Targeting neuropilin-1 abolishes anti-PD-1-upregulated regulatory T cells and synergizes with 4-1BB agonist for liver cancer treatment.

BACKGROUND AIMS: Regulatory T cells (Tregs) are an obstacle to PD-1 blockade-mediated antitumor efficacy. However, the behaviors of Tregs response to anti-PD-1 in HCC and the characteristics of Tregs tissue adaptation from peripheral lymphoid tissues to the tumor are still unclear. APPROACH RESULTS: Here, we determine that PD-1 monotherapy potentially augments the accumulation of tumor CD4 + Tregs. Mechanistically, anti-PD-1 mediates Tregs proliferation in lymphoid tissues rather than in the tumor. Increased peripheral Tregs burden replenishes intratumoral Tregs, raising the ratio of intratumoral CD4 + Tregs to CD8 + T cells. Subsequently, single-cell transcriptomics revealed that neuropilin-1 (Nrp-1) supports Tregs migration behavior, and the genes of Crem and Tnfrsf9 regulate the behaviors of the terminal suppressive Tregs. Nrp-1 + 4-1BB - Tregs stepwise develop to the Nrp-1 - 4-1BB + Tregs from lymphoid tissues into the tumor. Moreover, Treg-restricted Nrp1 depletion abolishes anti-PD-1-upregulated intratumoral Tregs burden and synergizes with the 4-1BB agonist to enhance the antitumor response. Finally, a combination of the Nrp-1 inhibitor and the 4-1BB agonist in humanized HCC models showed a favorable and safe outcome and evoked the antitumor effect of the PD-1 blockade. CONCLUSION: Our findings elucidate the potential mechanism of anti-PD-1-mediated intratumoral Tregs accumulation in HCC and uncover the tissue adaptation characteristics of Tregs and identify the therapeutic potential of targeting Nrp-1 and 4-1BB for reprogramming the HCC microenvironment.

Lenvatinib enhances antitumor immunity by promoting the infiltration of TCF1+ CD8+ T cells in HCC via blocking VEGFR2.

Lenvatinib is the favorable treatment for advanced hepatocellular carcinoma (HCC), and it is currently undergoing phase III clinical trials. However, the specific effects of lenvatinib on PD1+ CD8+ T cells in HCC microenvironment have not been systematically studied. Here, we established an orthotopic hepa1-6 mouse model treated with lenvatinib to investigate CD8+ T cells' role in the tumor and spleen. We found an increasing proportion of TCF-1+ in PD1+ CD8+ T cells and proliferation of PD1+ CD8+ T cells after lenvatinib treatment. Meanwhile, lenvatinib treatment upregulated the expression of granzyme B on PD1+ CD8+ T cells both in vitro and in vivo. Lenvatinib activated the endogenous mTOR pathway of exhausted CD8+ T cells, and mTOR pathway blockade eliminated the antitumor effect of lenvatinib and function of PD1+ CD8+ T cells. The effects of the mTOR pathway on PD1+ CD8+ T cells after lenvatinib treatment were mediated by VEGFR2 inhibition. Overall, our work provides insight into the mechanism of lenvatinib's antitumor efficacy through exhausted CD8+ T cells in HCC treatment.

Gestational cholestasis induced intrauterine growth restriction through triggering IRE1α-mediated apoptosis of placental trophoblast cells.

