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Abstract Background Tactile and mechanical pain are crucial to our interaction with the environment, yet the underpinning molecular mechanism is still elusive. Endophilin A2 (EndoA2) is an evolutionarily conserved protein that is documented in the endocytosis pathway. However, the role of EndoA2 in the regulation of mechanical sensitivity and its underlying mechanisms are currently unclear. Methods Male and female C57BL/6 mice (8–12 weeks) and male cynomolgus monkeys (7–10 years old) were used in our experiments. Nerve injury-, inflammatory-, and chemotherapy-induced pathological pain models were established for this study. Behavioral tests of touch, mechanical pain, heat pain, and cold pain were performed in mice and nonhuman primates. Western blotting, immunostaining, co-immunoprecipitation, proximity ligation and patch-clamp recordings were performed to gain insight into the mechanisms. Results The results showed that EndoA2 was primarily distributed in neurofilament-200-positive (NF200+) medium-to-large diameter dorsal root ganglion (DRG) neurons of mice and humans. Loss of EndoA2 in mouse NF200+ DRG neurons selectively impaired the tactile and mechanical allodynia. Furthermore, EndoA2 interacted with the mechanically sensitive ion channel Piezo2 and promoted the membrane trafficking of Piezo2 in DRG neurons. Moreover, as an adaptor protein, EndoA2 also bound to kinesin family member 5B (KIF5B), which was involved in the EndoA2-mediated membrane trafficking process of Piezo2. Loss of EndoA2 in mouse DRG neurons damaged Piezo2-mediated rapidly adapting mechanically activated currents, and re-expression of EndoA2 rescued the MA currents. In addition, interference with EndoA2 also suppressed touch sensitivity and mechanical hypersensitivity in nonhuman primates. Conclusions Our data reveal that the KIF5B/EndoA2/Piezo2 complex is essential for Piezo2 trafficking and for sustaining transmission of touch and mechanical hypersensitivity signals. EndoA2 regulates touch and mechanical allodynia via kinesin-mediated Piezo2 trafficking in sensory neurons. Our findings identify a potential new target for the treatment of mechanical pain.

Exchange programs have been found to have a positive impact on regional economic growth, university cooperation, and student academic, and professional development. However, there has been limited analysis of the factors that influence the quality of these programs. In this study, a mixed-methods approach was used to examine regional exchange programs in China. Qualitative research was conducted using Octopus Big Data Crawler software, Timdream.org software, and 33 interviews to develop a questionnaire that covers student expectations, service quality, and satisfaction. Using SPSS 22 and Amos 21, the study analyzed the interactive relationships among expectations, perceived service quality, and satisfaction in the context of regional education integration based on a sample of 246 questionnaires. The results demonstrate that perceived service quality plays an intermediary role in the relationship between expectations and satisfaction, with students' perceptions of learning quality and interpersonal quality at the receiving university influencing their satisfaction levels. This study provides important insights into the factors that impact the quality of regional exchange programs in China.

Abstract Hydroxycarboxylic acids are crucial metabolic intermediates involved in various physiological and pathological processes, some of which are recognized by specific hydroxycarboxylic acid receptors (HCARs). HCAR2 is one such receptor, activated by endogenous β-hydroxybutyrate (3-HB) and butyrate, and is the target for Niacin. Interest in HCAR2 has been driven by its potential as a therapeutic target in cardiovascular and neuroinflammatory diseases. However, the limited understanding of how ligands bind to this receptor has hindered the development of alternative drugs able to avoid the common flushing side-effects associated with Niacin therapy. Here, we present three high-resolution structures of HCAR2-Gi1 complexes bound to four different ligands, one potent synthetic agonist (MK-6892) bound alone, and the two structures bound to the allosteric agonist compound 9n in conjunction with either the endogenous ligand 3-HB or niacin. These structures coupled with our functional and computational analyses further our understanding of ligand recognition, allosteric modulation, and activation of HCAR2 and pave the way for the development of high-efficiency drugs with reduced side-effects.

