Ulcerative colitis (UC) is regarded as a recurring inflammatory disorder of the gastrointestinal tract, for which treatment approaches remain notably limited. In this study, we demonstrated that ginseng polysaccharides (GPs) could alleviate the development of dextran sulfate sodium (DSS)-induced UC as reflected by the ameliorated pathological lesions in the colon. GPs strikingly suppressed the expression levels of multiple inflammatory cytokines, as well as significantly inhibited the infiltration of inflammatory cells. Microbiota-dependent investigations by virtue of 16S rRNA gene sequencing, antibiotic treatment and fecal microbiota transplantation illustrated that GPs treatment prominently restored intestinal microbial balance predominantly through modulating the relative abundance of Lactobacillus. Additionally, GPs remarkably influenced the levels of microbial tryptophan metabolites, diminished the intestinal permeability and strengthened intestinal barrier integrity via inhibiting the 5-HT/HTR3A signaling pathway. Taken together, the promising therapeutic potential of GPs on the development of UC predominantly hinges on the capacity to suppress the expression of inflammatory cytokines as well as to influence Lactobacillus and microbial tryptophan metabolites.

Background: One critical component of the immune system that prevents breast cancer cells from forming distant metastasis is natural killer (NK) cells participating in immune responses to tumors. Ginsenoside Rh2 (GRh2) as one of the major active ingredients of ginseng has been employed in treatment of cancers, but the function of GRh2 in modulating the development of breast cancer remains elusive. Purpose: This study was to dissect the effect of GRh2 against breast cancer and its potential mechanisms associated with NK cells, both in vitro and in vivo. Methods: MDA-MB-231 and 4T1 cells were used to establish in situ and hematogenous mouse models. MDA-MB-231 and MCF-7 were respectively co-cultured with NK92MI cells or primary NK cells in vitro. Anti-tumor efficacy of GRh2 was verified by immunohistochemistry (IHC), Cell Counting Kit-8 (CCK8), high resolution microcomputed tomography (micro-CT) scanning of lungs and hematoxylin and eosin (H&E) staining. Lactate dehydrogenase (LDH) cytotoxicity assay, flow cytometry, in vivo depletion of NK cells, enzyme-linked immunosorbent assay (ELISA), western blot, quantitative reverse transcription polymerase chain reaction (qRT-PCR), immunofluorescence and cell transfection were performed for investigating the anti-tumor mechanisms of GRh2. Molecular docking, microscale thermophoresis (MST) and cellular thermal shift assay (CETSA) were employed to determine the binding between endoplasmic reticulum protein 5 (ERp5) and GRh2. Results: We demonstrated that GRh2 exerted prominent impacts on retarding the growth and metastasis of breast cancer through boosting the cytotoxic function of NK cells, as validated by the elevated release of perforin, granzyme B and interferon-gamma (IFN-gamma). Mechanistical studies revealed that GRh2 was capable of diminishing the expression of ERp5 and GRh2 directly bound to ERp5 in MDA-MB-231 cells as well as on a recombinant protein level. GRh2 prevented the formation of soluble MICA (sMICA) and upregulated the expression level of MICA in vivo and in vitro. Importantly, the reduced lung metastasis of breast cancer by GRh2 was almost abolished upon the depletion of NK cells. Moreover, GRh2 was able to insert into the binding pocket of ERp5 directly. Conclusion: We firstly demonstrated that GRh2 played a pivotal role in augmenting NK cell activity by virtue of modulating the NKG2D-MICA signaling axis via directly binding to ERp5, and may be further optimized to a therapeutic agent for the treatment of breast cancer.

