BACKGROUND & AIMS: :We previously reported that caffeine-assisted chemotherapy improved the treatment of malignant bone and soft tissue tumours such as osteosarcoma. Caffeine affects tumour cells through various pathways, including phosphatase and tensin homolog deleted on chromosome 10 (PTEN), AKT, Bcl-2-associated X protein (BAX), caspase-3 and p53, and has therefore been indicated as being useful for the treatment of malignant tumours. Here, the effects of caffeine on the proliferation of HOS osteosarcoma cells were assessed by WST-8 assay, and the effects on the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), mammalian target of rapamycin (mTOR) and mitogen-activated protein kinase (MAPK) pathways were assessed by western blot analyses. Caffeine inhibited proliferation of HOS cells and suppressed NF-κB, AKT, mTOR/S6K and ERK activities. Our results support those from previous studies relating to the use of caffeine in the treatment of osteosarcoma.

背景与目标： ：我们之前曾报道过咖啡因辅助的化疗改善了恶性骨和软组织肿瘤（如骨肉瘤）的治疗。咖啡因通过多种途径影响肿瘤细胞，包括在10号染色体（PTEN），AKT，Bcl-2相关X蛋白（BAX），caspase-3和p53缺失的磷酸酶和张力蛋白同源物，因此已被证明可用于恶性肿瘤的治疗。在这里，通过WST-8分析评估了咖啡因对HOS骨肉瘤细胞增殖的影响，以及对活化的B细胞（雷帕霉素的哺乳动物靶标）的核因子κ-轻链增强子（NF-κB）的影响（通过蛋白质印迹分析评估了mTOR）和有丝分裂原激活的蛋白激酶（MAPK）途径。咖啡因可抑制HOS细胞增殖，并抑制NF-κB，AKT，mTOR / S6K和ERK活性。我们的研究结果支持先前有关使用咖啡因治疗骨肉瘤的研究。

BACKGROUND & AIMS: INTRODUCTION:In the present study, we aimed to explore the functional role of Pellino-1 (Peli1) in inducing neovascularization after myocardial infarction (MI) and hindlimb ischemia (HLI) using Peli1 global knockout mice (Peli1-/-). Recently we have shown that Peli1, an E3 ubiquitin ligase, induce angiogenesis and improve survivability, with decreased necrosis of ischemic skin flaps. METHODS:Peli1fl/fl and Peli1-/- mice were subjected to either permanent ligation of the left anterior descending coronary artery (LAD) or sham surgery (S). Tissues from the left ventricular risk area were collected at different time points post-MI. In addition, Peli1fl/fl and Peli1-/- mice were also subjected to permanent ligation of the right femoral artery followed by motor function scores, Doppler analysis for blood perfusion and immunohistochemical analysis. RESULTS:Global Peli1 knockout exacerbated myocardial dysfunction, 30 and 60 days after MI compared to wild type (WT) mice as measured by echocardiogram. In addition, Peli1-/- mice also showed decreased motor function scores and perfusion ratios compared with Peli1fl/fl mice 28 days after the induction of HLI. The use of Peli1 in adenoviral gene therapy following HLI in CD1 mice improved the perfusion ratio at 28 days compared to Ad.LacZ-injected mice. CONCLUSION:These results suggest new insights into the protective role of Peli1 on ischemic tissues and its influence on survival signaling.

背景与目标： 简介：在本研究中，我们旨在探讨Pellino-1（Peli1）在使用Peli1整体敲除小鼠（Peli1-/-）诱导心肌梗死（MI）和后肢缺血（HLI）后新生血管形成中的功能。最近，我们发现Peli1，一种E3泛素连接酶，可诱导血管生成并提高生存能力，并减少缺血性皮瓣的坏死。
方法：对Peli1fl / fl和Peli1-/-小鼠进行永久性结扎左冠状动脉前降支（LAD）或假手术（S）。在MI后的不同时间点收集来自左心室风险区的组织。此外，还对Peli1fl / fl和Peli1-/-小鼠进行永久性右股动脉结扎，然后进行运动功能评分，多普勒血流灌注分析和免疫组化分析。
结果：通过超声心动图测量，与野生型（WT）小鼠相比，MI后30天和60天，整体Peli1基因敲除加重了心肌功能障碍。此外，诱导HLI后28天，与Peli1fl / fl小鼠相比，Peli1-/-小鼠还显示出降低的运动功能评分和灌注比。与注射Ad.LacZ的小鼠相比，在CD1小鼠中进行HLI后在腺病毒基因治疗中使用Peli1可以改善28天的灌注率。
结论：这些结果提示了Peli1对缺血组织的保护作用及其对生存信号的影响的新见解。