Epidemiological and animal experimental studies suggest an association between gestational cholestasis and intrauterine growth restriction (IUGR). Here, we explored the mechanism through which gestational cholestasis induced IUGR. To establish gestational cholestasis model, pregnant mice were subcutaneously injected with 17α-Ethynylestradiol (E2) on gestational day 13 (GD13)-GD17. Some pregnant mice were intraperitoneally injected with 4µ8C on GD13-GD17. The results found that the apoptosis of trophoblast cells was elevated in placentas of mice with gestational cholestasis and in deoxycholic acid (DCA)-treated human trophoblast cell lines and primary mouse trophoblast cells. Correspondingly, the levels of placental cleaved caspase-3 and Bax were increased, while placental Bcl2 level was decreased in mice with gestational cholestasis and in DCA-treated trophoblast cells. Further analysis found that placental IRE1α pathway was activated in mice with gestational cholestasis and in DCA-treated trophoblast cells. Interestingly, 4µ8C, an IRE1α RNase inhibitor, significantly inhibited caspase-3 activity and apoptosis of trophoblast cells in vivo and in vitro. Importantly, 4µ8C rescued gestational cholestasis-induced placental insufficiency and IUGR. Furthermore, a case-control study demonstrated that placental IRE1α and caspase-3 pathways were activated in cholestasis cases. Our results provide evidence that gestational cholestasis induces placental insufficiency and IUGR may be via triggering IRE1α-mediated apoptosis of placental trophoblast cells.

Trends in insufficient physical activity among adults in China 2010-18: a population-based study.

BACKGROUND: The global prevalence of insufficient physical activity (PA) was reported to be 27.5% in 2016, and there were stable levels of insufficient PA worldwide between 2001 and 2016. The global target of a 10% reduction in insufficient PA by 2025 will not be met if the trends remain. The relevant data for trends in China were still scarce. This study aimed to determine nationwide temporal trends in insufficient PA among adults in China from 2010 to 2018. METHODS: 645 903 adults aged 18 years or older were randomly selected from four nationally representative cross-sectional surveys of the China Chronic Disease and Risk Factor Surveillance conducted in 2010, 2013, 2015, and 2018. PA was measured using the Global Physical Activity Questionnaire. Temporal changes in insufficient PA prevalence and participation of domain-specific moderate- to vigorous-intensity PA (MVPA) were analyzed using logistic regression. RESULTS: From 2010 to 2018, the age-adjusted prevalence of insufficient PA in China increased from 17.9% (95% confidence interval 16.3% to 19.5%) in 2010 to 22.3% (20.9% to 23.8%) in 2018 (P for trend < 0.001). By age group, with a significant increase in insufficient PA in adults aged 18-34 years (P for trend < 0.001), which rose more rapidly than in adults aged ≥ 35 years (P for interaction < 0.001). Insufficient PA has increased significantly among adults engaged in agriculture-related work, non-manual work, and other manual work (all P for trend < 0.05). And among the occupational groups, those engaged in agriculture-related work had the fastest increase (P for interaction = 0.01). The percentage of adults participating in work-related MVPA decreased from 79.6% (77.8% to 81.5%) to 66.8% (64.9% to 68.7%) along with a decrease in time spent on work-related MVPA, while percentages of adults participating in recreation-related MVPA increased from 14.2% (12.5% to 15.9%) to 17.2% (16.0% to 18.4%) (all P for trend < 0.05). CONCLUSIONS: Among Chinese adults, an increasing trend was found in insufficient PA from 2010 to 2018, with more than one-fifth of adults failing to achieve the recommendation of adequate PA. More targeted PA promotion strategies should be developed to improve population health.

A study on the impact of the number of family generations on intergenerational support for centenarians: a study in a Chinese 'longevity city'.

BACKGROUND: As a natural source of support for the elderly, the family is an important channel for achieving a sense of security, happiness, and worthiness in old age. In this study, we analysed the characteristics of intergenerational support in families of centenarians and explored the impact of the number of family generations on intergenerational support. METHODS: We conducted a cross-sectional survey between April 2020 and January 2021 among 62 elderly people aged 99+ in Rugao, China, one of six 'longevity cities' in the world. Assisted by the researchers, centenarians completed questionnaires with details pertaining to general demographics, intergenerational support, and other aspects. We used a logistic regression model to analyse the influence of the number of family generations on intergenerational support that the centenarians received with respect to economic, living, and emotional aspects. RESULTS: Centenarians were primarily recipients of care in their families, and received intergenerational support mainly for their declined physical functions and limited self-care ability. The study results revealed that the greater the number of generations comprising the family, the greater was the intergenerational life care and emotional comfort provided for centenarians by the family. CONCLUSIONS: In this study, we found a positive effect of the number of family generations on intergenerational support for centenarians. The government and society should promote the tradition of respecting, caring for, and honouring the elderly while paying close attention to the dynamic changes in the family structure of centenarians in promoting high-quality and sustainable development of the people, economy, and society.

