狼疮性肾炎患者肾组织中胎源性微嵌合体的表达及其临床意义

目的：探讨胎源性微嵌合体（FMc）与狼疮性肾炎（LN）的相关性。方法：收集生育过男胎的24例LN和24例肾小球轻微病变（GML）的女性患者肾活检组织,采用荧光原位杂交技术（FISH）检测细胞中的Y染色体,并收集患者孕育史、临床和实验室资料进行分析。结果：在9例LN和3例GML的肾组织中发现Y染色体阳性嵌合细胞,两组发生率的差异有统计学意义（P<0.05）,但在LN组,嵌合细胞的出现与蛋白尿、血清肌酐、抗双链DNA抗体、抗核抗体、补体C3、C4水平及SLEDAI评分等之间均无相关（P>0.05）。结论：FMc在女性LN患者肾组织中的发生频率高于对照组GML患者,但FMc的存在很可能与LN的发病、疾病活动性和进展性无关。     

肝细胞生长因子和基质金属蛋白酶-9在过敏性紫癜中表达的意义

目的:探讨肝细胞生长因子(HGF)和基质金属蛋白酶-9(MMP-9)在儿童过敏性紫癜(HSP)中表达的意义,探寻肾脏损害预测的敏感指标。方法:选取HSP与过敏性紫癜肾炎(HSPN)患儿各30例,并选取同期体检的健康儿童30名作为对照组,采用酶联免疫吸附试验法(ELISA)检测3组血清和尿液中的HGF及血清MMP-9含量,并采用比浊法检测24 h尿蛋白,分析HGF、MMP-9与24 h尿蛋白的相关性。结果:HSP组、HSPN组血清、尿液HGF,血清MMP-9及尿蛋白定量均高于对照组(P0.05);HSPN组中Ⅱ~Ⅳ期患者血清HGF及Ⅰ~Ⅳ期尿液HGF、Ⅲ~Ⅳ期MMP-9及Ⅰ~Ⅳ期尿蛋白定量均高于HSP组(P0.05);血清及尿液HGF与血清MMP-9无明显相关性(r=0.014,0.027,P0.05);血清HGF与尿蛋白定量无明显相关性(r=0.032,P0.05);尿液HGF、血清MMP-9与尿蛋白定量均呈正相关(r=0.412、0.302,P0.05)。结论:尿液HGF、MMP-9均参与HSP、HSPN病情的发展,尿液HGF、MMP-9水平的监测有助于评估HSPN的病情及预后,但HGF与MMP-9之间无明显相关性。     

同种异体淋巴细胞致狼疮肾炎小鼠模型的研究

目的　制备狼疮肾炎小鼠模型。方法　采用昆明小鼠同种异体脾淋巴细胞经Lectin刺激后输入小鼠腹腔皮下的方法。结果　成功地制备了系膜增生型狼疮肾炎小鼠模型 ,通过光镜、免疫荧光、电镜观察其病理形态、并检测了血清抗ds -DNA抗体、补体以及尿蛋白 ,证实了该模型的可靠性。结论　制备的系膜增生型狼疮肾炎小鼠模型有实用价值     

Specific recognition of a DNA immunogen by its elicited antibody

DNA recognition by antibodies is a key feature of autoimmune diseases, yet model systems with structural information are very limited. The monoclonal antibody ED-10 recognizes one of the strands of the DNA duplex used in the immunogenic complex. Modifications of the 5' end decrease the binding affinity and short oligonucleotides retain high binding affinity. We determined crystal structures for the Fab bound to a 6-mer oligonucleotide containing the specific sequence that raised the antibody and compared it with the unliganded Fab. Only the first two bases from the 5' end (dTdC) display electron density and we observe four key hydrogen bonds at the interface. The thymine ring is stacked between TrpH50 and TrpH95, and the cytosine ring is packed against TyrL32. Upon DNA binding, TyrH97 and TrpH95 rearrange to allow subnanomolar binding affinity, five orders of magnitude higher than other reported complexes, possibly because of having gone through affinity maturation. This structure represents the first bona fide antibody DNA immunogen complex described in atomic detail.     

Knockdown of TRIM27 expression suppresses the dysfunction of mesangial cells in lupus nephritis by FoxO1 pathway

TRIM27 (tripartite motif-containing 27) is a member of the TRIM (tripartite motif) protein family and participates in a variety of biological processes. Some research has reported that TRIM27 was highly expressed in certain kinds of carcinoma cells and tissues and played an important role in the proliferation of carcinoma cells. However, whether TRIM27 takes part in the progression of lupus nephritis (LN) especially in cells proliferation remains unclear. Our study revealed that the overexpression of TRIM27 was observed in the kidneys of patients with LN, lupus mice and mesangial cells exposed to LN plasma which correlated with the proliferation of mesangial cells and ECM (extracellular matrix) deposition. Downregulation of TRIM27 expression suppressed the proliferation of mesangial cells and ECM accumulation in MRL/lpr mice and cultured human mesangial cells (HMCs) by regulating the FoxO1 pathway. Furthermore, the overexpression of FoxO1 remarkably decreased HMCs proliferation level and ECM accumulation in LN plasma-treated HMCs. In addition, the protein kinase B (Akt) signal pathway inhibitor LY294002 significantly reduced the expression of TRIM27 and inhibited the dysfunction of mesangial cells. These above data suggested that TRIM27 mediated abnormal mesangial cell proliferation in kidney of lupus and might be the potential target for treating mesangial cell proliferation of lupus nephritis.     

