自体黑色素细胞培植疗法治疗白癜风及其机制研究

目的 探讨自体黑色素细胞培植疗法对于治疗白癜风的临床效果.方法 选取我院2013年2月到2014年2月期间收治63例患者为研究对象,对这些患者主要采用自体黑色素细胞培植疗法,3个月后对所有患者的康复情况进行总结.蛋白质印迹法(Western blot)检测治疗前后病变皮肤XRCC1和HSP-90表达变化.结果 通过自体黑色素细胞培植疗法治疗以后,38例(60.3％)患者痊愈,10例(15.9％)患者病情明显好转,需要继续观察,15例(23.8％)患者病情无明显变化甚至有所恶化,需配合其他方法继续治疗.和对照组相比,治疗前XRCC1表达水平显著增加,而HSP-90表达水平降低;治疗后XRCC1表达水平降低,而HSP-90表达水平略升高.结论 自体黑色素细胞培植疗法治疗白癜风具有较好的疗效,其治疗效果与XRCC1和HSP-90密切相关.     

Immunopathogenesis of vitiligo

Vitiligo is a common depigmenting disorder which may have devastating psychological and social consequences and is characterized by the presence of circumscribed white macules in the skin due to the destruction of melanocytes in the epidermis. Various hypotheses have been proposed to explain the pathomechanisms involved in this disease, and studies have shown the participation of autoimmune processes in the pathogenesis of vitiligo. Cellular and humoral immunities have been implicated in the development of vitiligo and their role continues to be investigated. Peripheral blood and skin biopsies of patients with vitiligo show that T-cells, mononuclear cells, various pro-inflammatory cytokines, and auto-antibodies can damage melanocytes. Further research is required to determine whether autoimmunity is the main mechanism of vitiligo or only a consequence.     

Apoptosis of melanocytes in vitiligo results from antibody penetration

Vitiligo is a rather common disease characterized by depigmentation of skin and mucosae due to the loss of melanocytes, most likely as a result of autoimmune phenomena. In this study we demonstrated apoptotic markers in residual melanocytes in skin biopsies of patients with vitiligo, as well as the presence of serum antibodies to melanocyte-specific antigens in the vast majority of patients. Moreover, we were able to prove that serum IgG antibodies from vitiligo patients, but not from healthy controls, were capable to penetrate into cultured melanocytes in vitro, and trigger them to engage in apoptosis. Our results are consonant with the theory that melanocyte-specific antibodies are responsible for the deletion of melanocytes through antibody penetration and apoptosis.     

Cytokines: the yin and yang of vitiligo pathogenesis

Introduction: Dysregulation of melanocyte function is associated with vitiligo, an idiopathic autoimmune hypopigmentary skin disorder, caused by the selective destruction of melanocytes. Cytokines, the key mediators of immune response, which are pivotal in maintaining immune homeostasis, are crucial in vitiligo pathogenesis. Several studies indicate that there is an imbalance between pro- and anti-inflammatory cytokines in the skin and serum of vitiligo patients.

Areas covered: In this comprehensive review, we have summarized the correlation of cytokine imbalance and vitiligo pathogenesis, its role in melanocyte biology, and its impact on vitiligo treatment. We have integrated various published reports on the levels of major cytokines from skin and serum samples of vitiligo patients. We have also discussed the role of endoplasmic reticulum and oxidative stress on cytokine imbalance and vice versa leading to destruction of melanocytes.

Expert commentary: The review reflects that dysregulation of cytokines is multifactorial, ranging from genetic predisposition to altered protein expression relevant to vitiligo pathogenesis. We emphasize that cytokine imbalance in systemic and skin microenvironment plays a crucial role in vitiligo pathogenesis and has promising potential as therapeutic targets for vitiligo.     

调节性T细胞在白癜风发病中的研究进展

白癜风是一种由于黑色素细胞受损导致色素脱失的常见色素障碍性皮肤病,目前其发病机制尚未明确.近年来研究表明,调节性T细胞与白癜风的发病密切相关.在近年研究中,认为调节性T细胞的消耗引起细胞毒性T细胞的激活从而破坏黑色素细胞,调节性T细胞的数量和/或功能的变化也会诱导黑色素细胞的凋亡.因而研究调节性T细胞在白癜风中的作用机制及其应用具有重要意义.     

