Dental pulpal nerve fibers express ionotropic adenosine triphosphate (ATP) receptors, suggesting that ATP signaling participates in the process of dental nociception. In this study, we investigated if the principal enzymes responsible for extracellular ATP hydrolysis, namely, nucleoside triphosphate diphosphohydrolases (NTPDases), are present in human dental pulp. Immunohistochemical and immunofluorescence experiments showed that NTPDase2 was predominantly expressed in pulpal nerve bundles, Raschkow’s nerve plexus, and in the odontoblast layer. NTPDase2 was expressed in pulpal Schwann cells, with processes accompanying the nerve fibers and projecting into the odontoblast layer. Odontoblasts expressed the gap junction protein, connexin43, which can form transmembrane hemichannels for ATP release. NTPDase2 was localized close to connexin43 within the odontoblast layer. These findings provide evidence for the existence of an apparatus for ATP release and degradation in human dental pulp, consistent with the involvement of ATP signaling in the process of dentin sensitivity and dental pain.
The splicing of pre-mRNA can be constitutive or alternative. The resulting transcripts may play significant roles in plant development and responses to environmental changes. Wild soybean, as a wild relative of cultivated soybean, has excellent traits such as alkali tolerance and is an ideal material for studying stress mechanisms and mining resistant genes. In this study, we identified a splicing factor GsSCL30a of the SR family that depends on the GsSnRK1 protein kinase. Y2H and BiFC assays verified the interaction between GsSnRK1 and GsSCL30a. In vitro phosphorylation experiments confirmed that GsSnRK1 could phosphorylate GsSCL30a, and six phosphorylation sites of GsSCL30a by GsSnRK1 were determined. qRT-PCR results showed that the expression level of GsSCL30a was highest in wild soybean leaves, and its transcription level was upregulated under alkali stress. Splicing factor reporter analysis found that GsSCL30a could self-splice the third intron, which contains multiple conserved regions of the GAAG motif. In addition, the phosphorylation of GsSCL30a by GsSnRK1 promoted its self-splicing. Y2H experiments proved that there is a physical interaction between GsSCL30a and the U1 snRNP-specific protein U1-70K, indicating that GsSCL30a plays an important role at the 5' splicing site. We overexpressed GsSCL30a-GsSnRK1 in soybean hairy roots. It was found that OE# GsSCL30a-GsSnRK1 could synergistically enhance the tolerance of soybean to alkali stress. Our study revealed the response mechanism of GsSCL30a in soybean under alkali stress conditions, that is, GsSCL30a relies on GsSnRK1 for phosphorylation modification during the alkali stress process, which affects the activity of the splicing factor and then regulates the plant's response to alkali stress, providing a new idea for improving the stress resistance ability of crops.
Non-invasive positive-pressure ventilation (NIPPV) has been demonstrated to exhibit a cardioprotective function in a rat model of myocardial infarction (MI). However, the mechanism underlying NIPPV-mediated MI progression requires further investigation. We aimed to investigate the effectiveness and corresponding mechanism of NIPPV in an acute MI-induced heart failure (HF) rat model. Thirty each of healthy wild type (WT) and apoptosis signal-regulating kinase 1 (ASK-1)-deficient rats were enrolled in this study. MI models were established via anterior descending branch ligation of the left coronary artery. The corresponding data indicated that NIPPV treatment reduced the heart infarct area, myocardial fibrosis degree, and cardiac function loss in MI rats, and ameliorated apoptosis and reactive oxygen species (ROS) levels in the heart tissue. Furthermore, the expression level of ASK-1 level, a key modulator of the ROS-induced extrinsic apoptosis pathway, was upregulated in the heart tissues of MI rats, but decreased after NIPPV treatment. Meanwhile, the downstream cleavage of caspase-3, caspase-9, and PARP, alongside p38 phosphorylation and FasL expression, exhibited a similar trend to that of ASK-1 expression. The involvement of ASK-1 in NIPPV-treated MI in ASK-1-deficient rats was examined. Although MI modeling indicated that cardiac function loss was alleviated in ASK-1-deficient rats, NIPPV treatment did not confer any clear efficiency in cardiac improvement in ASK-1-knockdown rats with MI modeling. Nonetheless, NIPPV inhibited ROS-induced extrinsic apoptosis in the heart tissues of rats with MI by regulating ASK-1 expression, and subsequently ameliorated cardiac function loss and MI-dependent pathogenic changes in the heart tissue.
