The intestinal epithelium is maintained by stem cells at the crypt base, however, acute or chronic inflammation can severely disrupt stem cell function and tissue homeostasis. To characterize mucosal inflammation and tissue damage, we evaluated time-course gene expression changes using a dextran sulfate sodium (DSS)-induced mouse model of colitis. By applying normalization, Z-score transformation, and spline curve modeling, we trace the temporal expression dynamics of key intestinal genes. This computational approach offer insights into the molecular responses during inflammation and may help identifying new biomarkers for chronic inflammatory diseases.
Methods in molecular biology (Clifton, N.J.)Cewen Chen, Qiaochu Fu, Chao Gao, Shinya Tanaka, Masamichi Imajo
Colorectal cancer (CRC) is a major malignancy with significant global implications for human health. The development of a reliable and pathologically relevant orthotopic CRC model is essential for advancing our understanding of its molecular mechanisms and for developing more effective therapeutic interventions. However, the construction of such models is fraught with challenges primarily because of the technical complexities involved in the transplantation of CRC cells into the intestinal epithelium. In this chapter, we describe a recently developed method for generating an orthotopic CRC model within the mouse cecal epithelium. This model enables tumor development and progression within a natural tissue microenvironment and facilitates complex interactions between tumor cells and surrounding normal cells, thereby replicating the intratumor heterogeneity of CRC. All procedures in this method are visualized under stereomicroscopic observation, leading to the efficient engraftment of CRC cells and subsequent tumor development. The establishment of this reliable orthotopic CRC model, which mimics tumor development in a more natural microenvironment, offers new opportunities to explore the molecular mechanisms underlying CRC and to assess novel anticancer therapies in pathologically relevant contexts.
Colorectal cancer (CRC) remains a leading cause of cancer-related morbidity and mortality worldwide. Recent advancements in cancer research emphasize the importance of personalized medicine, necessitating robust preclinical models that closely mimic the tumor microenvironment. Colorectal cancer organoids have emerged as a pivotal tool in this domain, providing a platform that preserves the cellular heterogeneity and allows high-throughput drug testing. However, organoids do not recapitulate the complexity of the tumor ecosystem, its architecture, and stromal interactions characteristic of native tumors. Here, we describe a model to maintain CRC tissue explants in culture. These explants enable the assessment of therapeutic responses ex vivo. Furthermore, they may offer insights into tumor-immune interactions and facilitate the evaluation of novel immunotherapeutic agents. The continued refinement of CRC tissue explant models promises to enhance our understanding of tumor biology and improve the predictive accuracy of preclinical studies, ultimately contributing to more effective and personalized treatment strategies for colorectal cancer patients.
Reviews of physiology, biochemistry and pharmacologyUndurti N Das
Obesity and MASLD (NAFLD) are chronic, low-grade systemic inflammatory diseases resulting from complex interactions among diet, adipose tissue, gut microbiota, liver, muscle, pancreas, and the brain. Central to this integrative framework are AA and other polyunsaturated fatty acids (PUFAs), which regulate inflammation, insulin sensitivity, neuroendocrine signaling, and energy homeostasis.Excess consumption of energy-dense foods rich in saturated and trans fats leads to adipocyte hypertrophy, altered membrane fluidity, increased antigenicity, and enhanced recruitment of macrophages and T cells into adipose tissue. These changes promote the sustained release of pro-inflammatory cytokines TNF-α, IL-6, MIF, and CRP, contributing to insulin resistance, metabolic syndrome, and MASLD (NAFLD). In contrast, adequate availability of GLA, DGLA, AA, EPA, and DHA improves cell membrane dynamics and serves as a substrate for anti-inflammatory and pro-resolving lipid mediators, including lipoxins, resolvins, protectins, maresins, and nitrolipids, which suppress inflammatory signaling and enhance adiponectin production.Abdominal obesity can be considered a localized glucocorticoid excess state driven by increased adipose 11β-hydroxysteroid dehydrogenase-1 (11β-HSD-1) activity, linking visceral fat accumulation to insulin resistance, altered PPAR signaling, and chronic inflammation. Suppression of 11β-HSD-1, increased PUFA intake, and exercise can ameliorate these abnormalities.Gut microbiota and their short-chain fatty acids influence inflammation, leptin signaling, incretin secretion, and lipid metabolism, while PUFAs-including AA-activate gut-brain-liver neural circuits via vagal pathways and G protein-coupled receptors to regulate satiety, insulin secretion, and hepatic glucose production. This chapter also emphasizes the role of hypothalamic factors, particularly BDNF, in integrating nutritional, hormonal, and inflammatory signals.In summary, AA and related PUFAs are critical modulators of inflammation resolution, neuroendocrine control, and metabolic balance, offering a unifying mechanistic framework for the prevention and management of obesity and its associated metabolic disorders.
Methods in molecular biology (Clifton, N.J.)Samna Sagadevan, Paloma Ordóñez-Morán
Organoids and fibroblasts derived from patient tissue serve as physiologically meaningful in vitro models to investigate tissue biology, diseases, and treatment responses. In this study, we present a robust protocol for the simultaneous isolation and long-term culture of patient-derived organoids (PDOs) and fibroblasts from both healthy and colorectal cancer samples. The workflow uses enzymatic dissociation followed by efficient separation into epithelial and stromal cell fractions. Organoids are embedded in Matrigel or Matrigel-Collagen I mixtures to support three-dimensional growth, whereas fibroblasts are maintained on conventional two-dimensional culture dishes. This dual-culture approach facilitates a broad range of downstream applications.
Methods in molecular biology (Clifton, N.J.)Brinda Balasubramanian
Single-cell RNA sequencing (scRNA-seq) has revolutionized the ability to resolve cellular heterogeneity within complex tissues, enabling the identification of discrete cell states. Here, we present an in silico analytical pipeline designed to characterize intestinal stem cells (ISC), transit-amplifying (TA) progenitors, and BEST4⁺ enterocyte precursors from human scRNA-seq datasets, with a focus on inflammatory contexts such as inflammatory bowel disease (IBD). The pipeline integrates dataset acquisition, quality control, normalization, dimensionality reduction, unsupervised clustering, and cell type annotation using a reference cell atlas. We implemented iterative subsetting and re-clustering of ISC and TA compartments to identify inflammation-associated subpopulations and epithelial biomarkers. While demonstrated in the context of IBD, this computational framework is broadly applicable to other tissues and pathological conditions where stem/progenitor dynamics underpin disease progression and tissue repair.