Abnormal expression of MHC class II and ICAM-1 by melanocytes in vitiligo

The relationship between damage to cutaneous melanocytes and antimelanocyte autoimmunity in vitiligo is unclear. We have demonstrated abnormal expression of MHC class II molecules by perilesional melanocytes in 13/21 patients with vitiligo and a six-fold increase in the number expressing the intercellular adhesion molecule ICAM-1. These molecules have important roles in normal antigen presentation and activation of helper T lymphocytes, and their expression by melanocytes may contribute to the abnormal immune response in vitiligo. MHC class II is not expressed by melanocytes in psoriasis and is unlikely to be induced in vitiligo by cytokines released from activated non-melanocyte-specific T lymphocytes.     

白癜风患者外周血T淋巴细胞异常活化

本文以人类白细胞抗原ＤＲ位点（ＨＬＡ－ＤＲ）及白细胞介素２受体亚单位Ｐ５５和Ｐ７５（ＩＬ－２Ｒ，Ｐ５５和ＩＬ－２Ｒ，Ｐ７５）的表达为Ｔ淋巴细胞活化的指标，用双色免疫标记流式细胞法对白癜风患者外周血Ｔ淋巴细胞（ＶＰＢＴＬ）进行分析。实验发现：ＨＬＡ－ＤＲ在ＶＰＢＴＬ及其亚群Ｔ辅助细胞（Ｔｈ，ＣＤ４＾＋细胞）和Ｔ抑制细胞（Ｔｓ，ＣＤ８＾＋细胞）的表达均明显高于正常人，分别为：４９．６２％比１４．５４％     

微小RNA活性分析法筛选前列腺癌去势抵抗转化相关微小RNA的实验研究

目的 探讨微小RNA(miRNA)活性分析法筛选前列腺癌去势抵抗转化相关miRNA的效率。方法 应用miRNA活性分析法筛选出在前列腺癌去势抵抗过程中潜在的发挥活性作用的miRNAs。培养人激素敏感型前列腺癌LNCAP细胞(对照组)及人去势抵抗型前列腺癌C4-2细胞(C4-2组)、PC-3细胞(PC-3组)、DU-145细胞(DU-145组),提取各组细胞总RNA,采用实时荧光定量PCR(qPCR)检测miRNAs,比较各组细胞miRNAs的表达情况。结果 应用miRNA活性分析法,依据筛选流程,共选出9个差异表达的miRNAs,分别为miR-1、miR-122、miR-218、miR-145、miR-155、miR-210、miR-197、 miR-346、let-7b。采用qPCR检测这9个miRNAs,结果显示,7个miRNAs在两种不同状态下的前列腺癌细胞中存在差异表达;在不同去势抵抗型前列腺癌细胞中,miR-210、miR-197、miR-346、miR-218均明显高表达,而miR-122、miR-145、let-7b均明显低表达。结论 采用miRNA活性分析法筛选前列腺癌去势抵抗转化相关miRNA,有着较高的准确性与可信度;其具体转化过程还需进一步证实。     

基于基因芯片技术结合CIBERSORT反卷积算法研究骨关节炎的免疫机制及潜在中药靶向预测

目的:探究骨关节炎(OA)的免疫机制,挖掘其潜在干预中药。方法:通过GEO数据库获取OA滑膜组织相关基因探针,以正常人群滑膜组织为对照组,采用R软件识别差异表达基因并进行功能相关分析,采用STRING数据库对差异基因进行蛋白网络互作(PPI)分析,并筛选核心靶基因,通过CIBERSORT反卷法计算免疫细胞浸润情况及相关性,采用COREMINE数据库对显著富集的免疫相关生物学过程及核心靶基因进行中药预测。结果:共筛选出716个差异基因,其中上调基因382个,下调基因334个;差异基因PPI涉及IL-6、CXCL8、JUN、VEGFA、IL-1β、MMP9、ITGB2、FOS、APOB、CXCL12 10个核心靶基因;GO结果显示,上调基因与免疫炎症反应关系最为密切;免疫细胞浸润矩阵分析显示,浆细胞、M0型巨噬细胞和未活化的肥大细胞在OA滑膜组织中含量较高,而未活化的CD4记忆T细胞、活化的NK细胞、活化的肥大细胞在OA滑膜组织中含量降低;免疫细胞间相关性分析显示,OA未活化CD4记忆T细胞与活化的肥大细胞呈正相关,而未活化的肥大细胞与活化的肥大细胞呈负相关。COREMINE预测发现,青风...     