BACKGROUND & AIMS: BACKGROUND:Vaccinium oldhamii (V. oldhamii) has been reported to exert a variety of the pharmacological properties such as anti-oxidant activity, anti-cancer activity, and inhibitory activity of α-amylase and acetylcholinesterase. However, the anti-inflammatory activity of V. oldhamii has not been studied. In this study, we aimed to investigate anti-inflammatory activity of the stem extracts from V. oldhamii, and to elucidate the potential mechanisms in LPS-stimulated RAW264.7 cells. METHODS:Cell viability was evaluated by MTT assay. The determination of NO and PGE2 production was performed using Griess reagent and Prostaglandin E2 ELISA Kit, respectively. The change of mRNA or protein level was evaluated by RT-PCR and Western blot. RESULTS:Among VOS, VOL and VOF, the inhibitory effect of NO and PGE2 production induced by LPS was highest in VOS treatment. Thus, VOS was selected for the further study. VOS dose-dependently blocked LPS-induced NO and PGE2 production by inhibiting iNOS and COX-2 expression, respectively. VOS inhibited the expression of pro-inflammatory cytokines such as IL-1β, IL-6 and TNF-α. In addition, VOS suppressed TRAP activity and attenuated the expression of the osteoclast-specific genes such as NFATc1, c-FOS, TRAP, MMP-9, cathepsin K, CA2, OSCAR and ATPv06d2. VOS inhibited LPS-induced NF-κB signaling activation through blocking IκB-α degradation and p65 nuclear accumulation. VOS inhibited MAPK signaling activation by attenuating the phosphorylation of ERK1/2, p38 and JNK. Furthermore, VOS inhibited ATF2 phosphorylation and blocked ATF2 nuclear accumulation. CONCLUSIONS:These results indicate that VOS may exert anti-inflammatory activity by inhibiting NF-κB and MAPK/ATF2 signaling. From these findings, VOS has potential to be a candidate for the development of chemopreventive or therapeutic agents for the inflammatory diseases.

背景与目标： 背景：越橘越桔（V. oldhamii）具有多种药理特性，例如抗氧化活性，抗癌活性以及α-淀粉酶和乙酰胆碱酯酶的抑制活性。然而，尚未研究oldhamii的抗炎活性。在这项研究中，我们旨在调查V. oldhamii茎提取物的抗炎活性，并阐明LPS刺激的RAW264.7细胞的潜在机制。
方法：采用MTT法评价细胞活力。使用Griess试剂和前列腺素E2 ELISA试剂盒分别测定NO和PGE2的产生。通过RT-PCR和Western印迹评估mRNA或蛋白质水平的变化。
结果：在VOS，VOL和VOF中，LPS诱导的NO和PGE2生成的抑制作用在VOS处理中最高。因此，选择了VOS进行进一步研究。 VOS分别通过抑制iNOS和COX-2表达来剂量依赖性地阻断LPS诱导的NO和PGE2的产生。 VOS抑制促炎细胞因子如IL-1β，IL-6和TNF-α的表达。此外，VOS抑制了TRAP活性并减弱了破骨细胞特异性基因（如NFATc1，c-FOS，TRAP，MMP-9，组织蛋白酶K，CA2，OSCAR和ATPv06d2）的表达。 VOS通过阻止IκB-α降解和p65核蓄积来抑制LPS诱导的NF-κB信号激活。 VOS通过减弱ERK1 / 2，p38和JNK的磷酸化来抑制MAPK信号激活。此外，VOS抑制了ATF2的磷酸化并阻止了ATF2的核积累。
结论：这些结果表明VOS可能通过抑制NF-κB和MAPK / ATF2信号传导发挥抗炎活性。根据这些发现，VOS有潜力成为炎症性疾病的化学预防剂或治疗剂开发的候选者。

BACKGROUND & AIMS: :Exosomes are key mediators of intercellular communication and play a role in the pathogenesis and progression of cancer. Exosomes in circulating body fluids serve as molecular markers for cancer diagnosis. This study aimed to investigate the role of exosomal microRNA (miR)-1910-3p in breast cancer and determine its clinical diagnostic value. MiR-1910-3p promoted proliferation and migration of breast cancer cells in vitro and in vivo. In vitro, exosomes enriched in miR-1910-3p transferred miR-1910-3p to mammary epithelial cells and breast cancer cells, promoting proliferation and migration, inhibiting apoptosis, and inducing autophagy. In vivo, exosomes enriched in miR-1910-3p promoted the proliferation and migration of breast cancer cells. MiR-1910-3p downregulated myotubularin-related protein 3, activated the NF-κB and wnt/β-catenin signaling pathway, and promoted breast cancer progression. Serum miR-1910-3p in exosomes was an effective diagnostic marker that improved the sensitivity of breast cancer diagnosis when used in combination with the traditional tumor marker CA153. In conclusion, breast cancer cell-derived exosomes promoted the growth, metastasis, and autophagy of breast cancer cells by transferring miR-1910-3p. MiR-1910-3p in serum exosomes may serve as a novel molecular marker for breast cancer diagnosis.

背景与目标： ：外来体是细胞间通讯的关键介质，在癌症的发病机理和进程中发挥作用。循环体液中的外来体可作为癌症诊断的分子标记。这项研究旨在调查外泌体微小RNA（miR）-1910-3p在乳腺癌中的作用，并确定其临床诊断价值。 MiR-1910-3p在体外和体内均可促进乳腺癌细胞的增殖和迁移。在体外，富含miR-1910-3p的外泌体将miR-1910-3p转移至乳腺上皮细胞和乳腺癌细胞，从而促进增殖和迁移，抑制细胞凋亡并诱导自噬。在体内，富含miR-1910-3p的外泌体促进了乳腺癌细胞的增殖和迁移。 MiR-1910-3p下调了肌微管蛋白相关蛋白3，激活了NF-κB和wnt /β-catenin信号通路，并促进了乳腺癌的进展。外泌体中的血清miR-1910-3p是有效的诊断标记物，当与传统的肿瘤标记物CA153结合使用时，可提高乳腺癌诊断的敏感性。总之，乳腺癌细胞衍生的外来体通过转移miR-1910-3p促进了乳腺癌细胞的生长，转移和自噬。血清外泌体中的MiR-1910-3p可以作为乳腺癌诊断的新型分子标记。

