Quantitative Analysis of the Global Proteome in Peripheral Blood Mononuclear Cells from Patients with New‐Onset Psoriasis

Psoriasis is a common chronic autoimmune skin disease involving the activation of T cells. To explore the proteomic signature of peripheral blood mononuclear cells, a quantitative analysis of their global proteome was conducted in samples from Chinese patients with new‐onset psoriasis (n = 31) and healthy controls (n = 32) using an integrated quantitative approach with tandem mass tag labeling and LC–MS/MS. Protein annotation, unsupervised hierarchical clustering, functional classification, functional enrichment and cluster, and protein–protein interaction analyses were performed. A total of 5178 proteins were identified, of which 4404 proteins were quantified. The fold‐change cutoff was set at 1.2 (patients vs controls); 335 proteins were upregulated, and 107 proteins were downregulated. The bioinformatics analysis indicated that the differentially expressed proteins were involved in processes related to the activation of immune cells including the nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB) pathway, cellular energy metabolism, and proliferation. Three upregulated proteins and two phosphorylated proteins in the NF‐κB pathway were verified or identified by Western blotting. These results confirm that the NF‐κB pathway is critical to psoriasis. In addition, many differentially expressed proteins identified in this study have never before been associated with psoriasis, and further studies on these proteins are necessary.     

人外周血淋巴细胞凋亡与年龄及p21~(WAF1)的关系

 分别从老年人和新生儿外周血中分离淋巴细胞 ,用形态观察、DNA断裂电泳图谱、流式细胞仪分析凋亡峰等手段检测其在体外培养过程中发生凋亡的情况 .结果表明 ,不论是自发凋亡还是用1 0 mmol/L的丁酸钠诱导其凋亡 ,老年人淋巴细胞的凋亡率均高于新生儿组 .进一步检测淋巴细胞凋亡时 p2 1 WAF1的表达变化 ,结果表明老年人淋巴细胞 p2 1 WAF1的下调幅度明显大于新生儿组 .提示老年人淋巴细胞凋亡率高与其 p2 1 WAF1表达容易下调有关 .     

细胞因子诱导外周血单个核细胞的转录和蛋白表达研究

采用干扰素-γ、抗CD3单克隆抗体和IL-2体外诱导扩增外周血单个核细胞成为cIK细胞,并于诱导培养前及培养第15d时分别收集细胞样本。在对培养前后细胞的增殖、形态及表面标志变化检测的同时。提取总蛋白进行定量、双向电泳和银染。利用ImageMasterTM软件对培养前后表达相同和不同的蛋白质点进行分析,并选择其中24个蛋白质点进行质谱鉴定。对于部分培养前后具有代表性的蛋白,进一步采用qPCR技术分析其的转录情况。结果表明,培养前后细胞的蛋白质组学特征是完全不同的,相同表达蛋白点主要与基因的转录因子和细胞骨架相关,诱导后特异表达蛋白主要与细胞生长、增殖相关。虽然在转录与蛋白水平上呈现出部分负相关现象,由于蛋白质组才是基因表达的最终形式,结合蛋白差异研究结果提示,经细胞因子诱导后,CIK细胞的大量扩增与细胞内蛋白表达改变相关。     