卵巢癌关键基因的筛选及其与免疫细胞浸润的相关性分析

目的 利用生物信息学方法筛选卵巢癌 (Ovarian Cancer,OV) 关键基因并分析及其与免疫细胞浸润的相关性。 方法 从 GEO 检索并下载 3 组 OV 数据集,通过 GEO2R 在线分析工具筛选出 OV 组织中差异基因 (Differentially Expressed Genes,DEGs),并利用Metascape数据库对DEGs进行基因本体论 (Gene Ontology,GO) 功能分析和京都基因与基因组百科全书 (Kyoto Encyclopedia of Genes and Genomes,KEGG) 通路富集分析;使用 STRING 数据库分析出 DEGs 的 PPI 网络后,再把结果导入 Cytoscape软件,利用其中的 CytoHubba插件提供的 MCC 算法最终得到排名前十的关键基因;然后利用 Kaplan-Meier Plotter数据库和 GEPIA 对关键基因进行生存分析和共表达分析,得出相关性最高的一组基因再分析其 mRNA 和蛋白表达情况;最后利用TISIDB数据库探讨关键基因与OV免疫细胞浸润之间的关联。结果 本研究共得到161个DEGs, 其GO功能主要与细胞对转化生长因子β刺激的反应、P53类介导的DNA损伤反应及因信号转导造成的细胞周期阻滞和蛋白激酶结合等相关;KEGG通路主要富集在癌症通路、P53信号通路、补体和凝血级联以及PI3K-Akt等信号通路上。通过Cy‐ toscape筛选的 10个关键基因中,仅 CCNB1、HMMR、KIAA0101、AURKA、CEP55、ECT2和 TOP2A对 OV患者预后有显著影响,而其中 CCNB1 和 AURKA 在 OV 中表达相关性最高,且其 mRNA 和蛋白均高表达,并不利于患者预后;CCNB1 与 AURKA的表达与Th2细胞、Act-CD4细胞的表达丰度呈正相关,与嗜酸性粒细胞呈负相关。结论 CCNB1和AURKA是OV的关键基因,且与OV微环境中免疫细胞相关,可作为OV的早期确诊和治疗生物标志物。     

Serum homocysteine, vitamin B12, folic acid levels and methylenetetrahydrofolate reductase (MTHFR) gene polymorphism in vitiligo

The aim of this study was to determine serum vitamin B12, folic acid and homocysteine (Hcy) levels as well as MTHFR (C677, A1298C) gene polymorphisms in patients with vitiligo, and to compare the results with healthy controls. Forty patients with vitiligo and 40 age and sex matched healthy subjects were studied. Serum vitamin B12 and folate levels were determined by enzyme-linked immunosorbent assay. Plasma Hcy levels and MTHFR polymorphisms were determined by chemiluminescence and real time PCR methods, respectively. Mean serum vitamin B12 and Hcy levels were not significantly different while folic acid levels were significantly lower in the control group. There was no significant relationship between disease activity and vitamin B12, folic acid and homocystein levels. No significant difference in C677T gene polymorphism was detected. Heterozygote A1298C gene polymorphism in the patient group was statistically higher than the control group. There was no significant relationship between MTHFR gene polymorphisms and vitamin B12, folic acid and homocysteine levels. In conclusion, vitamin B12, folate and Hcy levels are not altered in vitiligo and MTHFR gene mutations (C677T and A1298C) do not seem to create susceptibility for vitiligo.     

