N) of twenty blood samples was taken from healthy people from the Islamia University Bahawalpur, Pakistan. Data from all groups were collected from January 1, 2018 to March 31, 2019. The structure of Hb was studied by FTIR spectroscopy and impact of glucose on the fine structure of HbA1C was estimated.Results:Hb secondary structure erythrocyte parameters were altered by changing glucose concentrations. From FTIR spectra of all three groups it was found that Hb structure was slightly altered in group A, but significantly changed in group B (P < 0.05). There was an increase in β-sheet structure and a reduction in α-helix structure at elevated levels of HbA1C (group B) in type 2 diabetes.Conclusion:We suggest that higher level of glycation reflected by increased HbA1C might be a contributing factor to structural changes in Hb in type 2 diabetic patients. FTIR spectroscopy can be a novel technique to find pathogenesis in type 2 diabetes.
简介:摘要目的探讨非衍生化串联质谱法(MS/MS)中十四烯酰基肉碱(C14∶1)/十二烯酰基肉碱(C12∶1)在新生儿筛查极长链酰基辅酶A脱氢酶缺乏症(VLCADD)中的应用价值,并探索筛查VLCADD的最佳指标组合方案。方法回顾性分析2014年1月至2021年12月南京市新生儿疾病筛查中心和苏州市新生儿疾病筛查中心17例MS/MS筛查发现并经酰基辅酶A脱氢酶极长链(ACADVL)基因确诊的VLCADD患儿和423 507名经MS/MS筛查正常的新生儿资料。采用受试者工作特征(ROC)曲线分别确定所有新生儿组、足月新生儿组和正常出生体重新生儿组C14∶1/C12∶1筛查VLCADD的截断值。同时将不同指标单独或联合组成5个结果判读方案,分别为方案1(C14∶1/C12∶1)、方案2(C14∶1)、方案3(C14∶1+C14∶1/C2+C14∶1/C16)、方案4(C14∶1/C12∶1+C14∶1)和方案5(C14∶1/C12∶1+C14∶1+C14∶1/C2+C14∶1/C16),并计算各方案的检出率、假阳性率、阳性预测值,采用χ²检验比较其筛查效率。结果所有新生儿组、足月新生儿组和正常出生体重新生儿组C14∶1/C12∶1的截断值均为2.80。5个判读方案筛查VLCADD的检出率均为17/17。其中方案1假阳性率[26.15‰(11 075/423 524)]最高,阳性预测值[0.15%(17/11 092)]最低。方案4(方案5)假阳性率最低,为[0.02‰(10/423 524)],阳性预测值最高,为[62.96%(17/27)],与方案1、方案2、方案3比较,假阳性率(χ²值分别为302.30、11 191.50和32.06)和阳性预测值(χ²值分别为102.51、3 485.61和13.83)差异均有统计学意义(P均<0.001)。结论C14∶1/C12∶1是新生儿筛查VLCADD有效的辅助判读指标,推荐C14∶1/C12∶1+C14∶1为非衍生化串联质谱法判读VLCADD的最佳筛查指标组合。
简介:摘要目的探讨1例结节性硬化症(tuberous sclerosis complex, TSC)患者可能的遗传学病因。方法应用高通量测序技术、多重连接探针扩增技术和Sanger测序技术对1例疑似结节性硬化症患者行基因变异分析,并在家系其他成员及100名正常对照中进行验证;通过反转录-PCR及Sanger测序技术确定该变异可能导致的mRNA转录变化。结果测序结果显示该家系中先证者发生了TSC2基因的c.2355+1G>C杂合变异,cDNA的序列分析表明该变异导致TSC2基因在第21外显子3′端插入62个碱基序列,这种剪切位点变化导致蛋白翻译提前终止,预测产生截短蛋白。结论TSC2基因c.2355+1G>C变异可能是该患者的致病原因。本研究结果不仅进一步丰富了TSC2基因的变异谱,同时为产前诊断和胚胎植入前遗传学检测的开展提供了理论基础。
简介:摘要目的探讨紫丁香苷对脂多糖(LPS)诱导肌管细胞萎缩的保护作用及其分子机制。方法诱导C2C12成肌细胞分化为肌管细胞后,按随机数字表法分为空白组、模型组、紫丁香苷组。模型组与紫丁香苷组培养基内加入终浓度为200 ng/ml 的LPS进行干预;紫丁香苷组培养基内同时加入10 μmol/L紫丁香苷干预24 h。采用细胞计数(CCK-8)法检测各组细胞活力,Griess试剂检测细胞上清液NO释放量;ELISA法检测上清液TNF-α水平;Western blot法检测NF-κB、过氧化物酶体增殖物激活受体γ1(peroxisome proliferators-activated receptors γ1,PPARγ1)、肌球蛋白重链(myosin heavy chain,MyHC)表达情况。结果与模型组比较,10、100 μmol/L紫丁香苷组细胞增殖率[(101.08±8.92)%、(79.53±5.19)%比(69.07±7.16)%]升高(P<0.01或P<0.05),其中10 μmol/L的紫丁香苷效果更佳。与模型组比较,干预6 h紫丁香苷组NO[(2.92±0.33)μmol/L比(3.57±0.41)μmol/L]水平降低(P<0.01),干预24 h紫丁香苷组细胞TNF-α[(2.73±0.29)pg/ml比(4.15±0.29)pg/ml]水平降低(P<0.01),细胞NF-κB[(0.95±0.24)比(1.16±0.28)]蛋白表达降低(P<0.05),MyHC[(0.79±0.15)比(0.70±0.16)]蛋白表达升高(P<0.05)。结论紫丁香苷可抑制LPS诱发的炎症反应,提高肌管细胞活力,拮抗LPS对肌管细胞的损伤作用。