肺癌患者外周血T淋巴细胞分型与抗核抗体之间的关系研究

摘要 目的：研究肺癌患者外周血T淋巴细胞分型与抗核抗体之间的关系。方法：选择2019年1月到2021年6月在我院接受治疗的肺癌患者81例作为研究组,并选择同期健康志愿者81例作为对照组,检测并比较两组患者外周血CD4⁺、CD8⁺和CD4⁺/CD8⁺淋巴细胞比例,以及抗核抗体血清滴度。比较不同抗核抗体、年龄、性别、TNM分期、肿瘤分化程度以及病理类型肺癌患者外周血CD4⁺、CD8⁺和CD4⁺/CD8⁺淋巴细胞比例。结果：（1）肺癌患者外周血CD4⁺和CD4⁺/CD8⁺淋巴细胞比例显著低于对照组,而CD8⁺淋巴细胞比例显著高于对照组（P<0.05）;（2）III+IV肺癌患者外周血CD4⁺、和CD4⁺/CD8⁺淋巴细胞比例均显著低于I+II肺癌患者,而CD8⁺淋巴细胞比例均显著高于I+II肺癌患者（P<0.05）;（3）小细胞肺癌患者外周血CD4⁺、和CD4⁺/CD8⁺淋巴细胞比例均显著低于非小肺癌患者,而CD8⁺淋巴细胞比例均显著高于非小肺癌患者（P<0.05）;（4）肺癌患者抗核抗体血清滴度显著高于对照组（P<0.05）;（5）抗核抗体阳性患者CD4⁺和CD4⁺/CD8⁺淋巴细胞亚群比例均显著低于抗核抗体阴性患者,而CD8⁺淋巴细胞亚群比例显著高于抗核抗体阴性患者（P<0.05）。结论：肺癌患者外周血T淋巴细胞亚群表达异常,并且其表达水平可能与抗核抗体滴度有关。     

胶片吸附的激光显微切割技术及其在蛋白质组研究中的应用

组织显微切割在特定细胞亚群的比较研究中起着重要作用.新近发展的胶片吸附激光显微切割技术能从复杂的生物组织中快速准确地分离纯化出特定细胞亚群.在这项技术中,透明的乙烯乙酸乙烯基酯热塑性胶片覆盖于病理组织切片表面,CO₂激光束特异性作用于目的细胞群表面的胶片,胶片对细胞较强的吸附作用使得选择性自动移取目的细胞群成为可能.目前,这项技术已在蛋白质组分析中获得成功应用,并有望对其产生较深远的影响.     

食管癌患者放射治疗中照射体积和时间与外周血淋巴细胞绝对值的相关性研究

摘要 目的：探究照射体积和时间与食管癌患者外周血淋巴细胞绝对值的相关性。方法：本研究方案将纳入2019年1月~2019年12月蚌埠医学院第一附属医院放疗科收治的放疗或同步放化疗食管癌患者84例,其中单独放疗患者24例,同步放化疗患者60例,采用血液细胞分析仪测定患者放疗期间每周复查外周血白细胞（WBC）、中性粒细胞（N）、淋巴细胞（L）、血红蛋白（HB）及血小板（PLT）计数等指标。Pearson相关性分析照射时间、剂量及体积与外周血指标之间的相关性。结果：食管癌放疗患者,包括同步放化疗及单纯放疗亚组,在治疗1-6周,照射时间与外周血指标均无相关性（P>0.05）。但在放疗第5-6周,患者放疗剂量与WBC、N、L、HB呈负相关（P<0.05）,同步放化疗亚组患者照射剂量与WBC、N、L、HB呈负相关（P<0.05）。在治疗1-4周,不同照射剂量下各梯度照射剂量对应照射体积与外周血指标均无相关性（P>0.05）。但在第5-6周时,患者不同梯度照射剂量下各照射体积与WBC、N呈负相关（P<0.05）,同时在20Gy-60Gy照射剂量,尤其20Gy和30Gy照射剂量下照射体积与L呈负相关（P<0.05）。同步放化疗亚组患者不同照射剂量下各照射体积与WBC、N呈负相关（P<0.05）,同时在20Gy-60Gy照射剂量下照射体积与L呈负相关（P<0.05）,而且在60Gy照射剂量下照射体积与HB呈负相关（P<0.05）。结论：放疗患者特别是同步放化疗亚组患者照射体积、照射剂量与食管癌患者外周血淋巴细胞计数成负相关,基线淋巴细胞与食管癌患者外周血淋巴细胞计数成正相关,而照射时间与食管癌患者外周血淋巴细胞计数无相关性。     