Abstract Background The vast majority of patients with ROS1 positive non‐small cell lung cancer (NSCLC) derive clinical benefit from currently approved ROS1 therapies, including crizotinib and entrectinib. However, a small proportion of patients treated with ROS1 inhibitors fail to derive any clinical benefit and demonstrate rapid disease progression. The biological mechanisms underpinning intrinsic resistance remain poorly understood for oncogene‐driven cancers. Methods We generated a patient‐derived cell line, CUTO33, from a ROS1 therapy naive patient with CD74‐ROS1+ NSCLC, who ultimately did not respond to a ROS1 inhibitor. We evaluated a panel of ROS1+ patient‐derived NSCLC cell lines and used cell‐based assays to determine the mechanism of intrinsic resistance to ROS1 therapy. Results The CUTO33 cell line expressed the CD74‐ROS1 gene fusion at the RNA and protein level. The ROS1 fusion protein was phosphorylated at baseline consistent with the known intrinsic activity of this oncogene. ROS1 phosphorylation could be inhibited using a wide array of ROS1 inhibitors, however these inhibitors did not block cell proliferation, confirming intrinsic resistance in this model and consistent with the patient's lack of response to a ROS1 inhibitor. CUTO33 expressed high levels of AXL, which has been associated with drug resistance. Combination of an AXL inhibitor or AXL knockdown with a ROS1 inhibitor partially reversed resistance. Conclusions In summary, we demonstrate that AXL overexpression is a mechanism of intrinsic resistance to ROS1 inhibitors.

Morphine is a frequently used analgesic that activates the mu-opioid receptor (MOR), which has prominent side effects of tolerance. Although the inefficiency of morphine in inducing the endocytosis of MOR underlies the development of morphine tolerance, currently, there is no effective therapy to treat morphine tolerance. In the current study, we aimed to develop a monoclonal antibody (mAb) precisely targeting MOR and to determine its therapeutic efficacy on morphine tolerance and the underlying molecular mechanisms. We successfully prepared a mAb targeting MOR, named 3A5C7, by hybridoma technique using a strategy of deoxyribonucleic acid immunization combined with cell immunization, and identified it as an immunoglobulin G mAb with high specificity and affinity for MOR and binding ability to antigens with spatial conformation. Treatment of two cell lines, HEK293T and SH-SY5Y, with 3A5C7 enhanced morphine-induced MOR endocytosis via a G protein-coupled receptor kinase 2 (GRK2)/β-arrestin2-dependent mechanism, as demonstrated by immunofluorescence staining, flow cytometry, Western blotting, coimmunoprecipitation, and small interfering ribonucleic acid (siRNA)-based knockdown. This mAb also allowed MOR recycling from cytoplasm to plasma membrane and attenuated morphine-induced phosphorylation of MOR. We established an in vitro morphine tolerance model using differentiated SH-SY5Y cells induced by retinoic acid. Western blot, enzyme-linked immunosorbent assays, and siRNA-based knockdown revealed that 3A5C7 mAb diminished hyperactivation of adenylate cyclase, the in vitro biomarker of morphine tolerance, via the GRK2/β-arrestin2 pathway. Furthermore, in vivo hotplate test demonstrated that chronic intrathecal administration of 3A5C7 significantly alleviated morphine tolerance in mice, and withdrawal jumping test revealed that both chronic and acute 3A5C7 intrathecal administration attenuated morphine dependence. Finally, intrathecal electroporation of silencing short hairpin RNA illustrated that the in vivo anti-tolerance and anti-dependence efficacy of 3A5C7 was mediated by enhanced morphine-induced MOR endocytosis via GRK2/β-arrestin2 pathway. Collectively, our study provided a therapeutic mAb, 3A5C7, targeting MOR to treat morphine tolerance, mediated by enhancing morphine-induced MOR endocytosis. The mAb 3A5C7 demonstrates promising translational value to treat clinical morphine tolerance.

Abstract The hierarchical equations of motion (HEOM) approach can describe the reduced dynamics of a system simultaneously coupled to multiple bosonic and fermionic environments. The complexity of exactly describing the system-environment interaction with the HEOM method usually results in time-consuming calculations and a large memory cost. Here, we introduce an open-source software package called HierarchicalEOM.jl: a Julia framework integrating the HEOM approach. HierarchicalEOM.jl features a collection of methods to compute bosonic and fermionic spectra, stationary states, and the full dynamics in the extended space of all auxiliary density operators (ADOs). The required handling of the ADOs multi-indexes is achieved through a user-friendly interface. We exemplify the functionalities of the package by analyzing a single impurity Anderson model, and an ultra-strongly coupled charge-cavity system interacting with bosonic and fermionic reservoirs. HierarchicalEOM.jl achieves a significant speedup with respect to the corresponding method in the Quantum Toolbox in Python (QuTiP), upon which this package is founded.

