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A rapid and sensitive high-performance liquid chromatography-high-resolution orbitrap mass spectrometry method was developed for the simultaneous screening of 354 organic poisons and metabolites in blood and urine, including drugs, medications, pesticides, rodenticides, veterinary drugs, alkaloids, and mycotoxins with a multi-toxicant chromatography-mass spectrometry information library. The method and library showed good prospects in clinical poisoning screening and forensic toxicological identification. Blood and urine samples were extracted successively with ethyl acetate in acidic and alkaline conditions; then, the extract was blown to nearly dry by nitrogen gas and redissolved with methanol-aqueous solution (v:v, 50:50), and the dissolved solution was analyzed by LC-MS/MS after filtering. Precursor ions' m/z was set for identification, retention time, fragment ions, and isotopic pattern which were used for confirmation. No interference peaks were found in the blank samples, showing good specificity. The LODs of toxicants in urine and blood were 1.00×10

Background. The aim of this study was to identify prognostic fatty acid metabolism lncRNAs and potential molecular targeting drugs in uveal melanoma through integrated bioinformatics analysis. Methods. In the present study, we obtained the expression matrix of 309 FAM-mRNAs and identified 225 FAM-lncRNAs by coexpression network analysis. We then performed univariate Cox analysis, LASSO regression analysis, and cross-validation and finally obtained an optimized UVM prognosis prediction model composed of four PFAM-lncRNAs (AC104129.1, SOS1-IT1, IDI2-AS1, and DLGAP1-AS2). Results. The survival curves showed that the survival time of UVM patients in the high-risk group was significantly lower than that in the low-risk group in the train cohort, test cohort, and all patients in the prognostic prediction model ( P < 0.05 ). We further performed risk prognostic assessment, and the results showed that the risk scores of the high-risk group in the train cohort, test cohort, and all patients were significantly higher than those of the low-risk group ( P < 0.05 ), patient survival decreased and the number of deaths increased with increasing risk scores, and AC104129.1, SOS1-IT1, and DLGAP1-AS2 were high-risk PFAM-lncRNAs, while IDI2-AS1 were low-risk PFAM-lncRNAs. Afterwards, we further verified the accuracy and the prognostic value of our model in predicting prognosis by PCA analysis and ROC curves. Conclusion. We identified 24 potential molecularly targeted drugs with significant sensitivity differences between high- and low-risk UVM patients, of which 13 may be potential targeted drugs for high-risk patients. Our findings have important implications for early prediction and early clinical intervention in high-risk UVM patients.

Background. Primary open-angle glaucoma (POAG) is the most common type of glaucoma. The potential influence of some DEGs on the progression of POAG was still incomplete. In this study, we integrated transcriptome data with clinical data to investigate the relationship between them in POAG patients. Methods. The gene expression profile (GSE27276) from Gene Expression Omnibus (GEO) was used to identify DEGs. The LIMMA package of R was used to identify the DEGs (Diboun et al., 2006). The adjusted P values (adj P value) were calculated instead to avoid the appearance of false-positive results. Genes with |log2 fold change (FC)| larger than 1 and adj P value < 0.01 were taken as DEGs between PH and PC samples. GO (Gene Ontology) function and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analyses of the DEGs were performed. Protein-protein interactions (PPIs) of the DEGs were constructed. Results. A total of 182 DEGs were identified through our analysis, of which 119 genes were upregulated and 63 genes were downregulated. GO enrichment analysis illustrated that these DEGs were mostly enriched into haptoglobin binding, antioxidant activity, and organic acid binding. KEGG enrichment analysis illustrated that these DEGs were mostly enriched into Staphylococcus aureus infection. The most significant module was identified by MCODE consists of 8 DEGs, and BCL11A is the seeded gene. The second most significant module consists of 5 DEGs, and IL1RN is the seeded gene. Conclusion. Our results demonstrate the potential influence of some DEGs on the progression of POAG, providing a comprehensive bioinformatics analysis of the pathogenesis, which may contribute to future investigation into the molecular mechanisms and biomarkers.

BackgroundOur previous study showed that transmembrane tumor necrosis factor alpha (tmTNF-α) is overexpressed in primary breast cancers including triple-negative breast cancers (TNBCs). Chimeric antigen receptor engineered-T (CAR-T) cells have been successfully used mainly in B-cell malignancies.MethodsWe generated CAR-T cells targeting tmTNF-α but not secreted tumor necrosis factor alpha and assessed the antitumor effect of the CAR-T cells on tmTNF-α-expressing breast cancer cells in vitro and in vivo.ResultsOur tmTNF-α CAR-T cells showed potent cytotoxicity against tmTNF-α-expressing breast cancer cells but not tmTNF-α-negative tumor cells with increased secretion of interferon gamma (IFN-γ) and interleukin (IL)-2 in vitro. In tmTNF-α-overexpressing TNBC-bearing mice, the tmTNF-α CAR-T therapy induced evident tumor regression, prolonged survival and increased serum concentrations of IFN-γ and IL-2. However, we found thattmTNF-α induced programmed death-ligand 1 (PD-L1) expression through the p38 pathway via TNF receptor (TNFR) and through the NF-κB and AKT pathways via outside-to-inside (reverse) signaling, which might limit the efficacy of the CAR-T cell therapy. Blockage of the PD-L1/programmed death-1 (PD-1) pathway by PD-1 monoclonal antibody significantly enhanced the antitumor effect of the tmTNF-α CAR-T cell therapy in vitro and in vivo, and the combination was effective for antiprimary tumors and had a tendency to increase the antimetastasis effect of the CAR-T cell therapy.ConclusionOur findings suggest a potent antitumor efficacy of the tmTNF-α CAR-T cells that can be enhanced by anti-PD-L1/PD-1 because high PD-L1 expression in TNBC was induced by the tmTNF-α signaling, indicating a promising individual therapy for tmTNF-α-positive breast cancers including TNBC.