哮喘患者诱导痰和外周血CD3+CD56+NKT 细胞的检测及意义

目的:研究CD3+CD56+NKT细胞在哮喘患者急性发作期诱导痰和外周血中的比例改变,并探讨其临床意义.方法:以28例哮喘急性发作期患者为研究组,22名正常人作为对照组,采用二色直接荧光素标记法和多参数流式细胞仪检测诱导痰和外周血CD3+CD56+NKT细胞的比例,同时检测外周血IL-4、Ig-E及INF-γ等水平.结果:哮喘患者急性发作期诱导痰和外用血CD3+CD56+NKT细胞明显高于健康对照组(P<0.01).哮喘患者急性发作期外周血IL-4、Ig-E及INF-γ等水平明显高于健康对照组(P<0.05).哮喘患者外周血中CD3+CD56+NKT细胞比例与IL-4、Ig-E及INF-γ升高成正相关.结论:哮喘患者急性发作期诱导痰和外周血中的CD3+CD56+NKT细胞明显增高,CD3+CD56+NKT细胞可能通过调节IL-4、Ig-E及INF-γ等细胞因子从而在哮喘的发病机制发挥重要作用.     

The connective connection: interstitial cells of Cajal (ICC) and ICC-like cells establish synapses with immunoreactive cells. Electron microscope study in situ

We present transmission electron microscope (TEM) evidence that ICC and ICC-like cells frequently establish close contacts (synapses) with several types of immunoreactive cells (IRC): lymphocytes, plasma cells, eosinophils, basophils, macrophages and mast cells. Such synapses were found in various organs: human mammary gland and myometrium, as well as rat stomach, gut, bladder and uterus. Specimens were observed by conventional TEM on ultrathin sections. Based on morphometric analyses and computer-aided 3-D reconstructions from serial sections, we propose an operational definition of ICC-IRC synapses: cell-to-cell close contacts where the two cells are separated by only approximately 15 nm, equivalent to twice the plasmalemmal thickness. Two types of such synapses were found: (i) uniform ('plain') synapses (PS). close contact extending for >200 nm, and (ii) multi-contact ('kiss and run') synapses (MS)--with multiple, focal, close-contact points alternating with regions of wider intermembrane distance. For instance, a typical PS between a rat bladder ICC-like cell and an eosinophil was 2.48 microm long and 11+/-4 nm wide. By contrast, a MS synapse in rat myometrium (between an ICC-like cell and an eosinophil) was 8.64 microm long and had 13 contact points. The synaptic cleft measured 15+/-8 nm at contact points and approximately 100 nm or more in wider areas. These synapses are different from gap junctions usually seen between ICC and between ICC and smooth muscle cells. We previously proposed that ICC-like cells might represent stromal progenitor cells, participate in juxtacrine/paracrine signaling and play a role in immune surveillance. The nanoscopic distances between the two contiguous membranes suggest a juxtacrine cell-to-cell signaling (chemical synapse), via juxtacrinins, a specific case of phenomenins. However, the (micro)vesicles found in the synaptic cleft may correspond to an exosome-based mechanism.     

Chromosomal Aberrations (CA), Sister Chromatid Exchanges (SCE), and High-Frequency Cells (HFC) Frequencies in Peripheral Blood Lymphocytes of Tunisian Gasoline Truck Loaders

Gasoline constitutes a mixture of chemicals that contain well-known genotoxicants. Thus, chronic occupational exposure to gasoline may be considered to possess genotoxic risk. In this study, the frequencies of total chromosomal aberrations (TCA), aberrant cells (Ab.c.), sister chromatid exchanges (SCE), high-frequency cells (HFC), and high-frequency cell individual (HFI) were investigated in peripheral blood lymphocytes from 17 gasoline-exposed workers (10 smokers and 7 non-smokers) and 22 unexposed reference subjects (12 smokers and 10 non-smokers). The exposed subjects were gasoline truck loaders at a gasoline company from Tunis City, north of Tunisia. The results indicate multiple CA, such as dicentrics (DIC), chromatid breaks (SB), and chromosome breaks (DB). A significant difference was observed in TCA and Ab.c. frequencies between exposed and unexposed groups (p < 0.01). A significant difference was found in frequencies of SCE (p < 0.01) and HFI (p < 0.05) between exposed and unexposed groups. SCE and TCA frequencies of smokers were found to be significantly higher than those of non-smokers in both groups. There was an interaction between gasoline exposure and smoking habit for TCA (p = 0.020), but not for SCE. Our findings indicate that gasoline truck loaders were under risk of significant cytogenetic damage that was enhanced by their smoking habit.     