Methods in molecular biology (Clifton, N.J.)Mohamed Ali Hussein, Anwar Abdelnaser
.: Nonalcoholic fatty liver disease (NAFLD) is a significant global health concern, impacting roughly 25% of people and leading to chronic liver conditions. It involves excess fat accumulation in the liver without significant alcohol intake and can develop into nonalcoholic steatohepatitis (NASH), fibrosis, or cirrhosis. While liver biopsy remains the gold standard for diagnosis, its invasive nature and associated risks restrict its routine use. Noninvasive biomarkers, such as serum ALT, AST, and various composite scores, are available; however, their clinical usefulness is often limited by variable sensitivity and specificity across different populations and disease stages. To overcome these limitations, this chapter offers a comprehensive, reproducible protocol for identifying and clinically validating circulating long noncoding RNA (lncRNA) biomarkers for NAFLD and NASH. The workflow integrates bioinformatic analysis of four Gene Expression Omnibus (GEO) transcriptomic datasets (two human and two murine cohorts) with network-based inference to construct a NAFLD-related lncRNA-miRNA-mRNA coregulatory network. This is followed by candidate prioritization based on cross-dataset evidence and a literature review. Candidate lncRNAs are then experimentally validated in patient-derived blood samples using quantitative PCR (qPCR), and their diagnostic performance is quantified using receiver operating characteristic (ROC) analysis, both as individual markers and multi-lncRNA panels. Circulating lncRNAs are detected in diverse biofluids, remain stable under standard preanalytical conditions, and are often tissue-specific. This integrated approach facilitates the development of more precise, scalable, and noninvasive biomarkers for NAFLD/NASH. The chapter further emphasizes essential translational steps, including preanalytical standardization, analytical validation, and validation in independent patient cohorts with relevant clinical endpoints.
Cell plasticity is a major driver of colorectal cancer (CRC), as tumors hijack intestinal stem cell regenerative programs to create a dynamic, bidirectional axis between complementary stem-like states. Cancer stem-like cells (CSCs) display hallmark stem cell properties, including self-renewal, multipotency, controlled differentiation, and quiescence, and are now recognized as key orchestrators of CRC biology. They can sustain tumor initiation and long-term growth, seed metastases, enable periods of dormancy, and promote resistance to chemotherapy. As a result, therapeutically dismantling CSC populations and their supportive niches offers a compelling strategy to improve treatment responses and ultimately patient survival.
Methods in molecular biology (Clifton, N.J.)William Dalleywater
Colonic inflammation induces profound alterations in the intestinal mucosa that are evident at both macroscopic and microscopic levels, including disruption of epithelial barrier integrity, crypt fission, and immune cell infiltration. Recent advances in single-cell and spatial RNA sequencing have greatly expanded our understanding of the cellular diversity of the colonic mucosa, enabling pathological features to be directly linked to underlying cellular and molecular mechanisms. This chapter demonstrates the application of single-cell RNA sequencing (scRNA-seq) analysis techniques to interrogate stem cell dynamics in the context of inflammatory bowel disease. Using publicly available datasets, the chapter provides a step-by-step workflow implemented in Python, covering data access, loading, integration of multiple datasets, and initial preprocessing. Cells are mapped to large single-cell atlas references to infer cell identities, followed by a pseudobulk analysis strategy to assess inflammation-associated changes in cell phenotypes. Finally, trajectory inference approaches are applied to explore potential mechanisms governing the specification and modulation of key cell types during inflammation. Accompanied by an online resource containing fully annotated scripts, this chapter offers guided instruction in contemporary scRNA-seq analysis workflows. It is intended as an accessible introduction for researchers seeking to develop practical skills in single-cell data analysis that can be readily applied to their own biological questions.
Pakistan journal of pharmaceutical sciencesOrcun Can, Abdullah Sakin
BACKGROUND: Esophageal adenocarcinoma is associated with poor survival despite advances in systemic therapy. Readily available serum biomarkers may improve risk stratification; however, their combined role in biomarker-guided treatment decisions remains insufficiently investigated. OBJECTIVES: To evaluate the prognostic significance of combined baseline carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) levels and explore their utility for biomarker-based risk stratification in patients with esophageal adenocarcinoma receiving systemic chemotherapy. METHODS: This retrospective real-world cohort study included 38 patients with histologically confirmed esophageal adenocarcinoma who received systemic chemotherapy at a tertiary referral center between January 2016 and December 2021. Patients were classified into three exploratory biomarker-defined risk groups according to baseline serum CEA and CA19-9 levels. Overall survival was evaluated using Kaplan-Meier analysis and Cox proportional hazards regression, while treatment outcomes were explored across biomarker-defined groups and chemotherapy regimens. RESULTS: Elevated baseline CEA and CA19-9 levels were associated with shorter overall survival. Patients with simultaneous elevation of both biomarkers demonstrated the poorest survival outcomes (multivariable hazard ratio = 2.22). Differences in survival were observed across chemotherapy regimen groups within biomarker-defined categories; however, these comparisons were exploratory and may have been influenced by treatment-selection bias and unmeasured clinical factors. Patients with distal or gastroesophageal junction adenocarcinoma showed more favorable outcomes than those with cervical or thoracic tuxzmors. CONCLUSION: Combined assessment of baseline CEA and CA19-9 may provide a practical and inexpensive approach for biological risk stratification in esophageal adenocarcinoma. However, these findings do not establish that the biomarkers can guide chemotherapy selection. Prospective multicenter studies with independent validation are required to determine their prognostic and predictive value before biomarker-guided treatment selection can be recommended in routine clinical practice.