BACKGROUND & AIMS: :Studying and understanding the mechanism of inflammation in nucleus pulposus is the key to understand and prevent intervertebral disc degeneration. We propose a model of mechanical sensitive ion channel Piezo1 mediated inflammation of nucleus pulposus cells. Piezo1 can up-regulate the level of interleukin-1β (IL-1β) in nucleus pulposus cells once it is activated. It is suggested that Piezo1 may mediate inflammation by activating Nod-like receptor protein 3 (NLRP3) inflammasome to accelerate the production and maturation of IL-1β. The primary objective of this study was to explore whether Piezo1 activates NLRP3 inflammasome in nucleus pulposus cells. Piezo1 sensitization was induced by mechanical stretch following which we analyzed the priming and assembly of NLRP3 inflammasome and also studied if the downstream Ca2+/NF-κB pathway mediated this activation in nucleus pulposus cells. The expression of Piezo1 and NLRP3 inflammasome increased in a time-dependent manner upon mechanical stretch. Piezo1 activation promoted NLRP3 inflammasome assembly, which was demonstrated by the upregulation of caspase-1 activation and IL-1β production. Transfection of Piezo1-siRNA reversed this process. The inhibition of Ca2+/NF-κB pathway reduced Piezo1-dependent activation of NLRP3 inflammasome. Thus, we propose that activation of NLRP3 inflammasome in nucleus pulposus cells mediated by Piezo1 through the Ca2+/NF-κB pathway is a novel pathogenesis for the progress of intervertebral disc degeneration. As per our knowledge this is the first study which has provided evidence linking Piezo1-mediated inflammation in nucleus pulposus cells with the production of NLRP3 inflammasome.

背景与目标： 研究和了解髓核炎症的机制是了解和预防椎间盘退变的关键。我们提出了机械敏感离子通道Piezo1介导的髓核细胞炎症模型。一旦激活，Piezo1可以上调髓核细胞中白介素-1β（IL-1β）的水平。推测Piezo1可能通过激活Nod样受体蛋白3（NLRP3）炎症小体来促进IL-1β的产生和成熟，从而介导炎症。这项研究的主要目的是探讨Piezo1是否激活髓核细胞中的NLRP3炎性小体。机械拉伸诱导Piezo1致敏，随后我们分析了NLRP3炎性小体的引发和组装，还研究了下游Ca2 /NF-κB通路是否在髓核细胞中介导了这种活化。机械拉伸后，Piezo1和NLRP3炎性小体的表达以时间依赖性方式增加。 Piezo1激活促进了NLRP3炎性小体组装，这通过caspase-1激活和IL-1β产生的上调来证明。 Piezo1-siRNA的转染逆转了这一过程。 Ca2 /NF-κB途径的抑制减少了NLRP3炎性小体的Piezo1依赖性激活。因此，我们认为，Piezo1通过Ca2 /NF-κB途径介导的髓核细胞中NLRP3炎性小体的激活是椎间盘退变进展的新发病机制。据我们所知，这是第一项研究，提供了将髓核细胞中Piezo1介导的炎症与NLRP3炎性体产生联系起来的证据。

BACKGROUND & AIMS: :Dietary nutrients shape complex interactions between hosts and their commensal gut bacteria, further promoting flexibility in host-microbiota associations that can drive nutritional symbiosis. However, it remains less clear if diet-dependent host signaling mechanisms also influence these associations. Using Drosophila, we show here that nuclear factor κB (NF-κB)/Relish, an innate immune transcription factor emerging as a signaling node linking nutrient-immune-metabolic interactions, is vital to adapt gut microbiota species composition to host diet macronutrient composition. We find that Relish is required within midgut enterocytes to amplify host-Lactobacillus associations, an important bacterial mediator of nutritional symbiosis, and thus modulate microbiota composition in response to dietary adaptation. Relish limits diet-dependent transcriptional inducibility of the cap-dependent translation inhibitor 4E-BP/Thor to control microbiota composition. Furthermore, maintaining cap-dependent translation in response to dietary adaptation is critical to amplify host-Lactobacillus associations. These results highlight that NF-κB-dependent host signaling mechanisms, in coordination with host translation control, shape diet-microbiota interactions.

背景与目标： 饮食营养素塑造宿主与其肠道菌之间复杂的相互作用，从而进一步促进宿主微生物群关联的灵活性，从而推动营养共生。然而，尚不清楚饮食依赖性宿主信号传导机制是否也影响这些关联。使用果蝇，我们在这里显示核因子κB（NF-κB）/ Relish，一种先天免疫转录因子，作为连接营养物-免疫-代谢相互作用的信号节点而出现，对于适应肠道微生物群组成来容纳饮食中的大量营养素组成至关重要。我们发现，中肠肠上皮细胞内需要Relish来放大宿主-乳酸杆菌，营养共生的重要细菌介体，从而调节微生物群组成以适应饮食适应。津津有味地限制了帽依赖性翻译抑制剂4E-BP / Thor的饮食依赖性转录诱导性，以控制微生物群的组成。此外，响应于饮食适应性而维持帽依赖性翻译对于扩增宿主-乳酸杆菌协会是至关重要的。这些结果表明，与宿主翻译控制协同作用，依赖于NF-κB的宿主信号传导机制影响了饮食-微生物群落的相互作用。

