A novel doxorubicin/CTLA-4 blocker co-loaded drug delivery system improves efficacy and safety in antitumor therapy.

Doxorubicin's antitumor effectiveness may be constrained with ineffective tumor penetration, systemic adverse effects, as well as drug resistance. The co-loading of immune checkpoint inhibitors and doxorubicin into liposomes can produce synergistic benefits and address problems, including quick drug clearance, toxicity, and low drug penetration efficiency. In our previous study, we modified a nanobody targeting CTLA-4 onto liposomes (LPS-Nb36) to be an extremely potent CTLA-4 signal blocker which improve the CD8+ T-cell activity against tumors under physiological conditions. In this study, we designed a drug delivery system (LPS-RGD-Nb36-DOX) based on LPS-Nb36 that realized the doxorubicin and anti-CTLA-4 Nb co-loaded and RGD modification, and was applied to antitumor therapy. We tested whether LPS-RGD-Nb36-DOX could targets the tumor by in vivo animal photography, and more importantly, promote cytotoxic T cells proliferation, pro-inflammatory cytokine production, and cytotoxicity. Our findings demonstrated that the combination of activated CD8+ T cells with doxorubicin/anti-CTLA-4 Nb co-loaded liposomes can effectively eradicate tumor cells both in vivo and in vitro. This combination therapy is anticipated to have synergistic antitumor effects. More importantly, it has the potential to reduce the dose of chemotherapeutic drugs and improve safety.

Establishing a protected area network in Xinlong with other effective area-based conservation measures.

Protected areas (PAs) are pivotal to biodiversity conservation, yet their efficacy is compromised by insufficient funding and management. So-called other effective area-based conservation measures (OECMs) present a paradigm shift and address PA limitations. Such measures can expand conservation areas, enhance connectivity, and improve the existing system. To assess the conservation status of biodiversity in Tibetan cultural areas in China, we investigated the spatial distribution of wildlife vulnerable to human disturbance (large- and medium-sized mammals and terrestrial birds) in Xinlong, a traditional Tibetan cultural area. In particular, we compared a PA (Xionglongxi Nature Reserve) and OECMs targeting species conservation. We also investigated the relationship of wildlife with human temporal and spatial activities. The OECMs complemented areas not covered by PA, especially in rich understory biodiversity regions. More species in OECMs tolerated human presence than species in the PA. Existing biodiversity reserves failed to cover areas of high conservation value in Tibet and offered limited protection capacity. Expanding PAs and identifying OECMs improved Xinlong's system by covering most biodiversity hotspots. Building on the tradition of wildlife conservation in Tibet, harnessing OECMs may be an effective means of augmenting biodiversity conservation capacity. We recommend further evaluation of OECMs effectiveness and coverage in Tibetan area as a way to enhance the current PA system.

自然保护地(protected areas, PAs)被认为是生物多样性保护的最重要且最有效的措施之一。然而, 由于资金不足和管理缺失等因素, 自然保护地体系的有效性被大幅度削弱。"其他有效的基于区域的保护措施(other effective area­based conservation measures, OECMs)"引起了基于区域的保护范式转变, 能够有效弥补PAs的不足。OECMs能够有效实现保护面积的扩大, 加强保护地之间的连通性, 完善现有保护地体系。基于此, 为进一步了解中国藏文化地区生物多样性的保护情况, 评估PAs和OECMs在物种保护上的差异。我们以中国新龙县为例, 调查了当地大中型哺乳动物和地栖鸟类这类容易受人为干扰的野生动物的空间分布和与人类互作的时空活动关系。结果表明, 在保护区无法覆盖的区域, OECMs能够提供有效的补充, 如林下区域的生物多样性保护。在OECMs范围内, 更多的物种能够在时空活动上容忍人类的存在, 尤其是猎物物种。在当前生物多样性优先保护区无法涵盖藏区高保护价值区域以及现有保护区保护能力有限的情况下, 藏区野生动物的保护需要采取更多针对性的措施。通过扩大保护区和确定OECMs区域, 能够覆盖新龙绝大部分生物多样性热点地区。鉴于藏区的野生动物保护传统, 借助OECMs的力量是完善和提高藏区生物多样性保护保护能力的有效手段。我们建议未来进一步评估藏区的OECMs的有效性及覆盖情况, 完善自然保护地体系。 基于其他有效的区域保护措施的中国新龙县保护地体系构建.

The Minimal Impact of Anthropogenic Disturbances on the Spatial Activities of Leopard Cats in Xinlong, China.