BACKGROUND: Ulcerative colitis (UC) is a chronic inflammatory bowel disease impacting quality of life. Combination oral and topical mesalazine is guideline-recommended for left-sided UC, but real-world data remain limited. OBJECTIVES: This retrospective study evaluated the efficacy and safety of mesalazine enteric-coated tablets plus a retention enema in patients with mild-to-moderate E2-type (left-sided) active UC. METHODS: Patients with mild to moderate E2-type active UC treated at a single center from January 2021 to December 2022 were retrospectively screened. After applying inclusion and exclusion criteria, 88 patients were included and assigned to either the study group (mesalazine enteric-coated tablets 1.0 g four times daily plus mesalazine retention enema 4.0 g once daily) or the control group (oral mesalazine enteric-coated tablets 1.0 g four times daily alone) based on clinical availability and patient preference. Treatment duration was 30 days. Outcomes included clinical efficacy (using modified local criteria and Mayo score components), symptom relief and time to improvement, endoscopic findings (Mayo Endoscopic Subscore), and adverse events. RESULTS: The "effective" rate was significantly higher in the study group (77.27% vs. 34.09%; OR=6.58, P<0.001). Symptom relief rate was 97.73% vs. 72.73% (OR=13.44, P=0.002). Symptom improvement times were shorter in the study group (all P<0.05). Post-treatment Mayo Endoscopic Subscore was lower in the study group (1.14±0.41 vs. 2.05±0.65; mean difference -0.91, P<0.001). Ulcer and congestion/edema incidences were lower in the study group (6.82% and 18.18% vs. 25.00% and 47.73%). Adverse events were mild and comparable between groups (13.64% vs. 11.36%, P=0.746). CONCLUSION: Mesalazine enteric-coated tablets combined with retention enema demonstrated superior efficacy compared to oral monotherapy for short-term symptom relief and mucosal healing in patients with mild to moderate E2-type active UC, with a favorable safety profile. Larger prospective studies with longer follow-up are needed to confirm these findings and evaluate long-term outcomes.
Pakistan journal of pharmaceutical sciencesKai Liang, Xiangyu Zhang, Yin Yang
BACKGROUND: Gastric cancer (GC) remains a leading cause of cancer-related mortality worldwide and effective therapeutic targets remain limited. OBJECTIVES: This study aimed to investigate the expression and functional role of miR-326 in gastric cancer, identify its direct target gene and elucidate the underlying molecular mechanisms involving the PI3K/Akt signaling pathway and the regulation of inflammatory cytokines. METHODS: miR-326 expression was detected by qRT-PCR in 56 paired GC tissues and adjacent normal tissues, as well as in GC cell lines (SGC-7901, MKN-45, AGS) and normal GES-1 cells. CCK-8 assays, flow cytometry, Transwell migration/invasion assays and ELISA were performed to assess cell proliferation, apoptosis, migration/invasion and cytokine secretion. Dual-luciferase reporter assay and Western blotting were used to validate the target gene and pathway activation. RESULTS: miR-326 was significantly upregulated in GC tissues and cell lines. ETS1 was identified as a direct target of miR-326. Overexpression of miR-326 reduced ETS1 expression, activated the PI3K/Akt pathway (increased p-PI3K and p-AKT), promoted cell proliferation, migration and invasion, inhibited apoptosis and induced an imbalance of inflammatory cytokines (increased IL-6 and TNF-α; decreased IL-10 and IL-17). Co-overexpression of ETS1 or treatment with the PI3K inhibitor LY294002 reversed these effects. CONCLUSIONS: miR-326 promotes malignant phenotypes in gastric cancer cells by targeting ETS1, activating the PI3K/Akt signaling pathway, and inducing dysregulation of inflammatory cytokines. The miR-326/ETS1/PI3K/Akt axis may serve as a potential diagnostic biomarker and therapeutic target for gastric cancer.
Pakistan journal of pharmaceutical sciencesMengyao Wang, Qingmiao Zhou
BACKGROUND: Acute pancreatitis (AP) is a common acute abdominal disease with high mortality in moderate and severe cases. Integrated Chinese and Western medicine therapy has promising clinical application prospects. OBJECTIVES: This study investigated the efficacy and safety of Tongfu Qinghua decoction enema combined with Ruyi Jinhuang powder external application for AP and its therapeutic effects across different age groups. METHODS: A total of 100 AP patients from October 2023 to August 2025 were randomly divided into observation and control groups (50 cases each). The control group received conventional Western medicine and the observation group received additional combined Chinese medicine therapy. Outcomes including hospital stay, symptom relief, inflammatory and pancreatic injury markers, clinical efficacy and adverse reactions were compared, with subgroup analysis of patients aged 18-40, 41-60 and 61-75 years. RESULTS: The observation group had significantly shorter hospital stay, faster symptom relief and gastrointestinal recovery (P<0.05). Post-treatment inflammatory and pancreatic markers improved significantly and the total effective rate was higher (P<0.05), with no significant difference in adverse reactions (P>0.05). Benefits were consistent across all age subgroups, with younger patients recovering faster and elderly patients still achieving significant improvement. CONCLUSION: This combined therapy is effective and safe for AP patients aged 18-75 years, significantly improving clinical outcomes and worthy of clinical promotion.
Drug development researchAkinade K Adetola, Regina N Ugbaja, Olubisi E Adeyi, Funmilola C Thomas, Johnson A Adeyemo, Oluwafemi A Odesola
Hepatocellular carcinoma (HCC) is a major contributor to cancer-related mortality worldwide and is closely linked to oxidative stress and progressive hepatic dysfunction. This study investigated the effects of plumbagin on antioxidant defense systems and hepatic function markers in an N-nitrosodiethylamine-thioacetamide (NDEA-TAA)-induced experimental model of HCC and compared its efficacy with that of sorafenib. Male Wistar rats were assigned to five groups: normal control, plumbagin-only, HCC control, HCC treated with plumbagin, and HCC treated with sorafenib. Hepatocarcinogenesis was induced using NDEA-TAA, after which antioxidant parameters including reduced glutathione (GSH), glutathione-S-transferase (GST), glutathione peroxidase (GPx), catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA) were evaluated alongside hepatic function markers alanine aminotransferase (ALT) and albumin (ALB). The HCC control group exhibited significant reductions in GSH, GST, GPx, CAT, and SOD activities, accompanied by elevated MDA and ALT levels and decreased ALB concentrations (p ≤ 0.05), indicating severe oxidative stress and hepatic injury. Treatment with plumbagin significantly restored antioxidant enzyme activities, reduced lipid peroxidation, and improved hepatic function relative to the untreated HCC group. These effects were comparable to, and in certain parameters exceeded, those observed following sorafenib administration. The findings demonstrate that plumbagin possesses substantial antioxidative and hepatoprotective properties capable of mitigating oxidative damage and improving liver function in experimental HCC. These findings support further investigation of plumbagin as a redox-modulating candidate in HCC and a basis for future studies incorporating combination treatment, molecular validation and translational models.