BACKGROUND & AIMS: :Carpesium macrocephalum (CM) Fr. et Sav. (Compositae) has been used in Chinese folk medicine as an analgesic, hemostatic, antipyretic, and to suppress inflammatory conditions. In the present study we aimed to provide scientific evidence for the anti-inflammatory properties of CM extract and evaluate the intrinsic mechanisms involved in both in vitro and in vivo experimental models. In in vitro findings, CM significantly inhibited the LPS-stimulated release of proinflammatory mediators such as nitric oxide, tumor necrosis factor-alpha, prostaglandin E2, and interleukin-6 in RAW264.7 macrophages in a concentration-dependent fashion. The attenuation of inflammatory responses in LPS-activated RAW264.7 cells by CM was closely associated with the suppression of nuclear factor-kappa B (NF-κB) phosphorylation, IκB-α degradation, and phosphorylation of Akt. CM treatment also attenuated the phosphorylation of STAT through TRIF dependent pathways in LPS-activated RAW264.7 cells. In vivo studies revealed that CM extract concentration dependently suppressed the acetic acid-induced vascular permeability in mice. Considering the data obtained regulation of multiple signaling mechanisms involving TRIF and Akt/NF-κB pathways might be responsible for the potent anti-inflammatory action of CM, substantiating its traditional use in inflammatory diseases.

背景与目标： ：大头翁（CM）Fr.等（菊科）在中国民间医学中已用作止痛，止血，解热和抑制炎性疾病的药物。在本研究中，我们旨在为CM提取物的抗炎特性提供科学依据，并评估涉及体内和体外实验模型的内在机制。在体外研究中，CM以浓度依赖性方式显着抑制RAW264.7巨噬细胞中LPS刺激的促炎性介质如一氧化氮，肿瘤坏死因子-α，前列腺素E2和白介素6的释放。 CM在LPS激活的RAW264.7细胞中减轻炎症反应与抑制核因子-κB（NF-κB）磷酸化，IκB-α降解和Akt磷酸化密切相关。 CM处理还通过LPS激活的RAW264.7细胞中的TRIF依赖性途径减弱了STAT的磷酸化。体内研究表明，CM提取物的浓度可依赖性地抑制乙酸诱导的小鼠血管通透性。考虑到所获得的数据，涉及TRIF和Akt /NF-κB途径的多种信号传导机制的调控可能是CM强大的抗炎作用，从而证实了其在炎性疾病中的传统用途。

BACKGROUND & AIMS: :The morbidity and mortality of primary liver cancer is one of the highest amongst all cancers. Deficiency of effective treatment and characteristics of cancer metastasis are believed to be responsible for this situation, thus a great demand is required for new agent development. Polyphyllin II (PP2), an important steroidal saponin extracted from Rhizoma Paris, has emerged as a potential anti-cancer agent, but the effects of PP2 in liver cancers and its underlying mechanisms remain unexplored. In our study, we found that PP2 could remarkably suppress the proliferation of two liver cancer cell lines, HepG2 and BEL7402, resulting in significant cell death. Besides, low doses of PP2 have displayed properties that inhibit cellular motility and invasion of liver cancer cells. In addition, we have found that PP2-mediated cofilin activity suppression was implicated in the inhibition of liver cancer cell motility. Decreased expression of two major hydrolytic enzymes (MMP2/MMP9), through the AKT/NF-κB signaling pathway may also be also responsible for this process. Rescue experiments done with either non-phosphorylatable mutant cofilin-1 (S3A) transfection or an activator of the AKT pathway significantly reversed the inhibition effects of PP2 on liver cancer cells. Taken together, we report a potential agent for liver cancer treatment and reveal its underlying mechanisms.

背景与目标： ：原发性肝癌的发病率和死亡率是所有癌症中最高的之一。人们认为有效治疗的缺乏和癌症转移的特征是造成这种情况的原因，因此对新药的开发有很大的需求。 Polyphyllin II（PP2）是从巴黎大根茎中提取的一种重要的甾体皂苷，已成为一种潜在的抗癌剂，但PP2在肝癌中的作用及其潜在机制尚待探索。在我们的研究中，我们发现PP2可以显着抑制两种肝癌细胞系HepG2和BEL7402的增殖，从而导致大量细胞死亡。此外，低剂量的PP2具有抑制细胞运动和侵袭肝癌细胞的特性。另外，我们发现PP2介导的cofilin活性抑制与肝癌细胞运动的抑制有关。通过AKT /NF-κB信号通路减少的两种主要水解酶（MMP2 / MMP9）的表达也可能是这一过程的原因。用不可磷酸化突变体cofilin-1（S3A）转染或AKT途径激活剂进行的抢救实验显着逆转了PP2对肝癌细胞的抑制作用。综上所述，我们报告了一种潜在的肝癌治疗药物，并揭示了其潜在机制。

BACKGROUND & AIMS: :Immune activation and inflammation are closely associated with the development of depression. Pioglitazone (PIO), a peroxisome proliferator-activated receptor gamma (PPAR-γ) agonist, has exhibited antidepressant-like effects in a couple of studies. However, the underlying mechanisms are far from being fully elucidated. The study aimed to investigate the effects of PIO on depression-like behaviors induced by lipopolysaccharide (LPS) and to explore the possible underlying mechanisms. The results showed that PIO pretreatment attenuated the depression-like behaviors in mice challenged with intracerebroventricular (i.c.v.) LPS administration. Moreover, Western blot analysis revealed the effects of PIO on inhibiting activation of the nuclear factor kappa B/interleukin 6/signal transducer and activator of transcription 3 (NF-κB/IL-6/STAT3) pathway, improving down-regulation of the cAMP response-element-binding protein/brain derived neurotrophic factor (CREB/BDNF) pathway, as well as regulating disturbed expression of proteins involved in central serotonergic neurotransmission following LPS administration. The beneficial effects of PIO, at both the behavioral and molecular level, were significantly inhibited by the PPAR-γ specific antagonist GW9662. In summary, our data reveals for the first time that the modulation of the NF-κB/IL-6/STAT3 and CREB/BDNF pathways, as well as the potential impact on central serotonergic neurotransmission, may be involved in the PPAR-γ-dependent effects of PIO on depression-like behaviors induced by LPS. Additionally, our findings may provide a novel therapeutic target for the treatment of depression-like behaviors in patients with inflammatory status.