Tumor cell extravasation across endothelial barrier has been recognized as a pivotal event in orchestrating metastasis formation. This event is initiated by the interactions of extravasating tumor cells with endothelial cells (ECs). Therefore, targeting the crosstalk between tumor cells and ECs might be a promising therapeutic strategy to prevent metastasis. In this study, we demonstrated that Rh1, one of the main ingredients of ginseng, hindered the invasion of breast cancer (BC) cells as well as diminished the permeability of ECs both in vitro and in vivo, which was responsible for the attenuated tumor cell extravasation across endothelium. Noteworthily, we showed that ECs were capable of inducing the epithelial-mesenchymal transition (EMT) and invadopodia of BC cells that are essential for tumor cell migration and invasion through limiting the nuclear translocation of hematopoietically expressed homeobox (HHEX). The decreased nuclear HHEX paved the way for initiating the CCL20/CCR6 signaling axis, which in turn contributed to damaged endothelial junctions, uncovering a new crosstalk mode between tumor cells and ECs. Intriguingly, Rh1 inhibited the kinase activity of casein kinase II subunit alpha (CK2α) and further promoted the nuclear translocation of HHEX in the BC cells, which resulted in the disrupted crosstalk between chemokine (C-C motif) ligand 20 (CCL20) in the BC cells and chemokine (C-C motif) receptor 6 (CCR6) in the ECs. The prohibited CCL20-CCR6 axis by Rh1 enhanced vascular integrity and diminished tumor cell motility. Taken together, our data suggest that Rh1 serves as an effective natural CK2α inhibitor that can be further optimized to be a therapeutic agent for reducing tumor cell extravasation.

Supplementary Figure 6 from Toll-like Receptor 9 Activation of Signal Transducer and Activator of Transcription 3 Constrains Its Agonist-Based Immunotherapy

Supplementary Figure 1 from Toll-like Receptor 9 Activation of Signal Transducer and Activator of Transcription 3 Constrains Its Agonist-Based Immunotherapy

Supplementary Figure 2 from Toll-like Receptor 9 Activation of Signal Transducer and Activator of Transcription 3 Constrains Its Agonist-Based Immunotherapy

Numerous pharmacological effects of quercetin have been illustrated, including antiinflammation, antioxidation, and anticancer properties. In recent years, the antioxidant activity of quercetin has been extensively reported, in particular, its impacts on glutathione, enzyme activity, signaling transduction pathways, and reactive oxygen species (ROS). Quercetin has also been demonstrated to exert a striking antiinflammatory effect mainly by inhibiting the production of cytokines, reducing the expression of cyclooxygenase and lipoxygenase, and preserving the integrity of mast cells. By regulating oxidative stress and inflammation, which are regarded as two critical processes involved in the defense and regular physiological operation of biological systems, quercetin has been validated to be effective in treating a variety of disorders. Symptoms of these reactions have been linked to degenerative processes and metabolic disorders, including metabolic syndrome, cardiovascular, neurodegeneration, cancer, and nonalcoholic fatty liver disease. Despite that evidence demonstrates that antioxidants are employed to prevent excessive oxidative and inflammatory processes, there are still concerns regarding the expense, accessibility, and side effects of agents. Notably, natural products, especially those derived from plants, are widely accessible, affordable, and generally safe. In this review, the antioxidant and antiinflammatory abilities of the active ingredient quercetin and its application in oxidative stress-related disorders have been outlined in detail.

Supplementary Figure 7 from Toll-like Receptor 9 Activation of Signal Transducer and Activator of Transcription 3 Constrains Its Agonist-Based Immunotherapy

Supplementary Figure 4 from Toll-like Receptor 9 Activation of Signal Transducer and Activator of Transcription 3 Constrains Its Agonist-Based Immunotherapy

Supplementary Figure 5 from Toll-like Receptor 9 Activation of Signal Transducer and Activator of Transcription 3 Constrains Its Agonist-Based Immunotherapy

Objective: The objective of this study was to investigate the mechanism by which total ginseng extract (TGE) inhibits the progression of lung cancer through network pharmacology and experimentation. Materials and Methods: A Lewis lung carcinomas (LLC) model was established by injecting cancer cells through the tail vein and through administering different doses of TGE. The infiltrated immune cells into the microenvironment of lung cancer metastasis through flow cytometry were evaluated, and the messenger RNA (mRNA) expression levels of various immune cell-related chemokines were determined using quantitative reverse transcription–polymerase chain reaction. Therapeutic targets and signaling pathways of TGE in nonsmall cell lung cancer were investigated using systematic pharmacology and virtual docking. Immunoblotting was performed to determine the impacts of TGE on migration-related proteins. Results: Flow cytometry showed that 1.82 g/kg TGE increased the infiltrated T cells and inhibited the recruitment of myeloid cells, which was caused by decreased mRNA expression of chemokines after TGE treatment. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that the delayed progression of lung cancer by TGE might be related to the promotion of lung cancer cell apoptosis-associated signaling pathways. The virtual docking results indicated that the active components of ginseng are directly bound to apoptosis-related proteins. Immunoblotting showed that TGE inhibited tumor metastasis by regulating the expression of migration-related proteins. Conclusions: The study reveals the potential mechanism of ginseng extract in the treatment of lung cancer progression and provides a reliable basis for its clinical application.