BackgroundEnzymes with dedicated cofactor preference are essential for advanced biocatalysis and biomanufacturing. However, directed evolution of an enzyme to switch its cofactor preference is often hindered by the lack of efficient and affordable method for screening as the cofactor per se or the substrate can be prohibitively expensive. Here, we developed a growth-based selection platform to identify nonnatural cofactor-dependent oxidoreductase mutants.ResultsThe growth-based selection platform was designed by coupling with nonnatural cofactor-dependent phosphite dehydrogenase (Pdh) mediated the conversion of non-metabolizable phosphite into phosphate in the culture media. Thus, Pdh variant that strongly favors nicotinamide cytosine dinucleotide (NCD), a NAD analogue, the feasibility of this strategy was successfully demonstrated using derived NCD-active malic enzyme as well as for the directed evolution of NCD synthetase in Escherichia coli.ConclusionsHere, we built a phosphite-based screening platform for identification of enzymes favoring nonnatural cofactor NCD. In the future, once Pdh variants favoring other biomimetic or nonnatural cofactors are available this selection platform may be readily redesigned to attain new enzyme variants with anticipated cofactor preference, providing opportunities to further expand the chemical space of redox cofactors in chemical biology and synthetic biology.

Soluble tumor necrosis factor-α (sTNF-α) plays an important role in colitis-associated cancer (CAC); however, little is known about transmembrane TNF-α (tmTNF-α). Here, we observed an increase in sTNF-α mainly in colitis tissues from an azoxymethane/dextran sodium sulfate (DSS)-induced CAC mouse model whereas tmTNF-α levels were chiefly increased on epithelial cells at the tumor stage. The ratio of intracolonic tmTNF-α/sTNF-α was negatively correlated with the levels of pro-inflammatory mediators (IL-1β, IL-6, and NO) and M1 macrophages but positively correlated with the infiltration of myeloid-derived suppressor cells, regulatory T cells, and the level of the anti-inflammatory cytokine IL-10, suggesting an anti-inflammatory effect of tmTNF-α. This effect of tmTNF-α was confirmed again by the induction of resistance to LPS in colonic epithelial cell lines NCM460 and HCoEpiC through the addition of exogenous tmTNF-α or transfection of the tmTNF-α leading sequence that lacks the extracellular segment but retains the intracellular domain of tmTNF-α. A tmTNF-α antibody was used to block tmTNF-α shedding after the first or second round of inflammation induction by DSS drinking to shift the time window of tmTNF-α expression ahead to the inflammation stage. Antibody treatment significantly alleviated inflammation and suppressed subsequent adenoma formation, accompanied by increased apoptosis. An antitumor effect was also observed when the antibody was administered at the malignant phase of CAC. Our results reveal tmTNF-α as a novel molecular marker for malignant transformation in CAC and provide a new insight into blocking the pathological process by targeting tmTNF-α processing.

Proanthocyanidins (PAs) are major antioxidant flavonoids that play a key role in protecting plants against adverse environmental stress, but the transcriptional regulation of PAs synthesis in rose has not been fully investigated. Here, we report the functional characterization of RcMYBPA2 from rose (Rosa chinensis). RcMYBPA2 expression was induced by wounding, methyl viologen and salicylic acid. Overexpression of RcMYBPA2 in tobacco (Nicotiana tabacum) led to increased PAs concentrations, as well as enhanced tolerance to oxidative stress in comparison with the wild type (WT). Of special note, the transgenic plants exhibited lower levels of reactive oxygen species (ROS) than the WT, accompanied by higher levels of antioxidant enzyme activity under oxidative conditions. The ROS scavenging ability of the transgenic plants was compromised by inhibitors of antioxidant enzymes. In addition, a range of several stress-responsive genes was also up-regulated in the transgenic tobacco under oxidative stresses. Taken together, it is demonstrated that RcMYBPA2 is a positive regulator of the PAs biosynthetic pathway and participates in oxidative tolerance partly via modulating ROS scavenging ability and stress-responsive genes expression. Overexpression of RcMYBPA2 in tobacco led to increased PA content, as well as enhanced tolerance to oxidative stress via scavenging ROS and modulating expression of stress-responsive genes.