背景与目标： 免疫激活和炎症与抑郁症的发展密切相关。吡格列酮（PIO）是一种过氧化物酶体增殖物激活的受体γ（PPAR-γ）激动剂，在两项研究中显示出抗抑郁样作用。但是，根本的机制远未得到充分阐明。这项研究旨在调查PIO对脂多糖（LPS）诱导的抑郁样行为的影响，并探讨可能的潜在机制。结果表明，PIO预处理可减轻经脑室内（i.c.v.）LPS给药攻击的小鼠的抑郁样行为。此外，蛋白质印迹分析揭示了PIO对抑制核因子κB/白介素6 /信号转导子和转录激活子3（NF-κB/ IL-6 / STAT3）通路的激活，改善了cAMP的下调响应元素结合蛋白/脑源性神经营养因子（CREB ​​/ BDNF）途径，以及在LPS给药后调节参与中枢5-羟色胺能神经传递的蛋白的表达紊乱。 PIO在行为和分子水平上的有益作用均被PPAR-γ特异性拮抗剂GW9662显着抑制。总而言之，我们的数据首次揭示了NF-κB/ IL-6 / STAT3和CREB ​​/ BDNF信号通路的调节，以及对中枢5-羟色胺能神经传递的潜在影响，可能与PPAR-γ-有关。 PIO对LPS诱导的抑郁样行为的依赖效应。此外，我们的发现可能为炎症状态患者的抑郁症样行为的治疗提供新的治疗目标。

BACKGROUND & AIMS: :The aim of the present study was to investigate the involvement of B cell‑activating factor (BAFF) in the pathogenesis of IgA nephropathy by activating the tumor necrosis factor receptor‑associated factor 6 (TRAF6)/NF‑κB signaling pathway in glomerular mesangial cells. For the clinical analysis, blood, urine and kidney tissue samples were collected from 58 patients diagnosed with primary IgA nephropathy by renal biopsy. For the in vitro study, glomerular mesangial cells were divided into five groups: Control (con)‑short hairpin RNA (shRNA) (control group); con‑shRNA + BAFF (20 ng/ml); con‑shRNA + BAFF + BAFF‑RFc chimera protein (500 µg/ml); TRAF6‑shRNA; and TRAF6‑shRNA + BAFF (20 ng/ml). For the in vivo experiments, 60 Sprague‑Dawley rats were randomly divided into four groups: Con‑small interfering RNA (siRNA) (control group); con‑siRNA + IgA (IgA nephropathy group), BAFF‑RFc chimera protein (2 µg/ml) + IgA, and TRAF6‑siRNA (0.2 µM) + IgA. Reverse transcription‑quantitative PCR was performed to evaluate the mRNA expression levels of TRAF6, connective tissue growth factor (CTGF), fibronectin (FN) and NF‑κBP65. Western blot analysis was used to detect the protein expression levels of TRAF6, FN, CTGF and phosphorylated‑NF‑κBP65 in glomerular mesangial cells and kidney tissues. The results revealed that plasma BAFF levels were positively correlated with the severity of pathological damage in patients with IgA nephropathy. In vitro, BAFF induced the mRNA and protein expression of TRAF6, CTGF, FN and NF‑κBP65 in glomerular mesangial cells. After the BAFF‑RFc chimera protein was added to inhibit the binding of BAFF and BAFF‑receptor (‑R), this effect was reduced. In vivo, inhibition of the effects of BAFF via injection with the BAFF‑R Fc chimera protein reduced kidney damage in rats suffering from IgA nephropathy. The effect on the expression of signaling pathway‑associated proteins was also alleviated. In conclusion, BAFF enhanced the expression of fibroblast factors in the kidneys by activating the TRAF6/NF‑κB signaling pathway.

背景与目标： ：本研究的目的是通过激活肾小球系膜中的肿瘤坏死因子受体相关因子6（TRAF6）/NF-κB信号通路，研究B细胞激活因子（BAFF）在IgA肾病发病机制中的参与细胞。为了进行临床分析，收集了58例经肾活检诊断为原发性IgA肾病的患者的血液，尿液和肾脏组织样本。对于体外研究，将肾小球系膜细胞分为五组：对照组（con）-短发夹RNA（shRNA）（对照组）；肾小球系膜细胞（shRNA）。 con‑shRNA BAFF（20 ng / ml）； con‑shRNA BAFF BAFF‑RFc嵌合蛋白（500 µg / ml）； TRAF6-shRNA；和TRAF6‑shRNA BAFF（20 ng / ml）。对于体内实验，将60只Sprague-Dawley大鼠随机分为四组：超小型干扰RNA（siRNA）（对照组）； con‑siRNA IgA（IgA肾病组），BAFF‑RFc嵌合蛋白（2 µg / ml）IgA和TRAF6‑siRNA（0.2 µM）IgA。进行逆转录定量PCR评估TRAF6，结缔组织生长因子（CTGF），纤连蛋白（FN）和NF-κBP65的mRNA表达水平。 Western blot分析用于检测肾小球系膜细胞和肾组织中TRAF6，FN，CTGF和磷酸化的NF-κBP65的蛋白表达水平。结果显示，血浆BAFF水平与IgA肾病患者的病理损害严重程度呈正相关。在体外，BAFF诱导肾小球系膜细胞中TRAF6，CTGF，FN和NF-κBP65的mRNA和蛋白表达。加入BAFF‑RFc嵌合蛋白以抑制BAFF和BAFF受体（‑R）结合后，这种作用减弱了。在体内，通过注射BAFF‑R Fc嵌合蛋白抑制BAFF的作用可减少IgA肾病大鼠的肾脏损害。也减轻了对信号通路相关蛋白表达的影响。总之，BAFF通过激活TRAF6 / NF‑κB信号通路增强了肾脏中成纤维细胞因子的表达。

