Vitamin D3 reverses immune tolerance and enhances the cytotoxicity of effector T cells in coal pneumoconiosis.

Coal worker's pneumoconiosis (CWP) is a common occupational disease that coal miners are highly susceptible due to long-term exposure to coal dust particles (CDP). CWP can induce the accumulation of immune cells surrounding the bronchioles and alveoli in the lungs, resulting in pulmonary fibrosis and compromised immune function. Using single-cell RNA sequencing (scRNA-Seq), our previous studies disclose that CDP exposure triggers heterogeneity of transcriptional profiles in mouse pneumoconiosis, while Vitamin D3 (VitD3) supplementation reduces CDP-induced cytotoxicity; however, the mechanism by which how VitD3 regulates immune status in coal pneumoconiosis remains unclear. In this study, we elucidated the heterogeneity of pulmonary lymphocytes in mice exposed to CDP and demonstrated the therapeutic efficacy of VitD3 using scRNA-Seq dataset. The validation of key lymphocyte markers and their functional molecules was performed using immunofluorescence. The results demonstrated that VitD3 increased the number of naive T cells by modulating CD4 + T cell differentiation and decreased the number of Treg cells in CDP-exposed mice, thereby enhancing the cytotoxic activity of CD8 + effector T cells. These effects markedly alleviated lung fibrosis and symptoms. Taken together, the mechanism by which VitD3 regulates the functions of lymphocytes in CWP provides a new perspective for further research on the prevention and treatment of CWP.

Multifunctional gold nanorods in low-temperature photothermal interactions for combined tumor starvation and RNA interference therapy.

Collateral damage to healthy tissue, uneven heat distribution, inflammatory diseases, and tumor metastasis induction hinder the translation of high-temperature photothermal therapy (PTT) from bench to practical clinical applications. In this report, a multifunctional gold nanorod (GNR)-based nanosystem was designed by attaching siRNA against B7-H3 (B7-H3si), glucose oxidase (GOx), and hyaluronic acid (HA) for efficient low-temperature PTT. Herein, GOx can not only exhaust glucose to induce starvation therapy but also reduce the heat shock protein (HSP), realizing the ablation of tumors without damage to healthy tissues. Evidence shows that B7-H3, a type I transmembrane glycoprotein molecule, plays essential roles in growth, metastasis, and drug resistance. By initiating the downregulation of B7-H3 by siRNA, siRNA-GOx/GNR@HA NPs may promote the effectiveness of treatment. By targeting cluster of differentiation 44 (CD44) and depleting B7-H3 and HSPs sequentially, siRNA-GOx/GNR@HA NPs showed 12.9-fold higher lung distribution than siRNA-GOx/GNR NPs. Furthermore, 50% of A549-bearing mice in the siRNA-GOx/GNR NPs group survived over 50 days. Overall, this low-temperature phototherapeutic nanosystem provides an appropriate strategy for eliminating cancer with high treatment effectiveness and minimal systemic toxicity. STATEMENT OF SIGNIFICANCE: To realize efficient tumor ablation under mild low-temperature (42-45 â„ƒ) and RNA interference simultaneously, here we developed a multifunctional gold nanorod (GNR)-based nanosystem (siRNA-GOx/GNR@HA NPs). This nanoplatform can significantly inhibit tumor cell proliferation and induce cell apoptosis by downregulation of HSP90α, HSP70, B7-H3, p-AKT, and p-ERK and upregulation of cleaved caspase-9 at mild low-temperature due to its superior tumor homing ability and the combined effect of photothermal effect, glucose deprivation-initiated tumor starvation, and B7-H3 gene silence effect. It is believed that this multifunctional low-temperature photothermal nanosystem with efficient and specific anticancer properties, shows a potential application in clinical tumor treatment.

Vitamin D3 reduces hippocampal NR2A and anxiety in nicotine withdrawal mice.

Nicotine withdrawal symptoms, mainly anxiety, cause high level of relapse rate after quitting smoking. Vitamin D supplementation has shown its potential for the prevention and treatment of anxiety disorders; however, neurobiological studies about the effect of vitamin D on nicotine withdrawal-induced anxiety are limited. To investigate the effect and molecular mechanism of vitamin D3 supplement by dietary on anxiety-like behavior during nicotine withdrawal, male C57/BL6 mice were divided into four groups: vehicle, nicotine only, vitamin D3 only, and nicotine plus vitamin D3. Mice were administrated with nicotine in drinking water (200 µg/mL), and vitamin D3 in feed for 6 weeks. During nicotine withdrawal, vitamin D3-treated mice showed significantly less anxiety-like behavior by an open-field test and marble buried test that performed an increase in the duration of the central zone and a decrease buried marble, respectively. Moreover, vitamin D3 supplementation attenuated the hippocampal NR2A expression on both protein and mRNA levels in nicotine and vitamin D3-treated mice. Our data showed that dietary supplementation with vitamin D3 ameliorated nicotine withdrawal-induced anxiety, which may be related to downregulation of NR2A expression in hippocampus. Vitamin D3 may provide a new dietary intervention with the easy access for smoking cessation.