Translational Systems Approaches to the Biology of Inflammation and Healing

Inflammation is a complex, non-linear process central to many of the diseases that affect both developed and emerging nations. A systems-based understanding of inflammation, coupled to translational applications, is therefore necessary for efficient development of drugs and devices, for streamlining analyses at the level of populations, and for the implementation of personalized medicine. We have carried out an iterative and ongoing program of literature analysis, generation of prospective data, data analysis, and computational modeling in various experimental and clinical inflammatory disease settings. These simulations have been used to gain basic insights into the inflammatory response under baseline, gene-knockout, and drug-treated experimental animals for in silico studies associated with the clinical settings of sepsis, trauma, acute liver failure, and wound healing to create patient-specific simulations in polytrauma, traumatic brain injury, and vocal fold inflammation; and to gain insight into host-pathogen interactions in malaria, necrotizing enterocolitis, and sepsis. These simulations have converged with other systems biology approaches (e.g., functional genomics) to aid in the design of new drugs or devices geared towards modulating inflammation. Since they include both circulating and tissue-level inflammatory mediators, these simulations transcend typical cytokine networks by associating inflammatory processes with tissue/organ impacts via tissue damage/dysfunction. This framework has now allowed us to suggest how to modulate acute inflammation in a rational, individually optimized fashion. This plethora of computational and intertwined experimental/engineering approaches is the cornerstone of Translational Systems Biology approaches for inflammatory diseases.     

Comprehensive analysis of tumor immune infiltration associated with endogenous competitive RNA networks in lung adenocarcinoma

Cancer immunotherapy has achieved unprecedented success in the treatment of cancer. However, different patients have different responses to immunotherapy. More and more studies have shown that tumor immune heterogeneity has an important influence on the prognosis of cancer. Therefore, understanding the clinical impact of tumor immune infiltration and the regulatory mechanism of RNA molecules is crucial for exploring the pathogenesis of lung adenocarcinoma (LUAD) and the development of immunotherapy protocols.The endogenous competitive RNA hypothesis provides new ideas for studying immune heterogeneity. Therefore, by using the method of immune genomics, this article explores the relationship between immune infiltration and prognosis of patients with lung adenocarcinoma, and found that B-cell immune infiltration highly affects the survival of patients. Through differential analysis, differential mRNAs, lncRNAs and miRNAs were extracted, and 318 differentially expressed mRNAs related to B cell immunity were screened by correlation analysis, and prognosis of patients with COX risk regression model was predicted and analyzed. Through multiple database searches, an immune-related ceRNA regulatory network was constructed, containing 3 key mRNAs, 4 miRNAs, and 50 lncRNAs. Three mRNAs and most miRNAs, lncRNAs, are significantly associated with LUAD prognosis. Bioinformatics analysis of the network showed that LINC00337 may up-regulate the expression of PBK and KIF23 through competitive binding of has-mir-373 and has-mir-519d. The competitive binding of has-mir-373 and has-mir-372 can up-regulate the expression of SLC7A11. The interaction between these RNAs may have an important regulatory role in the immune infiltration in lung adenocarcinoma, thereby affecting the patient's prognosis and immunotherapy efficacy.     

HLA-DRB1等位基因与吉林地区汉族白癜风和寻常型银屑病相关性研究

目的：探讨HLA-DRB1等位基因与吉林地区汉族白癜风和银屑病的相关性。方法：采用聚合酶链反应-序列特异性引物（PCR-SSP）分型技术,对82例汉族白癜风患者和80例寻常型银屑病患者HLA-DRB1等位基因进行检测,与86名汉族健康人群进行对照。结果：①白癜风患者组HLA-DRB1＊07和HLA-DRB1＊12等位基因频率比对照组明显增高（P〈0.05）;②银屑病患者组HLA-DRB1＊07等位基因频率高于对照组,HLA-DRB1＊01等位基因频率低于对照组（P〈0.05）。结论：①HLA-DRB1＊07和HLA-DRB1＊12等位基因可能是吉林地区汉族白癜风的易感基因或与易感基因相连锁;②HLA-DRB1＊07等位基因可能是寻常型银屑病的易感基因,HLA-DRB1＊01可能为寻常型银屑病的保护性基因。     

Association analysis of class II cytokine and receptor genes in vitiligo patients