The root of Codonopis bulleynana Forest ex Diels (cbFeD), a tonic food widely used in Yunnan Province of China, was found to have a wide range of pharmacological effects. The present study was designed to investigate the anti-fibrotic effect of water extracts of cbFeD in chronic liver injury mice model induced by carbon tetrachloride (CCl4) and to explore its underlying mechanisms. Phytochemical analysis revealed multiple components were present in the water extract of cbFeD and the compounds were mostly enriched in organic acid and its derivatives, flavone, amino acid derivatives, nucleotide and its derivatives, carbohydrates etc. Treatment with cbFeD significantly attenuated liver injury and fibrosis in CCl4-administered mice evidenced by improved liver histology, ameliorated apoptosis of hepatocytes, and decreased transaminase levels in the serum. Decreased activities of superoxide dismutase (SOD) and catalase (CAT) were markedly reversed upon treatment with cbFeD while levels of malondialdehyde (MDA) and glutathione (GSH) were significantly restored towards normal values. cbFeD also suppressed intrahepatic inflammatory cell infiltration and Kupffer cell activation. Furthermore, our study revealed an inhibitory effect of cbFeD on hepatic stellate cells (HSCs) activation both in vitro and in vivo. In conclusion, cbFeD could exert a protective role against liver fibrosis in mice model induced by CCl4 that is comparable to the positive control silymarin and might be developed into a promising anti-fibrotic drug.

Tumor necrosis factor-alpha (TNF-α) plays an important pathogenic role in cardiac hypertrophy and heart failure (HF); however, anti-TNF is paradoxically negative in clinical trials and even worsens HF, indicating a possible protective role of TNF-α in HF. TNF-α exists in transmembrane (tmTNF-α) and soluble (sTNF-α) forms. Herein, we found that TNF receptor 1 (TNFR1) knockout (KO) or knockdown (KD) by short hairpin RNA or small interfering RNA (siRNA) significantly alleviated cardiac hypertrophy, heart dysfunction, fibrosis, and inflammation with increased tmTNF-α expression, whereas TNFR2 KO or KD exacerbated the pathological phenomena with increased sTNF-α secretion in transverse aortic constriction (TAC)- and isoproterenol (ISO)-induced cardiac hypertrophy in vivo and in vitro, respectively, indicating the beneficial effects of TNFR2 associated with tmTNF-α. Suppressing TNF-α converting enzyme by TNF-α Protease Inhibitor-1 (TAPI-1) to increase endogenous tmTNF-α expression significantly alleviated TAC-induced cardiac hypertrophy. Importantly, direct addition of exogenous tmTNF-α into cardiomyocytes in vitro significantly reduced ISO-induced cardiac hypertrophy and transcription of the pro-inflammatory cytokines and induced proliferation. The beneficial effects of tmTNF-α were completely blocked by TNFR2 KD in H9C2 cells and TNFR2 KO in primary myocardial cells. Furthermore, we demonstrated that tmTNF-α displayed antihypertrophic and anti-inflammatory effects by activating the AKT pathway and inhibiting the nuclear factor (NF)-κB pathway via TNFR2. Our data suggest that tmTNF-α exerts cardioprotective effects via TNFR2. Specific targeting of tmTNF-α processing, rather than anti-TNF therapy, may be more useful for the treatment of hypertrophy and HF., In contrast to detrimental effects of soluble tumor necrosis factor-alpha (TNF-α) via TNFR1, this study shows that transmembrane TNF-α protects the heart by suppressing pressure overload-induced cardiac hypertrophy and inflammation via TNFR2. Targeting tmTNF-α processing may be more useful than TNF-antagonist for treatment of hypertrophy and heart failure.

Loan cancellation may cause loss for our company, understanding its relevant factors and predicting possible cancellation will help the business decision in the future. For the loan transactions predicted as highly likely to be cancelled, the company can give close monitor to prevent any unfavorable situation. &nbsp

Endoplasmic reticulum stress (ERS), which refers to a series of adaptive responses to the disruption of endoplasmic reticulum (ER) homeostasis, occurs when cells are treated by drugs or undergo microenvironmental changes that cause the accumulation of unfolded/misfolded proteins. ERS is one of the key responses during the drug treatment of solid tumors. Drugs induce ERS by reactive oxygen species (ROS) accumulation and Ca2+ overload. The unfolded protein response (UPR) is one of ERS. Studies have indicated that the mechanism of ERS-mediated drug resistance is primarily associated with UPR, which has three main sensors (PERK, IRE1α, and ATF6). ERS-mediated drug resistance in solid tumor cells is both intrinsic and extrinsic. Intrinsic ERS in the solid tumor cells, the signal pathway of UPR-mediated drug resistance, includes apoptosis inhibition signal pathway, protective autophagy signal pathway, ABC transporter signal pathway, Wnt/β-Catenin signal pathway, and noncoding RNA. Among them, apoptosis inhibition is one of the major causes of drug resistance. Drugs activate ERS and its downstream antiapoptotic proteins, which leads to drug resistance. Protective autophagy promotes the survival of solid tumor cells by devouring the damaged organelles and other materials and providing new energy for the cells. ERS induces protective autophagy by promoting the expression of autophagy-related genes, such as Beclin-1 and ATG5–ATG12. ABC transporters pump drugs out of the cell, which reduces the drug-induced apoptosis effect and leads to drug resistance. In addition, the Wnt/β-catenin signal pathway is also involved in the drug resistance of solid tumor cells. Furthermore, noncoding RNA regulates the ERS-mediated survival and death of solid tumor cells. Extrinsic ERS in the solid tumor cells, such as ERS in immune cells of the tumor microenvironment (TME), also plays a crucial role in drug resistance by triggering immunosuppression. In immune system cells, ERS in dendritic cells (DCs) and myeloid-derived suppressor cells (MDSCs) influences the antitumor function of normal T cells, which results in immunosuppression. Meanwhile, ERS in T cells can also cause impaired functioning and apoptosis, leading to immunosuppression. In this review, we highlight the core molecular mechanism of drug-induced ERS involved in drug resistance, thereby providing a new strategy for solid tumor treatment.

