Effects of ambient particulate matter on vascular tissue: a review

ABSTRACT

Fine and ultra-fine particulate matter (PM) are major constituents of urban air pollution and recognized risk factors for cardiovascular diseases. This review examined the effects of PM exposure on vascular tissue. Specific mechanisms by which PM affects the vasculature include inflammation, oxidative stress, actions on vascular tone and vasomotor responses, as well as atherosclerotic plaque formation. Further, there appears to be a greater PM exposure effect on susceptible individuals with pre-existing cardiovascular conditions.     

通窍活血汤含药脑脊液对OGD/R损伤大鼠BMECs的保护作用

目的:探讨通窍活血汤含药脑脊液对氧糖剥夺/复糖复氧(OGD/R)损伤大鼠脑微血管内皮细胞(BMECs)的保护作用及潜在机制。方法:通过酶消化法提取原代BMECs,并将细胞随机分为6组,分别为正常组,OGD/R组,通窍活血汤(TQHXT)组(20%),尼莫地平(NMDP)组(10μmol·L~(-1)),卡博替尼(BMS)组(1μmol·L~(-1))和合用药组。除正常组外,其余各组细胞在氧糖剥夺2 h后迅速复糖复氧24 h进行OGD/R造模并分组给药。采用细胞免疫荧光染色法鉴定BMECs,观察OGD/R损伤大鼠BMECs的形态学和超微结构改变并检测细胞跨膜电阻(TEER)值变化。用试剂盒检测细胞内一氧化氮(NO)水平、乳酸脱氢酶(LDH)活性、活性氧(ROS)荧光强度和组织型纤溶酶原激活因子(tPA)含量。采用流式细胞术检测细胞内钙离子浓度及细胞凋亡,观察血管新生标记因子CD34的表达,用蛋白免疫印迹法(Western blot)检测细胞中紧密连接蛋白(ZO-1),血管内皮生长因子(VEGF),黏着斑激酶(FAK)和桩蛋白(Paxillin)蛋白的表达情况。结果:与正常组比较,OGD/R组细胞皱缩、变圆,细胞TEER值和细胞中ZO-1蛋白表达显著降低,细胞中NO,LDH及ROS水平显著升高,tPA含量显著降低,细胞中钙离子浓度和细胞凋亡显著增加,细胞中CD34有所表达,VEGF,FAK和Paxillin蛋白表达显著升高(P0. 01);与OGD/R组比较,TQHXT组细胞损伤明显改善,细胞TEER值和细胞中ZO-1蛋白表达显著升高,细胞中NO,LDH及ROS含量显著降低,tPA含量显著升高,细胞中钙离子浓度和细胞凋亡显著减少,细胞中CD34表达增加,VEGF,FAK和Paxillin蛋白表达显著升高(P0. 05,P0. 01)。结论:通窍活血汤含药脑脊液对OGD/R损伤大鼠BMECs具有保护作用,该保护作用可能是通过VEGF/VEGF受体2(R2)/FAK/Paxillin信号通路促进血管生成来发挥作用。     

B cell and B cell-related pathways for novel cancer treatments

B cells are recognized as the main effector cells of humoral immunity which suppress tumor progression by secreting immunoglobulins, promoting T cell response, and killing cancer cells directly. Given these properties, their anti-tumor immune response in the tumor micro-environment (TME) is of great interest. Although T cell-related immune responses have become a therapeutic target with the introduction of immune checkpoint inhibitors, not all patients benefit from these treatments. B cell and B cell-related pathways (CCL19, −21/CCR7 axis and CXCL13/CXCR5 axis) play key roles in activating immune response through humoral immunity and local immune activation via tertiary lymphoid structure (TLS) formation. However they have some protumorigenic works in the TME. Thus, a better understanding of B cell and B cell-related pathways is necessary to develop effective cancer control. In this review, we summarize recent evidences regarding the roles of B cell and B cell-related pathways in the TME and immune response and discuss their potential roles for novel cancer treatment strategies.     

基于蛋白质组学分析色胺酮抑制小鼠乳腺癌的作用机制

目的:采用非标记定量(Label-free)蛋白质组学技术研究色胺酮抗小鼠体内乳腺癌的作用机制。方法:采用超高效液相色谱-质谱联用技术检测色胺酮抗小鼠乳腺癌的表达蛋白,选择Ionoptics nano UPLC C18色谱柱(0.075 mm×250 mm,1.6μm),流动相0.1%甲酸水溶液-0.1%甲酸乙腈溶液梯度洗脱,正离子模式,扫描范围m/z 100～1 700,使用MaxQuant 1.6.5.0进行数据库检索。采用Label-free高分辨质谱的蛋白质组学技术筛选4T1乳腺癌小鼠模型组与色胺酮(100 mg·kg~(-1))口服给药组之间的差异表达蛋白,进行色胺酮抗乳腺癌的蛋白质组学研究。结果:共鉴定出3 997个蛋白质,其中有2 911个蛋白可定量。模型组与色胺酮组共750个差异表达蛋白,其中286个蛋白上调,464个蛋白下调。基因本体分析表明,这些差异表达蛋白主要参与增殖、细胞迁移、凋亡、免疫、血管生成和炎症调节等生物学过程。京都基因与基因组百科全书通路分析进一步表明,这些蛋白主要集中于T细胞受体,B细胞受体,Toll样受体,核转录因子-κB(NF-κB),Ras蛋白,白细胞介素-17,肿瘤坏死因子,磷脂酰肌醇3-激酶/蛋白激酶B(PI3K-Akt)和丝裂原活化蛋白激酶(MAPK)等信号通路。结论:与色胺酮抗4T1乳腺癌作用密切相关的差异表达蛋白包括上调蛋白白细胞分化抗原14(CD14),前列腺素G/H合酶2(PTGS2),泛素蛋白连接酶E3和下调蛋白CD44,70 kDa热休克蛋白1A(HSPA1A),巨噬细胞移动抑制因子(MIF),NF-κB,核糖体蛋白S6激酶α-4(RPS6KA4)和高迁移率族蛋白B1(HMGB1),提示色胺酮主要通过调节肿瘤炎症微环境来达到抑制小鼠乳腺癌的作用。     