Comparative genomic analysis of high-altitude adaptation for Mongolia Mastiff,Tibetan Mastiff,and Canis Lupus

Tibetan Mastiff has adapted to the extreme environment of the Qinghai-Tibetan Plateau. Yet, the underlying mechanisms of its high-altitude-adaptation and origin remains elusive. Here, we generated the draft genomes of Mongolia Mastiff, Tibetan Mastiff, and Canis Lupus. The phylogenetic tree uncovered that Tibetan Mastiff and Mongolia Mastiff were derived from Canis Lupus species. The comparative genomic analyses identified that the expansion of gene families related to DNA repair and damage response, and contraction related to ATPase activity revealed the genetic adaptations of Tibetan Mastiff and Canis Lupus to high altitude. In addition, the Tibetan Mastiff and Canis Lupus had signals of positive selection for genes involved in fatty-acid α/β- oxidation for highland adaptation. Notably, the positively selected TERT of Tibetan Mastiff should be an adaptive trait for correcting DNA damage. These findings suggested that the Tibetan Mastiff and Canis Lupus evolves basic strategies for adaptation to high altitude.     

A quantitative lateral flow assay to detect complement activation in blood

Complement is a major effector arm of the innate immune system that responds rapidly to pathogens or altered self. The central protein of the system, C3, participates in an amplification loop that can lead to rapid complement deposition on a target and, if excessive, can result in host tissue damage. Currently, complement activation is routinely monitored by assessing total C3 levels, which is an indirect and relatively insensitive method. An alternative approach would be to measure downstream C3 activation products such as C3a and iC3b. However, in vitro activation can produce falsely elevated levels of these biomarkers. To circumvent this issue, a lateral flow immunoassay system was developed that measures iC3b in whole blood, plasma, and serum and avoids in vitro activation by minimizing sample handling. This assay system returns results within 15 min and specifically measures iC3b while having minimal cross-reactivity to other C3 split products. While evaluating the potential of this assay, it was observed that circulating iC3b levels can distinguish healthy individuals from those with complement activation-associated diseases. This tool is engineered to provide an improved method to assess complement activation at point of care and could facilitate studies to monitor disease progression in a variety of inflammatory conditions.     

Serum protein oxidation and apolipoprotein CIII levels in people with systemic lupus erythematosus with and without nephritis

Increased oxidative stress is a hallmark of the autoimmune disease systemic lupus erythematosus (SLE). This study compares serum protein oxidation levels in SLE patients without and with renal involvement (lupus nephritis); the latter have a significantly poorer prognosis. Similar increases in protein carbonyls and decreases in protein thiols were observed in both SLE groups compared to controls. Protein carbonyl distribution, determined by Western blotting of 2D gels, was similar in both SLE groups, suggesting factors other than oxidation also play a role in SLE complications. 2D electrophoresis examined the serum proteome further. Six proteins were significantly decreased in non-renal SLE patients compared to controls; five were identified by mass spectrometry, including one isoform of pro-atherogenic apoCIII. Total apoCIII levels (assessed by ELISA) in lupus nephritis patients were significantly elevated compared to controls or non-renal SLE patients. Thus, levels of oxidized proteins and apoCIII may be useful biomarkers in SLE studies.     

狼疮肾炎患者外周血单个核细胞钙调神经磷酸酶活性的检测及CD40L表达变化

目的:检测活动性狼疮肾炎(LN)患者外周血单个核细胞(PBMC)钙调神经磷酸酶(calcineurin,CaN)活性及其与PBMC CD40L表达的关系.方法:体外培养活动性LN患者PBMC,应用发色底物法检测胞浆CaN活性,流式细胞仪检测细胞CD40L的表达.结果:①在单纯培养情况下,正常对照组和LN组PBMC均出现一定量CaN活化,活动性LN组显著高于正常对照组(46.08±5.58 nrmmol/mg pro vs 8.81±3.61nmol/mg pro,P＜0.01);在PMA Ionomycin刺激下,各组CaN活性均升高,活动性LN组CaN活性明显高于正常对照组(69.34±12.59 nmol/mg provs 37.12±11.57 hmol/mg pro,P＜0.01);②LN患者PBMC在单纯培养和PMA Ionomycin刺激时CD40L蛋白和mRNA表达均显著高于相应的对照组(P＜0.01);③在单纯培养和PMA Ionomycin刺激时,FK506对LNPBMC表达CD40L蛋白和mRNA均有显著抑制作用(P＜0.01).结论:LN患者PBMC存在CaN过度活化;LN患者PBMC高效表达CD40L与其CaN过度活化密切相关,通过阻断CaN活化可调控CD40-CD40L共刺激信号途径的活化.