Acetylsalicylic acid, also known as aspirin, is often used in clinical antipyretic, analgesic and antiplatelet therapy. Aspirin can cause numerous side effects in the gastrointestinal (GI) tract, ranging from unpleasant GI symptoms without gastric mucosal lesions to ulcer bleeding and even death. However, recent studies have found that aspirin can significantly prevent GI tumors. Despite impressive advances in cancer research, screening and treatment options, GI tumors remain a leading cause of death worldwide. Prevention is a far better option than treatment for tumors. Therefore, the present review assesses the pros and cons of aspirin on the GI tract and, on this the basis, the appropriate dose of aspirin to protect it.

The root of Codonopis bulleynana Forest ex Diels (cbFeD), a tonic food widely used in Yunnan Province of China, was found to have a wide range of pharmacological effects. The present study was designed to investigate the anti-fibrotic effect of water extracts of cbFeD in chronic liver injury mice model induced by carbon tetrachloride (CCl

Background: Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of cells that expand in cancer, inflammation, and infection and negatively regulate inflammation and the immune response. Heart failure (HF) is a complex clinical syndrome wherein inflammation induction and incomplete resolution can potentially contribute to HF development and progression. However, the role of MDSCs in HF remains unclear. Methods: The percentage of MDSCs in patients with HF and in mice with pressure overload–induced HF using isoproterenol infusion or transverse aortic constriction (TAC) was detected by flow cytometry. The effects of MDSCs on isoproterenol- or TAC-induced HF were observed on depleting MDSCs with 5-fluorouracil (50 mg/kg) or gemcitabine (120 mg/kg), transferring purified MDSCs, or enhancing endogenous MDSCs with rapamycin (2 mg·kg −1 ·d −1 ). Hypertrophic markers and inflammatory factors were detected by ELISA, real-time polymerase chain reaction, or Western blot. Cardiac functions were determined by echocardiography and hemodynamic analysis. Results: The percentage of human leukocyte antigen-D–related (HLA-DR) − CD33 + CD11b + MDSCs in the blood of patients with HF was significantly increased and positively correlated with disease severity and increased plasma levels of cytokines, including interleukin-6, interleukin-10, and transforming growth factor–β. Furthermore, MDSCs derived from patients with HF inhibited T-cell proliferation and interferon-γ secretion. Similar results were observed in TAC- and isoproterenol-induced HF in mice. Pharmaceutical depletion of MDSCs significantly exacerbated isoproterenol- and TAC-induced pathological cardiac remodeling and inflammation, whereas adoptive transfer of MDSCs prominently rescued isoproterenol- and TAC-induced HF. Consistently, administration of rapamycin significantly increased endogenous MDSCs by suppressing their differentiation and improved isoproterenol- and TAC-induced HF, but MDSC depletion mostly blocked beneficial rapamycin-mediated effects. Mechanistically, MDSC-secreted molecules suppressed isoproterenol-induced hypertrophy and proinflammatory gene expression in cardiomyocytes in a coculture system. Neutralization of interleukin-10 blunted both monocytic MDSC- and granulocytic MDSC–mediated anti-inflammatory and antihypertrophic effects, but treatment with a nitric oxide inhibitor only partially blocked the antihypertrophic effect of monocytic MDSCs. Conclusions: Our findings revealed a cardioprotective role of MDSCs in HF by their antihypertrophic effects on cardiomyocytes and anti-inflammatory effects through interleukin-10 and nitric oxide. Pharmacological targeting of MDSCs by rapamycin constitutes a promising therapeutic strategy for HF.

Background Nearly one-third of the population worldwide is estimated to have latent tuberculosis infection (LTBI), which represents a vast reservoir for a constant source of tuberculosis (TB) transmission. It has been suggested that cynomolgus macaques are less susceptible to Mycobacterium tuberculosis (M.tb) infection than rhesus macaques, we examined M.tb infection of Chinese cynomolgus macaques. Methods Eight Chinese cynomolgus macaques were infected with M.tb Erdman strain with a small [25 colony forming unit (CFU)] or large dose (500 CFU) via bronchoscopy. The infected animals were monitored for symptoms and examined by chest X-ray, computed tomography (CT), tuberculin skin test (TST), and enzyme-linked immunospot (ELISPOT). Results Based on TST conversion and the specific immune responses to M.tb antigens, all animals were successfully infected. Half of the animals developed active infection and died within 15 months postinfection. The other four animals were grouped with latent M.tb infection because of positive TST but few clinical signs and pathological changes of TB during the course of this study. Interestingly, a challenge with a large dose of M.tb also induced latent infection. Similar to the changes that occur with human TB patients, the animals with active infection exhibited weight loss, cough and typical TB pathological changes, including caseous granulomas, cavities, consolidation, lipid pneumonia, pleural effusion, lymphadenopathy and bacterial burden in lungs and other organs. Conclusions The low dose of M.tb was sufficient to cause both active and latent M.tb infection in cynomolgus macaques of Chinese origin.

