Response inhibition impairment related to altered frontal-striatal functional connectivity in insomnia disorder: A pilot and non-clinical study.

BACKGROUND: It is not clear whether and how insomnia disorder (ID) impairs response inhibition ability. Fronto-striatal functional connectivity (FC) plays a critical role in response inhibition and is found be abnormal in patients with ID. In this study, we examined whether insomnia symptoms impair response inhibition in a large non-clinical sample and whether impaired response inhibition is related to abnormal fronto-striatal FC. METHODS: One hundred and fifteen young ID patients and 160 age and sex-matched healthy controls (HC) underwent resting-state functional magnetic response imaging scans and performed the stop-signal task (SST). Performance of SST, Gray Matter Volumes (GMVs), and connections of brain regions related to fronto-striatal circuits was compared between groups. Further examined the association between response inhibition impairment and fronto-striatal FC. RESULTS: The behavioral results showed that patients with ID had significantly longer stop-signal reaction time (SSRT) compared with the HC, reflecting the impaired response inhibition among IDs. Brain imaging results showed IDs had decreased GMVs of the Right Superior Frontal (SFG) and left Supplementary Motor area (SMA). Seed-based FC results showed that compared to HC, the ID showed decreased FC between left SMA and left Paracentral lobule, left SMA and right SMA, and right SFG and right Orbital Middle Frontal gyrus, and increased FC between right SFG and right putamen. Meanwhile, the FC between right SFG and putamen was positively correlated with SSRT in IDs. CONCLUSIONS: The current study found significantly impaired response inhibition among ID and this impairment may be related to abnormal fronto-striatal FC in ID.

Traumatic Iliac Arteriovenous Fistula Treated With the Amplatzer Vascular Plug II: A Case Report and Literature Review.

Traumatic iliac arteriovenous fistula is a rare complication of vascular injury. Open surgical repair has an incidence of postoperative complications. In recent years, endovascular treatment has shown better efficacy. We report a 62-year-old female AVF patient with a stab injury history of more than 16 years. Computed tomography angiography (CTA) revealed a large arteriovenous fistula between the right internal iliac artery and the common iliac vein. After considering the patient's relevant conditions, an endovascular approach was satisfactorily performed with the implantation of an Amplatzer Vascular Plug II to interrupt the abnormal vascular communication and maintain arterial and venous patency. The final control images showed closure of the arteriovenous communication.

Spatial heterogeneity of tumor microenvironment influences the prognosis of clear cell renal cell carcinoma.

BACKGROUND: Clear cell renal cell carcinoma (ccRCC) is an immunologically and histologically diverse tumor. However, how the structural heterogeneity of tumor microenvironment (TME) affects cancer progression and treatment response remains unclear. Hence, we characterized the TME architectures of ccRCC tissues using imaging mass cytometry (IMC) and explored their associations with clinical outcome and therapeutic response. METHODS: Using IMC, we profiled the TME landscape of ccRCC and paracancerous tissue by measuring 17 markers involved in tissue architecture, immune cell and immune activation. In the ccRCC tissue, we identified distinct immune architectures of ccRCC tissue based on the mix score and performed cellular neighborhood (CN) analysis to subdivide TME phenotypes. Moreover, we assessed the relationship between the different TME phenotypes and ccRCC patient survival, clinical features and treatment response. RESULTS: We found that ccRCC tissues had higher levels of CD8+ T cells, CD163- macrophages, Treg cells, endothelial cells, and fibroblasts than paracancerous tissues. Immune infiltrates in ccRCC tissues distinctly showed clustered and scattered patterns. Within the clustered pattern, we identified two subtypes with different clinical outcomes based on CN analysis. The TLS-like phenotype had cell communities resembling tertiary lymphoid structures, characterized by cell-cell interactions of CD8+ T cells-B cells and GZMB+CD8+ T cells-B cells, which exhibited anti-tumor features and favorable outcomes, while the Macrophage/T-clustered phenotype with macrophage- or T cell-dominated cell communities had a poor prognosis. Patients with scattered immune architecture could be further divided into scattered-CN-hot and scattered-CN-cold phenotypes based on the presence or absence of immune CNs, but both had a better prognosis than the macrophage/T-clustered phenotype. We further analyzed the relationship between the TME phenotypes and treatment response in five metastatic ccRCC patients treated with sunitinib, and found that all three responders were scattered-CN-hot phenotype while both non-responders were macrophage/T-clustered phenotype. CONCLUSION: Our study revealed the structural heterogeneity of TME in ccRCC and its impact on clinical outcome and personalized treatment. These findings highlight the potential of IMC and CN analysis for characterizing TME structural units in cancer research.

A Blockchain-Based Trustworthy Model Evaluation Framework for Deep Learning and Its Application in Moving Object Segmentation.