Influences of blood sampling procedures on basal hypothalamic-pituitary-adrenal hormone levels and leukocyte values in rhesus macaques (Macaca mulatta)

Abstract: The influences of housing location and temporal factors on the results of blood sampling were examined to determine their contributions to levels of hypothalamic-pituitary-adrenal hormones, as well as leukocyte subset counts from peripheral blood in rhesus monkeys. Differences in housing location and the amount of room disturbance associated with blood sampling have a significant impact on cell counts, but not on ACTH or Cortisol levels.     

溃疡性结肠炎大鼠模型的建立及外周血NK细胞活性的改变

目的　通过建立Wistar大鼠溃疡性结肠炎 (UC)模型 ,探讨外周血NK细胞活性的变化和发病机理的关系。方法　采用饮用Dextransulfatesodium (DSS)水溶液 ,复制UC动物模型 ,随机分为 4组 ,即模型组 :每天饮用生理盐水 6ml(简称M) ;锌治疗组 (Zn组 ) :每天 2次灌肠给予硫酸锌 180mg kg ;硒治疗组 (Se组 ) :每天给予硒力口服液 (含硒 6 0 μg ) 6ml;锌 +硒治疗组 :(ZS组 )每天给予上述剂量硫酸锌 硒力口服液 6ml;均给予 15d ,另设正常对照组 (C组 ) ,每天饮用生理盐水 6ml。用微量乳酸脱氢酶 (LDH)释放法测定各组外周血NK细胞活性。采用火焰原子吸收法测定结肠粘膜锌含量。采用荧光分光光度计比色法测定结肠粘膜硒含量。结果　DSS所致大鼠UC模型简单易行 ,与人类病变相类似。模型组外周血NK细胞活性较正常组降低 (P <0 0 5 ) ,经硒治疗后则有明显回升 (P<0 0 5 )。应用锌、硒治疗后结肠粘膜局部相应微量元素的含量明显提高 (P <0 0 1)。结论　经硒治疗后NK细胞活性明显升高 ,微量元素 (锌和硒 )对UC动物模型有一定的疗效     

应用流式细胞术分析林麝外周血 CD4+、CD8+ T淋巴细胞

通过对圈养林麝(Moschusberezovskii)外周血淋巴细胞CD4~+、CD8~+亚群的检测,探讨林麝细胞免疫功能状态,并探索应用流式细胞仪分析其淋巴细胞亚群的方法,为研究林麝重大疾病的病理机制及诊断方法提供科学依据。本研究选取健康林麝和患呼吸道疾病林麝各5头,以双色流式细胞术检测其外周血淋巴细胞CD4~+、CD8~+亚群的含量,并进行比较。结果显示,羊源CD4、CD8的流式荧光抗体能够标记林麝细胞并有效检测;患病林麝与健康林麝相比,外周血CD4~+细胞含量无差异(P 0.05),CD8~+细胞含量则显著降低(P 0.01),CD4~+/CD8~+比值显著增高(P 0.01)。结果表明,患呼吸系统炎性疾病的林麝其外周血淋巴细胞CD8~+亚群变化显著,检测淋巴细胞亚群对林麝疾病的诊断有重要意义。     

Toll-like receptors (TLRs): An old family of immune receptors with a new face in cancer pathogenesis

In the dark path of tumorigenesis, the more carefully the cancer biology is studied, the more brilliant answers could be given to the countless questions about its orchestrating derivers. The identification of the correlation between Toll-like receptors (TLRs) and different processes involved in carcinogenesis was one of the single points of blinding light highlighting the interconnection between the immune system and cancer. TLRs are a wide family of single-pass membrane-spanning receptors that have developed through the evolution to recognize the structurally conserved molecules derived from microorganisms or damaged cells. But this is not everything about these receptors as they could orchestrate several downstream signalling pathways leading to the formation or suppression of cancer cells. The present review is tempted to provide a concise schematic about the biology and the characters of TLRs and also summarize the major findings of the regulatory role of TLRs and their associated signalling in the pathogenesis of human cancers.     