For distance education, the application of computer network and software technology is very important to open teaching and management. This paper mainly studies the application of blockchain technology in the field of Open University distance education. Blockchain technology, as a new type of decentralized trust structure technology, provides necessary support for distance open education in teaching resource utilization, teaching process control, teaching effect evaluation and teaching management, and provides an effective foundation for teachers and students to participate in distance teaching practice together. This paper adopts a hierarchical network architecture and hierarchical storage structure, which has good scalability and node transparency, provides support for user service strategy in large-scale applications.

Achieving stable nitrite accumulation at low temperature continues to be a challenging problem in partial nitrification (PN). This study proposes a strategy to achieve stable PN of thickened sludge at low temperatures through selective inactivation of enzymes by intermittent thermal treatment. This method was verified using a sequencing batch reactor (SBR) operated at 9 ± 1 °C in full nitrification mode. After thermally treating the activated sludge at 35 °C for 2 days, which was the optimal condition for selective inactivation of enzymes determined by batch tests, an average nitrite accumulation rate (NAR) of 90.18 ± 5.74% was rapidly achieved and the SBR was stably operated at 9 ± 1 °C for 80 days. This was consistent with a significant decrease of the specific nitrite uptake rate (SNUR), but no a reduction, or even an increase, of the specific ammonia uptake rate (SAUR). Also, there was no remarkable change in the nitrifying bacterial community except for a slight increase in both ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB). In addition, after thermal treatment at 35 °C for 2 days, the nitrite oxidoreductase (NOR) activity was significantly decreased, while those of ammonia monooxygenase (AMO) and hydroxylamine oxidoreductase (HAO) showed an increasing trend. Thus, the achievement of stable PN at low temperatures with this strategy may be due to the selective inactivation of NOR, rather than NOB washout.

With the rapid development of the information age, it has promoted the development of all sectors of society, including education, to a certain extent. It is no longer a problem to use modern educational technology in teaching. In the teaching of computer vision media simulation technology courses, the use of modern educational technology can effectively enhance the teaching effect. The purpose of this article is to study the application of computer vision media simulation technology in distance education of the new generation of labour productivity. This topic uses the data collection method and action research method to study the effect of multimedia education teaching in changing the learning style of high school students, and conducts questionnaire surveys of students before and after the experiment, and analyses the differences in the survey results. The analysis results it shows that computer vision media simulation technology has improved experimental skills to varying degrees. Among them, it is believed that the biggest improvement is the standardization of operation (26%). The second only to the operation skills is the expansion of students’ knowledge, accounting for 25%.

// Meng Zhang 1, * , Jing Wang 1, * , Lingwei Jia 2 , Jin Huang 1 , Cheng He 1 , Fuqing Hu 2 , Lifei Yuan 1 , Guihua Wang 2 , Mingxia Yu 1 and Zhuoya Li 1 1 Department of Immunology, Basic Medical College, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, P.R. China 2 Molecular Medical Center, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, P.R. China * These authors have contributed equally to this work Correspondence to: Zhuoya Li, email: zhuoyali@mails.tjmu.edu.cn Keywords: activation-induced cell death, transmembrane TNF-α, secretory TNF-α, reverse signaling, apoptosis Received: June 29, 2016 Accepted: June 20, 2017 Published: July 10, 2017 ABSTRACT Secretory tumor necrosis factor-alpha (sTNF-α) is known to mediate activation- induced cell death (AICD). However, the role of tmTNF-α in AICD is still obscure. Here, we demonstrated that tmTNF-α expression significantly increased accompanied with enhanced apoptosis during AICD in Jurkat and primary human T cells. Knockdown or enhancement of tmTNF-α expression in activated T cells suppressed or promoted AICD, respectively. Treatment of activated T cells with exogenous tmTNF-α significantly augmented AICD, indicating that tmTNF-α as an effector molecule mediates AICD. As tmTNF-α can function as a receptor, an anti-TNF-α polyclonal antibody was used to trigger reverse signaling of tmTNF-α. This antibody treatment upregulated the expression of Fas ligand, TNF-related apoptosis-inducing ligand and tmTNF-α to amplify AICD, and promoted activated T cells expressing death receptor 4, TNF receptor (TNFR) 1 and TNFR2 to enhance their sensitivity to AICD. Knockdown of TNFR1 or TNFR2 expression totally blocked tmTNF-α reverse signaling increased sensitivity to sTNF-α- or tmTNF-α-mediated AICD, respectively. Our results indicate that tmTNF-α functions as a death ligand in mediation of AICD and as a receptor in sensitization of activated T cells to AICD. Targeting tmTNF-α in activated T cells may be helpful in facilitating AICD for treatment of autoimmune diseases.

