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Abstract The efficacy of chimeric antigen receptor T (CAR-T) cells for solid tumors remains unsatisfactory due to the limited tumor infiltration and immunosuppressive microenvironment. To overcome these limitations, the genetically engineered recombinant oncolytic adenoviruses (OAVs) that conditionally replicate in tumor cells were developed to modify the tumor microenvironment (TME) to facilitate CAR-T-mediated tumor eradication. Here in the present study, a novel recombinant OAV carrying CCL5, IL12, and IFN-γ controlled by Ki67 promoter was constructed (named AdKi67-C3). The antitumor activity of AdKi67-C3 was tested in vitro and in vivo by using mono administration or combing with CAR-T cells targeting B7H3. It proved that CCL5 expressed by AdKi67-C3 indeed induced more CAR-T migration in vitro and CAR-T infiltration in tumor mass in vivo. Meanwhile, cytokines of IFN-γ and IL12 secreted by AdKi67-C3-infected tumor cells significantly promoted proliferation and persistence of CAR-T cells in vitro and in vivo. In tumor-bearing xenograft mouse models of kidney, prostate or pancreatic cancer, local pretreatment with AdKi67-C3 dramatically enhanced CAR-T cell efficacy and eliminated local and distant tumors. More importantly, mice achieving complete tumor regression resisted to re-challenge with the same tumor cells, suggesting establishment of long-term antitumor immune response. Therefore, OAVs armored with cytokines could be developed as a bioenhancer to defeat the immunosuppressive microenvironment and improve therapeutic efficacy of CAR-T in solid tumors.

Chronic hepatitis B virus (HBV) infection is a leading cause of hepatocellular carcinoma. However, the function and mechanism of the effect of HBV on host protein ubiquitination remain largely unknown. We aimed at characterizing whether and how HBV promotes self-replication by affecting host protein ubiquitination. In this study, we identified UBXN7, a novel inhibitor for nuclear factor kappa B (NF-κB) signaling, was degraded via interaction with HBV X protein (HBx) to activate NF-κB signaling and autophagy, thereby affecting HBV replication. The expression of UBXN7 was analyzed by Western blot and quantitative reverse transcription polymerase chain reaction in HBV-transfected hepatoma cells and HBV-infected primary human hepatocytes (PHHs). The effects of UBXN7 on HBV replication were analyzed by using in vitro and in vivo assays, including stable isotope labeling by amino acids in cell culture (SILAC) analysis. Changes in HBV replication and the associated molecular mechanisms were analyzed in hepatoma cell lines. SILAC analyses showed that the ubiquitination of UBXN7 was significantly increased in HepG2.2.15 cells compared with control cells. After HBV infection, HBx protein interacted with UBXN7 to promote K48-linked ubiquitination of UBXN7 at K99, leading to UBXN7 degradation. On the other hand, UBXN7 interacted with the ULK domain of IκB kinase β through its ubiquitin-associating domain to facilitate its degradation. This in turn reduced NF-κB signaling, leading to reduced autophagy and consequently decreased HBV replication.

Although chimeric antigen receptor T cell (CAR-T) therapy has been successful for hematological malignancies, it is less effective for solid tumors. The primary reason is that the immune microenvironment restricts CAR-T cells from infiltrating and proliferating in tumors. Oncolytic virotherapy has emerged as a novel immunogenic therapy to augment antitumor immune response. Here we combined an oncolytic adenovirus carrying decorin with a CAR-T targeting carbonic anhydrase IX (CAIX) to perform the antitumor activity for renal cancer cells. We found that OAV-Decorin combined with CAIX-CAR-T exhibited significantly reduced tumor burden, altered the composition of extracellular matrix (ECM) by inhibiting the distribution of collagen fibers, decreased the expression of TGF-β in tumor cells, enhanced IFN-γ secretion, and obtained higher numbers of CAR-T cells. The combination treatment modality showed prolonged mice survival. The intratumoral injection of OAV-Decorin into tumor-bearing immunocompetent mice activated the inflammatory immune status and resulted in tumor regression. These data supported further investigation of the combination of OAV-Decorin and CAIX-CAR-T cells in solid tumors.

The microstructure of machined superficial layer significantly affects the mechanical properties and service life of mechanical components. Herein, to obtain a machined surface with excellent properties, a comprehensive application of theoretical calculations and test analysis research methods was conducted to investigate the influence mechanism of machining on performance and quality of machined superficial layer of high-strength alloy steel during machining, and reveal the intrinsic correlation between microstructure evolution, quality and performance of machined superficial layer. The results indicate that the machining can achieve gradient microstructure on the machined surface, which can improve the performance of machined surface greatly. From the surface to the inside region, the layers are respectively the recrystallized layer where compact nano-sized equiaxed grains are located, the rheological layer with clusters of high-density sub-grain structures, and the distorted layer with residual distorted grains. The maximum strain of superficial layer in the machined state is 2.81 times that of the original state. The grains are evidently refined and the grain size is reduced by 49.03%. The dislocation density is enhanced by 100.69%, while the number of small-angle grain boundaries (SAGB) and other substructures increase sharply, being 5.26 times that in the original state. By performance testing, the machining substantially enhances the hardness of the superficial layer of high-strength alloy steel with certain rise in toughness simultaneously, which is exactly the result of the combined effect of fine-grain strengthening and dislocation strengthening caused by the machining process above.