The habitat plays a crucial role in ensuring the survival of wildlife. However, the increasing disturbances caused by human activities present a substantial threat to habitats, especially for species such as the leopard cat (Prionailurus bengalensis), which is a significant small predator. Currently, research on leopard cats predominantly focuses on low-altitude regions within its distribution range, leaving plateau areas understudied. To enhance our understanding of the impact of human disturbances on leopard cat habitats, we undertook a study employing infrared camera trappings to monitor leopard cats' activity in Xinlong of southwestern China between 2015 and 2023. We analyzed the spatial distribution and habitat suitability of the leopard cats by utilizing ensemble species distribution models (ESDMs). Moreover, we employed two-species occupancy models to investigate the spatial interaction between leopard cats and human disturbances. The results indicated that (1) the potential suitable habitat area for leopard cats encompassed approximately 1324.93 km2 (14.3%), primarily located along the banks of Yalong river. (2) The distribution of suitable habitat was predominantly influenced by competitors, specifically the yellow-throated marten (YTM), accounting for 52.4% of the influence, as well as environmental factors such as distance to water (DTW) at 12.0% and terrain roughness index (TRI) at 10.0%. Human interference, including cattle presence (4.6%), distance to road (DTD, 4.9%), and distance to settlement (DTS, 3.5%), had a limited impact on the habitat distribution. (3) Within a 5 km radius, habitat suitability increased with proximity to human settlements. (4) Leopard cats exhibited spatial independence from humans and domestic cattle (species interaction factor (SIF) = 1.00) while avoiding domestic horses (SIF = 0.76 ± 0.03). The relatively minor impact of human disturbances in Xinlong could be attributed to traditional cultural practices safeguarding wildlife and the leopard cat's environmental adaptability. We recommend establishing a novel conservation paradigm based on the living dynamics of wildlife communities in Xinlong, thereby offering a more targeted approach to biodiversity preservation in the future.

Immunometabolism changes in fibrosis: from mechanisms to therapeutic strategies.

Immune cells are essential for initiating and developing the fibrotic process by releasing cytokines and growth factors that activate fibroblasts and promote extracellular matrix deposition. Immunometabolism describes how metabolic alterations affect the function of immune cells and how inflammation and immune responses regulate systemic metabolism. The disturbed immune cell function and their interactions with other cells in the tissue microenvironment lead to the origin and advancement of fibrosis. Understanding the dysregulated metabolic alterations and interactions between fibroblasts and the immune cells is critical for providing new therapeutic targets for fibrosis. This review provides an overview of recent advances in the pathophysiology of fibrosis from the immunometabolism aspect, highlighting the altered metabolic pathways in critical immune cell populations and the impact of inflammation on fibroblast metabolism during the development of fibrosis. We also discuss how this knowledge could be leveraged to develop novel therapeutic strategies for treating fibrotic diseases.

Integrated analysis of Dendrobium nobile extract Dendrobin A against pancreatic ductal adenocarcinoma based on network pharmacology, bioinformatics, and validation experiments.

Background: Dendrobium nobile (D. nobile), a traditional Chinese medicine, has received attention as an anti-tumor drug, but its mechanism is still unclear. In this study, we applied network pharmacology, bioinformatics, and in vitro experiments to explore the effect and mechanism of Dendrobin A, the active ingredient of D. nobile, against pancreatic ductal adenocarcinoma (PDAC). Methods: The databases of SwissTargetPrediction and PharmMapper were used to obtain the potential targets of Dendrobin A, and the differentially expressed genes (DEGs) between PDAC and normal pancreatic tissues were obtained from The Cancer Genome Atlas and Genotype-Tissue Expression databases. The protein-protein interaction (PPI) network for Dendrobin A anti-PDAC targets was constructed based on the STRING database. Molecular docking was used to assess Dendrobin A anti-PDAC targets. PLAU, one of the key targets of Dendrobin A anti-PDAC, was immunohistochemically stained in clinical tissue arrays. Finally, in vitro experiments were used to validate the effects of Dendrobin A on PLAU expression and the proliferation, apoptosis, cell cycle, migration, and invasion of PDAC cells. Results: A total of 90 genes for Dendrobin A anti-PDAC were screened, and a PPI network for Dendrobin A anti-PDAC targets was constructed. Notably, a scale-free module with 19 genes in the PPI indicated that the PPI is highly credible. Among these 19 genes, PLAU was positively correlated with the cachexia status while negatively correlated with the overall survival of PDAC patients. Through molecular docking, Dendrobin A was found to bind to PLAU, and the Dendrobin A treatment led to an attenuated PLAU expression in PDAC cells. Based on clinical tissue arrays, PLAU protein was highly expressed in PDAC cells compared to normal controls, and PLAU protein levels were associated with the differentiation and lymph node metastatic status of PDAC. In vitro experiments further showed that Dendrobin A treatment significantly inhibited the proliferation, migration, and invasion, inducing apoptosis and arresting the cell cycle of PDAC cells at the G2/M phase. Conclusion: Dendrobin A, a representative active ingredient of D. nobile, can effectively fight against PDAC by targeting PLAU. Our results provide the foundation for future PDAC treatment based on D. nobile.

