Background: Evidence supports the effectiveness and safety of open-source automated insulin delivery (AID) in patients with type 1 diabetes. However, evidence regarding the clinical application of open-source AID in perioperative patients with type 2 diabetes remains limited. Methods: This was an open-label, single-center, exploratory pilot randomized controlled trial (RCT) with parallel groups. Patients with diabetes (excluding type 1 diabetes mellitus) scheduled for elective surgery were randomly assigned to the closed-loop group (open-source hybrid closed-loop AID system) or the control group (conventional insulin pump). The primary outcome was the percentage of time in the target glucose range (TIR, 3.9-10.0 mmol/L). Other efficacy and safety outcomes were also compared between the groups. Results: A total of 49 participants were included and randomized to the closed-loop group (n = 25) or the control group (n = 24). Participants underwent abdominal, orthopedic, thoracic surgery, or neurosurgery during hospitalization. Patients in the closed-loop group had significantly higher TIR than patients in the control group (76.4 ± 14.1% vs. 61.2 ± 20.0%, p = 0.005). Compared with the control group, the closed-loop group also exhibited a 15.6 percentage point reduction in time above range (TAR, >10 mmol/L) without increasing time below range (TBR, <3.9 mmol/L). There were no episodes of severe hypoglycemia (<2.2 mmol/L) or diabetic ketoacidosis in either group. Conclusions: This study demonstrates that in patients with diabetes undergoing elective surgery, the open-source hybrid closed-loop AID system provides better glycemic control than conventional insulin pump therapy.

Cancer cells undergo metabolic reprogramming to meet the demands of rapid proliferation, survival and chemotherapy resistance. Targeting cancer-specific metabolic vulnerabilities offers a compelling strategy for therapeutic intervention. Owing to the Warburg effect and the unique tumor microenvironment, pancreatic ductal adenocarcinoma (PDAC) cells exhibit a high demand for glucose to sustain their energy metabolism. Here, we identify a novel regulatory mechanism controlling the cell surface abundance of glucose transporter 1 (GLUT1), mediated by RAB8A-dependent vesicular trafficking. RAB8A, a member of the RAS oncogene family, enhances GLUT1 membrane localization and thereby increases glucose uptake in PDAC cells. Mechanistically, we demonstrate that ubiquitin-specific peptidase 20 (USP20) negatively regulates RAB8A activation by selectively removing K48-linked polyubiquitin chains from its inactive form. Functional assays in vitro and in vivo validate the tumor-suppressive role of the USP20-RAB8A signaling axis. Furthermore, using primary PDAC cells derived from KPC (KrasG12D/+; Trp53 R172Hflox/flox; Pdx1-Cre) mice, we show that dual knockdown of Rab8a and Glut1 markedly attenuates tumor-promoting effects driven by oncogenic Kras and Trp53 loss. Collectively, our findings reveal that the USP20-RAB8A-GLUT1 axis regulates glucose uptake and metabolic reprogramming in PDAC, thereby inhibiting tumor growth and metastasis. Targeting this signaling axis provides a novel insight into metabolic therapy for pancreatic cancer.

Gallbladder cancer (GBC) remains a challenging malignancy with a poor prognosis largely due to its highly metastatic nature and lack of effective treatment options. In this study, we investigated the role of N6-methyladenosine (m6A) modification and its regulatory factors, METTL3 and METTL14, in GBC. Our results showed that m6A levels as well as the expression of METTL3 and METTL14 were significantly downregulated in GBC tissues compared to normal gallbladder tissues. In vitro experiments showed that manipulation of METTL3 and METTL14 expression regulated GBC cell migration and invasive ability, and the liver metastasis model of nude mice further demonstrated the involvement of m6A modification in regulating GBC metastasis. Further investigation identified miR-146a-5p as a downstream target regulated by m6A, with tumour-suppressive effects on GBC cell migration and invasion. Overall, our findings provide new insights into the role of m6A modification and its regulation on microRNA in GBC pathogenesis and offer a potential strategy for the treatment of GBC.