Comprehensive analysis of the relationship between competitive endogenous RNA (ceRNA) networks and tumor infiltrating-cells in hepatocellular carcinoma

BackgroundThe innovation of immune checkpoint blockade (ICB) represents a promising shift in the treatment of advanced hepatocellular carcinoma (HCC). However, response to ICB has varied largely due to the high tumor heterogeneity and complex tumor microenvironment (TME). The competitive endogenous RNA (ceRNA) network also plays an important role in tumor occurrence and progression, but its relation with tumor-infiltrating immune cells (TICs) remains largely unexplored in HCC. The overriding objective of our study was thus to construct a prognosis-related risk model and to further evaluate the relationship between ceRNA networks and TICs.MethodsDifferentially expressed gene (DEG) analysis was performed to identify the differentially expressed RNAs. Lasso and multivariable Cox regression analyses were used to construct risk models, which were assessed by the area under the receiver operating characteristic curve (AUC of ROC) and Kaplan-Meier (K-M) curves. Then, a single-sample gene set enrichment analysis (ssGSEA) algorithm was adopted to dissect the TICs in HCC samples. Nomograms were constructed and calibration curves were used to verify the discrimination and accuracy of the nomograms. Finally, integration analysis was performed to validate the correlation of ceRNA and TICs.ResultsIn the study, 7 differentially expressed RNAs [5 messenger RNA s (mRNAs) and 2 micro RNAs (miRNAs)] were incorporated to construct a ceRNA risk model. The AUC of the 1-, 3-, and 5-year overall survival (OS) were 0.784, 0.685, and 0.691 respectively. Likewise, 7 types TICs were in the TICs signature model and the AUC of the 1-, 3-, and 5-year OS were 0.706, 0.731, and 0.721 respectively. The integration analysis showed that 7 pairs of mRNA-TICs and 1 pair of miRNA-TICs had a close relation (all correlation coefficients >0.2, P<0.001).ConclusionsThrough constructing two risk models based on ceRNA network and TICs, we identified the hub RNAs and key TICs in the progression and prognosis of HCC, and further explored the relationship between ceRNA and TME. Importantly, targeting these hub RNAs may facilitate the remodeling of the TME and be a potential therapeutic alternative to enhancing the response to ICB, thus improving the prognosis of HCC patients.     

Vitiligo surgery: A journey from tissues via cells to the stems!

Depigmented patches in vitiligo, a common dermatosis, cause a great psychological distress to the patients. Hence, apart from halting the disease process, the strategies to impart normal skin colour to these white patches carry an important role in the management of vitiligo. Surgical procedures are often required for stable vitiligo lesions not responding to medical therapies. It involves “shuffling” of melanocytes from the pigmented skin to the depigmented areas. During the last fifty years, the vitiligo surgery has evolved from tissue transplantation via cellular transplantation to reach a stage where the use of stem cells or immunomodulatory cells is contemplating. We would like to depict this wonderful journey of vitiligo surgery through this viewpoint.     

多囊卵巢综合征患者血清外泌体中miR-184的表达水平及其对卵巢颗粒细胞增殖的影响

目的研究多囊卵巢综合征(PCOS)患者血清外泌体中miR-184的表达水平及其对卵巢细胞颗粒增殖的影响。方法回顾性选取2018年2月至2020年4月期间盘锦辽油宝石花医院收治的60例PCOS患者作为PCOS组,另纳入同期30名健康成年女性作为对照组,提取2组受试者血清外泌体。应用实时荧光定量聚合酶链式反应(qRT-PCR)法检测2组受试者血清外泌体中及人卵巢颗粒细胞KGN和人正常卵巢上皮细胞IOSE80中的miR-184的相对表达。转染miR-184抑制剂的KGN细胞设为miR-184抑制剂组,转染抑制剂对照物的KGN细胞设为对照组,不做任何处理的KGN细胞设为空白组。应用MTT法检测各组KGN细胞的增殖情况,应用平板克隆实验检测各组KGN细胞的克隆形成率。结果PCOS患者血清外泌体中和KGN细胞中的miR-184的相对表达量为4.23±0.49、5.81±0.73,显著高于对照组(1.32±0.21)和IOSE80组细胞(1.19±0.15),差异有统计学意义(P<0.05)。miR-184抑制剂组KGN细胞在24、48、72、96 h的吸光度值分别为0.20±0.05、0.22±0.07、0.26±0.08、0.29±0.07,均显著低于阴性对照组(0.31±0.09、0.52±0.11、0.78±0.12、0.96±0.18)和空白对照组(0.31±0.08、0.53±0.10、0.77±0.14、0.94±0.18),克隆形成率(26.78±5.98)%显著低于阴性对照组[(45.98±4.12)%]和空白对照组[(43.56±4.34)%],差异有统计学意义(P<0.05)。结论PCOS患者血清外泌体和人卵巢颗粒细胞KGN中miR-184呈现高表达,PCOS的发病可能与miR-184促进卵巢颗粒细胞增殖有关。     