Myeloid-derived suppressor cells (MDSCs) accumulated in tumor sites promote immune evasion. We found that TNFR deficiency-induced rejection of transplanted tumor was accompanied with markedly decreased accumulation of MDSCs. However, the mechanism(s) behind this phenomenon is not completely understood. Here, we demonstrated that TNFR deficiency did not affect the amount of MDSCs in bone marrow (BM), but decreased accumulation of Gr-1+CD11b+ MDSCs in the spleen and tumor tissues. The chemotaxis of Tnfr-/- MDSCs was prominently decreased in response to both tumor cell culture supernatants and tumor tissue homogenates from Tnfr-/- and wild-type mice, indicating an effect of TNFR signaling on chemokine receptor expression in MDSCs. We used real-time PCR to detect gene expression for several chemokine receptors in MDSCs from BM and found that CXCR4 was the most affected molecule at the transcriptional level in Tnfr-/- MDSCs. Neutralizing CXCR4 in wild-type MDSCs by a specific antibody blocked their chemotactic migration. Interestingly, it was tmTNF-α, but not sTNF-α, that induced CXCR4 expression in MDSCs. This effect of tmTNF-α was totally blocked in TNFR2-/- but not in TNFR1-/- MDSCs, and partially inhibited by PDTC or SB203580, an inhibitor of NF-κB or p38 MAPK pathway, respectively. Adoptive transfer of wild-type MDSCs restored MDSCs accumulation in tumors of Tnfr-/- mice, but this could be partially blocked by treatment with a CXCR4 inhibitor AMD3100. Our data suggest that tmTNF-α upregulates CXCR4 expression that promotes chemotaxis of MDSCs to tumor, and give a new insight into a novel mechanism by which tmTNF-α facilitates tumor immune evasion.

Tumor necrosis factor alpha (TNF-α), originally described as an anti-neoplastic cytokine, has been found, in apparent contradiction to its name, to play an important role in promoting the development and progression of malignant disease. Targeting TNF-α with TNF antagonists has elicited an objective response in certain solid tumors in phase I and II clinical trials. This review focuses on the relationship of TNF-α expressed by leukemia cells and adverse clinical features of leukemia. TNF-α is involved in all steps of leukemogenesis, including cellular transformation, proliferation, angiogenesis, and extramedullary infiltration. TNF-α is also an important factor in the tumor microenvironment and assists leukemia cells in immune evasion, survival, and resistance to chemotherapy. TNF-α may be a potent target for leukemia therapy.

Mechanistic target of rapamycin complex 1 (mTORC1) regulates CD8(+) T-cell differentiation and function. Despite the links between PI3K-AKT and mTORC1 activation in CD8(+) T cells, the molecular mechanism underlying mTORC1 activation remains unclear. Here, we show that both the kinase activity and the death domain of DAPK1 are required for maximal mTOR activation and CD8(+) T-cell function. We found that TCR-induced activation of calcineurin activates DAPK1, which subsequently interacts with TSC2 via its death domain and phosphorylates TSC2 to mediate mTORC1 activation. Furthermore, both the kinase domain and death domain of DAPK1 are required for CD8(+) T-cell antiviral responses in an LCMV infection model. Together, our data reveal a novel mechanism of mTORC1 activation that mediates optimal CD8(+) T-cell function and antiviral activity.

Transmembrane TNF-α (tmTNF-α) and secretory TNF-α (sTNF-α) display opposite effects in septic shock. Reducing tmTNF-α shedding can offset the detrimental effects of sTNF-α and increase the beneficial effect of tmTNF-α. We previously developed a monoclonal antibody that is specific for tmTNF-α and does not cross-react with sTNF-α. In this study, we show that this antibody can specifically suppress tmTNF-α shedding by competing with a TNF-α converting enzyme that cleaves the tmTNF-α ectodomain to release sTNF-α. This tmTNF-α antibody significantly inhibited LPS-induced secretion of interleukin (IL)-1β, IL-6, interferon-β, and nitric oxide by monocytes/macrophages, and protected mice from septic shock induced by lipopolysaccharide (LPS) or cecal ligation and puncture, while reducing the bacterial load. The mechanism associated with the protective effect of this tmTNF-α antibody involved promotion of LPS-induced toll-like receptor 4 (TLR4) internalization and degradation by recruiting Triad3A to TLR4. Moreover, the tmTNF-α antibody inhibited LPS-induced activation of nuclear factor-κB and interferon regulatory factor 3 pathways by upregulating expression of A20 and monocyte chemotactic protein-induced protein 1. Similarly, treatment of macrophages with exogenous tmTNF-α suppressed LPS/TLR4 signaling and release of proinflammatory cytokines, indicating that increased levels of tmTNF-α promoted by the antibody contributed to its inhibitory effect. Thus, use of this tmTNF-α antibody for specific suppression of tmTNF-α shedding may be a promising strategy to treat septic shock.