Abstract Hepatitis B virus (HBV) infection remains a major health issue worldwide and the leading cause of cirrhosis and hepatocellular carcinoma (HCC). It has been reported previously that HBV invasion can extensively alter transcriptome, the proteome of exosomes and host cell lipid rafts. The impact of HBV on host proteins through regulating their global post-translational modifications (PTMs), however, is not well studied. Viruses have been reported to exploit cellular processes by enhancing or inhibiting the ubiquitination of specific substrates. Nevertheless, host cell physiology in terms of global proteome and ubiquitylome has not been addressed yet. Here by using HBV-integrated HepG2.2.15 model cell line we first report that HBV significantly modify the host global ubiquitylome. As currently the most widely used HBV cell culture model, HepG2.2.15 can be cultivated for multiple generations for protein labeling, and can replicate HBV, express HBV proteins and secrete complete HBV Dane particles, which makes it a suitable cell line for ubiquitylome analysis to study HBV replication, hepatocyte immune response and HBV-related HCC progression. Our previous experimental results showed that the total ubiquitination level of HepG2.2.15 cell line was significantly higher than that of the corresponding parental HepG2 cell line. By performing a Ubiscan quantification analysis based on stable isotope labeling of amino acids in cell culture (SILAC) of HepG2.2.15 and HepG2 cell lines, we identified a total of 7188 proteins and the protein levels of nearly 19% of them were changed over 2-folds. We further identified 3798 ubiquitinated Lys sites in 1476 host proteins with altered ubiquitination in response to HBV. Our results also showed that the global proteome and ubiquitylome were negatively correlated, indicating that ubiquitination might be involved in the degradation of host proteins upon HBV integration. We first demonstrated the ubiquitination change of VAMP3, VAMP8, DNAJB6, RAB8A, LYN, VDAC2, OTULIN, SLC1A4, SLC1A5, HGS and TOLLIP. In addition, we described 5 novel host factors SLC1A4, SLC1A5, EIF4A1, TOLLIP and BRCC36 that efficiently reduced the amounts of secreted HBsAg and HBeAg. Overall, the HBV-mediated host proteome and ubiquitylome change we reported will provide a valuable resource for further investigation of HBV pathogenesis and host-virus interaction networks.

Hepatitis B virus (HBV) infection remains a major global health problem and the primary cause of cirrhosis and hepatocellular carcinoma (HCC). HBV intrusion into host cells is prompted by virus–receptor interactions in clathrin-mediated endocytosis. Here, we report a comprehensive view of the cellular endocytosis-associated transcriptome, proteome and ubiquitylome upon HBV infection. In this study, we quantified 273 genes in the transcriptome and 190 endocytosis-associated proteins in the proteome by performing multi-omics analysis. We further identified 221 Lys sites in 77 endocytosis-associated ubiquitinated proteins. A weak negative correlation was observed among endocytosis-associated transcriptome, proteome and ubiquitylome. We found 33 common differentially expressed genes (DEGs), differentially expressed proteins (DEPs), and Kub-sites. Notably, we reported the HBV-induced ubiquitination change of secretory carrier membrane protein (SCAMP1) for the first time, differentially expressed across all three omics data sets. Overexpression of SCAMP1 efficiently inhibited HBV RNAs/pgRNA and secreted viral proteins, whereas knockdown of SCAMP1 significantly increased viral production. Mechanistically, the EnhI/XP, SP1, and SP2 promoters were inhibited by SCAMP1, which accounts for HBV X and S mRNA inhibition. Overall, our study unveils the previously unknown role of SCAMP1 in viral replication and HBV pathogenesis and provides cumulative and novel information for a better understanding of endocytosis in response to HBV infection.

This study designed a new microgroove cutting tool to machine Inconel 718 and focused on the effect of microgroove structure on the cutting performance and chip morphology during the turning. A comparative analysis of the cutting force, cutting temperature, tool life, tool wear, and chip morphology of the microgroove cutting tool and the original cutting tool was conducted. The main cutting force and temperature of the microgroove cutting tool were reduced by 12% and 12.17%, respectively, compared with the original cutting tool. The microgroove cutting tool exhibited a significant improvement compared with the original cutting tool, which extended the tool life by up to 23.08%. Further, the microgroove cutting tool distorted the curl radius of the chips extensively. The experimental results showed that the microgroove structure can not only improve the tool life, but also improve the chip breaking effect.