To analyze the effects of mangiferin combined with bortezomib on the proliferation, invasion, apoptosis and autophagy of human Burkitt lymphoma Raji cells, as well as the expression of CXC chemokine receptors (CXCRs) family, and explore the molecular mechanism between them to provide scientific basis for basic research and clinical work of Burkitt lymphoma.Raji cells were intervened with different concentrations of mangiferin and bortezomib alone or in combination, then cell proliferation was detected by CCK-8 assay, cell invasion ability was detected by Transwell chamber method, cell apoptosis was detected by Annexin V/PI double-staining flow cytometry, apoptosis, autophagy and Akt/mTOR pathway protein expression were detected by Western blot, and the expression changes of CXCR family was detected by real-time quantitative PCR (RT-qPCR).Different concentrations of mangiferin intervened Raji cells for different time could inhibit cell viability in a concentration- and time-dependent manner (r =-0.682, r =-0.836). When Raji cells were intervened by combination of mangiferin and bortezomib, compared with single drug group, the proliferation and invasion abilities were significantly decreased, while the apoptosis level was significantly increased (P <0.01). Mangiferin combined with bortezomib could significantly up-regulate the expression of pro-apoptotic protein Bax and down-regulate the expression of anti-apoptotic protein Bcl-2 after intervention in Raji cells. Caspase-3 was also hydrolyzed and activated, and then induced the apoptosis of Raji cells. Mangiferin combined with bortezomib could up-regulate the expression of LC3Ⅱ protein in Raji cells, and the ratio of LC3Ⅱ/LC3Ⅰ in cells was significantly up-regulated compared with single drug or control group (P <0.01). Mangiferin combined with bortezomib could significantly inhibit the phosphorylation levels of Akt and mTOR, inhibit the proliferation and invasion of Raji cells by inhibiting Akt/mTOR pathway, and induce cell autophagy and apoptosis. Mangiferin and bortezomib could down-regulate the expressions of CXCR4 and CXCR7 mRNA after single-agent intervention in Raji cells, and the down-regulations of CXCR4 and CXCR7 mRNA expression were more significant when the two drugs were combined (P <0.01). Mangiferin alone or combined with bortezomib had no significant effect on CXCR5 mRNA expression in Raji cells (P >0.05), while the combination of the two drugs could down-regulate the expression of CXCR3 (P <0.05).Mangiferin combined with bortezomib can synergistically inhibit the proliferation and invasion of Raji cells, and induce autophagy and apoptosis. The mechanism may be related to the inhibition of Akt/mTOR signaling pathway, down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bax, and the inhibition of the expression of CXCR family.Mangiferin联合硼替佐米对Burkitt淋巴瘤恶性生物学行为的调控机制及对CXCR表达的影响.分析芒果苷（mangiferin）联合硼替佐米对人Burkitt淋巴瘤Raji细胞增殖、侵袭、凋亡和自噬的作用及对CXC趋化因子受体家族（CXCR）表达的影响，并探究其间存在的分子机制，为Burkitt淋巴瘤基础研究与临床提供科学依据。.采用不同浓度Mangiferin、硼替佐米单药或联合干预Raji细胞，利用CCK-8法检测细胞增殖，Transwell小室法检测细胞侵袭能力，Annexin V/PI双染流式细胞术检测细胞凋亡，Western blot检测凋亡、自噬及Akt/mTOR通路蛋白表达情况，实时荧光定量PCR检测CXCR家族的表达变化。.不同浓度Mangiferin干预Raji细胞不同时间后，可抑制Raji细胞活力，且呈浓度及时间依赖性（r =-0.682，r =-0.836）；与单药组相比，Mangiferin联合硼替佐米干预Raji细胞时，细胞增殖与侵袭能力显著下降、凋亡水平显著升高（P <0.01）。Mangiferin联合硼替佐米干预Raji细胞后，可上调促凋亡蛋白Bax表达并显著下调抗凋亡蛋白Bcl-2的表达，同时亦使Caspase-3水解活化，继而诱导Raji细胞发生凋亡。Mangiferin联合硼替佐米干预Raji细胞后可上调LC3Ⅱ蛋白的表达，且细胞中LC3Ⅱ/LC3Ⅰ比值较单药或对照组显著上调（P <0.01）。Mangiferin联合硼替佐米可显著抑制Akt和mTOR的磷酸化水平，通过抑制Akt/mTOR通路来使Raji细胞增殖及侵袭受抑，并诱导细胞发生自噬与凋亡。Mangiferin及硼替佐米单药干预Raji细胞后，可下调 CXCR4、CXCR7 mRNA的表达，当两药联合时 CXCR4、CXCR7 mRNA表达下调更为显著（P <0.01）。Mangiferin单药或联合硼替佐米干预Raji细胞后 CXCR5 mRNA表达无显著变化（P >0.05），但两药联合时可使CXCR3 表达下调（P <0.05）。.Mangiferin联合硼替佐米能协同抑制Raji细胞增殖、侵袭，并诱导其发生自噬与凋亡，机制可能与抑制Akt/mTOR信号通路并通过下调抗凋亡蛋白Bcl-2和上调促凋亡蛋白Bax以及使CXCR家族表达受抑等有关。.