Model evaluation is critical in deep learning. However, the traditional model evaluation approach is susceptible to issues of untrustworthiness, including insecure data and model sharing, insecure model training, incorrect model evaluation, centralized model evaluation, and evaluation results that can be tampered easily. To minimize these untrustworthiness issues, this paper proposes a blockchain-based model evaluation framework. The framework consists of an access control layer, a storage layer, a model training layer, and a model evaluation layer. The access control layer facilitates secure resource sharing. To achieve fine-grained and flexible access control, an attribute-based access control model combining the idea of a role-based access control model is adopted. A smart contract is designed to manage the access control policies stored in the blockchain ledger. The storage layer ensures efficient and secure storage of resources. Resource files are stored in the IPFS, with the encrypted results of their index addresses recorded in the blockchain ledger. Another smart contract is designed to achieve decentralized and efficient management of resource records. The model training layer performs training on users' servers, and, to ensure security, the training data must have records in the blockchain. The model evaluation layer utilizes the recorded data to evaluate the recorded models. A method in the smart contract of the storage layer is designed to enable evaluation, with scores automatically uploaded as a resource attribute. The proposed framework is applied to deep learning-based motion object segmentation, demonstrating its key functionalities. Furthermore, we validated the storage strategy adopted by the framework, and the trustworthiness of the framework is also analyzed.

Effects of whole-body vibration on sensorimotor deficits and brain plasticity among people with chronic ankle instability: a study protocol for a single-blind randomized controlled trial.

BACKGROUND: Chronic ankle instability (CAI) is a form of musculoskeletal disease that can occur after a lateral ankle sprain, and it is characterized by pain, recurrent ankle sprains, a feeling of "giving way" at the ankle joint, and sensorimotor deficits. There has been increasing evidence to suggest that plastic changes in the brain after the initial injury play an important role in CAI. As one modality to treat CAI, whole-body vibration (WBV) has been found to be beneficial for treating the sensorimotor deficits accompanying CAI, but whether these benefits are associated with brain plasticity remains unknown. Therefore, the current study aims to investigate the effect of WBV on sensorimotor deficits and determine its correlation with plastic changes in the brain. METHODS: The present study is a single-blind randomized controlled trial. A total of 80 participants with CAI recruited from the university and local communities will be divided into 4 groups: whole-body vibration and balance training (WBVBT), balance training (BT), whole-body vibration (WBV), and control group. Participants will be given the WBV intervention (25-38 Hz, 1.3-2 mm, 3-time per week, 6-week) supervised by a professional therapist. Primary outcome measures are sensorimotor function including strength, balance, proprioception and functional performance. Brain plasticity will be evaluated by corticomotor excitability, inhibition, and representation of muscles, as measured by transcranial magnetic stimulation. Activation of brain areas will be assessed through functional near-infrared spectroscopy. Secondary outcome measures are self-reported functional outcomes involving the Cumberland Ankle Instability Tool and the Foot and Ankle Ability Measure. All tests will be conducted before and after the WBV intervention, and at 2-week follow-up. Per­protocol and intention-to-treat analysis will be applied if any participants withdraw. DISCUSSION: This is the first trial to investigate the role of brain plasticity in sensorimotor changes brought by WBV for individuals with CAI. As plastic changes in the brain have been an increasingly important aspect in CAI, the results of the current study can provide insight into the treatment of CAI from the perspective of brain plasticity. TRIAL REGISTRATION: Chinese Clinical Trial Registry (ChiCTR2300068972); registered on 02 March 2023.

International business in the digital age: Global strategies in a world of national institutions.

Digital technologies are changing how businesses strategize and organize internationally. They not only enable cost reduction in businesses crossing national boundaries but also enable novel types of products and business models. Yet, barriers to cross-border businesses persist or even re-emerge, such that the study of international business remains important in the digital age, but may have to shift focus. We argue that businesses operating internationally develop digital business strategies that are interdependent with their internationalization strategies. In doing so, they have to account for differences across national contexts including informal institutions, formal institutions, and resource endowments. We offer a conceptual framework linking external and internal antecedents to digital business and internationalization strategies. We focus in particular on three digital strategies: owning digital platforms, participating in digital platforms, and transforming traditional businesses for the digital world. On this basis, we discuss the contributions of the papers in this special issue and conclude by outlining an agenda for future research.

Les technologies numériques modifient la manière dont les entreprises élaborent leurs stratégies et s'organisent au niveau international. Elles permettent non seulement de réduire des coûts des entreprises transfrontalières, mais aussi de créer de nouveaux modèles économiques et types de produits. Pourtant, les obstacles aux entreprises transfrontalières persistent, voire réapparaissent, de sorte que la recherche en affaires internationales reste importante à l'ère du numérique, mais doit peut-être changer d'orientation. Nous argumentons que les entreprises opérant au niveau international développent des stratégies d'entreprise numériques qui sont interdépendantes de leurs stratégies d'internationalisation. Ce faisant, elles doivent tenir compte des différences entre les contextes nationaux, incluant les institutions informelles, les institutions formelles et les dotations en ressources. Nous proposons un cadre conceptuel reliant les antécédents externes et internes aux stratégies d'entreprise numériques et d'internationalisation. Nous nous focalisons en particulier sur trois stratégies numériques : posséder des plateformes numériques, participer à des plateformes numériques et transformer des affaires traditionnelles pour le monde numérique. Sur cette base, nous discutons les contributions des articles de ce numéro spécial, et concluons notre article en esquissant un programme de recherches futures.