This paper proposes a solution for the challenges faced in predicting the output of distributed solar power stations due to difficulties in obtaining and utilizing data. The proposed method is a multi-scale regional photovoltaic power generation forecasting approach that uses sequence coding reconstruction. Firstly, the reference sites were selected to enhance the data in time domain and frequency domain for the limited site dataset. Then, the forecasting model of sequence coding reconstruction is constructed based on convLSTM, and the attention mechanism is added in the decoding process, so that it can make full use of the spatio-temporal correlation characteristics of regional photovoltaic power and realize the rolling forecasting of regional photovoltaic power. The result shows that the proposed method can maintain a good multi-scale regional photovoltaic power forecasting effect under the condition of limited data amount and low data collection cost, the accuracy of the proposed model is increased compared with the naive forecasting method.

Live streaming e-commerce has emerged as a novel online marketing model. Drawing upon influencer marketing theory, this study examines the mechanisms through which influencers (live streamers) promote consumers’ purchase intention in the context of live streaming e-commerce. A sample of 449 valid questionnaires was utilized to test the proposed theoretical framework. The empirical research findings reveal that customer experience significantly and positively impacts both influencer trust and influencer attachment. Furthermore, trust and attachment established with live streamers are identified as two effective mechanisms influencing consumer decision-making. Notably, influencer attachment exhibits a stronger influence on consumer purchase intention compared to influencer trust. By comparing the effects of Taobao and Douyin live streamers on stimulating consumption and purchase intention, the study demonstrates that live streamers play a crucial mediating role between customer experience and consumer purchase intention. Specifically, the results indicate that consumer purchase intention influenced by top Taobao streamers is stronger than that of Douyin streamers, whereas influencer attachment for Taobao streamers is relatively weaker than that for Douyin streamers. These findings provide theoretical and managerial implications for platforms and live streamers seeking to stimulate robust purchase intentions among consumers by fostering attachment relationships. The establishment of an emotional connection between the live streamer and the audience proves particularly valuable in increasing purchase intention. This research contributes to the understanding of the underlying mechanisms driving consumer behavior in the context of live streaming e-commerce. It emphasizes the significance of customer experience, influencer trust, and influencer attachment as key drivers of consumer purchase intention. The findings offer valuable insights for platforms and live streamers to optimize their strategies and enrich user data labels in order to enhance consumer engagement and stimulate purchase intentions. Ultimately, this research contributes to the advancement of the live streaming e-commerce field, strengthens the application of data elements in live streaming e-commerce marketing, and guides effective decision-making by industry practitioners.

Abstract Background The mechanism of hepatitis B virus (HBV)‐induced carcinogenesis remains an area of interest. The accumulation of hepatitis B surface antigen in the endoplasmic reticulum (ER) of hepatocytes stimulates persistent ER stress. Activity of the unfolded protein response (UPR) pathway of ER stress may play an important role in inflammatory cancer transformation. How the protective UPR pathway is hijacked by cells as a tool for malignant transformation in HBV‐related hepatocellular carcinoma (HCC) is still unclear. Here, we aimed to define the key molecule hyaluronan‐mediated motility receptor (HMMR) in this process and explore its role under ER stress in HCC development. Methods An HBV‐transgenic mouse model was used to characterize the pathological changes during the tumor progression. Proteomics and transcriptomics analyses were performed to identify the potential key molecule, screen the E3 ligase, and define the activation pathway. Quantitative real‐time PCR and Western blotting were conducted to detect the expression of genes in tissues and cell lines. Luciferase reporter assay, chromatin immunoprecipitation, coimmunoprecipitation, immunoprecipitation, and immunofluorescence were employed to investigate the molecular mechanisms of HMMR under ER stress. Immunohistochemistry was used to clarify the expression patterns of HMMR and related molecules in human tissues. Results We found sustained activation of ER stress in the HBV‐transgenic mouse model of hepatitis‐fibrosis‐HCC. HMMR was transcribed by c/EBP homologous protein (CHOP) and degraded by tripartite motif containing 29 (TRIM29) after ubiquitination under ER stress, which caused the inconsistent expression of mRNA and protein. Dynamic expression of TRIM29 in the HCC progression regulated the dynamic expression of HMMR. HMMR could alleviate ER stress by increasing autophagic lysosome activity. The negative correlation between HMMR and ER stress, positive correlation between HMMR and autophagy, and negative correlation between ER stress and autophagy were verified in human tissues. Conclusions This study identified the complicated role of HMMR in autophagy and ER stress, that HMMR controls the intensity of ER stress by regulating autophagy in HCC progression, which could be a novel explanation for HBV‐related carcinogenesis.