Drug development researchLamia W Mohamed, Ahmed A Saadeldin, Ayman B Farag
Vascular endothelial growth factor receptor-2 (VEGFR-2) is a vital mediator of angiogenesis. Therefore, VEGFR-2 inhibition is considered a promising therapeutic target to combat cancer. In the present study, a series of 22 imidazo[4,5-b]pyridine-acrylonitrile-based derivatives was designed and synthesized. All compounds were evaluated for their VEGFR-2 inhibitory activity. Seven of the tested compounds showed high inhibitory activity against VEGFR-2 with IC50 (0.029-0.087 µM) compared to reference drug sorafenib IC50 = 0.091 µM. Four of these derivatives were selected for in vitro cytotoxic activity against breast cancer (MCF-7) and hepatocellular (HepG2) carcinoma cell lines, showing IC50 (0.073-0.196 µM) and (0.20-0.21 µM), respectively, relative to sorafenib IC50 0.16 and 0.19 µM, respectively. Compound 2f exhibiting a superior VEGFR-2 inhibition (IC50 = 0.029 µM compared to sorafenib IC50 = 0.091 µM) and cytotoxicity against MCF-7 and HepG2 (IC50 = 0.073 µM and IC50 = 0.09 µM, respectively), was subjected to cell cycle analysis, apoptosis assay, and molecular modeling studies, which strongly supported the results. It caused cell cycle arrest at G2/M phase by 13.67% and a promising apoptosis induction. 2f exhibited a promising binding score and pose inside the VEGFR-2 active sites. It also showed a good safety profile with a moderate selectivity index. Based on the prediction of physicochemical and pharmacokinetic properties for 2f, it showed excellent prediction results to be orally bioavailable.
Drug development researchFang-Fang Li, Can Cui, Zhou-Juan Zheng, Man-Yu Xiao, Si Li, Peng Xie, Xiang-Lan Piao
Hepatocellular carcinoma (HCC) is a common malignant tumor of the digestive system. The liver is the primary organ for cholesterol production and metabolism in the body. Abnormal cholesterol levels can promote the initiation and progression of liver cancer, as well as influence treatment and patient prognosis. Damulin A and damulin B, a pair of isomeric dammarane-type saponins isolated from heat-treated Gynostemma pentaphyllum, have been shown to inhibit HCC cell proliferation and migration, arrest the cell cycle at the G0/G1 phase, induce apoptosis, and reduce intracellular cholesterol levels in vitro. Damulin B exhibited more potent effects in these assays. RNA sequencing and Western blot analysis revealed that both compounds downregulate the expression of key cholesterol biosynthesis-related genes, namely isopentenyl-diphosphate delta isomerase 1 (IDI1) and geranylgeranyl diphosphate synthase 1 (GGPS1). However, damulin A uniquely increased the expression of other cholesterol pathway genes: farnesyl diphosphate synthase (FDPS), farnesyl-diphosphate farnesyltransferase 1 (FDFT1), and lanosterol synthase (LSS). These findings indicate that damulin A and damulin B regulate intracellular cholesterol biosynthesis through distinct mechanisms, which may account for their differential inhibitory effects on hepatocellular carcinoma.
BACKGROUND: miR-185 expression is dysregulated in various tumors, but its role in gastric cancer remains unclear. OBJECTIVES: To investigate miR-185 expression and clinical significance in gastric cancer, explore its effects on malignant phenotypes and preliminarily elucidate the underlying mechanism. METHODS: Real-time polymerase chain reaction (PCR) was used to detect miR-185 expression in 58 archived gastric cancer tissue samples and paired adjacent normal tissues obtained from a hospital biospecimen repository. In SGC-7901 and AGS cells, the effects of miR-185 modulation on proliferation, apoptosis, epithelial-mesenchymal transition (EMT) and the cell division cycle protein 42 (Cdc42)-neural Wiskott-Aldrich syndrome protein (N-WASP) pathway were assessed using 5-ethynyl-2'-deoxyuridine (EdU) assay, flow cytometry, Western blot and dual-luciferase reporter assay. RESULTS: miR-185 expression was significantly downregulated in gastric cancer tissues and correlated with lymph node metastasis and advanced stage. In vitro, miR-185 overexpression inhibited proliferation, induced apoptosis, increased E-cadherin expression and decreased Vimentin, N-cadherin, Snail, Cdc42 and neural WASP expression. A dual-luciferase reporter assay confirmed that cortactin (CTTN) is a direct target of miR-185. These findings were consistent in both cell lines. CONCLUSION: miR-185 is downregulated in gastric cancer and its low expression correlates with lymph node metastasis and advanced TNM stage. In vitro experiments indicate that upregulation of miR-185 suppresses gastric cancer cell proliferation, induces apoptosis and reverses epithelial-mesenchymal transition, possibly by targeting CTTN and thereby affecting Cdc42 and N-WASP expression. This single-center, small-sample (n=58) in vitro study lacks in vivo validation; the findings require confirmation in larger, multicenter cohorts. Nevertheless, this study provides the first experimental evidence for the miR-185/CTTN/Cdc42-N-WASP axis in gastric cancer.