Immunotherapy has been recognized as an effective and important therapeutic modality for multiple types of cancer. Nevertheless, it has been increasing recognized that clinical benefits of immunotherapy are less than expected as evidenced by the fact that only a small population of cancer patients respond favorably to immunotherapy. The structurally and functionally abnormal tumor vasculature is a hallmark of most solid tumors and contributes to an immunosuppressive microenvironment, which poses a major challenge to immunotherapy. In turn, multiple immune cell subsets have profound consequences on promoting neovascularization. Vascular normalization, a promising anti-angiogenic strategy, can enhance vascular perfusion and promote the infiltration of immune effector cells into tumors via correcting aberrant tumor blood vessels, resulting in the potentiation of immunotherapy. More interestingly, immunotherapies are prone to boost the efficacy of various anti-angiogenic therapies and/or promote the morphological and functional alterations in tumor vasculature. Therefore, immune reprograming and vascular normalization appear to be reciprocally regulated. In this review, we mainly summarize how tumor vasculature propels an immunosuppressive phenotype and how innate and adaptive immune cells modulate angiogenesis during tumor progression. We further highlight recent advances of anti-angiogenic immunotherapies in preclinical and clinical settings to solidify the concept that targeting both tumor blood vessels and immune suppressive cells provides an efficacious approach for the treatment of cancer.

Tumor vasculature is characterized by aberrant structure and function, resulting in immune suppressive profiles of tumor microenvironment (TME) through limiting immune cell infiltration into tumors. The defective vascular perfusion in tumors also impairs the delivery and efficacy of chemotherapeutic agents. Targeting abnormal tumor blood vessels has emerged as an effective therapeutic strategy to improve the outcome of chemotherapy and immunotherapy. In this study, we demonstrated that Salvianolic acid B (SalB), one of the major ingredients of Salvia miltiorriza elicited vascular normalization in the mouse models of breast cancer, contributing to improved delivery and response of chemotherapeutic agent cisplatin as well as attenuated metastasis. Moreover, SalB in combination with anti-PD-L1 blockade retarded tumor growth, which was mainly due to elevated infiltration of immune effector cells and boosted delivery of anti-PD-L1 into tumors. Mechanistically, tumor cell enhancer of zeste homolog 2 (Ezh2)-driven cytokines disrupted the endothelial junctions with diminished VE-cadherin expression, which could be rescued in the presence of SalB. The restored vascular integrity by SalB via modulating the interactions between tumor cells and endothelial cells (ECs) offered a principal route for achieving vascular normalization. Taken together, our data elucidated that SalB enhanced sensitivity of tumor cells to chemotherapy and immunotherapy through triggering tumor vascular normalization, providing a potential therapeutic strategy of combining SalB and chemotherapy or immunotherapy for patients with breast cancer.