Pancreatic cancer is the seventh leading cause of cancer death worldwide, which is demonstrated with remarkable resistance to radiotherapy and chemotherapy. The identification of prognosis signature and novel prognostic markers will facilitate patient stratification and an individualized precision therapy strategy. In this study, TCGA-PAAD was used to screen prognostic E3 ubiquitin ligases and establish prognostic signatures, and GEO database was used to verify the accuracy of prognostic signatures. Functional analysis, in vitro experiments and clinical cohort studies were used to analyze the function and prognostic efficacy of the target gene. An E3 ligase-based signature of 9 genes and the nomogram were developed, and the signature was proved to accurately predict the prognosis of patients with pancreatic cancer. WDR37 might be the most prognostic E3 ubiquitin ligase in pancreatic cancer, and the clinical cohort analyses suggested a tumor‐suppressive role. The results of functional analysis and in vitro experiments indicated that WDR37 may promote the degradation of TCP1 complex to inhibit tumor and improve immune cell infiltration. The E3 ligase-based signature accurately predicted the prognosis of patients with pancreatic cancer, so it can be used as a decision-making tool to guide the treatment of patients with pancreatic cancer. At the same time, WDR37, the main gene in E3PMP signature, can be used as the most prognostic E3 ubiquitin ligase in the treatment of pancreatic cancer.

Cholangiocarcinoma is one of the most lethal human cancers, and chemotherapy failure is a major cause of recurrence and poor prognosis. We previously demonstrated that miR-200 family members are downregulated in clinical samples of cholangiocarcinoma and inhibit cholangiocarcinoma tumorigenesis and metastasis. However, the role of differentially expressed miR-200b-3p in 5-fluorouracil chemosensitivity remains unclear. Here, we examined how miR-200b-3p modulates 5-fluorouracil chemosensitivity in cholangiocarcinoma. We observed that miR-200b-3p was associated with 5-fluorouracil sensitivity in cholangiocarcinoma and increased 5-fluorouracil-induced mitochondrial apoptosis in cholangiocarcinoma cells. Mechanistically, miR-200b-3p suppressed autophagy in cholangiocarcinoma cells to mediate 5-fluorouracil sensitivity. Further, we identified KLF4 as an essential target of miR-200b-3p in cholangiocarcinoma. Notably, the miR-200b-3p/ KLF4/autophagy pathway augmented the chemosensitivity of cholangiocarcinoma cells to 5-fluorouracil. Our findings underscore the key role of miR-200b-3p in chemosensitivity to 5-fluorouracil and highlight the miR-200b-3p/KLF4/autophagy axis as a potential therapeutic target for cholangiocarcinoma.

Pyroptosis is a lytic and inflammatory form of gasdermin protein-mediated programmed cell death that is typically initiated by inflammasomes. The inflammasome response is an effective mechanism for eradicating germs and cancer cells in the event of cellular injury. The gasdermin family is responsible for initiating pyroptosis, a process in which holes are made in the cell membrane to allow inflammatory chemicals to escape. Mounting evidence indicates that pyroptosis is critical for controlling the development of cancer. In this review, we provide a general overview of pyroptosis, examine the relationship between the primary elements of pyroptosis and tumors, and stress the necessity of pyroptosis-targeted therapy in tumors. Furthermore, we explore its dual nature as a double-edged sword capable of both inhibiting and facilitating the growth of cancer, depending on the specific conditions. Ultimately, pyroptosis is a phenomenon that has both positive and negative effects on tumors. Using this dual impact in a reasonable manner may facilitate investigation into the initiation and progression of tumors and offer insights for the development of novel treatments centered on pyroptosis.