Las tecnologías digitales están cambiando la forma en que las empresas formulan estrategias y se organizan internacionalmente. Estas no sólo permiten reducir costos en las empresas que cruzan las fronteras nacionales, sino que también habilitan tipos de productos y modelos de negocio novedosos. Con todo, las barreras a las empresas transfronterizas persisten o incluso vuelven a surgir, de modo que el estudio de los negocios internacionales sigue siendo importante en la era digital, pero puede que tenga un cambio de perspectiva. Sostenemos que las empresas que operan internacionalmente desarrollan estrategias de negocio digitales que son interdependientes de sus estrategias de internacionalización. Al hacerlo, deben tener en cuenta las diferencias entre los contextos nacionales, incluidas las instituciones informales, las instituciones formales y el patrimonio de recursos. Ofrecemos un marco conceptual que vincula los antecedentes externos e internos de las estrategias de negocio digital y de internacionalización. Nos centramos en particular en tres estrategias digitales: poseer plataformas digitales, participar en plataformas digitales y transformar los negocios tradicionales para el mundo digital. Sobre esta base, discutimos las contribuciones de los artículos de este número especial y concluimos esquematizando una agenda para futuras investigaciones.

Tecnologias digitais estão mudando como empresas criam estratégias e se organizam internacionalmente. Elas não apenas permitem a redução de custos em negócios que cruzam fronteiras nacionais, mas também permitem novos tipos de produtos e modelos de negócio. No entanto, barreiras para negócios transfronteiriços persistem ou até ressurgem, de modo que o estudo sobre negócios internacionais continua importante na era digital, mas pode precisar mudar o foco. Argumentamos que empresas que operam internacionalmente desenvolvem estratégias de negócios digitais que são interdependentes de suas estratégias de internacionalização. Ao fazê-lo, devem considerar diferenças entre contextos nacionais, incluindo instituições informais, instituições formais e dotações de recursos. Oferecemos um modelo conceptual que liga antecedentes externos e internos aos negócios digitais e às estratégias de internacionalização. Focamos particularmente em três estratégias digitais: possuir plataformas digitais, participar em plataformas digitais e transformar negócios tradicionais para o mundo digital. Com base nisso, discutimos as contribuições dos artigos nesta edição especial e concluímos delineando uma agenda para pesquisas futuras.

SIRT1 promotes glucolipid metabolic conversion to facilitate tumor development in colorectal carcinoma.

Background: Fatty acid oxidation (FAO) is a major alternate energy metabolism pathway in tumor cells subjected to metabolic stress caused by glucose deficiency during rapid progression. However, the mechanism of metabolic reprogramming between glycolysis and FAO in tumor cells is unknown. Therefore, identifying the metabolic glucolipid conversion hub in tumor cells is crucial. Methods: We used single-cell RNA sequencing (scRNA-Seq), RNA sequencing (RNA-Seq), The Cancer Genome Atlas (TCGA), and chromatin immunoprecipitation sequencing (ChIP-Seq) to predict the critical regulator and mechanism of metabolic glucolipid conversion in colorectal cancer (CRC) tumor cells. We used Seahorse metabolic analysis, immunoblotting, immunofluorescence, and immunohistochemical (IHC) technology to verify the prediction and mechanism of this regulator in cancer cell lines, a nude mouse xenograft model, and clinical CRC samples. Results: We demonstrated that sirtuin-1 (SIRT1) was upregulated in CRC cells in response to glucose deprivation and oxidative stress. SIRT1 was also a hub of metabolic glucolipid conversion. SIRT1 upregulation deacetylated ß-catenin, translocated it from the nucleus to the cytoplasm, attenuated glycolysis, and was positively correlated with fatty acid oxidation (FAO). Clinical analysis of SIRT1 expression in tumor tissues showed the SIRT1High profile was associated with poor prognosis in CRC patients. SIRT1 interference therapy significantly suppressed tumors in the mouse xenograft model. Conclusions: In hostile, glucose-deficient TMEs, SIRT1 is upregulated, and CRC cells transform the Warburg phenotype to FAO. SIRT1 indicates the frequency of glucolipid transformation and rapid tumor progression and is a promising therapeutic target of CRC.

Heat stress and excessive maturity of fruiting bodies suppress GABA accumulation by modulating GABA metabolism in Pleurotus ostreatus (Jacq. ex Fr.) P. Kumm.