Nationally determined contributions raised by Paris Agreement aim to control the temperature rise below 2 °C or even 1.5 °C at the end of the 21st century, compared to pre-industrial levels. However, the climate response of the Nationally Determined Contributions (NDCs) remains uncertain due to unstable policies and their credibility. In this study, we calculated the uncertainty of global temperature rise caused by uncertain NDCs and carbon-neutral policies and discussed the difficulty of policy implementation. The results show that there will be 8 GtC uncertainty in emission at the end of the 21st century, responsible for the temperature rise of 0.37 °C (1.73–2.10 °C). A delayed policy in emission reduction by major emitters would result in a temperature rise of over 2 °C, while under non-delay policy, the 2 °C target will be possibly achieved. Besides, low-emission countries would introduce a 30 GtC cumulative emission uncertainty at the end of the 21st century if there are no restrictions, leading to a 0.3 °C global warming uncertainty. Developed countries need more substantial reductions in carbon intensity to achieve their climate policies while developing countries are under less pressure. The reduction of carbon intensity requires the strengthening of technical and economic methods. This study provides a reference for the realization of emission policies and temperature rise targets.

Abstract The low-temperature thermoelectric performance of Bi-rich n-type Mg3(Bi,Sb)2 was limited by the electron transport scattering at grain boundaries, while removing grain boundaries and bulk crystal growth of Mg-based Zintl phases are challenging due to the volatilities of elemental reactants and their severe corrosions to crucibles at elevated temperatures. Herein, for the first time, we reported a facile growth of coarse-grained Mg3Bi2-x Sb x crystals with an average grain size of ~800 μm, leading to a high carrier mobility of 210 cm2 · V−1 · s−1 and a high z of 2.9 × 10−3 K−1 at 300 K. A $$\Delta$$ Δ T of 68 K at T h of 300 K, and a power generation efficiency of 5.8% below 450 K have been demonstrated for Mg3Bi1.5Sb0.5- and Mg3Bi1.25Sb0.75-based thermoelectric modules, respectively, which represent the cutting-edge advances in the near-room temperature thermoelectrics. In addition, the developed grain growth approach can be potentially extended to broad Zintl phases and other Mg-based alloys and compounds.

Abstract Bentonite is a key material for engineering barriers to prevent groundwater and nuclide migration in the multi-barrier system of high-level radioactive waste (HLW) geological disposal. However, its barrier property will be degraded under the action of iron corrosion products of steel disposal containers. In this paper, the effects of iron corrosion products on the degradation of bentonite structure and properties were investigated in the simulated environments for HLW geological disposal. The results showed that Fe2+/Fe3+ dissolved from iron powder could enter montmorillonite (Mt) interlayer and substituted part of Na+, which caused the decrease of the volume and interlayer spacing of Mt, and the structural integrity of Mt was destroyed. Macroscopically, the water absorption and swelling property of bentonite were significantly decreased. The degradation mechanism of Mt structure was mainly that Fe2+/Fe3+ generated by iron corrosion entered the interlayer domain of Mt to compensate for the interlayer charge deficit.