Objective: We aimed to investigate the subjective global assessment (SGA), symptoms and blood nutritional indicators in pancreaticobiliary tumor patients and the associated factors. Method: 153 pancreaticobiliary tumor patients’ information was collected on admission including SGA, symptoms, blood lymphocyte count (Lmy), hemoglobin (Hb), creatinine (Cr), total cholesterol (TC), total triacylglycerol (TG), serum C-reactive protein (CRP), globulin, albumin (Alb), prealbumin (PA), transferrin (Tf), retinol binding protein (RBP) and micro-nutrients (folic acid, vit B12, vit A, vit E, vit K, vit D, iron, zinc and copper). Associated factors were explored. Result: Advanced tumor stage, elevated CRP and CA 19-9 were risk factors for malnutrition by SGA; elevated CRP contributed to fatigue, anorexia and pain; Alb, PA and Tf decreased with age; men were associated with decrease of Lmy, folic acid and TC, and increase of vit D; advanced tumor stage was associated with elevated vit B12;elevated CRP was associated with decreased Hb, Alb, PA, Tf, RBP, vit A, vit D and increased globulin and copper; SGA was associated with decreased Lym, HB, Alb, PA and Tf; elevated bilirubin was associated with decreased Cr , Alb, vit K and vit D, and increased TC, TG, vit B12 and copper; Conclusion：Inflammation should be controlled to relieve the discomforts; old patients should give more protein; gender affects nutritional indicators; advanced cancer underestimates vitB12 deficiency. Inflammation overestimates the deficiency of serum protein, vit A, vit D, iron and zinc.Malnutrition by SGA increases the deficiency of Lym and serum protein. Obstructive jaundice increases the deficiency of Cr, Alb, vit D and vitK, and cause vitB12 elevation, hyperlipemia, and copper accumulation.

Inhibition of immunocyte infiltration and activation has been suggested to effectively ameliorate nonalcoholic steatohepatitis (NASH). Paired immunoglobulin-like receptor B (PirB) and its human ortholog receptor, leukocyte immunoglobulin-like receptor B (LILRB2), are immune-inhibitory receptors. However, their role in NASH pathogenesis is still unclear. Here, we demonstrate that PirB/LILRB2 regulates the migration of macrophages during NASH by binding with its ligand angiopoietin-like protein 8 (ANGPTL8). Hepatocyte-specific ANGPTL8 knockout reduces MDM infiltration and resolves lipid accumulation and fibrosis progression in the livers of NASH mice. In addition, PirB −/− bone marrow (BM) chimeras abrogate ANGPTL8-induced MDM migration to the liver. And yet, PirB ectodomain protein could ameliorate NASH by sequestering ANGPTL8. Furthermore, LILRB2-ANGPTL8 binding-promoted MDM migration and inflammatory activation are also observed in human peripheral blood monocytes. Taken together, our findings reveal the role of PirB/LILRB2 in NASH pathogenesis and identify PirB/LILRB2-ANGPTL8 signaling as a potential target for the management or treatment of NASH.

As the most abundant and conserved internal modification in eukaryote RNAs, N6-methyladenosine (m6A) is involved in a wide range of physiological and pathological processes. The YT521-B homology (YTH) domain-containing family proteins (YTHDFs), including YTHDF1, YTHDF2, and YTHDF3, are a class of cytoplasmic m6A-binding proteins defined by the vertebrate YTH domain, and exert extensive functions in regulating RNA destiny. Distinct expression patterns of the YTHDF family in specific cell types or developmental stages result in prominent differences in multiple biological processes, such as embryonic development, stem cell fate, fat metabolism, neuromodulation, cardiovascular effect, infection, immunity, and tumorigenesis. The YTHDF family mediates tumor proliferation, metastasis, metabolism, drug resistance, and immunity, and possesses the potential of predictive and therapeutic biomarkers. Here, we mainly summary the structures, roles, and mechanisms of the YTHDF family in physiological and pathological processes, especially in multiple cancers, as well as their current limitations and future considerations. This will provide novel angles for deciphering m6A regulation in a biological system.

Laparoscopic duodenum-preserving total pancreatic head resection (LDPPHRt) is used for treating benign or low-grade malignant tumors of the pancreatic head. However, preservation of the duodenum and biliary tract integrity remains challenging. We present a new approach for LDPPHRt and evaluate its feasibility and safety. From April 2020 to December 2020, 30 patients successfully underwent LDPPHRt using the intracapsular approach in our center. Their medical records were reviewed for relevant clinical characteristics, pathologic findings, postoperative complications, and survival. The median diameter of the lesions was 3.6 cm (range, 2.0–5.5 cm). The median operative time was 234.7 min (range, 195–310 min). The median blood loss was 66.7 ml (range, 20–250 ml). The morbidity rate was 26.7