Combination therapy with B7H3-redirected bispecific antibody and Sorafenib elicits enhanced synergistic antitumor efficacy.

Rationale: Current traditional treatment options are frequently ineffective to fight against ovarian cancer due to late diagnosis and high recurrence. Therefore, there is a vital need for the development of novel therapeutic agents. B7H3, an immune checkpoint protein, is highly expressed in various cancers, representing it a promising target for cancer immunotherapy. Although targeting B7H3 by bispecific T cell-engaging antibodies (BiTE) has achieved successes in hematological malignancies during recent years, attempts to use them for the treatment of solid cancers are less favorable, in part due to the heterogeneity of tumors. Sorafenib is an unselective inhibitor of multiple kinases currently being tested in clinical trials for several tumors, including ovarian cancer which showed limited activity and inevitable side effect for ovarian cancer treatment. However, it is able to enhance antitumor immune response, which indicates sorafenib may improve the efficiency of immunotherapy. Methods: We evaluated the expression of B7H3 in ovarian cancer using online database and validated its expression of tumor tissues by immunohistochemistry staining. Then, B7H3 expression and the effects of sorafenib on ovarian cancer cell lines were determined by flow cytometry. In addition, 2D and 3D ovarian cancer models were established to test the combined therapeutic effect in vitro. Finally, the efficiency of B7H3×CD3 BiTE alone and its combination with sorafenib were evaluated both in vitro and in vivo. Results: Our data showed that B7H3 was highly expressed in ovarian cancer compared with normal samples. Treatment with sorafenib inhibited ovarian cancer cell proliferation and induced a noticeable upregulation of B7H3 expression level. Further study suggested that B7H3×CD3 BiTE was effective in mediating T cell killing to cancer cells. Combined treatment of sorafenib and B7H3×CD3 BiTE had synergistic anti-tumor effects in ovarian cancer models. Conclusions: Overall, our study indicates that combination therapy with sorafenib and B7H3×CD3 BiTE may be a new therapeutic option for the further study of preclinical treatment of OC.

[Effect of cigarette smoke extract on RAW264. 7 cell proliferation, autophagy and its mechanism].

OBJECTIVE: To inquiry the effects of cigarette smoke extract(CSE) on RAW264. 7 cell proliferation, autophagy and its mechanism. METHODS: RAW264. 7 cell were used and divided into control, starvation and CSE group(2%, 3%, 4%, 5%CSE). CCK-8 was used to detect the toxic action of CSE on RAW264. 7 cell. Western blot and mRFP-GFP-LC3 cell fluorescence spot count were used to explore the function of CSE on RAW264. 7 cell autophagy and its mechanism. RESULTS: Compared with the control group, the result of CCK-8(0. 671 ± 0. 03、0. 746± 0. 10、0. 584 ± 0. 07、0. 588±0. 05) showed that CSE inhibit the proliferation of RAW 264. 7 cell on 24 hours, the difference was statistically significant(P < 0. 05). The outcomes of Western blot showed that, compared with the control group, LC3 B in the CSE group increased, difference in 6(6. 612 ± 0. 35)/12(4. 383 ± 1. 99)/24(5. 781 ± 0. 78) hours, while P62 decreased in 6(1. 815±0. 08)/12(4. 383±1. 99)/24(0. 414±0. 06) hours also different, P-mTOR(1. 744 ± 0. 15) and P-AKT(0. 376 ± 0. 03) decreased, the difference was statistically significant(P<0. 05), but Beclin1 was not significantly changed. The mRFP-GFP-LC3 cell fluorescence spot count showed that the green fluorescence spot(GFP)decreased and the red fluorescence spot(mRFP) remained stable in CSE group, combined mRFP-GFP is shown as yellow and red spots. CONCLUSION: CSE has toxic effect on cell proliferation and leads to RAW264. 7 cell autophagy enhanced through AKT/m TOR pathways.