BACKGROUND & AIMS: :Polyphyllin VII is an active compound isolated from Paris polyphylla, which is termed Chonglou in China. The present study was designed to investigate the underlying mechanisms of the antitumor effect of Polyphyllin VII in lung cancer cells. The cytotoxic effect of Polyphyllin VII in human lung cancer A549 cells was analyzed; the results revealed an IC50 value of 0.41±0.10 µM at 24 h. The associated mechanisms were investigated by phase‑contrast microscopy, fluorescence microscopy, flow cytometry and western blot analysis. Exposure of A549 cells to Polyphyllin VII resulted in apoptosis. Pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF‑κB, and wortmannin, an inhibitor of PI3K, both decreased the proportion of viable A549 cells in the presence of Polyphyllin VII. The ratio of apoptotic cells increased in the presence of wortmannin and PDTC. Western blot analysis revealed that PI3K, phosphorylated (p)‑PI3K, Akt, p‑Akt, NF‑κB and p‑NF‑κB were downregulated following treatment with Polyphyllin VII. Increased caspase‑3 activity, increased poly‑(ADP‑ribose) polymerase cleavage and a downregulation of inhibitor of caspase‑activated DNase were observed following treatment with Polyphyllin VII, and these effects were enhanced by either wortmannin or PDTC. The present results revealed that Polyphyllin VII was able to induce apoptotic cell death in A549 human lung cancer cells via inhibition of the PI3K/Akt and NF‑κB pathways.

背景与目标： ：Polyphyllin VII是一种从巴黎多叶草中分离出来的活性化合物，在中国被称为崇楼。本研究旨在研究Polyphyllin VII在肺癌细胞中抗肿瘤作用的潜在机制。分析了叶绿素VII在人肺癌A549细胞中的细胞毒性作用。结果显示24小时的IC50值为0.41±0.10 µM。通过相差显微镜，荧光显微镜，流式细胞仪和蛋白质印迹分析研究了相关的机制。将A549细胞暴露于聚叶绿素VII导致细胞凋亡。 NF‑κB的抑制剂吡咯烷二硫代氨基甲酸酯（PDTC）和PI3K的抑制剂渥曼青霉素都降低了在存在Polyphyllin VII的情况下存活的A549细胞的比例。在渥曼青霉素和PDTC存在下，凋亡细胞的比例增加。 Western印迹分析表明，用多聚叶绿素VII处理后，PI3K，磷酸化的（p）-PI3K，Akt，p-Akt，NF-κB和p-NF-κB被下调。在用Polyphyllin VII处理后，观察到caspase-3活性增加，聚（ADP-核糖）聚合酶裂解增加以及caspase激活的DNase抑制剂的下调，而渥曼青霉素或PDTC增强了这些作用。目前的结果表明，Polyphyllin VII能够通过抑制PI3K / Akt和NF-κB途径来诱导A549人肺癌细胞凋亡。

BACKGROUND & AIMS: :In mice, acquisition of Ly49 receptors characterizes one of the developmental stages of NK cells. We previously described a novel Ly49 promoter, Pro1, involved in Ly49 gene regulation in immature NK cells. Pro1 transcriptional activity requires a NF-kappaB binding site; however, only NF-kappaB/p50 binding to this element was observed. Cotransfection of NF-kappaB/p65 with Ly49g Pro1 in LNK cells induced a decrease in the transcriptional activity of the core promoter. Moreover, decreasing NF-kappaB/p65 protein expression by RNA interference increases Pro1 transcriptional activity. A high rate of NF-kappaB/p65 degradation in LNK cells correlates with Pro1 activity, since treatment with the proteasome inhibitor MG132 increased levels of NF-kappaB/p65 protein and decreased Pro1 activity. In addition, analysis of the Ly49 repertoire in NF-kappaB/p50 null mice reveals a decrease in the proportion of NK cells expressing a given Ly49 molecule. The defect in Ly49 expression is observed in the bone marrow and the spleen with a similar altered pattern of developmental stages in each tissue. The frequency of Ly49 expression in NF-kappaB/p52 null mice is slightly increased, indicating the specific role of NF-kappaB/p50 in Ly49 gene activation. These results suggest that NF-kappaB p50/p65 plays a major role in the initiation of Ly49 gene expression in NK cells.