Integrating Network Pharmacology and Bioinformatics to Explore the Effects of Dangshen (Codonopsis pilosula) Against Hepatocellular Carcinoma: Validation Based on the Active Compound Luteolin.

Purpose: This study aimed to explore the pharmacological mechanism of Dangshen (Codonopsis pilosula) against hepatocellular carcinoma (HCC) based on network pharmacology and bioinformatics, and to verify the anticancer effect of luteolin, the active ingredient of Codonopsis pilosula, on HCC cells. Methods: The effective compounds and potential targets of Codonopsis pilosula were established using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database. The genes related to HCC were obtained through the GeneCards database. The interactive genes were imported into the Visualization and Integrated Discovery database for Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) signal enrichment, and the hub genes were screened out. The Cancer Genome Atlas database was used to construct a prognosis model, and the prognosis and clinicopathological correlation were analyzed. In in vitro experiments, we verified the effects of luteolin, an active compound of Codonopsis pilosula, on the proliferation, cell cycle, apoptosis and migration of HCC cells. Results: A total of 21 effective compounds of Codonopsis pilosula and 98 potential downstream target genes were screened through the TCMSP database, and 1406 HCC target genes were obtained through the GeneCards database. Finally, 53 interacting genes between the two databases were obtained, among which, the 10 key node genes were CASP3, TP53, MDM2, AKT1, ESR1, BCL2L1, MCL1, HSP90AA1, CASP9, and CCND1, involving 77 typical GO terms and 72 KEGG signals. The Kaplan-Meier survival curve of the model group showed that the overall survival of the low-risk group was significantly higher than that of the high-risk group. Luteolin significantly inhibited the proliferation and migration of HCC cells, induced apoptosis, and increased the G2/M phase ratio. Mechanistically, luteolin significantly inhibited the phosphorylation of MAPK-JNK and Akt (Thr308) and subsequently led to upregulation of ESR1. Pharmacological inhibition of ESR1 with fulvestrant enhanced cell viability and migration and attenuated apoptosis. Conclusion: Codonopsis pilosula has potential for clinical development due to its anti-HCC properties. Luteolin, the effective component of Codonopsis pilosula, plays anti-HCC role through AKT- or MAPK-JNK signaling mediated ESR1.

Protective effects of endothelial progenitor cell microvesicles carrying miR­98­5p on angiotensin II­induced rat kidney cell injury.

With the increasing number of patients with hypertensive nephropathy worldwide, it has posed a major threat to health and studies on its treatment and pathogenesis are imminent. The present study investigated the mechanism through which microRNA (miR)-98-5p in microvesicles (MVs) secreted by endothelial progenitor cells (EPCs) is involved in the repair of angiotensin II (Ang II)-induced injury of rat primary renal kidney cells (PRKs). After isolation of rat renal cortical sections, PRKs were isolated by density gradient centrifugation and identified by immunofluorescence staining. Transmission electron microscopy identifies successful separation of Mvs. An in vitro cell injury model was constructed using Ang II. The Gene Expression Omnibus was used to analyze the differentially expressed genes between diabetic rats and normal rats, and the Kyoto Encyclopedia of Genes and Genomes was used to analyze the signaling pathways involved in these differentially expressed genes. Reverse transcription-quantitative PCR was used to analyze the effect of EPC-MVs on the expression of miRNA induced by Ang II, and the levels of target genes and signaling pathway-related proteins involved were analyzed by western blot. luciferase was used to detect the targeted binding of miR-98-5p to insulin-like growth factor 1 receptor (IGF1R). Enzyme-linked immunosorbent assay was used to analyze the effect of EPC-MVs on Ang II-induced oxidative stress and inflammation levels on PRKs. Cell Counting Kit-8 was used to analyze the effect of EPC-MVs on the cell viability of PRKs induced by Ang II. The results showed that treatment of PRKs with Ang II decreased cell viability, whereas oxidative stress and inflammation were increased. However, EPC-MVs alleviated Ang II-induced damage of the PRKs. During this process, the Ang-II-induced downregulation of miR-98-5p was reversed by EPC-MVs, so miR-98-5p may be a key factor regulating the action of EPC-MVs. Dual-luciferase assay confirmed that miR-98-5p targets IGF1R. It was subsequently demonstrated that EPC-MVs overexpressing miR-98-5p promoted phosphorylation of PI3K/Akt/endothelial nitric oxide synthase (eNOS), and inhibited the oxidative stress and inflammation in PRKs, which were reversed by the overexpression of IGF1R. In conclusion, the results of the present study demonstrated that EPC-MVs with high expression of miR-98-5p can activate the PI3K/Akt/eNOS pathway by regulating IGF1R, as well as protect PRKs from Ang II-induced oxidative stress, inflammation and inhibition of cell viability.