自身免疫性溶血性贫血(AIHA)患者的不同输血方法与效果

目的:探讨不同输血方法治疗自身免疫性溶血性贫血(autoimmune hemolytic anemia,AIHA)的效果。方法:2017年1月-2018年12月选择在本院血液科诊治的64例自身免疫性溶血性贫血患儿,根据输血方法的不同分为观察组与对照组,各32例。观察组给予洗涤红细胞输注治疗,对照组给予非洗涤红细胞(悬浮红细胞)输注治疗,记录两组输血效果。结果:治疗后4 w观察组的总有效率显著高于对照组(100.0%vs.87.5%,P0.05)。两组治疗后4 w的红细胞计数与血红蛋白都显著高于治疗前,且观察组显著高于对照组(P0.05)。观察组的吸氧、机械通气、住院时间都显著少于对照组(P0.05)。观察组治疗过程的过敏反应、发热反应、紫癜等不良反应发生率显著低于对照组(3.1%vs. 21.9%,P0.05)。结论:洗涤红细胞输注治疗自身免疫性溶血性贫血患儿能促进机体红细胞计数与血红蛋白恢复正常,减少不良反应的发生,提高治疗效果与促进患儿康复。     

自身免疫性肝病患者外周血Th1/Th2细胞的表达及其与肝功能的相关性研究

目的:探讨自身免疫性肝病(AILD)患者外周血中辅助T细胞1(Th1)和辅助T细胞2(Th2)的表达情况及其与肝功能的相关性。方法:选择42例AILD患者为实验组,11例健康体检者作为对照组,采用流式细胞术检测实验组和对照组外周血单个核细胞中Th1和Th2细胞在淋巴细胞亚群中所占百分率,并检测患者肝功能,分析Th1、Th2细胞水平与肝功能血清生化指标的相关性。结果:AILD组患者Th1细胞和Th2细胞百分率均高于健康对照组(P0.05);AILD活动期患者Th1和Th2细胞百分率明显高于健康对照组,AILD缓解期患者外周血淋巴细胞亚群中Th1细胞所占比例明显降低,Th2细胞百分率较健康人群高,Th1/Th2比值明显降低。AILD患者Th1细胞百分率与血清中TBIL、DBIL、GGT水平呈正相关关系(r分别为0.428、0.472、0.511,P值分别为0.010、0.004、0.002)。Th1/Th2比值与血清中TBIL、DBIL水平呈正相关关系(r分别为0.424、0.405,P值分别为0.011、0.016)。结论:AILD患者Th1细胞和Th2细胞百分率异常升高,且Th1细胞百分率及Th1/Th2比率与AILD患者肝功能指标具有良好的相关性。     

A new role for PGRP-S (Tag7) in immune defense: lymphocyte migration is induced by a chemoattractant complex of Tag7 with Mts1

PGRP-S (Tag7) is an innate immunity protein involved in the antimicrobial defense systems, both in insects and in mammals. We have previously shown that Tag7 specifically interacts with several proteins, including Hsp70 and the calcium binding protein S100A4 (Mts1), providing a number of novel cellular functions. Here we show that Tag7–Mts1 complex causes chemotactic migration of lymphocytes, with NK cells being a preferred target. Cells of either innate immunity (neutrophils and monocytes) or acquired immunity (CD4⁺ and CD8⁺ lymphocytes) can produce this complex, which confirms the close connection between components of the 2 branches of immune response.     