Rapid detection of carbamate pesticide residues using microchip electrophoresis combining amperometric detection.

The long-term consumption of food with pesticide residues has harmful effects on human health and the demand for pesticide detection technology tends to be miniaturized and instant. To this end, we demonstrated the first application of indirectly detecting two carbamate pesticides, metolcarb and carbaryl, by gold nanoparticle-modified indium tin oxide electrode in dual-channel microchip electrophoresis and amperometric detection (ME-AD) system. m-Cresol and α-naphthol were obtained after pesticide hydrolysis in alkaline solution, and then separated and detected by ME-AD. Parameters including the detection potential and running buffer concentration and pH were optimized to improve the detection sensitivity and separation efficiency. Under the optimal conditions, the two analytes were completely separated within 80 s. m-Cresol and α-naphthol presented a wide linear range from 1 to 100 µM, with limits of detection of 0.16 µM and 0.34 µM, respectively (S/N = 3). Moreover, the reliability of this system was demonstrated by analyzing metolcarb and carbaryl in spiked vegetable samples.

Identification and characterization of the lamprey cathepsin genes.

Cathepsins are key mammalian proteases that play an important role in the immune response. Several studies have revealed the versatile and critical functions of cathepsins. Here, we obtained ten kinds of cathepsin homologs and identified seven homologs with complete coding sequences. Phylogenetic analysis verified their identities and supported the classification of cathepsins into seven families, which is similar to other vertebrates. Tissue-specific expression analysis showed that all lamprey cathepsins (L-cathepsins) are present in the supraneural body (SB), kidney, gill, intestine, brain, heart, and liver, but their relative abundance varied among tissues. Additionally, we focused on the lamprey cathepsin L (L-cathepsin L) and used recombinant L-cathepsin L protein (rL-cathepsin L) to prepare anti rL-cathepsin L polyclonal antibodies, which were used to detect its distribution in lamprey tissues. The L-cathepsin L protein was primarily detected in the SB, kidney, gill, intestine, brain, and liver via western blot and immunohistochemistry assays. Importantly, quantitative real-time PCR (RT-PCR) revealed that the expression level of L-cathepsins mRNA significantly increased after exposure to three different stimuli (poly I:C, Staphylococcus aureus (S.a) and Vibro anguilarum (V.an)). This suggested that L-cathepsins may participate in defense processes. These results revealed that L-cathepsins may play key roles in the immune response to exogenous stimuli. The findings provide important information for future studies aiming to understand the molecular mechanisms underlying the immune response to pathogen invasion in lamprey.

Effects of tillage managements and maize straw returning on soil microbiome using 16S rDNA sequencing.

Agricultural practices could affect bacterial diversity and community structure by altering soil physical and chemical properties. Straw returning and tillage practices are widely used in agriculture, however, the effects of these agricultural practices on microbiomes are still unclear. In the present study, we compared the 18 bacterial communities of soil with different straw returning and tillage treatment combinations. The V3-V4 regions of the 16S ribosomal RNA were amplified and analyzed by high-throughput sequencing technology. The results showed that the bacterial communities were consistently dominated by Acidobacteria, Proteobacteria, Actinobacteria, and Chloroflexi. Short-term straw returning and tillage practices significantly altered the diversity, relative abundance and functions of the soil microbiome. Soil subjected to rotary tillage and straw returning (RTS) combination possessed the highest bacterial diversity and lowest ratio of G+/G- bacteria, indicating that RTS could be an efficient integrated management system to improve microbiome in the short term. Double verifications based on relative abundance and network analysis, revealed close relationships of Mycobacterium and Methylibium with RTS, indicating they could serve as biomarkers for RTS. Investigating microbial changes under different agricultural practices will provide valuable foundations for land sustainable utilization and increase crop yields.