Objective To explore the mechanism of transdifferentiation of human renal tubular epithelial cells induced by high glucose based on reactive oxygen species (ROS)/NF-κB signaling pathway. Methods HK-2 normal human proximal tubular epithelial cells were randomly divided into blank control group, osmotic pressure control group, high glucose group and pyrrolidine dithiocarbamate (PDTC) group. Cell morphology was observed by phase contrast microscope. Cell viability was measured by MTT assay. Flow cytometry was used to detect intracellular ROS content. Malondialdehyde (MDA) content and superoxide dismutase activity were tested by ELISA. Western blot analysis was used to examine the protein levels of NF-κBp65, phosphorylated IκBα (p-IκBα) and IKKα, monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1). The expressions of NF-κBp65, epithelial cadherin (E-cadherin) and alpha smooth muscle actin (α-SMA) were assessed by immunocytochemistry. Results In high glucose group, the cells became spindle-shaped or irregular, with radial edges, enlarged intercellular space, decreased refractive index and irregular arrangement. At the same time, the cell activity decreased with the prolongation of treatment time, and the cell activity in PDTC group was higher than that in high glucose group. Compared with the blank control group, the content of ROS and MDA increased and the activity of SOD decreased in the high glucose group, while the content of ROS and MDA decreased and the activity of SOD increased in the PDTC group compared with the high glucose group. Compared with the blank group, the protein levels of NF-κBp65, p-IκBα, IKKα, MCP-1 and ICAM-1 increased in the high glucose group, while the protein levels above decreased in the PDTC group compared with the high glucose group. Compared with the blank group, the proportion of positive cells of NF-κBp65 and α-SMA increased and the proportion of positive cells of E-cadherin decreased in the high glucose group. Compared with the high glucose group, the proportion of positive cells of NF-κBp65 and α-SMA decreased and the proportion of positive cells of E-cadherin increased in the PDTC group. Conclusion High glucose can induce epithelial-mesenchymal transition in HK-2 cells. Activation of NF-κB signaling pathway mediated by ROS participates in the above process, which can be blocked by PDTC.

Circular RNAs (CircRNAs), the endogenous long noncoding RNAs, unlike linear RNAs, are structurally continuous, covalently closed loops without 5’ cap or 3’ polyadenylated tail. High-throughput RNA sequencing has enabled the discovery of several endogenous circRNAs in different species and tissues. The circRNAs mainly act as sponges to cytoplasmic microRNA, aid in protein translation, or interact with RNA-binding proteins to generate RNA-protein complexes which control transcription. Recently, circRNAs have been reported to participate in cancer pathogenesis, particularly tumor metastasis in humans, mainly due to their frequent aberrant expression in cancers. However, the detail molecular mechanism of circRNAs activity in tumor metastasis is still elusive. Some specifically expressed circRNAs can potentially be used as biomarkers and therapeutic targets for tumor treatment. Further understanding of the network interactions and regulation of circRNAs is paving the way for the identification of better therapeutic strategies in tumor metastasis. In this mini review, we have summarized the current state of research on functions and mechanisms of novel circRNAs that regulate tumorigenesis and have evaluated the relationship between dysregulation of circRNAs and tumor metastasis.

Macroautophagy/autophagy is a conserved ubiquitous pathway that performs diverse roles in health and disease. Although many key, widely expressed proteins that regulate autophagosome formation followed by lysosomal fusion have been identified, the possibilities of cell-specific elements that contribute to the autophagy fusion machinery have not been explored. Here we show that a macrophage-specific isoform of the vacuolar ATPase protein ATP6V0D2/subunit d2 is dispensable for lysosome acidification, but promotes the completion of autophagy via promotion of autophagosome-lysosome fusion through its interaction with STX17 and VAMP8. Atp6v0d2-deficient macrophages have augmented mitochondrial damage, enhanced inflammasome activation and reduced clearance of Salmonella typhimurium. The susceptibility of atp6v0d2 knockout mice to DSS-induced colitis and Salmonella typhimurium-induced death, highlights the in vivo significance of ATP6V0D2-mediated autophagosome-lysosome fusion. Together, our data identify ATP6V0D2 as a key component of macrophage-specific autophagosome-lysosome fusion machinery maintaining macrophage organelle homeostasis and, in turn, limiting both inflammation and bacterial infection. Abbreviations: ACTB/β-actin: actin, beta; ATG14: autophagy related 14; ATG16L1: autophagy related 16-like 1 (S. cerevisiae); ATP6V0D1/2: ATPase, H+ transporting, lysosomal V0 subunit D1/2; AIM2: absent in melanoma 2; BMDM: bone marrow-derived macrophage; CASP1: caspase 1; CGD: chronic granulomatous disease; CSF1/M-CSF: colony stimulating factor 1 (macrophage); CTSB: cathepsin B; DSS: dextran sodium sulfate; IL1B: interleukin 1 beta; IL6: interleukin 6; IRGM: immunity-related GTPase family M member; KO: knockout; LAMP1: lysosomal-associated membrane protein 1; LC3: microtubule-associated protein 1 light chain 3; LPS: lipo-polysaccaride; NLRP3: NLR family, pyrin domain containing 3; PYCARD/ASC: PYD and CARD domain containing; SNARE: soluble N-ethylmaleimide-sensitive factor attachment protein receptor; SNAP29: synaptosomal-associated protein 29; SQSTM1/p62: sequestosome 1; STX17: syntaxin 17; TLR: toll-like receptor; TNF: tumor necrosis factor ; TOMM20: translocase of outer mitochondrial membrane 20; ULK1: unc-51 like kinase 1; VAMP8: vesicle-associated membrane protein 8; WT: wild type; 3-MA: 3-methyladenine