外周血中性粒细胞CD4（nCD4）在白血病合并细菌感染患者检测中的应用价值

目的:观察外周血中性粒细胞CD4(neutrophil CD4,nCD4)在白血病合并细菌感染患者检测中的准确度和灵敏度,评价其临床应用价值。方法:筛选我院在2016年1月至2019年1月期间收治的100例白血病患者作为研究对象并纳入研究组,其中合并47例合并细菌感染者纳入研究Ⅰ组,其余53例未合并细菌感染者纳入研究Ⅱ组,另外根据血培养检查结果将研究Ⅰ组分为阳性组(n=12)和阴性组(n=35),同时,筛选同时期进行健康体检者50例纳入对照组;对比降钙素原(Procalcitonin,PCT)、C-反应蛋白(C-reactive protein,CRP)及nCD4水平,并比较阳性组和阴性组的PCT、CRP、nCD4的诊断准确度和灵敏度。结果:研究组的PCT、CRP、nCD4高于对照组(P0.05),研究Ⅰ组的PCT、CRP、nCD4高于研究Ⅱ组(P0.05);研究Ⅰ组中,阳性组的nCD4荧光强度高于阴性组(P0.05);nCD4诊断白细胞细菌感染的准确度、灵敏度、特异度优于CRP及PCT诊断。结论:nCD4在白血病合并细菌感染患者检测中具有较好的临床参考价值,联合PCT、CRP检测能够显著提高检测准确性,可以作为诊断白血病合并细菌感染的诊断指标。     

Influence of In Vitro IL-2 or IL-15 Alone or in Combination with Hsp 70 Derived 14-Mer Peptide (TKD) on the Expression of NK Cell Activatory and Inhibitory Receptors on Peripheral Blood T Cells,B Cells and NKT Cells

Previous studies from Multhoff and colleagues reported that plasma membrane Hsp70 acts as a tumour-specific recognition structure for activated NK cells, and that the incubation of NK cells with Hsp70 and/or a 14-mer peptide derived from the N-terminal sequence of Hsp70 (TKDNNLLGRFELSG, TKD, aa 450–463) plus a low dose of IL-2 triggers NK cell proliferation and migration, and their capacity to kill cancer cells expressing membrane Hsp70. Herein, we have used flow cytometry to determine the influence of in vitro stimulation of peripheral blood mononuclear cells from healthy individuals with IL-2 or IL-15, either alone or in combination with TKD peptide on the cell surface expression of CD94, NK cell activatory receptors (CD16, NK2D, NKG2C, NKp30, NKp44, NKp46, NKp80, KIR2DL4, DNAM-1 and LAMP1) and NK cell inhibitory receptors (NKG2A, KIR2DL2/L3, LIR1/ILT-2 and NKR-P1A) by CD3+CD56+ (NKT), CD3+CD4+, CD3+CD8+ and CD19+ populations. NKG2D, DNAM-1, LAMP1 and NKR-P1A expression was upregulated after the stimulation with IL-2 or IL-15 alone or in combination with TKD in NKT, CD8+ T cells and B cells. CD94 was upregulated in NKT and CD8+ T cells. Concurrently, an increase in a number of CD8+ T cells expressing LIR1/ILT-2 and CD4+ T cells positive for NKR-P1A was observed. The proportion of CD8+ T cells that expressed NKG2D was higher after IL-2/TKD treatment, when compared with IL-2 treatment alone. In comparison with IL-15 alone, IL-15/TKD treatment increased the proportion of NKT cells that were positive for CD94, LAMP1 and NKRP-1A. The more potent effect of IL-15/TKD on cell surface expression of NKG2D, LIR1/ILT-2 and NKRP-1A was observed in B cells compared with IL-15 alone. However, this increase was not of statistical significance. IL-2/TKD induced significant upregulation of LAMP1 in CD8+ T cells compared with IL-2 alone. Besides NK cells, other immunocompetent cells present within the fraction of peripheral blood mononuclear cells were influenced by the treatment with low-dose interleukins themselves or in combination with hsp70 derived (TKD) peptide.