GABA is a health-promoting bioactive substance. Here, the GABA biosynthetic pathways were investigated, and then the dynamic quantitative changes in GABA and the expression levels of genes related to GABA metabolism under heat stress or at different developmental stages of fruiting bodies in Pleurotus ostreatus (Jacq. ex Fr.) P. Kumm were determined. We found that the polyamine degradation pathway was the main route of GABA production under growth normal condition. The accumulation of GABA and the expression of most genes related to GABA biosynthesis, including genes encoding glutamate decarboxylase (PoGAD-2), polyamine oxidase (PoPAO-1), diamine oxidase (PoDAO) and aminoaldehyde dehydrogenase (PoAMADH-1 and PoAMADH-2), were significantly suppressed by heat stress and the excessive maturity of fruiting bodies. Finally, the effects of GABA on the mycelial growth, heat tolerance and the morphogenesis and development of fruiting bodies were studied, the results showed that the deficiency of endogenous GABA inhibited the mycelial growth and primordial formation and aggravated heat damage, whereas exogenous application of GABA could improve thermotolerance and promote the development of fruiting bodies.

EDA-E7 Activated DCs Induces Cytotoxic T Lymphocyte Immune Responses against HPV Expressing Cervical Cancer in Human Setting.

Cervical cancer is a major cause of cancer death in women worldwide. Targeting human papillomavirus (HPV) viral oncoproteins E6 and E7 is a new strategy for cervical cancer immunotherapy and has been associated with resolution of HPV-induced lesions. How to efficiently induce T cell target killing of HPV infected cervical cancer is of great potential benefit for cervical cancer treatment. Fusion protein containing the extra domain A (EDA) from fibronectin, a natural ligand for Toll-like receptor 4 (TLR4), and HPVE7 (EDA-E7) has been shown to efficiently induce dendritic cells maturation and trigger specific antitumor CD8+ T cells response in mice. In this study, we constructed EDA-E7 fusion protein of human origin and tested its function in dendritic cell maturation as well as antitumor T cell response. We found that EDA-E7 could be efficiently captured by human PBMC derived dendritic cells (DCs) in vitro and induce DCs maturation. Importantly, this effect could work in synergy with the TLR ligand anti-CD40 agonist, polyinosinic-polycytidylic acid [poly (I:C)], R848, and CpG2216. EDA-E7 matured DCs could activate T cells and trigger an anti-tumor response in vitro. Single cell RNA sequencing and T cell targeted killing assay confirmed the activation of T cells by EDA-E7 matured DCs. Therefore, therapeutic vaccination with EDA-E7 fusion protein maybe effective for human cervical carcinoma treatment.

Effects of six weeks of sub-plateau cold environment training on physical functioning and athletic ability in elite parallel giant slalom snowboard athletes.

Background: Hypoxic and cold environments have been shown to improve the function and performance of athletes. However, it is unclear whether the combination of subalpine conditions and cold temperatures may have a greater effect. The present study aims to investigate the effects of 6 weeks of training in a sub-plateau cold environment on the physical function and athletic ability of elite parallel giant slalom snowboard athletes. Methods: Nine elite athletes (four males and five females) participated in the study. The athletes underwent 6 weeks of high intensity ski-specific technical training (150 min/session, six times/week) and medium-intensity physical training (120 min/session, six times/week) prior to the Beijing 2021 Winter Olympic Games test competition. The physiological and biochemical parameters were collected from elbow venous blood samples after each 2-week session to assess the athletes' physical functional status. The athletes' athletic ability was evaluated by measuring their maximal oxygen uptake, Wingate 30 s anaerobic capacity, 30 m sprint run, and race performance. Measurements were taken before and after participating in the training program for six weeks. The repeated measure ANOVA was used to test the overall differences of blood physiological and biochemical indicators. For indicators with significant time main effects, post-hoc tests were conducted using the least significant difference (LSD) method. The paired-samples t-test was used to analyze changes in athletic ability indicators before and after training. Results: (1) There was a significant overall time effect for red blood cells (RBC) and white blood cells (WBC) in males; there was also a significant effect on the percentage of lymphocytes (LY%), serum testosterone (T), and testosterone to cortisol ratio (T/C) in females (p < 0.001 - 0.015, η p 2 = 0 . 81 - 0 . 99 ). In addition, a significant time effect was also found for blood urea(BU), serum creatine kinase (CK), and serum cortisol levels in both male and female athletes (p = 0.001 - 0.029, η p 2 = 0 . 52 - 0 . 95 ). (2) BU and CK levels in males and LY% in females were all significantly higher at week 6 (p = 0.001 - 0.038), while WBC in males was significantly lower (p = 0.030). T and T/C were significantly lower in females at week 2 compared to pre-training (p = 0.007, 0.008, respectively), while cortisol (C) was significantly higher in males and females at weeks 2 and 4 (p (male) = 0.015, 0.004, respectively; p (female) = 0.024, 0.030, respectively). (3) There was a noticeable increase in relative maximal oxygen uptake, Wingate 30 s relative average anaerobic power, 30 m sprint run performance, and race performance in comparison to the pre-training measurements (p < 0.001 - 0.027). Conclusions: Six weeks of sub-plateau cold environment training may improve physical functioning and promote aerobic and anaerobic capacity for parallel giant slalom snowboard athletes. Furthermore, male athletes had a greater improvement of physical functioning and athletic ability when trained in sub-plateau cold environments.