Pakistan journal of pharmaceutical sciencesZhenhua Yin, Xiaolan Lu, Limei Bu
BACKGROUND: General anesthesia is essential for gastric cancer surgery, but delayed awakening increases postoperative risks and costs. Although morroniside is known for its neuroprotective properties, its role in anesthesia recovery has remained unclear. OBJECTIVES: To investigate whether morroniside accelerates awakening from propofol anesthesia and whether hypothalamic GABAergic neurons mediate this effect. METHODS: A gastric cancer mouse model was established. Mice were divided into seven groups: normal, model, vehicle control, light anesthesia, deep anesthesia, morroniside-treated and propofol-only. The loss and recovery of righting reflex (LORR/RORR) were measured. EEG recording, immunofluorescence staining for GABA and c-Fos and fiber photometry for Ca2+ signals in hypothalamic GABAergic neurons were performed. RESULTS: Morroniside significantly shortened RORR without affecting LORR. During anesthesia induction, Ca2+ signals in hypothalamic GABAergic neurons increased sharply, then decreased during maintenance and remained low during early recovery. Morroniside increased GABA concentrations in multiple brain regions, especially the hypothalamus and reduced the EEG δ-band power while increasing α-band power. It also decreased propofol-induced c-Fos expression. CONCLUSION: Morroniside accelerates awakening from propofol anesthesia in a gastric cancer mouse model, potentially by modulating hypothalamic GABAergic neuron activity and EEG oscillatory patterns. These findings support its potential as an adjuvant in anesthesia.
GliaLorena Morton, Luis E Villafuerte, Alejandra P Garza, Nina Lindemann, Elisa Wider-Eberspächer, Dunja Bruder, Ildiko R Dunay
Ulcerative colitis (UC) patients experience cycles of active gut inflammation and remission, with neuropsychiatric comorbidities persisting even during clinical remission. While the dextran sulfate sodium (DSS) mouse model was previously applied to study gut-brain interactions, those studies focused on acute protocols missing the chronic, relapsing-remitting nature of human UC, when patients continue to experience central nervous system symptoms. Thus, we employed a chronic DSS treatment regimen comprising three cycles of active intestinal inflammation followed by remission phases to investigate region-specific microglial dynamics and their functional consequences on neuronal synapses. Transient blood-brain barrier alteration was detected during active chronic inflammation that was resolved during remission. Cortical microglia exhibited sustained iNOS-enriched activation state during remission, which coincided with synaptic imbalance: VGLUT1+ glutamatergic synaptosomes increased significantly in remission, while VGAT+ GABAergic vesicles declined, alongside suppressed neuronal c-fos expression. Hippocampal microglia adopted an ARG1-dominant phenotype with enriched TREM2, P2Y12R, and F4/80 expression during remission, indicating a phagocytic, reparative state. Hippocampal synaptosome analysis revealed selective excitatory enhancement with preserved inhibitory markers. Morphological analysis confirmed region-specific remodeling: cortical microglia displayed delayed process elaboration during remission, while hippocampal responses varied from transient (CA1) to persistent (CA3) somatic hypertrophy. These findings establish that remission from peripheral inflammation does not fully restore brain immune homeostasis. Persistent, region-specific microglial reactivity with cortical pro-inflammatory states associated with synaptic dysfunction and hippocampal adaptive responses preserving circuit integrity provides a mechanistic understanding for neuropsychiatric comorbidities in UC and suggests that brain-targeted therapies may be required to address the full disease burden.
BACKGROUND: Xiaoshi Lipi formula, a traditional Chinese medicine, may alleviate Gastric precancerous lesion (GPL) progression through multi-target mechanisms. OBJECTIVES: To evaluate the protective effects of Xiaoshi Lipi formula on GPL and its underlying mechanisms. METHODS: GPL was induced in rats by N-methyl-N'-nitro-N-nitrosoguanidine and sodium deoxycholate. Animals were assigned to normal, model, positive control and low-, medium-, or high-dose Xiaoshi Lipi formula groups. Gastric pathology was assessed by HE and AB-PAS staining. Serum inflammatory cytokines (TNF-α, IL-6, IL-1β), gastric function markers (G-17, PG I, PG II, PGE2), oxidative stress indices (ROS, MDA, SOD, GSH), ferroptosis- and antioxidant-related proteins and apoptosis (TUNEL) were analyzed. RESULTS: Xiaoshi Lipi formula markedly alleviated gastric mucosal atrophy and intestinal metaplasia and improved gastric function. In the medium-dose group, G-17, PG I and PGE2 increased by ~90%, 70% and 62%, respectively, while PGⅡ decreased by ~40%. TNF-α, IL-6 and IL-1β were reduced by ~45%. ROS and MDA declined by ~48% and 50%, while SOD and GSH rose by ~38% and 83%. GPX4 and SLC7A11 were restored, Nrf2/HO-1 pathway activated and TUNEL-positive cells reduced by ~68%. Medium-dose effects were comparable to the positive control. CONCLUSION: Xiaoshi Lipi formula ameliorates GPL by targeting inflammation, oxidative stress and ferroptosis, supporting its potential in gastric cancer prevention.
PulmonologyCristina Aljama, Ane Lopez-Gonzalez, Hanan Tanash, Maria Torres-Duran, Alice M Turner, Carlota Rodriguez-Garcia, Angelo Guido Corsico, Catarina Guimarães, José…
BACKGROUND: Data on health utilities in Alpha-1 antitrypsin deficiency (AATD) are scarce. This study aimed to evaluate health-related quality of life (HRQoL) using the EQ-5D in a cohort of AATD Pi*ZZ individuals from the European AATD Research Collaboration (EARCO) registry and to identify factors associated with impaired health utilities. METHODS: Observational, cross-sectional, multicenter study including AATD patients with Pi*ZZ genotype enrolled up to April 2025. HRQoL was assessed using the EQ-5D questionnaire (index and visual analogue scale (VAS)). Sociodemographic data, smoking history, respiratory symptoms, comorbidities, lung function and exacerbations were collected. Multivariable generalised linear models were used to identify factors independently associated with EQ-5D index scores in the overall population and in patients with AATD-COPD. RESULTS: A total of 1710 Pi*ZZ individuals were analysed with a mean age 55.2 ± 13.8 years, 51% were male and 49.6% had COPD. Mean EQ-5D index and VAS were 0.82 ± 0.22 and 62.9 ± 25.9, respectively. Patients with COPD had significantly worse EQ-5D index (0.75 ± 0.24 vs 0.90 ± 0.16) and VAS scores (56.3 ± 24.4 vs 70.4 ± 26.8; both p < 0.001). In multivariable analyses, higher symptom burden (CAT, mMRC), long-term oxygen therapy, being unemployed, exacerbations, COPD, bronchiectasis and lower educational level were independently associated with worse EQ-5D scores in Pi*ZZ subjects. CONCLUSIONS: HRQoL was substantially impaired in Pi*ZZ individuals, particularly in those with COPD. Symptom burden and exacerbations were stronger determinants of EQ-5D than lung function alone in all Pi*ZZ patients. Patients with AATD-associated COPD exhibited an HRQoL impairment comparable to that observed in usual COPD populations, but at a younger age.