Comprehensive Analysis of the Carcinogenic Process, Tumor Microenvironment, and Drug Response in HPV-Positive Cancers.

Human papillomavirus (HPV) is a common virus, and about 5% of all cancers worldwide is caused by persistent high-risk HPV infections. Here, we reported a comprehensive analysis of the molecular features for HPV-related cancer types using TCGA (The Cancer Genome Atlas) data with HPV status. We found that the HPV-positive cancer patients had a unique oncogenic process, tumor microenvironment, and drug response compared with HPV-negative patients. In addition, HPV improved overall survival for the four cancer types, namely, cervical squamous cell carcinoma (CESC), head and neck squamous cell carcinoma (HNSC), stomach adenocarcinoma (STAD), and uterine corpus endometrial carcinoma (UCEC). The stronger activity of cell-cycle pathways and lower driver gene mutation rates were observed in HPV-positive patients, which implied the different carcinogenic processes between HPV-positive and HPV-negative groups. The increased activities of immune cells and differences in metabolic pathways helped explain the heterogeneity of prognosis between the two groups. Furthermore, we constructed HPV prediction models for different cancers by the virus infection score (VIS) which was linearly correlated with HPV load and found that VIS was associated with drug response. Altogether, our study reveals that HPV-positive cancer patients have unique molecular characteristics which help the development of precision medicine in HPV-positive cancers.

Protective effects of endothelial progenitor cell microvesicles on Ang II­induced rat kidney cell injury.

Chronic hypertension can lead to kidney damage, known as hypertensive nephropathy or hypertensive nephrosclerosis. Further understanding of the molecular mechanisms via which hypertensive nephropathy develops is essential for effective diagnosis and treatment. The present study investigated the mechanisms by which endothelial progenitor cells (EPCs) repair primary rat kidney cells (PRKs). ELISA, Cell Counting Kit­8 and flow cytometry assays were used to analyze the effects of EPCs or EPC­MVs on the oxidative stress, inflammation, cell proliferation, apoptosis and cycle of PRKs induced by AngII. A PRK injury model was established using angiotensin II (Ang II). After Ang II induction, PRK proliferation was decreased, apoptosis was increased and the cell cycle was blocked at the G1 phase before entering the S phase. It was found that the levels of reactive oxygen species and malondialdehyde were increased, while the levels of glutathione peroxidase and superoxide dismutase were decreased. Moreover, the levels of the inflammatory cytokines IL­1ß, IL­6 and TNF­α were significantly increased. Thus, Ang II damaged PRKs by stimulating oxidative stress and promoting the inflammatory response. However, when PRKs were co­cultured with EPCs, the damage induced by Ang II was significantly reduced. The current study collected the microvesicles (MVs) secreted by EPCs and co­cultured them with Ang II­induced PRKs, and identified that EPC­MVs retained their protective effect on PRKs. In conclusion, EPCs protect PRKs from Ang II­induced damage via secreted MVs.

Pathologic role of peptidyl-prolyl isomerase Pin1 in pulmonary artery remodeling.

Peptidyl-prolyl isomerase Pin1 is crucial for cell proliferation, but its role in pulmonary artery remodeling (PAR) is unclear. In the present study, we aimed to evaluate the expression and contribution of Pin1 in PAR. Treatment with Pin1 inhibitor Juglone or Pin1-specific siRNAs ameliorated the expression of Pin1 and proliferating cell nuclear antigen (PCNA) in human pulmonary artery smooth muscle cells (PASMCs) in vitro, and Juglone treatment arrested the cell cycle at the G1 phase. Treatment with transforming growth factor ß1 (TGF-ß1) also enhanced Pin1 expression and PASMC proliferation. Immunohistochemical staining revealed that Pin1 and PCNA expression levels were increased and positively correlated with each other in PAR samples from humans and monocrotaline-treated Sprague-Dawley rats; these proteins were mainly localized in arteries undergoing remodeling, as well as inflammatory cells, and hyperplastic bronchial epithelial cells. Intraperitoneal injection of Juglone also led to morphologic and hemodynamic changes in PAR rats. Additionally, PAR rats displayed higher serum and lung TGF-ß1 levels compared with controls, while administration of Juglone to PAR rats suppressed serum and lung TGF-ß1 levels. The findings in this study suggest that TGF-ß1 and Pin1 constitute a positive feedback loop, which plays an important role in the pathophysiology of PAR.