Ecosystem social responsibility in international digital commerce.

Despite the surge of interest in digital globalization, its social dimensions have received far less attention than deserved. The lack of conversation between the two prominent areas of IB research, digitalization, and corporate social responsibility, presents a valuable opportunity for extending the agenda Ioannou and Serafeim (J Int Bus Stud 43(9):834-864, 2012) pioneered a decade earlier. We briefly depict the organizational differences between multinational enterprises (MNEs) and multinational platforms (MNPs), followed by a closer look at how social responsibility of digital platforms might depart from our conventional understanding derived from MNEs. We then propose the notion of ecosystem social responsibility emphasizing social value co-creation before categorizing the main areas of social issues specific to MNPs. Based on these ideas, we derive several new insights into the social challenges faced by firms governing global platforms versus multidomestic platforms, respectively, as they serve international markets. Lastly, we discuss future research directions and, in particular, the implications for ecosystem sustainability.

Malgré le regain d'intérêt pour la globalisation numérique, ses dimensions sociales ont reçu beaucoup moins d'attention qu'elles ne le méritent. L'absence de conversation entre les deux principaux domaines de recherche en affaires internationales, à savoir la numérisation et la responsabilité sociale des entreprises, constitue une occasion précieuse d'étendre le programme que Ioannou et Serafeim (2012) ont lancé une décennie plus tôt. Nous décrivons brièvement les différences organisationnelles entre les entreprises multinationales (Multinational enterprises - MNEs) et les plates-formes multinationales (Multinational platforms - MNPs). Puis, nous examinons de plus près comment la responsabilité sociale des plates-formes numériques pourrait diverger de notre compréhension conventionnelle dérivée des MNEs. Nous proposons ensuite la notion de responsabilité sociale de l'écosystème en mettant l'accent sur la cocréation de valeur sociale, et ce avant de catégoriser les principaux domaines de problèmes sociaux propres aux MNPs. Sur la base de ces idées, nous élaborons plusieurs nouveaux renseignements sur les défis sociaux auxquels sont confrontées les entreprises qui régissent des plates-formes multidomestiques vs globales durant leurs exploitations des marchés internationaux. Enfin, nous discutons de futures directions de recherche et, en particulier, des implications pour la durabilité des écosystèmes.

Pese al auge del interés por la globalización digital, sus dimensiones sociales han recibido mucha menos atención de la que merecen. La falta de conversación entre las dos áreas prominentes de la investigación de negocios internacionales, la digitalización y la responsabilidad social corporativa, presenta una valiosa oportunidad para ampliar la agenda que Ioannou y Serafeim (2012) iniciaron una década antes. Describimos brevemente las diferencias organizacionales entre las empresas multinacionales (EMN) y las plataformas multinacionales (MNPs por sus iniciales en inglés), seguidas de un análisis más detallado de cómo la responsabilidad social de las plataformas digitales podría apartarse de nuestra comprensión convencional derivada de las empresas multinacionales. A continuación, proponemos la noción de responsabilidad social del ecosistema, que enfatiza la co-creación de valor social, antes de clasificar las principales áreas de los asuntos sociales específicos de las plataformas multinacionales. Sobre la base de estas ideas, derivamos varios aportes nuevos sobre los retos sociales a los que se enfrentan las empresas que dominan las plataformas globales versus las plataformas multidomésticas, respectivamente, cuando sirven a los mercados internacionales. Por último, discutimos las direcciones futuras de la investigación y, en particular, las implicaciones para la sostenibilidad del ecosistema.

Apesar do aumento do interesse pela globalização digital, suas dimensões sociais receberam muito menos atenção do que merecem. A falta de diálogo entre as duas proeminentes áreas de pesquisa em IB, digitalização e responsabilidade social corporativa, apresenta uma oportunidade valiosa para estender a agenda que Ioannou e Serafeim (2012) introduziram uma década antes. Descrevemos brevemente as diferenças organizacionais entre empresas multinacionais (MNEs) e plataformas multinacionais (MNPs), seguidas de um olhar mais atento sobre como a responsabilidade social de plataformas digitais pode se afastar do nosso entendimento convencional derivado de MNEs. Em seguida propomos a noção de responsabilidade social ecossistêmica enfatizando a cocriação de valor social antes de categorizar as principais áreas de questões sociais específicas de MNPs. Baseados nessas ideias, derivamos vários novos insights sobre os desafios sociais enfrentados pelas empresas que governam plataformas globais versus plataformas multidomésticas, respectivamente, ao atender mercados internacionais. Por fim, discutimos futuras direções de pesquisa e, particularmente, as implicações para a sustentabilidade ecossistêmica.