Diabetic nephropathy (DN) is a common disease, and patients often do not have satisfactory treatments. We investigated therapeutic effects of Fuxin Granules(FX) on DN and potential molecular mechanisms. We orally administered doses of FX to db/db mice for 10 weeks and measured total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol. H&E, PAS, Masson, and Oil Red O staining were used to observe the structure of kidneys and calculate indices of kidney function. We used pharmacological analysis to investigate potential mechanisms of FX. Relative mRNA and protein levels in the TGF-β1/Smad, TGF-β1/Smad, and VEGF/VEGFR2 pathways were examined. TC, TG, and LDL-C were markedly reduced, lipid accumulation was low, fibrosis reduced, kidney atrophy improved, kidney lipid droplet number significantly reduced, and glomerular filtration function improved by FX treatment. Multi-channel therapeutic effects in DN through the TGF-β1/Smad and VEGF/VEGFR2 signaling pathways occurred, and FX substantially reduced expression of TGF-β1 in the glomeruli. FX significantly inhibited TGF-β1, Smad2/3 total protein levels, Smad2/3 phosphorylation mRNA levels of TGF-β1, Smad2, and Smad3. eNOS, VEGFA, and VEGFR2 expression was regulated, levels of VEGFA and VEGFR2 were decreased, and FX increased eNOS. FX ameliorated symptoms of DN, resulting in marked improvement in hyperglycemia and hyperlipidemia and optimized structure and function of kidneys in db/db mice. FX efficacy was associated with the TGF-β1/Smad and VEGF/VEGFR2 signaling pathways. We verified this potential mechanism and hope that this study will provide benefits for the clinical treatment of DN.

目的 观察山海丹颗粒对勃起功能障碍(Erectile dysfunction,ED)疾病的改善作用,并基于前期网络药理学分析结果,探讨可能的相关机制.方法 选取140只Wistar大鼠作为实验动物,雌雄各半.采用双侧髂内动脉结扎的方法,用雄性大鼠建立血管性ED动物模型,分为正常组、模型组、西地那非组、疏肝益阳胶囊组、山海丹颗粒低剂量组、山海丹颗粒中剂量组、山海丹颗粒高剂量组,给药组灌胃给药8周,正常组和模型组给予等体积的双蒸水.考察各组大鼠性行为学指标,TUNEL染色检测睾丸内生精细胞凋亡情况,Western blot和qPCR检测阴茎组织的iNOS、eNOS、nNOS和p38表达水平.结果 和模型组相比,山海丹颗粒显著缩短了ED大鼠的扑捉潜伏期和射精潜伏期,减少睾丸组织内凋亡的生精细胞数量,增加iNOS、eNOS、nNOS表达,抑制p38磷酸化.结论 山海丹颗粒可有效改善ED模型大鼠的疾病状态,其治疗ED的作用机制可能是通过调控iNOS、eNOS、nNOS和p38来实现的.

Background: Natural killer (NK) cells play an indispensable role in anti-tumor immunity. TGF-β1 is the main accomplice of tumor immune escape, inhibiting tumor immunity mediated by NK cells. It is reported that Salvia miltiorrhiza can promote the immune killing effect of NK cells. In this study, Tanshinol, a water-soluble active component of Salvia miltiorrhiza, was used to investigate its effect on the inhibition of NK cell functions mediated by TGF-β1 in breast cancer. Methods: We constructed a mouse model of breast cancer by tail vein injection, H&E staining and ELISA were used to verify the role of TGF-β1 and the effects of Tanshinol on breast cancer and NK cells. In vitro, we used CCK8 and cytotoxicity assays to preliminarily evaluate the effect of Tanshinol on the anti-tumor effect of NK cells intervention by TGF-β1. We explored the killing activity of NK cells and related signal pathways by immunofluorescence imaging technology, RT-PCR, ELISA and flow cytometry. Also, Western blot, RT-PCR and immunofluorescence experiments were applied to investigate the expression level of the natural killer group 2 member D (NKG2D)-NKG2D ligands (NKG2DL) signal axis, and combined with immunoprecipitation, to detect the formation of NKG2D-DNAX-activating protein of 10 kD (DAP10) complex. Results: TGF-β1 played a role in promoting lung metastasis of breast cancer and inhibiting the secretion of cytotoxic mediators from NK cells, but Tanshinol could reverse it. High-dose Tanshinol also significantly optimized the survival rate of tumor-bearing mice. TGF-β1 could destroy the NKG2D-NKG2DL axis, down-regulate the expression and nuclear accumulation of p-smad2/3. Moreover, TGF-β1 inhibited the activation of PI3K-ERK1/2-PLCγ2 signaling pathway that is related to the degranulation of NK cells, and diminished the expression of degranulation marker CD107a and the release of anti-tumor cytotoxic killing medium of NK cells. However, Tanshinol was able to interfere with the negative regulation of TGF-β1 on the functions of NK cells, mainly through promoting the expression of NKG2D and its molecular chaperone DAP10, thereby propelling the formation of NKG2D-DAP10 complex. Conclusions: Collectively, Tanshinol enables NK cells to activate and release multiple killing mediators to carry out immune attacks on tumor cells.