Non-alcoholic steatohepatitis (NASH) is an advanced form of non-alcoholic fatty liver disease characterized by hepatosteatosis, liver cell injury, and inflammation. The pathogenesis of NASH involves dysregulated transcription of genes involved in critical processes in the liver, including metabolic homeostasis and inflammation. Chromatin immunoprecipitation (ChIP) utilizes antibody-mediated immunoprecipitation followed by the detection of associated DNA fragments via real-time PCR or high-throughput sequencing to quantitatively profile the interactions of proteins of interest with functional chromatin elements. Here, we present a detailed protocol to study the interactions of DNA and chromatin-associated proteins (e.g., transcription factors, co-activators, co-repressors, and chromatin modifiers) and modified histones (e.g., acetylated and methylated) in isolated primary mouse hepatocytes and mouse liver. The application of these methods can enable the identification of molecular mechanisms that underpin dysregulated hepatic processes in NASH.

Objective The aim of this study was to compare the short- and long-term outcomes of laparoscopic surgery (LS) and open surgery (OP) for perihilar cholangiocarcinoma (PHC) using a large real-world dataset in China. Methods Data of patients with PHC who underwent LS and OP from January 2013 to October 2018, across 10 centers in China, were extracted from medical records. A comparative analysis was performed before and after propensity score matching (PSM) in the LS and OP groups and within the study subgroups. The Cox proportional hazards mixed-effects model was applied to estimate the risk factors for mortality, with center and year of operation as random effects. Results A total of 467 patients with PHC were included, of whom 161 underwent LS and 306 underwent OP. Postoperative morbidity, such as hemorrhage, biliary fistula, abdominal abscess, and hepatic insufficiency, was similar between the LS and OP groups. The median overall survival (OS) was longer in the LS group than in the OP group (NA vs. 22 months; hazard ratio [HR] 1.19, 95% confidence interval [CI] 1.02–1.39, p = 0.024). Among the matched datasets, OS was comparable between the LS and OP groups (NA vs. 35 months; HR 0.99, 95% CI 0.77–1.26, p = 0.915). The mixed-effect model identified that the surgical method was not associated with long-term outcomes and that LS and OP provided similar oncological outcomes. Conclusions Considering the comparable long-term prognosis and short-term outcomes of LS and OP, LS could be a technically feasible surgical method for PHC patients with all Bismuth–Corlett types of PHC.

患者男性，39岁。因"腰痛8个月"入院。8个月前患者出现腰痛，伴乏力、肩颈痛，偶有心悸，无头痛、头昏，无四肢疼痛，至本院门诊就诊，测血钙2.69~2.78 mmol/L(正常值2.15~2.50 mmol/L，下同)，门诊以"高钙血症"收住入院。自起病以来，患者精神、睡眠、饮食欠佳，大小便正常，体力下降、体重下降约5 kg。既往史：否认特殊病史，否认家族遗传病史，其父有高钙血症病史，无任何症状。体格检查：体温36.5℃，脉搏128次/min，呼吸20次/min，血压120/92 mmHg(1 mmHg＝0.133 kPa)，神志清楚，精神可，无关节畸形等，查体未见明显特殊。

Abstract Background This study aimed to identify new protein markers that can evaluate the metastatic ability of Pancreatic neuroendocrine tumors (PanNETs) and predict prognosis together with Ki-67. Methods Multicentric series analysis was performed to evaluate the effect of metastasis on overall survival (N = 1,109). Single-cell RNA sequencing, proteomics (32 PanNETs and 10 normal pancreas tissues), and immunohistochemical staining (152 PanNETs) were conducted to screen proteins associated with PanNET metastatic capability. Associations between expression of screened proteins and survival were examined using Cox regression. In vitro experiments were performed to verify the function of candidate protein. A survival signature was constructed using recursive partitioning and survival forest analyses. Results The combination of G grade and metastasis predicted long-term survival more accurately though retrospective analysis. Single-cell RNA-seq highlighted the PanNET characteristics of metastasis that do not correspond with G grade and identified high metastatic capability subgroups in PanNET. Five proteins (PIN1, POSTN, SEMA4F, ASPN, and KCDT12), which may be related to the metastatic capability of PanNETs, were identified by proteomics. Random survival forest analysis showed that PIN1 could be a marker of PanNET metastatic ability. PIN1 overexpression promoted metastasis through LAMIN in PanNET cells. Finally, a survival signature with a consistency index of 0.921 and strong calibration was established, with good ability to predict patient survival. Conclusion In summary, we found that PIN1 can precisely evaluate the metastatic potential of PanNETs and, together with the proliferation marker Ki-67, can accurately predict the survival of PanNET patients.