CRISPR/Cas9-mediated endogenous TCR knockout enhances TCR-T cells targeted killing HPV16-positive cervical cancer cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：基于CRISPR/Cas9基因编辑技术制备无内源TCR的TCR-T细胞并鉴定其在体外杀伤HPV16阳性宫颈癌SiHa细胞的功能。方法：培养健康志愿者外周血CD8+ T细胞和Jurkat细胞，CRISPR/Cas9基因编辑技术敲除CD8+ T、Jurkat细胞的TCR基因，制备过表达转基因TCR的重组慢病毒，在敲除内源性TCR的CD8+ T和Jurkat细胞中用慢病毒过表达转基因TCR制备TCR-T细胞，多色FCM检测TCR-T细胞中TCR和CD3的表达水平，荧光素酶活性实验检测TCR-T细胞对HPV16阳性SiHa细胞的杀伤效率。结果：CRIPSR/Cas9基因编辑技术高效地敲除了外周血CD8+ T细胞和Jurkat细胞中的TRAC和TRBC基因，敲除效率分别为（81.4±4.5）%、（98.5±0.07）%，制备的无内源TCR的TCR-T细胞高效表达转基因TCR，在外周血CD8+ T和Jurkat细胞中表达率为（66.0±17.8）%、（97.3±2.6）%，敲除内源TRAC和TRBC基因有效增强CD8+ T和Jurkat细胞膜表达转基因TCR（均P<0.01），敲除内源TCR增强TCR-T细胞特异性杀伤HPV16阳性的SiHa细胞[（71.4±1.0）% vs （35.1±2.0）%，P<0.01）]。结论：无内源TCR的TCR-T细胞显著增强转基因TCR的表达和对HPV16阳性宫颈癌SiHa细胞的靶向杀伤能力，为提高TCR-T细胞的临床疗效提供了实验依据。

OTUD3 suppresses the mTORC1 signaling by deubiquitinating KPTN.

Background: Ubiquitination and deubiquitination modifications play pivotal roles in eukaryotic life processes, regulating protein dynamics via the ubiquitin-proteasome pathway. Dysregulation can impact disease development, including cancer and neurodegenerative disorders. Increasing evidence highlights their role in tumorigenesis, modulating key proteins. OTUD3, a deubiquitinase, stabilizes PTEN, suppressing tumor growth by inhibiting PI3K-AKT signaling. Yet, further OTUD3 substrates remain underexplored. Methods: We employed the In vivo ubiquitination assay to investigate the ubiquitination role of OTUD3 on KPTN within the cellular context. Additionally, CRISPR/Cas9 editing and Immunofluorescence were utilized to study the impact of OTUD3 on the mTOR signaling pathway in cells. Furthermore, Cell proliferation assay and NMR were employed to explore the effects of OTUD3 on cellular growth and proliferation. Results: OTUD3 serves as a deubiquitinase for KPTN. OTUD3 interacts with KPTN, facilitated by the OTU domain within OTUD3. Further investigations confirmed KPTN's ubiquitination modification, primarily at lysine residue 49. Ubiquitination experiments demonstrated OTUD3's ability to mediate KPTN's deubiquitination without affecting its protein levels. This suggests KPTN's ubiquitination is a function-regulated, non-degradable modification. Under various amino acid starvation or stimulation conditions, overexpressing OTUD3 reduces mTORC1 signaling activation, while knocking out OTUD3 further enhances it. Notably, OTUD3's regulation of mTORC1 signaling relies on its deubiquitinase activity, and this effect is observed even in PTEN KO cells, confirming its independence from PTEN, a reported substrate. OTUD3 also promotes GATOR1's lysosomal localization, a process requiring KPTN's involvement. Ultimately, OTUD3 affects cellular metabolic pool products by downregulating the mTORC1 pathway, significantly inhibiting tumor cell growth and proliferation. Discussion: Our experiments shed light on an alternative perspective regarding the intrinsic functions of OTUD3 in inhibiting tumor development. We propose a novel mechanism involving KPTN-mediated regulation of the mTORC1 signaling pathway, offering fresh insights into the occurrence and progression of tumor diseases driven by related genes. This may inspire new approaches for drug screening and cancer treatment, potentially guiding future therapies for relevant tumors.

Membrane-bound transcription factor TaNTL1 positively regulates drought stress tolerance in transgenic Arabidopsis.