Once an active distribution network of a cyber-physical system is in alert state, it is vulnerable to cross-domain cascading failures. It is necessary to transit the state of an active distribution network of cyber-physical system from an alert state to a normal state using a preventive control policy against cross-domain cascading failures. In fact, it is difficult to construct and analyze a preventive control policy via theoretical analysis methods or physical experimental methods. The theoretical analysis methods may not be accurate due to approximated models, and the physical experimental methods are expensive and time consuming for building prototypes. This paper presents a preventive control policy construction method based on a deep deterministic policy gradient idea (shorted as PCMD) to generate and optimize a preventive control policy with Artificial Intelligence (AI) technologies. It adopts the reinforcement learning technique to make full use of the available historical data to overcome the problems of high cost and low accuracy. Firstly, a preventive control model is designed based on the finite automaton theory, which can guide the data collection and learning policy selection. The control model considers the voltage stability, frequency stability, current overload prevention, and the control cost reduction as a feedback variable, without the specific power flow equations and differential equations. Then, after enough training, a local optimal preventive control policy can be constructed under the comparability condition among a fitted action-value function and a fitted policy function. The constructed preventive control policy contains some control actions to achieve a low cost and in accord with the principle of shortening a cross-domain cascading failures propagation sequence as far as possible. The PCMD is more flexible and closer to reality than the theoretical analysis methods and has a lower cost than the physical experimental methods. To evaluate the performance of the proposed method, an experimental case study, China Electric Power Research-Cyber-Physical System (shorted as CEPR-CPS), which comes from China Electric Power Research Institute, is carried out. The result shows that the effectiveness of preventive control policy construction with the PCMD is better than most current methods, such as the multi-agent method in terms of reducing the number of failure nodes and avoiding the state space explosion.

Abstract Background Heart failure (HF) with improved ejection fraction (EF, HFimpEF) is a distinct HF subtype, characterized by left ventricular (LV) reverse remodeling and myocardial functional recovery. Multiple cardiometabolic factors are implicated in this process. Epicardial adipose tissue (EAT), emerging as an endocrine and paracrine organ, contributes to the onset and progression of HF. However, the relation between EAT and the incidence of HFimpEF is still unclear. Methods A total of 203 hospitalized HF patients with reduced EF (HFrEF, LVEF ≤ 40%) who underwent coronary CT angiography (CCTA) during index hospitalization were consecutively enrolled between November 2011 and December 2022. Routine follow-up and repeat echocardiograms were performed. The incidence of HFimpEF was defined as (1) an absolute LVEF improvement ≥ 10% and (2) a second LVEF > 40% (at least 3 months apart). EAT volume and density were semiautomatically quantified on non-enhanced series of CCTA scans. Results During a median follow-up of 8.6 (4.9 ~ 13.3) months, 104 (51.2%) patients developed HFimpEF. Compared with HFrEF patients, HFimpEF patients had lower EAT volume (115.36 [IQR 87.08 ~ 154.78] mL vs. 169.67 [IQR 137.22 ~ 218.89] mL, P

We illustrated a rare case of malignant solitary fibrous tumor (MSFT) with epithelioid morphology in the occipital region of a 59-year-old female, in which a rare NAB2ex7-STAT6 exon15/16 double fusion subtype was detected by the Next-generation sequencing (NGS) and STAT6 immunohistochemistry (IHC) was diffusely and strongly positively expressed, without recurrence after 20 months of postoperative follow-up. The morphological and molecular genetic aspects and the differential diagnosis are described, and the relevant literature was assessed in order to broaden our understanding and diagnostic capability of this malignancy.

Background: The incidence rate and mortality of bladder cancer are increasing year by year. Interestingly, the commonly used metabolic regulatory drug metformin has been reported to have anti-tumor effect in recent years. Nevertheless, it keeps unclear whether the usage of metformin is beneficial or unbeneficial in treating bladder cancer. Thus, a meta-analysis was conducted to explore the long-term effect of metformin on the incidence of bladder cancer and OS, PFS, DSS and RFS in bladder cancer patients with T2DM.Method: We aim to collect evidence of the association between the usage of metformin and the incidence and treatment outcome of bladder cancer. We searched PubMed, Embase, Ovid Medline and Cochrane Library up to February 2021 to get effective literature reporting the effects of metformin in bladder cancer. The main outcomes were the protective effects of metformin on the incidence, overall survival (OS), recurrence-free survival (RFS), progression-free survival (PFS), and disease-specific survival (DSS) of bladder cancer. And OR (odds ratio) and HR (hazard ratio) with their 95%CI were pooled. Two independent researchers assessed the quality of included studies using the Newcastle-Ottawa Scale (NOS).Results: We involved 12 studies meeting the inclusion criteria, including a total of 1,552,773 patients. The meta-analysis showed that use of metformin could decrease the incidence (OR = 0.45, 95%CI = 0.37–0.56; p < 0.01) and prolong recurrence-free-survival (HR = 0.56, 95%CI = 0.41–0.76; p = 0.91) of bladder cancer. However, there were no significant protective effects in the overall survival (HR = 0.93, 95%CI = 0.67–1.28, p = 0.05), disease-specific-survival (HR = 0.73, 95%CI = 0.47–1.16; p = 0.01), and progression-free-survival (HR = 0.78, 95%CI = 0.53–1.15, p = 0.34).Conclusion: The results revealed that the usage of metformin could reduce the incidence of bladder cancer and prolong the prognosis of bladder cancer in T2DM patients, respectively. More prospective studies are needed to prove the protective role of metformin on bladder cancer.