The loss of melanocytes in vitiligo is mainly attributed to defective autoimmune mechanisms and lately autoinflammatory mediators have become more emphasized. Among these, a number of class II cytokines and their receptors have displayed altered expression patterns in vitiligo. Thus, we selected 30 SNPs from the regions of respective genes to be genotyped in Estonian case-control sample (109 and 328 individuals, respectively). For more precise analyses, patients were divided into subgroups based on vitiligo progression activity, age of onset, sex, occurrence of vitiligo among relatives, extent of depigmented areas, appearance of Köbner’s phenomenon, existence of halo nevi, occurrence of spontaneous repigmentation, and amount of thyroid peroxidase antibodies. No associations appeared in whole vitiligo group. In subgroups, several allelic and haplotype associations were found. The strongest involved SNPs rs12301088 (near IL26 gene), that was associated with familial vitiligo and existence of halo nevi, and rs2257167 (IFNAR1 gene), that was associated with female vitiligo. Additionally, haplotypes consisting of rs12301088 and rs12321603 alleles (IL26–IL22 genes), that were associated with familial vitiligo and existence of halo nevi. In conclusion, several genetic associations with vitiligo subphenotypes were revealed and functional explanations to these remain to be determined in respective studies.     

Relationship of macrophage migration inhibitory factor levels in PBMCs,lesional skin and serum with disease severity and activity in vitiligo vulgaris

Melanocyte loss in vitiligo vulgaris is believed to be an autoimmune process.Macrophage migration inhibitory factor (MIF) is involved in many autoimmune skindiseases. We determined the possible role of MIF in the pathogenesis of vitiligovulgaris, and describe the relationship between MIF expressions and diseaseseverity and activity. Serum MIF concentrations and mRNA levels in PBMCs weremeasured in 44 vitiligo vulgaris patients and 32 normal controls, using ELISAand real-time RT-PCR. Skin biopsies from 15 patients and 6 controls wereanalyzed by real-time RT-PCR. Values are reported as median (25th-75thpercentile). Serum MIF concentrations were significantly increased in patients[35.81 (10.98-43.66) ng/mL] compared to controls [7.69(6.01-9.03) ng/mL]. MIF mRNA levels were significantly higher in PBMCs frompatients [7.17 (3.59-8.87)] than controls [1.67(1.23-2.42)]. There was also a significant difference in MIF mRNA levels inPBMCs between progressive and stable patients [7.86 (5.85-9.13)vs 4.33 (2.23-8.39)] and in serum MIF concentrations[40.47 (27.71-46.79) vs 26.80 (10.55-36.07) ng/mL].In addition, the vitiligo area severity index scores of patients correlatedpositively with changes of both serum MIF concentrations (r = 0.488) andMIF mRNA levels in PBMCs (r = 0.426). MIF mRNA levels were significantlyhigher in lesional than in normal skin [2.43 (2.13-7.59)vs 1.18 (0.94-1.83)] and in patients in theprogressive stage than in the stable stage [7.52 (2.43-8.84)vs 2.13 (1.98-2.64)]. These correlations suggest thatMIF participates in the pathogenesis of vitiligo vulgaris and may be useful asan index of disease severity and activity.     

Evaluation of the effect and mechanism of action of local phenytoin in treatment of vitiligo

There are many theories explaining vitiligo such as genetic, autoimmune, neural, free radicals, biochemical, intrinsic defect, melanocytorrhagy, and convergent theories. Phenytoin is a widely used anticonvulsant, which is used in cutaneous medicine for treatment of ulcers and epidermolysis bullosa. The aim of this study is to assess the effectiveness of topical phenytoin gel in the treatment of vitiligo patients and explaining the underlying mechanism using immunohistochemistry for evaluation of HMB45, CD4, and CD8. Only 9 patients out of 28 experienced response to phenytoin in the form of dull, white color change and light brown color. Post-phenytoin treatment biopsies showed decreased density of inflammation, increased melanin and increased HMB45 positive cells together with an increased number of CD4 positive lymphocytes and decreased number of CD8 positive lymphocytes. These observations did not reach significant level (P > 0.05). A high percentage of CD4 positive lymphocytes was significantly associated with a long duration of vitiligo (p = 0.03) and segmental vitiligo type (p = 0.02). The current study applied phenytoin as 2% concentrated gel for 3 months, which is a relatively short duration without observed side effects throughout the period. These results indicate that topical phenytoin of low concentrations may have beneficial effects through immunomodulatory activity by affecting CD4 and CD8 counts and subsequently the ratio between them. Further studies are recommended to combine phenytoin with other antivitiligo agents as local corticosteroids or phototherapy to clarify if it could potentiate their effects.