Abstract Background Recently, periumbilical fat (PF) mass, an autologous material with a high survival rate, has been transplanted to treat sunken or dissatisfactory double eyelids. However, the intricate complications of PF grafts and associated reconstructive strategies are infrequently discussed. Methods During 3 years, 20 patients (33 eyes) with eyelid malformations caused by PF grafts into the orbital septum or on the surface of the levator aponeurosis underwent corrective blepharoplasty. We recorded patients’ subjective feelings and identified deformities from crease abnormalities, bloated appearance, and problems with the eyelid’s height. Then, we categorize them into three types based on their complexity: type I, swollen appearance; type II, obvious adhesion; type III, severe comprehensive damage. The relevant management included removing fat implants, releasing the adhesion, and rebuilding the physical structure according to the anatomic damage mechanism. The improvement effect was assessed with a satisfaction survey from patients and doctors at 6 months of follow-up. Results The swollen appearance was observed in 26 eyes (78.8%), an unsmooth double-eyelid line was in 23 eyes (69.7%), and the incidence of adhesion was in 22 eyes (66.7%). Following a comprehensive evaluation, 15 eyes (45.5%) and 13 (39.4%) were classified as type I and type II respectively. After the 6-month follow-up, 22 eyes (66.7%) showed exceptional aesthetic results, whereas only 2 eyes as type III had a poor outcome. Conclusions The deformities emerging from periumbilical fat into the upper eyelid are associated with the shape of the fat and the adhesion in tissues. Graft removal, adhesion release, and restoration of the natural anatomic structure can have positive outcomes.

Abstract Thoracic aortic aneurysms (TAAs) develop asymptomatically and are characterized by dilatation of the aorta. This is considered a life-threating vascular disease due to the risk of aortic rupture and without effective treatments. The current understanding of the pathogenesis of TAA is still limited, especially for sporadic TAAs without known genetic mutation. Sirtuin 6 (SIRT6) expression was significantly decreased in the tunica media of sporadic human TAA tissues. Genetic knockout of Sirt6 in mouse vascular smooth muscle cells accelerated TAA formation and rupture, reduced survival, and increased vascular inflammation and senescence after angiotensin II infusion. Transcriptome analysis identified interleukin (IL)-1β as a pivotal target of SIRT6, and increased IL-1β levels correlated with vascular inflammation and senescence in human and mouse TAA samples. Chromatin immunoprecipitation revealed that SIRT6 bound to the Il1b promoter to repress expression partly by reducing the H3K9 and H3K56 acetylation. Genetic knockout of Il1b or pharmacological inhibition of IL-1β signaling with the receptor antagonist anakinra rescued Sirt6 deficiency mediated aggravation of vascular inflammation, senescence, TAA formation and survival in mice. The findings reveal that SIRT6 protects against TAA by epigenetically inhibiting vascular inflammation and senescence, providing insight into potential epigenetic strategies for TAA treatment.

Abstract Background Pediatric interventional oncology (PIO) is a growing field intended to provide additional or alternative treatment options for pediatric patients with benign or malignant tumors. Large series of patients treated uniformly and subjected to rigorous endpoints for efficacy are not available. Methods We designed a collaborative initiative to capture data from pediatric patients with benign and malignant tumors who underwent a therapeutic interventional radiology procedure. Modified Response Evaluation Criteria in Solid Tumors (mRECIST) was utilized as a measure of radiologic response and data were collected regarding improvement in pain and functional endpoints. Cumulative incidence of progressive disease was calculated using both the treated site and the patient as the analytic unit. Findings Forty patients, 16 with malignant tumors and 24 with benign tumors, underwent a total of 88 procedures. Cryo‐ and radiofrequency ablation were the most frequently utilized techniques for both cohorts of patients. A complete or partial response, or prolonged disease stability, were achieved in approximately 40% of patients with malignant tumors and 60% of patients with benign tumors. No patients had progressive disease as their best response. Resolution of pain and improved mobility with return‐to‐baseline activity were demonstrated across patients from both cohorts. Only minor complications were experienced. Interpretation Interventional radiology‐guided interventions can serve as an alternative or complementary approach to the treatment of benign and malignant tumors in pediatric patients. Prospective, multi‐institutional trials are required to adequately study utility, treatment endpoints, and durability of response.