Drought negatively affects plant growth and development to cause major yield losses in crops. Transcription factors (TFs) play important roles in abiotic stress response signaling in plant. However, the biological functions of membrane-bound transcription factors (MTFs) in abiotic stress have rarely been studied in wheat. In this study, we identified a homologue of the maize ZmNTL1 gene in wheat, which was designated as TaNTL1. TaNTL1 is a NAC family MTF (NTM1-like, NTL proteins) encoding 481 amino acid residues with a transmembrane motif at the C-terminal. Quantitative results and expression profile analysis showed that TaNTL1 could respond to drought. We demonstrated the transcriptional activity of TaNTL1 and that it could specifically bind to NAC recognition cis-acting elements (NACBS). The full-length TaNTL1 protein localized in the plasma membrane and TaNTL1 lacking the transmembrane motif (TaNTL1-ΔTM) localized in the nucleus. TaNTL1 was proteolytically activated by PEG6000 and abscisic acid (ABA). Phenotypic and physiological analyses showed that overexpression transgenic Arabidopsis exhibited enhanced drought resistance, which was greater with TaNTL1-ΔTM than TaNTL1. Transient silencing of TaNTL1 significantly reduced the resistance to drought stress in wheat. Germination by the TaNTL1 and TaNTL1-ΔTM transgenic Arabidopsis seeds was also hypersensitive to ABA. Most of the stress-related genes in transgenic plants were upregulated under drought conditions. These results suggest that MTF TaNTL1 is a positive regulator of drought and it may function by entering the nucleus through cleavage.

RING E3 ubiquitin ligase TaSADR1 negatively regulates drought resistance in transgenic Arabidopsis.

Drought stress is an important factor that affects crop yields and quality. E3 ubiquitin ligase has crucial roles in the responses to abiotic stresses. However, few studies have investigated the role of E3 ubiquitin ligase during drought stress in wheat. In this study, we cloned and identified the orthologous gene of Oryza sativa Salt-, ABA- and Drought-Induced RING Finger Protein 1 (OsSADR1) in wheat (Triticum aestivum L.) called TaSADR1. TaSADR1 encodes a protein containing 486 amino acids with a C3HC4 type RING finger conserved domain at the N-terminal. We confirmed that TaSADR1 has an E3 ubiquitin ligase activity and it is located in the nucleus. High expression of TaSADR1 was induced by treatment with PEG6000 and abscisic acid (ABA). TaSADR1-overexpressing transgenic Arabidopsis plants exhibited decreased drought tolerance. Under drought stress, compared with the wild-type (WT) lines, TaSADR1-overexpressing transgenic Arabidopsis lines had lower proline and chlorophyll contents, and antioxidant enzyme activities (superoxide dismutase, peroxidase, and catalase), whereas the water loss rate, malondialdehyde content, and relative electrolyte leakage were higher. In addition, the overexpressing transgenic Arabidopsis lines were more sensitive to mannitol and ABA treatment at seed germination and during seedling growth. The expression levels of genes related to stress were downregulated under drought conditions in the transgenic plants. Our results demonstrate that TaSADR1 may negatively regulate drought stress responses by regulating the expression of stress-related genes.

CD9, a potential leukemia stem cell marker, regulates drug resistance and leukemia development in acute myeloid leukemia.

BACKGROUND: Leukemia stem cells (LSCs) are responsible for the initiation, progression, and relapse of acute myeloid leukemia (AML). Therefore, a therapeutic strategy targeting LSCs is a potential approach to eradicate AML. In this study, we aimed to identify LSC-specific surface markers and uncover the underlying mechanism of AML LSCs. METHODS: Microarray gene expression data were used to investigate candidate AML-LSC-specific markers. CD9 expression in AML cell lines, patients with AML, and normal donors was evaluated by flow cytometry (FC). The biological characteristics of CD9-positive (CD9+) cells were analyzed by in vitro proliferation, chemotherapeutic drug resistance, migration, and in vivo xenotransplantation assays. The molecular mechanism involved in CD9+ cell function was investigated by gene expression profiling. The effects of alpha-2-macroglobulin (A2M) on CD9+ cells were analyzed with regard to proliferation, drug resistance, and migration. RESULTS: CD9, a cell surface protein, was specifically expressed on AML LSCs but barely detected on normal hematopoietic stem cells (HSCs). CD9+ cells exhibit more resistance to chemotherapy drugs and higher migration potential than do CD9-negative (CD9-) cells. More importantly, CD9+ cells possess the ability to reconstitute human AML in immunocompromised mice and promote leukemia growth, suggesting that CD9+ cells define the LSC population. Furthermore, we identified that A2M plays a crucial role in maintaining CD9+ LSC stemness. Knockdown of A2M impairs drug resistance and migration of CD9+ cells. CONCLUSION: Our findings suggest that CD9 is a new biomarker of AML LSCs and is a promising therapeutic target.

Cathepsin C promotes breast cancer lung metastasis by modulating neutrophil infiltration and neutrophil extracellular trap formation.

Lung metastasis is the major cause of breast cancer-related mortality. The neutrophil-associated inflammatory microenvironment aids tumor cells in metastatic colonization in lungs. Here, we show that tumor-secreted protease cathepsin C (CTSC) promotes breast-to-lung metastasis by regulating recruitment of neutrophils and formation of neutrophil extracellular traps (NETs). CTSC enzymatically activates neutrophil membrane-bound proteinase 3 (PR3) to facilitate interleukin-1ß (IL-1ß) processing and nuclear factor κB activation, thus upregulating IL-6 and CCL3 for neutrophil recruitment. In addition, the CTSC-PR3-IL-1ß axis induces neutrophil reactive oxygen species production and formation of NETs, which degrade thrombospondin-1 and support metastatic growth of cancer cells in the lungs. CTSC expression and secretion are associated with NET formation and lung metastasis in human breast tumors. Importantly, targeting CTSC with compound AZD7986 effectively suppresses lung metastasis of breast cancer in a mouse model. Overall, our findings reveal a mechanism of how tumor cells regulate neutrophils in metastatic niches and support CTSC-targeting approaches for cancer treatment.