High mobility group box transcription factor 1 (HBP1) from Lampetra japonica affects cell cycle regulation.

High mobility group (HMG) box-containing protein 1 (HBP1) is a member of the HMG family of chromosomal proteins. Previous studies have shown that human HBP1 exhibits tumor-suppressor activity. Here, we identified a homologue of HBP1, L-hbp1, in Lampetra japonica. The L-hbp1 gene shared high sequence similarity with its homologues in jawed vertebrates, as shown by bioinformatics analyses. L-hbp1 contains a 1,584-bp open reading frame that encodes 527 amino acids. A pAdenox-L-HBP1 plasmid was constructed and transfected successfully in Raji cells, as revealed by real-time PCR. The overexpression of L-HBP1 reduced cell growth rates, inhibited G1 phase progression, decreased cyclin D1 and c-Myc protein expression, and increased p53 protein expression. Western blot and immunohistochemical assays showed that L-HBP1 was primarily distributed in the heart, kidney, gill and liver of lamprey. Cell cycle analysis revealed that decreased L-HBP1 expression in HBP1 morpholino oligonucleotide-transfected lamprey cells resulted in a decreased fraction of cells in the G1 phase and corresponding increases in the S and G2/M phases. Additionally, treatment of lamprey cardiac cells with pharmacological inhibitors of p38 MAP kinase released the cells from G1 arrest. Together, these results indicated that HBP1 expression in lamprey was correlated with the onset of mitotic arrest in these cells, which have implications for cell cycle regulation.

Investigation of lethal thresholds of nanosecond pulsed electric field in rabbit VX2 hepatic tumors through finite element analysis and verification with a single-needle bipolar electrode: A prospective strategy employing three-dimensional comparisons.

Pulsed electric field has emerged as a promising modality for the solid tumor ablation with the advantage in treatment planning, however, the accurate prediction of the lesion margin requires the determination of the lethal electric field (E) thresholds. Herein we employ the highly repetitive nanosecond pulsed electric field (RnsPEF) to ablate the normal and VX2 tumor-bearing livers of rabbits. The ultrasound-guided surgery is operated using the conventional double- and newly devised single-needle bipolar electrodes. Finite element analysis is also introduced to simulate the E distribution in the practical treatments. Two- and three-dimensional investigations are performed on the image measurements and reconstructed calcification models on micro-CT, respectively. Specially, an algorithm considering the model surface, volume and shape is employed to compare the similarities between the simulative and experimental models. Blood vessel injury, temperature and synergistic efficacy with doxorubicin (DOX) are also investigated. According to the three-dimensional calculation, the overall E threshold is 4536.4 ± 618.2 V/cm and the single-needle bipolar electrode is verified to be effective in tissue ablation. Vessels are well preserved and the increment of temperature is limited. Synergy of RnsPEF and DOX shows increased apoptosis and improved long-term tumor survival. Our study presents a prospective strategy for the evaluation of the lethal E threshold, which can be considered to guide the future clinical treatment planning for RnsPEF.

Separation and purification of flavonoid from Taxus remainder extracts free of taxoids using polystyrene and polyamide resin.