Although non-alcoholic steatohepatitis (NASH) can progress to liver cancer and liver failure, no FDA-approved drugs exist to treat NASH. Deciphering the molecular mechanisms underlying the pathogenesis of NASH will facilitate the development of effective treatments for NASH, and requires loss- or gain-of-function experimental approaches. While genetically modified animals provide important information about the function of a gene, adenovirus is a fast, effective, and versatile tool that allows transient knockdown, knockout, or overexpression of one or more genes of interest (GOIs) in primary hepatocytes in vitro and in mouse liver in vivo. In addition, adenovirus is a promising treatment method in preclinical animal models, including rodents and non-human primates, and is used in many clinical trials. Here, we describe a step-by-step protocol to generate adenovirus for basic medical research. We discuss critical steps during virus propagation and purification and provide notes about how to avoid common pitfalls.

AJCC TNM stage and WHO grade (G) are two widely used staging systems to guide clinical management for pancreatic neuroendocrine neoplasms (panNENs), based on clinical staging and pathological grading information, respectively. We proposed to integrate TNM stage and G grade into one staging system (TNMG) and to evaluate its clinical application as a prognostic indicator for panNENs. Accordingly, 5254 patients diagnosed with panNENs were used to evaluate and to validate the applicability of TNMG to panNENs. The predictive accuracy of TNMG system was compared with that of each separate staging/grading system. We found that TNM stage and G grade were independent risk factors for survival in both the Surveillance, Epidemiology, and End Result (SEER) and multicenter series. The interaction effect between TNM stage and G grade was significant. Twelve subgroups combining the TNM stage and G grade were proposed in the TNMG stage, which were classified into five stages TNMG. According to the TNMG staging classification in the SEER series, the estimated median survival for stages I, II, III, IV, and V were 203, 174, 112, 61, and 8 months, respectively. The predictive accuracy of TNMG stage was higher than that of TNM stage and G grade used independently. The TNMG stage classification was more accurate in predicting panNEN patient's prognosis than either the TNM stage or G grade.

Copper is an essential nutrient and a co-factor of numerous enzymes governing a wide range of intracellular processes. Copper deficiency has emerged to be associated with various lipid metabolism diseases, including non-alcoholic fatty liver disease (NAFLD). However, the molecular mechanisms of how copper regulates lipid metabolism and is sensed remain elusive. Here, we reveal that copper elevation caused by hepatic ceruloplasmin (CP) ablation enhances lipid catabolism by promoting the assembly of copper-load SCO1/AMPK complex. We report that overnutrition-mediated CP elevation results in hepatic copper loss, and that liver-specific CP ablation counteracts this reduction in copper levels and ameliorates NAFLD in mice. Mechanistically, SCO1 constitutively interacts with LKB1 even in the absence of copper, and copper-loaded SCO1 directly tethers LKB1 to AMPK, thereby activating AMPK and consequently promoting mitochondrial biogenesis and fatty acid oxidation in hepatocytes. Therefore, this study reveals an unexpected role for AMPK to sense copper alteration via SCO1 and uncovers a previously unidentified mechanism by which copper, as a signaling molecule, improves hepatic lipid catabolism, and indicates that targeting copper-AMPK signaling pathway ameliorates NAFLD development by modulating AMPK activity.