背景与目标： ：在小鼠中，Ly49受体的获得是NK细胞发育阶段的特征之一。我们先前描述了一种新型Ly49启动子Pro1，参与未成熟NK细胞中Ly49基因的调控。 Pro1转录活性需要一个NF-κB结合位点。然而，仅观察到NF-kappaB / p50与该元件的结合。 NF-κB/ p65与Ly49g Pro1在LNK细胞中的共转染诱导核心启动子转录活性的降低。此外，通过RNA干扰降低NF-κB/ p65蛋白表达可增加Pro1转录活性。 LNK细胞中较高的NF-κB/ p65降解率与Pro1活性相关，因为用蛋白酶体抑制剂MG132处理会增加NF-κB/ p65蛋白的水平并降低Pro1活性。另外，对NF-kappaB / p50无效小鼠中Ly49组成部分的分析揭示了表达给定Ly49分子的NK细胞比例的降低。 Ly49表达的缺陷在骨髓和脾脏中观察到，每个组织的发育阶段都有类似的变化模式。 NF-kappaB / p52缺失小鼠中Ly49表达的频率略有增加，表明NF-kappaB / p50在Ly49基因激活中的特定作用。这些结果表明，NF-kappaB p50 / p65在NK细胞中Ly49基因表达的启动中起主要作用。

BACKGROUND & AIMS: :The transcription factor NF-ĸB is a master regulator of the innate immune response and plays a central role in inflammatory diseases by mediating the expression of pro-inflammatory cytokines. Ubiquitination-triggered proteasomal degradation of DNA-bound NF-ĸB strongly limits the expression of its target genes. Conversely, USP7 (deubiquitinase ubiquitin-specific peptidase 7) opposes the activities of E3 ligases, stabilizes DNA-bound NF-ĸB, and thereby promotes NF-ĸB-mediated transcription. Using gene expression and synthetic peptide arrays on membrane support and overlay analyses, we found here that inhibiting USP7 increases NF-ĸB ubiquitination and degradation, prevents Toll-like receptor-induced pro-inflammatory cytokine expression, and represents an effective strategy for controlling inflammation. However, the broad regulatory roles of USP7 in cell death pathways, chromatin, and DNA damage responses limit the use of catalytic inhibitors of USP7 as anti-inflammatory agents. To this end, we identified an NF-ĸB-binding site in USP7, ubiquitin-like domain 2, that selectively mediates interactions of USP7 with NF-ĸB subunits but is dispensable for interactions with other proteins. Moreover, we found that the amino acids 757LDEL760 in USP7 critically contribute to the interaction with the p65 subunit of NF-ĸB. Our findings support the notion that USP7 activity could be potentially targeted in a substrate-selective manner through the development of noncatalytic inhibitors of this deubiquitinase to abrogate NF-ĸB activity.

背景与目标： 转录因子NF-ĸB是先天免疫反应的主要调节因子，通过介导促炎性细胞因子的表达在炎症疾病中起着重要作用。泛素触发的蛋白酶体降解的DNA结合的NF-κB强烈限制了其靶基因的表达。相反，USP7（去泛素酶泛素特异性肽酶7）对抗E3连接酶的活性，稳定DNA结合的NF-ĸB，从而促进NF-ĸB介导的转录。在膜支持和覆盖分析上使用基因表达和合成肽阵列，我们在这里发现抑制USP7可增加NF-ĸB泛素化和降解，防止Toll样受体诱导的促炎性细胞因子表达，并代表控制炎症的有效策略。但是，USP7在细胞死亡途径，染色质和DNA损伤反应中的广泛调节作用限制了USP7催化抑制剂作为抗炎药的使用。为此，我们在USP7中发现了一个泛素样结构域2的NF- B结合位点，该位点选择性介导了USP7与NF-ĸB亚基的相互作用，但对于与其他蛋白质的相互作用是必不可少的。此外，我们发现USP7中的氨基酸757LDEL760关键地促进了与NF-κB的p65亚基的相互作用。我们的发现支持以下观点：通过开发这种去泛素酶的非催化抑制剂来消除NF-κB活性，USP7活性可能以底物选择性的方式靶向。

BACKGROUND & AIMS: ETHNOPHARMACOLOGICAL RELEVANCE:Ulcerative colitis (UC) is tightly associated with inflammation response and oxidative stress. As a folk medicine applied in treatment of diarrhea, Bruguiera gymnorrhiza also possesses anti-inflammatory and anti-oxidative activities, which indicated that B. gymnorrhiza may exert anti-colitis effect. AIM OF THE STUDY:To investigate effect and mechanism of B. gymnorrhiza on experimental UC. MATERIALS AND METHODS:Aqueous extract of B. gymnorrhiza leaves (ABL) was used for investigation in the present study. Murine UC was established through access to 3% dextran sulfate sodium (DSS) for 7 days. Meanwhile, mice accepted treatment with ABL (25, 50, 100 mg/kg) or sulfasalazine (200 mg/kg) once daily. On the last day, disease activity index (DAI) including body weight loss, fecal character and degree of bloody diarrhea was evaluated, colon segments were obtained for length measurement and further analysis and feces were collected for intestinal microbiota analysis. RESULTS:ABL ameliorated DAI scores, colon length shortening and histopathological damage in DSS-induced colitis mice obviously. SOD activity, levels of MDA and GSH altered by colitis were restored remarkably after ABL treatment. ABL inhibited increases in levels of colonic COX-2, iNOS, TNF-α, IL-6, IL-1β, IL-4, IL-10 and IL-11 in colitis mice. Moreover, ABL prominently suppressed NF-κB p65 and IκB phosphorylation and down-regulated mRNA levels of COX-2, iNOS, TNF-α, IL-6 and IL-1β elevated by colitis. As shown in microbiota analysis, ABL modulated composition of intestinal microbiota of colitis mice. CONCLUSION:ABL exhibited protective effect against DSS-induced ulcerative colitis through suppressing NF-κB activation and modulating intestinal microbiota.