An efficient separation process of flavonoid from Taxus wallichiana var. mairei remainder extracts free of taxoids was developed in this study. AB-8 macroporous resin and polyamide resin offered the fine adsorption capacity, and its adsorption rate at 30°C fitted well to the Langmuir and Freundich isotherms. Resin dynamic adsorption and desorption experiments were conducted to optimize the separation process of total flavonoids from T. wallichiana var. mairei remainder extracts free of taxoids. The optimum parameters for adsorption by AB-8 resin were as follows: (1) the concentration of flavonoids in a sample solution of 5.61 mg/mL with a processing volume of 2 bed volume (BV) (60 mL); (2) for desorption, ethanol-water (80:20, v/v), with 6 BV as an eluent at a flow rate of 2 BV/h. After a one-run treatment with AB-8 resin, the content of flavonoids was increased 5.10-fold from 4.05 to 20.65%. The optimum parameters for adsorption by polyamide resin were as follows: processing volume of 2 BV (30 mL); for desorption, ethanol-water (70:30, v/v), with 8 BV as an eluent at a flow rate of 2 BV/h. After one-run treatment with polyamide resin, the content of total flavonoids increased from 20.65 to 65.21%. The method will provide a potential approach for large-scale separation and purification of flavonoid for its wide pharmaceutical use.

CD163 in Macrophages: A Potential Biomarker for Predicting the Progression of Diabetic Nephropathy based on Bioinformatics Analysis.

OBJECTIVE: This study aimed to identify the potential biomarkers in DN. METHODS: DN datasets GSE30528 and GSE47183 were downloaded from the Gene Expression Omnibus database. Immune cell infiltration was analyzed using CIBERSORT. Weighted gene co-expression network analysis (WGCNA) was performed to obtain the module genes specific to DN. The relevant genes were identified intersecting the module genes and differentially expressed genes (DEGs). The core genes were identified using the MCC algorithm in Cytoscape software. ROC and Pearson analyses alongside gene set enrichment analysis (GSEA) were performed to identify the key gene for the core genes. Finally, we performed the Spearman to analyze the correlation between key gene and glomerular filtration rate (GFR), serum creatinine (Scr), age and sex in DN. RESULTS: CIBERSORT analysis revealed the immune cell infiltration in the DN renal tissue and Venn identified 12 relevant genes. Among these, 5 core genes, namely TYROBP, C1QA, C1QB, CD163 and MS4A6A, were identified. Pearson analyses revealed that immune cell infiltration and expression of core genes are related. The key genes with high diagnostic values for DN were identified to be CD163 via ROC analyses. After Spearman correlation analysis, the expression level of CD163 was correlated with GFR (r =0.27), a difference that nearly reached statistical significance (P =0.058). However, there was no correlation between the level of CD163 and age (r =-0.24, P =0.09), sex (r =-0.11, P=0.32) and Scr (r=0.15, P=0.4). CONCLUSION: We found that CD163 in macrophages may be a potential biomarker in predicting and treating DN.

Comparison between preoperative hook-wire and liquid material localization for pulmonary nodules: a meta-analysis.

Introduction: Computed tomography (CT)-guided liquid material (LM) and hook-wire (HW) are usually localized for pulmonary nodules (PNs) before video-assisted thoracic surgery (VATS) resection, but the relative advantages of these 2 techniques remain uncertain. Aim: This meta-analysis was conceived to juxtapose the efficacy and safety of HW localization (HWL) and LM localization (LML), both guided by CT, for the preoperative localization of PNs. Material and methods: The PubMed, Web of Science, and Wanfang databases were searched to identify relevant studies published as of March 2023, after which pooled analyses of study outcomes were conducted. Results: A total of 7 studies were included in this meta-analysis from 142 relevant studies. These 7 studies included 551 patients (583 PNs) with CT-guided HWL and 551 patients (612 PNs) with LML. The successful localization rate was significantly higher in the LM group (LMG) than in the HW group (HWG) (p = 0.002). The LMG also exhibited significantly lower pooled total complication and lung haemorrhage rates than the HWG (p = 0.007 and 0.00001, respectively). Pooled localization duration, pneumothorax rates, and VATS procedure duration were comparable in both groups (p = 0.45, 0.15, and 0.74, respectively). Furthermore, the pooled postoperative hospital stay was significantly shorter in the LMG than in the HWG (p = 0.009). Significant heterogeneity was detected in the endpoints of localization duration and pneumothorax rate (I2 = 93% and 66%, respectively). Conclusions: CT-guided LML is safer and more successful than HWL for patients with PNs before VATS resection.