Annals of medicineHong-Yu Long, Yong-Mei Wu, Xue Li, Bai-Qing Chen
BACKGROUND: To compare the predictive performance of computed tomography (CT) body composition indices, anthropometric indices, and laboratory indices for gallstones occurrence, and to construct machine-learning models to improve performance. METHODS: The dual-center retrospective cohort enrolled patients who underwent initial abdominal CT between January 2017 and January 2023, had no gallstones detected, and completed at least 3 years of follow-up. The data analysis was performed in April 2026. They were divided into gallstone group and non‑gallstone group by follow‑up findings. A deep-learning tool, Body and Organ Analysis (BOA), was used to quantify fat, muscle, and bone at the level of the third lumbar vertebra. The area under the receiver operating characteristic curve (AUC) of these indices was compared with that of anthropometric and laboratory indices. Predictive models were developed in the training cohort. Model performance was evaluated using fivefold cross-validation and an independent test cohort. RESULTS: 1,944 patients were evaluated, including 1,437 in the training cohort (Center 1; median age, 63 years [25th-75th percentile, 55-72]; 699 males) and 507 in the test cohort (Center 2; median age, 63 years [55-71]; 266 males). In univariate analysis, neutrophil-to-lymphocyte ratio (NLR) showed the highest AUC (0.627, 95% confidence interval [CI]: 0.586-0.668). The extreme trees (ET) model performed best, with a test-set AUC of 0.772 (95% CI: 0.714-0.822). SHapley Additive exPlanations (SHAP) analysis identified the area ratio of subcutaneous to total fat as the most important feature. CONCLUSIONS: NLR was the best single predictor but had limited standalone utility. Among models integrating the three indicator categories, the ET performed best.
The global prevalence of Metabolic dysfunction-associated steatotic liver disease (MASLD) among persons with HIV (PWHIV) is estimated at 34%. This study examined the influence of gut microbial and metabolic alterations in PWHIV with MASLD. We conducted 16S rRNA sequencing on 60 fecal samples (24 from HIV group and 36 from HIV-MASLD group) to assess microbial composition, and employed LC/MS-based metabolomics on both fecal and plasma samples to identify variations in metabolites. An exploratory analysis was performed to compare plasma cytokines and oxidative stress markers between the two groups. Sequencing in the HIV-MASLD group showed lower Faecalibacterium and Anaerobutyricum, and higher Klebsiella, Escherichia, and Enterococcus. Metabolomic analysis revealed a reduction in anti-inflammatory metabolites (Dodecanedioic acid, 5-Phenylvaleric acid, Kynurenic acid) and antioxidant metabolites, coupled with an increase in the pro-inflammatory metabolite Gamma-Glutamyl-L-Putrescine in the HIV-MASLD group. And plasma analysis indicated a decline in 20-carboxy arachidonic acid. Additionally, Faecalibacterium and Anaerobutyricum correlated positively with anti-inflammatory metabolites, while Klebsiella, Enterobacter, and Citrobacter correlated negatively. Furthermore, Faecalibacterium and Anaerobutyricum also showed a moderate negative correlation with MDA levels. A random forest model using plasma metabolites showed the optimal discriminative ability for HIV-MASLD (AUC = 0.819 ± 0.0059). Differences in gut microbiota and metabolites in PWHIV may affect MASLD progression by influencing inflammation and oxidative stress. Modulating gut microbiota, such as adding Faecalibacterium and Anaerobutyricum, could be a treatment strategy for HIV-MASLD. Plasma metabolites have potential as noninvasive markers.
Gynecological endocrinology : the official journal of the International Society of Gynecological EndocrinologyXiangyan Ruan, Juan Du, Muqing Gu, Fengyu Jin, Jing Jin, Yanglu Li, Jiaojiao Cheng, Yanqiu Li, Weimin Kong, Qi Zhang, Zecheng Wang, Mingzhen Zhang, Anming Liu,…
OBJECTIVE: This article reports on the world's first live birth after ovarian tissue cryopreservation and transplantation (OTCT) in a patient with colon cancer. METHODS: A 29-year-old unmarried nulliparous patient underwent ovarian tissue cryopreservation before chemotherapy. After chemotherapy-induced premature ovarian insufficiency, she received the first transplantation of six thawed ovarian cortex strips. Despite normal ovarian function being maintained for over 2 years, she requested and underwent a second transplantation with another six strips to maximize her chances of pregnancy. RESULTS: After the first transplantation, she resumed spontaneous menstruation with restored ovarian function, which was remained functional for 28 months until the second transplantation. Following the second transplantation, she conceived spontaneously without assisted reproductive technology two months after grafting. Routine prenatal examinations showed normal fetal development without abnormalities. She delivered a healthy baby girl weighing 2785 g with Apgar scores of 8/9 at 1 and 5 min. CONCLUSIONS: This case demonstrates the feasibility and safety of OTCT for fertility and ovarian function preservation in patients with colon cancer.