Specific ablation of CD4+ T-cells promotes heart regeneration in juvenile mice.

Unlike adult cardiomyocytes, neonatal cardiomyocytes can readily proliferate that contributes to a transient regenerative potential after myocardial injury in mice. We have recently reported that CD4+ regulatory T-cells promote this process; however, the role of other CD4+ T-cell subsets as well as CD8+ T-cells in postnatal heart regeneration has been less studied. Methods: by comparing the regenerating postnatal day (P) 3 and the non-regenerating P8 heart after injury, we revealed the heterogeneity of CD4+ and CD8+ T-cells in the myocardium through single cell analysis. We also specifically ablated CD4+ and CD8+ T-cells using the lytic anti-CD4 and -CD8 monoclonal antibodies, respectively, in juvenile mice at P8 after myocardial injury. Results: we observe significantly more CD4+FOXP3- conventional T-cells in the P8 heart when compared to that of the P3 heart within a week after injury. Surprisingly, such a difference is not seen in CD8+ T-cells that appear to have no function as their depletion does not reactivate heart regeneration. On the other hand, specific ablation of CD4+ T-cells contributes to mitigated cardiac fibrosis and increased cardiomyocyte proliferation after injury in juvenile mice. Single-cell transcriptomic profiling reveals a pro-fibrotic CD4+ T-cell subset in the P8 but not P3 heart. Moreover, there are likely more Th1 and Th17 cells in the P8 than P3 heart. We further demonstrate that cytokines of Th1 and Th17 cells can directly reduce the proliferation and increase the apoptosis of neonatal cardiomyocytes. Moreover, ablation of CD4+ T-cells can directly or indirectly facilitate the polarization of macrophages away from the pro-fibrotic M2-like signature in the juvenile heart. Nevertheless, ablation of CD4+ T-cells alone does not offer the same protection in the adult heart after myocardial infarction, suggesting a developmental change of immune cells including CD4+ T-cells in the regulation of age-related mammalian heart repair. Conclusions: our results demonstrate that ablation of CD4+ but not CD8+ T-cells promotes heart regeneration in juvenile mice; and CD4+ T-cells play a distinct role in the regulation of heart regeneration and repair during development.

Coreceptor blockade targeting CD4 and CD8 allows acceptance of allogeneic human pluripotent stem cell grafts in humanized mice.

We have previously demonstrated that short-term coreceptor blockade with non-lytic monoclonal antibodies enables the long-term survival of fully allogeneic embryonic stem cell (ESC) transplants in mice. Here, we describe the use of Hu-PBL humanized mice to determine whether short-term coreceptor blockade with humanized anti-human CD4 and CD8 antibodies can achieve the same outcome towards human ESC derivatives. While control Hu-PBL mice rejected allogeneic hESC-derived transplants within weeks, mice treated with coreceptor blocking antibodies held their grafts for 7 weeks, the duration of the study. Rejection in the control mice was associated with demonstrable infiltrates of human CD45 white blood cells, predominantly of CD8 T-cells, whereas anti-CD4, but not anti-CD8 antibody treated mice showed remarkably reduced lymphocyte infiltration and prolonged allograft survival, indicating that the CD4+ T-cells were crucial to the rejection process. Our results give support to the principle that short-term blockade of T-cell co-receptors can achieve long-term acceptance of regenerative cell transplants in humans.

Genome-wide identification and characterization of late embryogenesis abundant protein-encoding gene family in wheat: Evolution and expression profiles during development and stress.

Late embryogenesis abundant (LEA) proteins are involved in plant stress responses and osmotic regulation, and they are accumulated in the late embryonic stage. There have been no previous genome-wide analyses of the LEA gene family members in wheat and its close relatives. In this study, 281, 53, 151, 89, 99, and 99 LEA genes were identified in wheat (Triticum aestivum), Triticum urartu, Triticum dicoccoides, Aegilops tauschii, barley, and Brachypodium distachyon, respectively. The wheat LEA gene family (TaLEA genes) was divided into eight subfamilies according to the conserved domains. All TaLEA genes contain very few introns (<3) and they are unevenly distributed on the 21 chromosomes. We identified 39 pairs of tandem duplication genes and 9 pairs of segmental duplication genes in the wheat LEA gene family. This proved that the tandem duplication and segmental duplication played an important role in the expansion of the TaLEA gene family. According to published transcriptome data and qRT-PCR analysis, the TaLEA genes exhibit different tissue expression patterns and they are regulated by various abiotic stresses, especially salt and cold stress. This study provides a comprehensive understanding of the wheat LEA gene family.