Benefits of SGLT2 inhibitors combining with renin-angiotensin-system blockers on cardiovascular outcomes in chronic kidney disease patients: A systemic review and meta-analysis.

BACKGROUND AND OBJECTIVE: Efficacy of sodium-glucose cotransporter 2 (SGLT2) inhibitors in combination with renin-angiotensin-system (RAS) blockers for CKD remains controversial. We conducted this meta-analysis to explore the effect of SGLT2 inhibitors combining with RAS blockers on cardiovascular outcomes in chronic kidney disease (CKD) patients. METHODS: We searched Embase, PubMed, Web of Science, and Cochrane Library databases with the following keywords. "Renal Insufficiency, Chronic" or "Diabetic Nephropathies" and "Sodium-glucose cotransporter 2 inhibitors". We included randomized controlled trials (RCTs) based on angiotensin-converting enzyme inhibitor (ACEI) or angiotensin II receptor blocker (ARB) therapy. The outcome events included cardiac and renal outcomes and other adverse events. This study is registered with PROSPERO: CRD42020218337. RESULTS: Ten RCTs including 16,983 CKD patients met the inclusion criteria. Compared with placebo plus RAS blockers, SGLT2 inhibitors plus RAS blockers significantly reduced cardiovascular mortality and heart failure-related hospitalization rates (RR=0.78, 95% CI: 0.66-0.91, p=0.002; RR=0.7, 95% CI: 0.61-0.8, p=0.000). We also performed trials sequential analysis (TSA) and the results indicated that our results are reliable. Additionally, it significantly reduced the 24-h urinary albumin excretion rate (24hUAE) and the creatinine elevation rate (WMD=-0.19, 95% CI: -0.24 to -0.14; RR=0.61, 95% CI: 0.51-0.74, p=0.000), delayed progression to end-stage renal disease (ESRD) (RR=0.69, 95% CI: 0.59-0.81, p=0.000). Further, it had no significant effect on the incidence of renal-related adverse events or renal-related mortality. Although it decreased the estimated glomerular filtration rate (eGFR) (WMD=-5.4, 95% CI: -7.24 to -3.57), this effect was reversible. CONCLUSIONS: These data provide a well-document testimonial of the benefits of the combined use of SGLT2 inhibitors and RAS blockers for cardiovascular and renal outcomes in CKD patients.

Low red to far-red light ratio promotes salt tolerance by improving leaf photosynthetic capacity in cucumber.

Soil salinity severely inhibits leaf photosynthesis and limits agricultural production. Red to far-red light ratio (R/FR) affects leaf photosynthesis under salt stress, however, its regulation mechanism is still largely unknown. This study investigated the effects of different R/FR on plant growth, gas exchange parameters, photosynthetic electron transport, Calvin cycle and key gene expression under salt stress. Cucumber seedlings were exposed to four treatments including 0 mM NaCl and R/FR=7 (L7, control), 0 mM NaCl and R/FR=0.7 (L0.7), 80 mM NaCl and R/FR=7 (H7) and 80 mM NaCl and R/FR=0.7 (H0.7) for 9 days in an artificial climate chamber. The results showed that compared to L7 treatment, H7 treatment significantly reduced relative growth rate (RGR), CO2 assimilation rate (P n), maximum photochemical efficiency PSII (F v/F m), most JIP-test parameters and total Rubisco activity, indicating that salt stress severely inhibited photosynthetic electron transport from PSII to PSI and blocked Calvin cycle in cucumber leaves. However, these suppressions were effectively alleviated by low R/FR addition (H0.7 treatment). Compared to H7 treatment, H0.7 treatment significantly increased RGR and P n by 209.09% and 7.59%, respectively, enhanced F v/F m, maximum quantum yield for primary photochemistry (φ Po), quantum yield for electron transport (φ Eo) and total Rubisco activity by 192.31%, 17.6%, 36.84% and 37.08%, respectively, and largely up-regulated expressions of most key genes involved in electron transport and Calvin cycle. In conclusion, low R/FR effectively alleviated the negative effects of salt stress on leaf photosynthesis by accelerating photosynthetic electron transport from PSII to PQ pool and promoting Calvin cycle in cucumber plants. It provides a novel environmentally friendly light-quality regulation technology for high efficiency salt-resistant vegetable production.