Dynamic programming for solving a simulated clinical scenario of sepsis resuscitation.

BACKGROUND: A major challenge in clinical research is population heterogeneity and we need to consider both historical response and current condition of an individual in considering medical decision making. The idea of precise medicine cannot be fully accounted for in traditional randomized controlled trials. Reinforcement learning (RL) is developing rapidly and has found its way into various fields including clinical medicine in which RL is employed to find an optimal treatment strategy. The key idea of RL is to optimize the treatment policy depending on the current state and previous treatment history, which is consistent with the idea behind dynamic programming (DP). DP is a prototype of RL and can be implemented when the system dynamics can be fully quantified. METHODS: The present article aims to illustrate how to perform DP algorithm in a clinical scenario of Sepsis resuscitation. The state transition dynamics are constructed in the framework of Markov Decision Process. The state space is defined by mean arterial pressure (MAP) and lactate; the action space is comprised of fluid administration and vasopressor. The implementation of policy evaluation, policy improvement and iteration are explained with R code. RESULTS: the DP algorithm was able to find the optimal treatment policy depending on the current states and previous conditions. The iteration process converged at finite steps. We defined several functions such as nextStep(), policyEval() and policy_iteration() to implement the DP algorithm. CONCLUSIONS: This article illustrates how DP can be used to solve a clinical problem. We show that DP is a potential useful tool to tailor treatment strategy to patients with different conditions/states. Potential audience of the paper are those who are interested in using DP for solving clinical problems with dynamic changing states.

Repurposing antitussive benproperine phosphate against pancreatic cancer depends on autophagy arrest.

Pancreatic cancer (PC) is one of the most common human malignancies worldwide and remains a major clinical challenge. Here, we found that benproperine phosphate (BPP), a cough suppressant, showed a significant anticancer effect on PC both in vitro and in vivo via the induction of autophagy-mediated cell death. Mechanistic studies revealed that BPP triggered AMPK/mTOR-mediated autophagy initiation and disturbed Ras-related protein Rab-11A (RAB11A)-mediated autophagosome-lysosome fusion, resulting in excessive accumulation of autophagosomes. Inhibition of autophagy or overexpression of RAB11A partially reversed BPP-induced growth inhibition in PC cells, suggesting that BPP might induce lethal autophagy arrest in PC cells. In conclusion, our results identify BPP as a potent antitumor agent for PC via the induction of autophagy arrest, therefore providing a new potential therapeutic strategy for the treatment of PC.

Improving Performance and Adaptivity of Anchor-Based Detector Using Differentiable Anchoring With Efficient Target Generation.

Most anchor-based object detection methods have adopted predefined anchor boxes as regression references. However, the proper setting of anchor boxes may vary significantly across different datasets, improperly designed anchors severely limit the performances and adaptabilities of detectors. Recently, some works have tackled this problem by learning anchor shapes from datasets. However, all of these works explicitly or implicitly rely on predefined anchors, limiting universalities of detectors. In this paper, we propose a simple learning anchoring scheme with an effective target generation method to cast off predefined anchor dependencies. The proposed anchoring scheme, named as differentiable anchoring, simplifies learning anchor shape process by adding only one branch in parallel with the existing classification and bounding box regression branches. The proposed target generation method, including the Lp norm ball approximation and the optimization difficulty-based pyramid level assignment approach, generates positive samples for the new branch. Compared with existing learning anchoring-based approaches, the proposed method doesn't require any predefined anchors, while tremendously improving performances and adaptiveness of detectors. The proposed method can be seamlessly integrated to Faster RCNN, RetinaNet, and SSD, improving the detection mAP by 2.8%, 2.1% and 2.3% respectively on MS COCO 2017 test-dev set. Moreover, the differentiable anchoring-based detectors can be directly applied to specific scenarios without any modification of the hyperparameters or using a specialized optimization. Specifically, the differentiable anchoring-based RetinaNet achieves very competitive performances on tiny face detection and text detection tasks, which are not well handled by the conventional and guided anchoring based RetinaNets for the MS COCO dataset.