背景与目标： 人类药理学联系：溃疡性结肠炎（UC）与炎症反应和氧化应激密切相关。作为一种用于治疗腹泻的民间药物，裸子草还具有抗炎和抗氧化的作用，这表明裸子叶芥子可能发挥抗结肠炎的作用。
研究目的：探讨裸露芽孢杆菌对实验性UC的作用及其机制。
材料与方法：本研究以裸子叶败血菜叶（ABL）的水提物为研究对象。通过使用3％的葡聚糖硫酸钠（DSS）7天来建立小鼠UC。同时，小鼠每天接受一次ABL（25、50、100 mg / kg）或柳氮磺胺吡啶（200Âmg/ kg）的治疗。在最后一天，评估包括体重减轻，粪便特性和血性腹泻程度在内的疾病活动指数（DAI），获得结肠段以进行长度测量并进一步分析，并收集粪便用于肠道菌群分析。
结果：ABL可明显改善DSS诱导的结肠炎小鼠的DAI评分，结肠长度缩短和组织病理学损伤。 ABL治疗后，结肠炎改变了SOD活性，MDA和GSH水平。 ABL抑制结肠炎小鼠结肠COX-2，iNOS，TNF-α，IL-6，IL-1β，IL-4，IL-10和IL-11的水平增加。此外，ABL显着抑制了由结肠炎引起的NF-κBp65和IκB磷酸化，并下调了COX-2，iNOS，TNF-α，IL-6和IL-1β的mRNA水平。如微生物群分析所示，ABL调节了结肠炎小鼠肠道微生物群的组成。
结论：ABL通过抑制NF-κB活化和调节肠道菌群对DSS诱导的溃疡性结肠炎具有保护作用。

BACKGROUND & AIMS: :Stroke is a leading global cause of mortality and disability. However, the pathogenesis that contributes to stroke has not been fully understood. The tripartite motif (TRIM)-containing proteins usually exhibit essential regulatory roles during various biological processes. TRIM8 is a RING domain-containing E3 ubiquitin ligase, playing crucial roles in regulating inflammation and apoptosis. In the present study, we reported that TRIM8 expression was significantly induced in the peri-infarct cortex area of mice after stroke onset. TRIM8 siRNA in vivo transfection resulted in the attenuated cognitive impairments in mice with cerebral ischaemia-reperfusion (IR) injury. In addition, TRIM8 knockdown was neuroprotective, as evidenced by the reduced infarct area, decreased neurological deficit score and down-regulated number of TUNEL-positive cells in the peri-infarct area. Moreover, TRIM8 inhibition obviously repressed glial fibrillary acidic protein (GFAP) expression in peri-hematoma cortex and hippocampus. Furthermore, inflammation induced by cerebral IR injury was highly restrained by TRIM8 knockdown in serum, peri-infarct area and hippocampus, which were along with the remarkable decreases in the phosphorylated expression of IκB kinase alpha (IKKα), inhibitory κB α (IκBα) and nuclear factor kappa B (NF-κB). Moreover, TRIM8 knockdown significantly reduced apoptosis in hippocampus of mice with cerebral IR injury by reducing Caspase-3 cleavage. The in vitro experiment confirmed the neuroprotective role of TRIM8-knockdown in regulating cerebral IR injury. Intriguingly, we found that TRIM8 over-expression-promoted inflammatory response and apoptosis could be markedly attenuated by the inactivation of NF-κB signaling through pre-treatment of JSH-23 or QNZ in lipopolysaccharide (LPS)-incubated astrocytes (ASTs). Therefore, TRIM8 positively regulated cerebral IR injury by activating NF-κB pathway to enhance inflammation and apoptosis. Targeting TRIM8 could provide feasible therapeutic treatment for stroke.

背景与目标： ：中风是导致死亡和残疾的全球主要原因。但是，尚未完全了解导致中风的发病机理。包含三重基序（TRIM）的蛋白质通常在各种生物学过程中表现出必不可少的调节作用。 TRIM8是含RING域的E3泛素连接酶，在调节炎症和凋亡中起关键作用。在本研究中，我们报道了中风发作后在小鼠的梗死周围皮层区域中TRIM8表达被显着诱导。 TRIM8 siRNA体内转染导致脑缺血再灌注（IR）损伤的小鼠的认知功能减退。此外，TRIM8敲低具有神经保护作用，如梗死面积减少，神经功能缺损评分降低和梗死周围区域TUNEL阳性细胞数量下调所证明。此外，TRIM8抑制明显抑制血肿周围皮层和海马中的胶质纤维酸性蛋白（GFAP）的表达。此外，血清，梗死周围区域和海马中的TRIM8抑制可高度抑制脑IR损伤引起的炎症，同时IκB激酶α（IKKα），抑制性κBα（IκBα）和IκB激酶的磷酸化表达显着降低。核因子κB（NF-κB）。此外，通过减少Caspase-3裂解，TRIM8敲低显着降低了脑IR损伤小鼠海马的细胞凋亡。体外实验证实了TRIM8-knockdown在调节脑IR损伤中的神经保护作用。有趣的是，我们发现，通过脂多糖（LPS）培养的星形胶质细胞（ASTs）中JSH-23或QNZ的预处理，NF-κB信号的失活可以显着减弱TRIM8过表达促进的炎症反应和细胞凋亡。因此，TRIM8通过激活NF-κB通路来增强炎症和细胞凋亡，从而积极调节脑IR损伤。靶向TRIM8可以为中风提供可行的治疗方法。