Predictions of genotoxic potential,mode of action,molecular targets,and potency via a tiered multiflow® assay data analysis strategy

The in vitro MultiFlow® DNA Damage Assay multiplexes γH2AX, p53, phospho-histone H3, and polyploidization biomarkers into a single flow cytometric analysis. The current report describes a tiered sequential data analysis strategy based on data generated from exposure of human TK6 cells to a previously described 85 chemical training set and a new pharmaceutical-centric test set (n = 40). In each case, exposure was continuous over a range of closely spaced concentrations, and cell aliquots were removed for analysis following 4 and 24 hr of treatment. The first data analysis step focused on chemicals' genotoxic potential, and for this purpose, we evaluated the performance of a machine learning (ML) ensemble, a rubric that considered fold increases in biomarkers against global evaluation factors (GEFs), and a hybrid strategy that considered ML and GEFs. This first tier further used ML output and/or GEFs to classify genotoxic activity as clastogenic and/or aneugenic. Test set results demonstrated the generalizability of the first tier, with particularly good performance from the ML ensemble: 35/40 (88%) concordance with a priori genotoxicity expectations and 21/24 (88%) agreement with expected mode of action (MoA). A second tier applied unsupervised hierarchical clustering to the biomarker response data, and these analyses were found to group certain chemicals, especially aneugens, according to their molecular targets. Finally, a third tier utilized benchmark dose analyses and MultiFlow biomarker responses to rank genotoxic potency. The relevance of these rankings is supported by the strong agreement found between benchmark dose values derived from MultiFlow biomarkers compared to those generated from parallel in vitro micronucleus analyses. Collectively, the results suggest that a tiered MultiFlow data analysis pipeline is capable of rapidly and effectively identifying genotoxic hazards while providing additional information that is useful for modern risk assessments—MoA, molecular targets, and potency. Environ. Mol. Mutagen. 60:513–533, 2019. © 2019 Wiley Periodicals, Inc.     

Stem cells for babies and their surgeons: The future is now

Pediatric surgeons are ideal allies for the translation of basic science including stem cell therapies. In the spirit of Robert E. Gross, of applying creative solutions to pediatric problems with technical expertise, we describe the impending cellular therapies that may be derived from stem and progenitor cells. Understanding the types and capabilities of stem and progenitor cells is important for pediatric surgeons to join and facilitate progress for babies. We are developing an induced pluripotent stem cell therapy for enteric neuropathies such as Hirschsprung disease that might be helpful for children in the near future. Our goals, which we hope to share with other surgeons and scientists, include working to establish safe clinical trials and meeting regulatory standards in a thoughtful way that balances patients need and unknown risks.     

Platelets are important for the development of immune tolerance: Possible involvement of TGF‐β in the mechanism

Platelets have diverse roles in immune processes in addition to their key functions in haemostasis and thrombosis. Some studies imply that platelets may be possibly related to the immune tolerance induction. However, the role of platelets in the development of immune tolerance is not fully understood. The purpose of this study was to investigate the role of platelets in the development of regulatory mechanisms responsible for cutaneous inflammation using a mouse model of low zone tolerance (LZT). Mice were treated with 2,4,6‐trinitro‐1‐chlorobenzene (TNCB) 8 times every other day for tolerance induction with administration of anti‐platelet antibody or control antibody during the tolerance induction phase every 3 days. After the treatment for the tolerance induction, mice were sensitized and then challenged with TNCB. The contact hypersensitivity (CHS) was significantly decreased at 24 hours after challenge in the mice with LZT than in those without LZT. Platelet depletion via administration of anti‐platelet antibody reversed the inhibition of CHS and reduced the frequency of Foxp3⁺ Tregs in the inflamed skin and draining lymph nodes in mice with LZT. In addition, repeated low‐dose skin exposure resulted in elevated plasma levels of transforming growth factor (TGF)‐β1. Interestingly, platelet depletion reduced plasma TGF‐β1 levels of mice with LZT. Furthermore, the CHS response was reduced by administration of recombinant TGF‐β1 during platelet depletion in mice with LZT. Administration of anti‐TGF‐β antibody reversed the inhibition of the CHS responses. These results suggest that platelets are involved in the induction of immune tolerance via the release of TGF‐β1.