ATP is involved in neurosensory processing, including nociceptive transduction. Thus, ATP signaling may participate in dentin hypersensitivity and dental pain. In this study, we investigated whether pannexins, which can form mechanosensitive ATP-permeable channels, are present in human dental pulp. We also assessed the existence and functional activity of ecto-ATPase for extracellular ATP degradation. We further tested if ATP is released from dental pulp upon dentin mechanical or thermal stimulation that induces dentin hypersensitivity and dental pain and if pannexin or pannexin/gap junction channel blockers reduce stimulation-dependent ATP release. Using immunofluorescence staining, we demonstrated immunoreactivity of pannexin 1 and 2 in odontoblasts and their processes extending into the dentin tubules. Using enzymatic histochemistry staining, we also demonstrated functional ecto-ATPase activity within the odontoblast layer, subodontoblast layer, dental pulp nerve bundles, and blood vessels. Using an ATP bioluminescence assay, we found that mechanical or cold stimulation to the exposed dentin induced ATP release in an in vitro human tooth perfusion model. We further demonstrated that blocking pannexin/gap junction channels with probenecid or carbenoxolone significantly reduced external dentin stimulation-induced ATP release. Our results provide evidence for the existence of functional machinery required for ATP release and degradation in human dental pulp and that pannexin channels are involved in external dentin stimulation-induced ATP release. These findings support a plausible role for ATP signaling in dentin hypersensitivity and dental pain.
The design and optimization of the actuator layout for a 1.5-meter scale adaptive grating was performed using an integrated finite-element-analysis and genetic-optimization model. The optimization process, criteria, and an optimized design will be presented.
Porous Si (PSi) is shown to support and promote primary osteoblast growth (shown in the figure), protein-matrix synthesis, and mineralization. It is also shown that the osteoconductivity of PSi and other cellular responses can be controlled by altering the micro-/ nanoarchitecture of the porous interface. This material has the potential to be a functional biomaterial with both osteoconductivity and drug- delivery functions.
Purinergic signaling is involved in pain generation and modulation in the nociceptive sensory nervous system. Adenosine triphosphate (ATP) induces pain via activation of ionotropic P2X receptors while adenosine mediates analgesia via activation of metabotropic P1 receptors. These purinergic signaling are determined by ecto-nucleotidases that control ATP degradation and adenosine generation. Using enzymatic histochemistry, we detected ecto-AMPase activity in dental pulp, trigeminal ganglia (TG) neurons, and their nerve fibers. Using immunofluorescence staining, we confirmed the expression of ecto-5'-nucleotidase (CD73) in trigeminal nociceptive neurons and their axonal fibers, including the nociceptive nerve fibers projecting into the brainstem. In addition, we detected the existence of CD73 and ecto-AMPase activity in the nociceptive lamina of the trigeminal subnucleus caudalis (TSNC) in the brainstem. Furthermore, we demonstrated that incubation with specific anti-CD73 serum significantly reduced the ecto-AMPase activity in the nociceptive lamina in the brainstem. Our results indicate that CD73 might participate in nociceptive modulation by affecting extracellular adenosine generation in the trigeminal nociceptive pathway. Disruption of TG neuronal ecto-nucleotidase expression and axonal terminal localization under certain circumstances such as chronic inflammation, oxidant stress, local constriction, and injury in trigeminal nerves may contribute to the pathogenesis of orofacial neuropathic pain.
Summary Most splicing factors are extensively phosphorylated but their physiological functions in plant salt resistance are still elusive. We found that phosphorylation by SnRK1 kinase is essential for SRRM1L nuclear speckle formation and its splicing factor activity in plant cells. In Arabidopsis, loss‐of‐function of SRRM1L leads to the occurrence of alternative pre‐mRNA splicing events and compromises plant resistance to salt stress. In Arabidopsis srrm1l mutant line, we identified an intron‐retention Nuclear factor Y subunit A 10 ( NFYA10 ) mRNA variant by RNA‐Seq and found phosphorylation‐dependent RNA‐binding of SRRM1L is indispensable for its alternative splicing activity. In the wild‐type Arabidopsis, salt stress can activate SnRK1 to phosphorylate SRRM1L, triggering enrichment of functional NFYA10.1 variant to enhance plant salt resistance. By contrast, the Arabidopsis srrm1l mutant accumulates nonfunctional NFYA10.3 variant, sensitizing plants to salt stress. In summary, this work deciphered the molecular mechanisms and physiological functions of SnRK1‐SRRM1L‐NFYA10 module, shedding light on a regulatory pathway to fine‐tune plant adaptation to abiotic stress at the post‐transcriptional and post‐translational levels.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)