Drug deliveryEliska Kopecna, Leona Raskova Kafkova, Petr Kosztyu, Kristyna Sloupenska, Katerina Zachova, Alica Cutkova, Jozef Skarda, Jana Vaculova, Milan Kuchar, Natalya P…
Interleukin 23 (IL-23) is a key mediator of immune-mediated intestinal inflammation and plays an important role in the pathogenesis of inflammatory bowel disease. Inhibition of IL-23 signaling has therefore been validated as a therapeutic target in IBD. Here, we tested small binding proteins of non-immunoglobulin origin targeting either the IL-23 cytokine p19 subunit or the IL-23 receptor subunit IL-23R, designated ILP and REX binders, respectively. ILP317 and REX115 variants of formerly identified protein collections were tested in murine experimental colitis. Lactococcus lactis bacteria engineered for ILP317 or REX115 expression were administered to experimental mice via yogurt-coated food pellets before and during dextran sulfate sodium (DSS) and trinitrobenzenesulfonic acid (TNBS)-induced colitis. To compare both protein blockers, ILP317 and REX115 binders were administered intraperitoneally as soluble recombinant proteins. General health status, colon macroscopic and histological changes, mucosal tissue cell infiltration, local and systemic inflammatory cytokines were analyzed. In DSS- and TNBS-induced colitis, intraperitoneal administration of soluble ILP317 or REX115 reduced disease severity, including disease activity index, colon shortening, and mucosal inflammatory cell infiltration. Oral administration of recombinant L. lactis expressing ILP317 also showed consistent protective effects, whereas oral L. lactis expressing REX115 produced less pronounced effects in several experimental settings. Overall, ILP317 appeared more effective than REX115 when delivered orally by recombinant L. lactis. Oral administration of non-colonizing L. lactis expressing an IL-23p19 inhibitor may represent a promising preclinical strategy for the development of novel, non-invasive approaches to modulate intestinal inflammation.
Annals of medicineChaochao Wang, Huiting Rao, Xuefei Zhang, Guibiao Zhang, Guodong Xia, Li Zhang
BACKGROUND: Fatty liver disease is frequently identified during routine health examinations, but practical strategies for risk stratification in adults with thyroid dysfunction remain limited. This study evaluated metabolic dysfunction-associated fatty liver disease (MAFLD) prevalence across thyroid function categories and developed health examination-based risk stratification models. METHODS: This retrospective cross-sectional study included 46,311 adults who underwent routine health examinations from November 2019 to February 2025. Participants were classified as euthyroid, hypothyroid, or hyperthyroid. MAFLD was defined as ultrasound-confirmed hepatic steatosis plus overweight/obesity, diabetes, or metabolic dysregulation. Logistic regression and restricted cubic spline analyses assessed thyroid-related associations. Least absolute shrinkage and selection operator regression and multivariable logistic regression were used to develop models for adults with hypothyroidism or hyperthyroidism. Discrimination, calibration, and utility for post-examination risk stratification were evaluated. RESULTS: Among 46,311 participants, 42,978 were euthyroid, 2,645 had hypothyroidism, and 688 had hyperthyroidism. MAFLD prevalence was 33.0%, 35.5%, and 19.9%, respectively. FT3 was positively associated with MAFLD, whereas FT4 was inversely associated; TSH was not independently associated after full adjustment. The hypothyroid model retained age, BMI, ALT, TG, and HDL-C, with AUCs of 0.906 and 0.901 in the training and validation sets. The hyperthyroid model retained age, BMI, FT4, ALT, and HDL-C, with AUCs of 0.918 and 0.932. Both models showed good calibration and potential utility for post-examination risk stratification. CONCLUSIONS: Routine health examination indicators can stratify fatty liver risk among adults with thyroid dysfunction and may help identify individuals requiring targeted liver-metabolic follow-up. External validation and prospective evaluation are needed.
Following the publication of the above article, a concerned reader has contacted the Editorial Office to draw attention to the fact that there appeared to be discrepancies between the numbers of patients reported with low‑ or high‑ZNF703 expression in Fig. 3 on p. 1074: Fig. 3A reported 70 patients with low ZNF703 expression and 68 patients with high expression; however, the stage‑stratified Kaplan‑Meier curves showed the following: In Fig. 3B, stages I‑IIIA: 57 low‑expression and 20 high‑expression patients, and in Fig. 3C, stages IIIB‑IV: 20 low‑expression and 41 high‑expression patients (and so 70 vs. 68 patients in Fig. 3A, and 77 vs. 61 patients in Fig. 3B and C). In addition, the AJCC stage‑specific counts reported in Table I also showed internal inconsistencies in terms of the overall numbers of low‑ and high‑expression patients, and finally, an additional discrepancy was noted for the definition of the high expression group: The Materials and methods section stated that an immunohistochemical score of ≥6 was classified as high expression, whereas the Table I column heading defined high expression as '>6'. To clarify these apparent discrepancies, the authors should resolve the following: i) provide the actual numbers of low‑ and high‑expression patients in each AJCC stage group; ii) clarify whether the cutoff used for the Kaplan‑Meier analyses was ≥6 or >6; iii) provide the exact patient numbers used to generate Fig. 3A, B and C; and iv) clarify whether the counts, totals and percentages in Table I require correction. Finally, upon performing an independent analysis of the data in this paper in the Editorial Office, it came to light that, regarding the wound healing assay data shown in Fig. 6A on p. 1076, the '24 h / Control' and '24 h / NC siRNA' panels showed an overlapping section of data, such that data which were intended to show the results of experiments performed under different conditions had apparently been derived from the same original source. The authors have been contacted by the Editorial Office to offer an explanation for the apparent anomalies in the description of the data, and the incorrect assembly of data in Fig. 6, in this paper, and we are awaiting their response. Owing to the fact that the Editorial Office has been made aware of this potential issue concerning the scientific integrity of this paper, we are issuing an Expression of Concern to notify readers of this potential problem while the Editorial Office continues to investigate this matter further. [Oncology Reports 32: 1071‑1077, 2014; DOI: 10.3892/or.2014.3313].
Biometrical journal. Biometrische ZeitschriftVinicius F Calsavara
In survival analysis, addressing non-proportional hazards and the presence of long-term survivors poses substantial methodological challenges, as traditional models often fail to capture these features adequately. To overcome these limitations, a novel long-term survival model is proposed that integrates the Yang and Prentice framework with a defective Gompertz distribution. The proposed model provides a robust approach for analyzing time-to-event data with non-proportional hazards and a cure fraction. Comprehensive simulation studies and an application to colorectal cancer data illustrate its flexibility in capturing time-varying hazard ratios and long-term survivors, making it a useful alternative to existing methods.