肿瘤血管和淋巴管生成是肿瘤生长、进展和转移的必要条件,且血管生成与肿瘤浸润免疫细胞息息相关.一方面,免疫细胞依赖于血管内皮细胞上的黏附分子而渗入肿瘤组织,并在其中表现出抗肿瘤的特性.另一方面,免疫细胞可以通过分泌趋化因子和细胞因子等多种途径调节肿瘤血管和淋巴管生成,因而其在肿瘤血行转移过程中起重要作用.此外,免疫细胞促肿瘤血行转移的作用凸显出了抗血管生成疗法的不足之处,所以就免疫细胞和血管新生之间的联系,同时靶向免疫细胞和肿瘤新生血管可以提高治疗效果.基于此,该文对肿瘤转移中免疫细胞对血管和淋巴管的作用以及相关免疫抗血管疗法进行综述,以期为后续研究提供参考.

免疫检查点抑制剂(immune checkpoint inhibitor,ICI)通过阻断免疫负调节信号激活机体的抗肿瘤免疫应答,临床试验表明,ICI仅对部分晚期癌症患者有效.而通过阻断肿瘤血管生成常用的抗血管生成药物尽管可以抑制肿瘤的生长,对患者的生存期却未体现出改善,且存在耐药等应用局限.肿瘤免疫反应与血管生成密切相关,同时,肿瘤血管生成高度依赖于免疫抑制微环境.近来一些研究表明,ICI联合血管生成抑制剂不但可以减轻其治疗耐药性,且疗效优于任何单一疗法.联合疗法加强血管正常化和免疫重编程之间的正反馈循环,调节肿瘤免疫微环境以诱导机体持久的抗肿瘤免疫反应.该文对ICI联合抗血管生成疗法的最新进展展开论述,以期为后续研究提供新的思路与借鉴.

肿瘤组织内部含有丰富的新生血管,这些血管分布不均一、无规律且分支紊乱.血管生成素(angiopoietin,Ang)/酪氨酸激酶受体Tie轴介导肿瘤新生血管的稳定成熟过程.Angl主要发挥促血管稳定作用,Ang2在肿瘤血管内皮细胞(endothelial cells,ECs)中高表达,可使血管结构和功能异常.血管的渗漏为循环肿瘤细胞侵袭和转移提供了机会.靶向Ang/Tie轴纠正肿瘤血管的异常状态可促进其正常化,联合化疗药物或免疫疗法可发挥抗肿瘤协同作用.本文就Ang/Tie轴在肿瘤血管生成和转移中的作用进行概述,旨在为临床治疗肿瘤提供新的思路与策略.

气学理论是中医基础理论的重要组成部分.用现代生物技术阐明气学理论的科学内涵,对于发展中医学具有积极意义.近几十年,国内外有学者先后对气的物质本质和气学理论的内涵提出了自己的观点:早期对气本质的认识集中于气的物理特性;近年多从现代生物学角度阐释气的科学内涵,如从生物电、一氧化氮、水蒸气、新陈代谢、Ca2+等角度探讨气的物质形式,认为气与基因表达、细胞通讯等密切相关,也有从脑电能量、自由能、神经血管单元角度阐释气的本质.目前从现代医学角度对气功能的阐释还很少,有人认为气虚导致心脏微血管通透性增加、血浆白蛋白外漏属于气不固摄;有人认为补气是增强机体免疫功能.然而迄今为止,现代医学尚未能完全阐明气学理论的科学内涵,多数停留在理论阐释而没有实验验证,研究也尚未能得到学界的公认.笔者以为,不论气的本质究竟是什么,其功能的发挥离不开机体组成的基本结构——细胞及亚细胞结构.因此,从细胞和亚细胞着手,研究气的功能和本质,不失为一条可行之路.