A pan-cancer analysis of the oncogenic role of Holliday junction recognition protein in human tumors.

Although cell-based or animal-based research evidence support the association of Holliday junction recognition protein (HJURP) with cancers, no pan-cancer investigation has been reported. The datasets of Gene Expression Omnibus database along with The Cancer Genome Atlas project were used to evaluate the expression of HJURP in various types of tumors. HJURP is overexpressed in a considerable number of cancers, and some changes in DNA methylation and genetic alterations are discovered in some types of tumors, such as kidney-related and adrenal gland-related tumors. Based on PrognoScan and gene expression profiling interactive analysis (GEPIA), the elevated expression of HJURP worsened the survival time of individuals with cancer. The biological general repository for interaction datasets (BioGRID) and The database for annotation, visualization and integrated discovery (DAVID) were used to establish the functional molecular network. It revealed that the cell cycle and p53 signaling pathway are the key molecular mechanisms that HJURP promotes carcinogenesis. The nomograms between HJURP and clinical pathological factors based on the Cox proportional hazards model showed a good prognostic performance in kidney carcinoma, hepatocellular carcinoma, and lung adenocarcinoma. Our first pan-cancer study provides a relatively profound insights into the oncogenic roles of HJURP across different tumors.

Development and validation of a nomogram for predicting mild cognitive impairment in middle-aged and elderly people.

BACKGROUND: Mild cognitive impairment (MCI) is a clinical cognitive impairment state between dementia and normal aging. Early identification of MCI is beneficial, and it can delay the development of dementia. We aimed to develop and validate a prediction model to predict MCI of middle-aged and elderly people (aged 45 years and over). METHODS: According to 478 middle-aged and elderly people (48-85 years old) from a cross-sectional study, we developed and validated a predictive nomogram. The least absolute shrinkage and selection operator (LASSO) regression model and multivariate logistic regression analysis were used to select variables and develop a prediction model. The performance of the nomogram was evaluated in terms of its discriminative power, calibration, and decision curve analysis (DCA). RESULTS: The predictive nomogram was composed of the following: age, gender, education level, residence, and reading. The model showed good discrimination power (area under receiver-operating characteristic (ROC) curve was 0.8704) and good calibration. Similar results were seen in 10-fold cross-validation. The nomogram showed clinically useful in DCA analysis. CONCLUSION: This predictive nomogram provides researchers with a practical tool for predicting MCI. The variables included in this nomogram were readily available. The population used for this nomogram was middle-aged and elderly people.

Disulfiram/Copper Induces Immunogenic Cell Death and Enhances CD47 Blockade in Hepatocellular Carcinoma.