Purpose: Cancer stem cells (CSCs) initiate and maintain tumorigenesis due to their unique pluripotency.However, pancreatic stem cell gene signatures are not completely revealed yet.Here, we isolated pancreatic cancer stem cells (P-CSCs) and exploited their distinct genome-wide mRNA and miRNA expression profiles using microarrays.Methods: CD24 + CD44 + ESA + cells were isolated from two pancreatic xenograft cells by the flow cytometry and identified the stem cell-like properties by the tumor formation, self-renew and chemoresistance.Microarrays and qRT-PCR were used to exploit their distinct Genome-wide mRNA and miRNA expression profiles.The function and candidate target genes of key microRNA were detected after Ectopic restoration in the pancreatic cancer cell lines MIA Paca-2 (CSC high ) and BxPC-3 (CSC low ).Results: In this study, we isolated P-CSCs from two xenografts cells.Genome-wide profiling experiments showed 479 genes and 15 microRNAs specifically expressed in the P-CSCs, including genes involved in TGF-β and p53 signaling pathways and particularly miR-146b-3p as the most significantly downregulated miRNA.We confirmed miR-146b-3p as a downregulated signature in pancreatic cancer tissues and cell line MIA Paca-2 (CSC high ) cells.Ectopic restoration of miR-146b-3p expression with pre-miR reduced cell proliferation, induced apoptosis, increased G1 phase and reduced S phase in cell cycle in MIA Paca-2 (CSC high ), but not in BxPC-3 (CSC low ).Re-expression of miR-146b-3p with lentivirus significantly inhibited tumorigenicity in vivo in MIA Paca-2, but slightly in BxPC-3.Furthermore, we demonstrated that miR-146b-3p directly targeted MAP3K10 and might activate Hedgehog pathway as well through DYRK2 and GLI2.Conclusions: These results suggest that P-CSCs have distinct gene expression profiles.MiR-146b-3p inhibits proliferation and induced apoptosis in P-CSCs high cells lines by targeting MAP3K10.Targeting P-CSCs specific genes may provide novel strategies for therapeutic purposes.

BACKGROUND:The American Joint Committee on Cancer (AJCC) made improvements for staging pancreatic neuroendocrine tumors (pNETs) in its 8th Edition; however, multicenter studies were not included. METHODS:We collected multicenter datasets (n = 1,086, between 2004 and 2018) to validate the value of AJCC 8 and other coexisting staging systems through univariate and multivariate analysis for well-differentiated (G1/G2) pNETs. RESULTS:Compared to other coexisting staging systems, AJCC 7 only included 12 (1.1%) patients with stage III tumors. Patients with European Neuroendocrine Tumor Society (ENETS) stage IIB disease had a higher risk of death than patients with stage IIIA (hazard ratio [HR]: 4.376 vs. 4.322). For the modified ENETS staging system, patients with stage IIB disease had a higher risk of death than patients with stage III (HR: 6.078 vs. 5.341). According to AJCC 8, the proportions of patients with stage I, II, III, and IV were 25.7%, 40.3%, 23.6%, and 10.4%, respectively. As the stage advanced, the median survival time decreased (NA, 144.7, 100.8, 72.0 months, respectively), and the risk of death increased (HR: II = 3.145, III = 5.925, and IV = 8.762). CONCLUSION:These findings suggest that AJCC 8 had a more reasonable proportional distribution and the risk of death was better correlated with disease stage.

The elucidation of the mechanisms whereby the liver maintains glucose homeostasis is crucial for the understanding of physiological and pathological states. Here, we show a novel role of hepatic transcriptional co-activator with PDZ-binding motif (TAZ) in the inhibition of glucocorticoid receptor (GR). TAZ is abundantly expressed in pericentral hepatocytes and its expression is markedly reduced by fasting. TAZ interacts via its WW domain with the ligand-binding domain of GR to limit the binding of GR to the GR response element in gluconeogenic gene promoters. Therefore, liver-specific TAZ knockout mice show increases in glucose production and blood glucose concentration. Conversely, the overexpression of TAZ in mouse liver reduces the binding of GR to gluconeogenic gene promoters and glucose production. Thus, our findings demonstrate that hepatic TAZ inhibits GR transactivation of gluconeogenic genes and coordinates gluconeogenesis in response to physiological fasting and feeding.