ICE1 (Inducer of CBF Expression 1), a MYC-like basic helix-loop-helix (bHLH) element for the transcribing process, is critical to plant cold response. However, the physiological function and regulatory mechanism of the ICE1 in woody flower plants remain unclear. In the present study, the functional characterization of RmICE1 from wild rose (Rosa multiflora Thunb.) was elucidated. The transcription levels of RmICE1 were affected by dehydration, salt and cold treatments. RmICE1 exhibited transcriptional activity, and the N-terminal was critical to its activity. RmICE1 over-expressed in tobacco improved the cold tolerance by higher proline content, lower electrolyte leakages (EL) and malondialdehyde (MDA) in contrast to wild-type (WT) plants at cold temperatures. Meantime, transgenic tobacco accumulated a significantly smaller amount of reactive oxygen species (ROS), complying with enhanced activity and expression levels of antioxidant enzymes after cold treatment in contrast to the WT. Besides, six genes stress-responsive genes in the transgenic lines exhibited higher expression levels of than the WT before and after cold stress. On the whole, as revealed from the mentioned data, RmICE1 could promote cold tolerance, probably, at least partially, because of the modulating process of ROS scavenging and the up-regulating process of stress-responsive genes.

Introduction Zenker's diverticulum (ZD) refers to a pouch-like structure similar to the esophageal lumen formed from the herniation of the esophageal mucosa; this structure makes it difficult for food to pass through the esophagus to the stomach. The development of endoscopic technology has made minimally invasive surgical treatments for ZD possible. Patient concerns A female 72-year-old patient was admitted to our hospital due to recurrent dysphagia for more than 5 years. A 62-year-old female patient underwent a gastroscopic examination due to recurrent dysphagia for 10 years and aggravated dysphagia accompanied by bad breath for 1 year. Diagnosis A significant diverticulum with food residue at the entrance of the esophagus was found on gastroscopy in both cases. Interventions After completing a relevant examination and excluding surgical contraindications, both patients underwent submucosal tunneling endoscopic septum division. Outcomes Both patients were discharged after symptoms alleviated on postoperative day 4. A 3-month follow-up gastroscopy showed the disappearance of the diverticulum and recovery of the esophageal anatomical structure. No symptom relapse was found at the 6-month follow-up assessment. Conclusion Submucosal tunneling endoscopic septum division has become the most common minimally invasive treatment option. It is efficient and safe for relieving symptomatic ZD in the short term.

Macrophages perform key functions in tissue homeostasis that are influenced by the local tissue environment. Within the tumor microenvironment, tumor-associated macrophages can be altered to acquire properties that enhance tumor growth. Here, we found that lactate, a metabolite found in high concentration within the anaerobic tumor environment, activated mTORC1 that subsequently suppressed TFEB-mediated expression of the macrophage-specific vacuolar ATPase subunit ATP6V0d2. Atp6v0d2(–/–) mice were more susceptible to tumor growth, with enhanced HIF-2α–mediated VEGF production in macrophages that display a more protumoral phenotype. We found that ATP6V0d2 targeted HIF-2α but not HIF-1α for lysosome-mediated degradation. Blockade of HIF-2α transcriptional activity reversed the susceptibility of Atp6v0d2(–/–) mice to tumor development. Furthermore, in a cohort of patients with lung adenocarcinoma, expression of ATP6V0d2 and HIF-2α was positively and negatively correlated with survival, respectively, suggesting a critical role of the macrophage lactate/ATP6V0d2/HIF-2α axis in maintaining tumor growth in human patients. Together, our results highlight the ability of tumor cells to modify the function of tumor-infiltrating macrophages to optimize the microenvironment for tumor growth.

Tumor necrosis factor (TNF)-α exists in two bioactive forms, a 26-kDa transmembrane form (tmTNF-α) and a 17-kDa soluble form (sTNF-α). sTNF-α has been recognized as a key regulator of hepatitis; however, serum sTNF-α disappears in mice during the development of severe liver injury, and high levels of serum sTNF-α do not necessarily result in liver damage. Interestingly, in a mouse model of acute hepatitis, we have found that tmTNF-α expression on Kupffer cells (KCs) significantly increases when mice develop severe liver injury caused by lipopolysaccharide (LPS)/D-galactosamine (D-gal), and the level of tmTNF-α expression is positively related to the activity of serum transaminases. Therefore, we hypothesized that KC-expressed tmTNF-α constitutes a pathomechanism in hepatitis and have explored the role of tmTNF-α in this disease model. Here, we have compared the impact of KCs(tmTNFlow) and KCs(tmTNFhigh) on acute hepatitis in vivo and ex vivo and have further demonstrated that KCs(tmTNFhigh), rather than KCs(tmTNFlow), not only exhibit an imbalance in secretion of pro- and anti-inflammatory cytokines, favoring inflammatory response and exacerbating liver injury, but also induce hepatocellular apoptosis via tmTNF-α and the expression of another pro-apoptotic factor, Fas ligand. Our data suggest that KC(tmTNFhigh) is a major contributor to liver injury in LPS/D-gal-induced hepatitis.