Some chemotherapeutic agents have been found to enhance antitumor immunity by inducing immunogenic cell death (ICD). The combination of disulfiram (DSF) and copper (Cu) has demonstrated anti-tumor effects in a range of malignancies including hepatocellular carcinoma (HCC). However, the potential of DSF/Cu as an ICD inducer and whether it can enhance the efficacy of the immune checkpoint blockade in HCC remains unknown. Here, we showed that DSF/Cu-treated HCC cells exhibited characteristics of ICD in vitro, such as calreticulin (CRT) exposure, ATP secretion, and high mobility group box 1 (HMGB1) release. DSF/Cu-treated HCC cells elicited significant immune memory in a vaccination assay. DSF/Cu treatment promoted dendritic cell activation and maturation. The combination of DSF/Cu and CD47 blockade further facilitated DC maturation and subsequently enhanced CD8+ T cell cytotoxicity. Mechanically, DSF/Cu promoted the nuclear accumulation and aggregation of nuclear protein localization protein 4 (NPL4) to inhibit the ubiquitin-proteasome system; thus, inducing endoplasmic reticulum (ER) stress. The inhibition of NPL4 induced ICD-associated damage-associated molecular patterns. Collectively, our findings demonstrated that DSF/Cu-induced ICD-mediated immune activation in HCC enhanced the efficacy of CD47 blockade.

Combination with Toll-like receptor 4 (TLR4) agonist reverses GITR agonism mediated M2 polarization of macrophage in Hepatocellular carcinoma.

The glucocorticoid-induced tumor necrosis factor receptor (GITR) agonistic antibody (DTA-1) has been proved to elicit robust immune response in various kinds of tumors. However, only a few of the HCC patients could benefit from it, and the mechanism of DTA-1 resistance remains unknown. Here, we measured GITR expression in different immunocytes in HCC microenvironment, and we observed that tumor-infiltrating regulatory T cells (Ti-Tregs) significantly expressed GITR, which were associated with poor prognosis. Meanwhile, we analyzed the variation of tumor-infiltrating immune components and associated inflammation response after DTA-1 treatment in orthotopic liver cancer model of mice. Surprisingly, DTA-1 treatment reduced the infiltration of Tregs but failed to activate CD8+ T cells and elicit antitumor efficacy. In particular, DTA-1 treatment enforced alternative M2 polarization of macrophage, and macrophage depletion could enhance DTA-1-mediated antitumor efficacy in HCC. Mechanistically, macrophage M2 polarization attributed to the IL-4 elevation induced by Th2 immune activation in the treatment of DTA-1, resulting in DTA-1 resistance. Furthermore, Toll-like receptor 4 (TLR4) agonist could diminish the macrophage (M2) polarization and reverse the M2-mediated DTA-1 resistance, eliciting robust antitumor effect in HCC. Our finding demonstrated that the TLR4 agonist synergized with DTA-1 was a potential strategy for HCC treatment.

Di-(2-ethylhexyl) phthalate induces testicular endoplasmic reticulum stress and germ cell apoptosis in adolescent mice.

Di-(2-ethylhexyl) phthalate (DEHP) is a male reproductive toxicant. This research is aimed at investigating the effect of pubertal DEHP exposure on testicular endoplasmic reticulum (ER) stress and germ cell apoptosis. Five-week-old male mice were orally administered with DEHP (0, 0.5, 50, or 500 mg/kg/day) for 35 days. Testis weight and sperm count were reduced in mice exposed to 500 mg/kg/day DEHP. The number of seminiferous tubules in stages VII-VIII, mature seminiferous tubules, was reduced and the number of seminiferous tubules in stages IX-XII, immature seminiferous tubules, was elevated in mice treated with 500 mg/kg/day DEHP. Numerous apoptotic germ cells were observed in mouse seminiferous tubules exposed to 50 and 500 mg/kg/day DEHP. Moreover, cleaved caspase-3 was elevated in mouse testes exposed to 500 mg/kg/day DEHP. In addition, Bcl-2 was reduced and Bax/Bcl-2 was elevated in mouse testes exposed to 500 mg/kg/day DEHP. Additional experiment showed that GRP78, an ER molecular chaperone, was downregulated in mouse testes exposed to 500 mg/kg/day DEHP. Testicular p-IRE-1α, p-JNK, and CHOP, three markers of ER stress, were upregulated in mice exposed to 500 mg/kg/day DEHP. These results suggest that pubertal exposure to high doses of DEHP induces germ cell apoptosis partially through initiating ER stress in testes.