Glioblastoma (GBM) is a lethal cancer with limited therapeutic options. Dendritic cell (DC)-based cancer vaccines provide a promising approach for GBM treatment. Clinical studies suggest that other immunotherapeutic agents may be combined with DC vaccines to further enhance antitumor activity. Here, we report a GBM case with combination immunotherapy consisting of DC vaccines, anti-programmed death-1 (anti-PD-1) and poly I:C as well as the chemotherapeutic agent cyclophosphamide that was integrated with standard chemoradiation therapy, and the patient remained disease-free for 69 months. The patient received DC vaccines loaded with multiple forms of tumor antigens, including mRNA-tumor associated antigens (TAA), mRNA-neoantigens, and hypochlorous acid (HOCl)-oxidized tumor lysates. Furthermore, mRNA-TAAs were modified with a novel TriVac technology that fuses TAAs with a destabilization domain and inserts TAAs into full-length lysosomal associated membrane protein-1 to enhance major histocompatibility complex (MHC) class I and II antigen presentation. The treatment consisted of 42 DC cancer vaccine infusions, 26 anti-PD-1 antibody nivolumab administrations and 126 poly I:C injections for DC infusions. The patient also received 28 doses of cyclophosphamide for depletion of regulatory T cells. No immunotherapy-related adverse events were observed during the treatment. Robust antitumor CD4+ and CD8+ T-cell responses were detected. The patient remains free of disease progression. This is the first case report on the combination of the above three agents to treat glioblastoma patients. Our results suggest that integrated combination immunotherapy is safe and feasible for long-term treatment in this patient. A large-scale trial to validate these findings is warranted.

Abstract Forest musk deer (Moschus berezovskii) are currently a threatened species under conservation, and the development of captive populations is restricted by health problems. To evaluate the application potential of interferon (IFN)-ω in the prevention and control of forest musk deer disease, 5 forest musk deer IFN-ω (fmdIFNω) gene sequences were successfully obtained by homologous cloning method for the first time. FmdIFNω5 was selected and recombinant fmdIFNω protein (rIFNω) was successfully expressed by pGEX-6P-1 plasmid and E. coli expression system. The obtained protein was used to stimulate forest musk deer lung fibroblasts cells FMD-C1 to determine its regulatory effect on interferon-stimulated genes (ISGs). In addition, an indirect ELISA method based on anti-rIFNω serum was established to detect endogenous IFN-ω levels in 8 forest musk deer. The results showed that there were 18 amino acid differences among the 5 fmdIFNω subtypes, all of which had the basic structure to exert the activity of type I IFN and were close to Cervus elaphus IFN-ω in the phylogenetic tree. The protein expressed was 48 kDa, and the transcription levels of all ISGs were increased in FMD-C1 cells stimulated by rIFNω, and the amount of transcription accumulation was time-dependent. Meanwhile, Anti-rIFNω serum of mice could react with both rIFNω and forest musk deer serum, and the OD450nm value of forest musk deer serum with the most obvious symptoms was the highest, suggesting that the level of natural IFN-ω in different forest musk deer could be monitored by the rIFNω-based ELISA method. These results indicate that fmdIFNω has the potential as an antiviral drug and an early indication of innate immunity, which is of great significance for the prevention and control of forest musk deer diseases.

Abstract Background Limited data have been used to evaluate the efficacy of immunotherapy in metastatic colorectal cancer (mCRC). Furthermore, potential markers that can be used to identify responding patients and to further improve efficacy have not been fully explored. Methods and Results In our study, we included a total of 97 patients with mCRC, who each received programmed death‐1 (PD‐1) inhibitor‐based combination therapy at our center. All 12 hypermutated patients benefited from immunotherapy, with median progression‐free survival (mPFS) reaching 28.3 months, regardless of liver metastasis. The objective response rate (ORR) of non‐hypermutated patients was 16.5% (14/85), with an mPFS of 4.0 months. For non‐hypermutated patients, multivariate analysis revealed that the combination of liver metastasis and baseline lesion number significantly stratified response and survival. The lesion‐based analysis indicated that the lymph node was the most responsive, followed by the peritoneum and lung, with liver metastasis being the least responsive. None of the patients (0/7) with negative programmed ligand‐1 (PD‐L1) expression responded, and positive PD‐L1 expression may serve as a biomarker (mPFS 5.7 vs. 2.2 months, p = 0.002) that can be used to further guide treatment in non‐hypermutated mCRC with liver metastasis (CRLMs). Conclusion Patients with hypermutated mCRC benefited significantly from immunotherapy, whereas the non‐hypermutated cohort with liver metastasis and numerous lesions showed less benefit. The lesion sites reflected varying levels of efficacy, among which PD‐L1 potentially cooperated to guide the immunotherapy of CRLMs.