Peritoneal dialysis (PD)-associated peritonitis is a leading cause of technique failure. We aimed to develop and internally validate a Cox regression‑based nomogram for predicting peritonitis‑free survival in incident PD patients. This single-center retrospective cohort study included 372 patients who underwent peritoneal dialysis between June 2018 and December 2024, with follow-up through December 2025. LASSO‑Cox regression was used to select predictors from 23 candidate variables. A nomogram was constructed to predict 6‑, 12‑, and 24‑month peritonitis‑free survival. Model performance was assessed by concordance index (C‑index), time‑dependent ROC, calibration curves, and decision curve analysis. Over a median follow‑up of 26.3 months, 92 (35.4%) and 34 (30.4%) peritonitis events occurred in the training and validation cohorts, respectively. Nine predictors (education, start age, BMI, hemoglobin, potassium, sodium, albumin, calcium, phosphorus) were retained. Independent predictors were lower education (≤9 vs >9 years: HR 1.93, 95% CI 1.03-3.64), lower hemoglobin (HR 0.99, 0.97-1.00), lower albumin (HR 0.92, 0.87-0.97), and lower phosphorus (HR 0.53, 0.29-0.99). The C‑index was 0.707 (95% CI 0.650-0.765). Time‑dependent ROCs in the training cohort were 75.8%, 77.5%, and 69.7% at 6, 12, and 24 months; in the validation cohort, they were 77.7%, 63.8%, and 58.4%, respectively. Calibration was satisfactory at 6 and 12 months (Brier scores 6.0%-11.1%), with wider clinical net benefit at 12-24 months. This nomogram using nine routinely available variables demonstrated moderate discrimination and satisfactory calibration for predicting peritonitis‑free survival in PD patients. While promising, the model requires external validation in larger, multicenter cohorts to confirm its generalizability before routine clinical use.
Journal of medical economicsMichael Dore, Derek W Ebner, A Mark Fendrick, Chris Estes, Gustavus Aranda, Amber Coleman, Vahab Vahdat, Jordan J Karlitz, Tiago Rua, Frank van Hees, Paul J Li…
AIM: Multi-target stool DNA (mt-sDNA) and fecal immunochemical testing (FIT) are home-based options for colorectal cancer (CRC) screening. Next-generation mt-sDNA has enhanced sensitivity for CRC and advanced precancerous lesions compared with FIT and is not subject to the same capacity constraints as colonoscopy. This analysis evaluated the economic and clinical implications of increasing the proportional utilization of next-generation mt-sDNA. METHODS: A budget impact model was developed from the validated CRC-AIM microsimulation model. The model simulated a cohort of 1 M average-risk, screen-eligible adults over a 3-year time horizon from an integrated delivery network perspective. The status quo scenario assumed an initial utilization of 20% next-generation mt-sDNA, 20% FIT, and 60% colonoscopy screening. In a hypothetical increased next-generation mt-sDNA scenario, its utilization expanded to 35% within 3 years, while FIT and colonoscopy utilization decreased to 15% and 50%, respectively. Test performance characteristics, real-world adherence, and cost inputs (i.e. screening test and programs, colonoscopy adverse events, CRC treatment) were derived from published literature. Multiple scenario analyses were conducted. RESULTS: Among 1 M members, total costs in the status quo scenario were $1.058B and with increased next-generation mt-sDNA use were $1.046B, generating an estimated savings of $12.5 M (-$0.35 per member per month). Increasing next-generation mt-sDNA improved the CRC screening rate by 1.1%. Overall, 10,467 fewer screening colonoscopies and 1,912 more follow-up colonoscopies after a positive stool-based test were performed. An additional 17 CRCs were screening-detected, 11 fewer were clinically detected, and there was one less CRC-related death. Increased next-generation mt-sDNA utilization remained cost-saving with all scenario analyses. CONCLUSIONS: In this budget impact model, shifting a small proportion of current FIT and colonoscopy utilization to next-generation mt-sDNA for average-risk CRC screening is estimated to be cost-saving. Increasing the next-generation mt-sDNA utilization also improves the screening rate and clinical outcomes while optimizing availability of capacity-limited colonoscopy resources.
NMR in biomedicineNaama Lev-Cohain, Jacob Sosna, Rachel Katz-Brull
Hyperpolarized MRI is a medical imaging technology that has been used so far only in focused clinical studies. This technology enables metabolic imaging without ionizing radiation and is combined with standard-of-care MRI protocols. Unlike metabolic imaging performed with radiolabeled materials (e.g., PET and SPECT), hyperpolarized MRI enables differentiation between the injected metabolic substrate and its product(s). In this way, the number of imaging contrasts that may be attained is relatively large. The purpose of this review is two-fold: to provide coherent terminology for the various contrasts obtained in HP-MRI to facilitate further development in medicine and to highlight its potential contribution to liver nodule characterization. We reviewed 33 hyperpolarized MRI papers published from 2013 to 2024 that describe clinical studies in human subjects and patients, and characterized the metabolic contrast provided by this technology across various organs and in cancer tumors. We have further reviewed 20 preclinical studies of hyperpolarized MRI in the liver to evaluate the potential of this technology for early detection and characterization of hepatocellular carcinoma. Most clinical studies have used a single hyperpolarized substrate, namely, pyruvate, labeled with carbon-13 at position 1 ([1-13C]pyruvate). Even with this single substrate, the number of possible imaging contrasts is high, making the interpretation of these images challenging. We aimed to explain these imaging contrasts and provide a consistent nomenclature for them that will serve the medical community. Preclinical MRI studies using hyperpolarized substrates suggest the potential to characterize liver nodules in patients. In summary, hyperpolarized MRI is a promising clinical imaging modality in oncology; however, the small number of patients investigated so far precludes addressing possible biological confounding factors. Further studies are required for improving tumor characterization using this technology, and studies in liver imaging aimed at early detection of HCC are warranted.
The Journal of experimental medicineJiaojiao Zhang, Tian Zhao, Hong Lei, Rui Gan, Xin Liu, Yuanyuan Meng, Dongfeng Ren, Jingyi Su, Yinmei Tang, Jiaxing Sun, Wen Li, Yongze Chen, Yelin Zhang, Xuya…
The cGAS-STING pathway plays a critical role in antiviral defense and inflammatory homeostasis. While posttranslational regulation of STING is well characterized, the transcriptional networks directly governing Sting1 expression remain unclear. Here, we identify the transcription factor GA-binding protein alpha (GABPA) as a critical regulator of STING-dependent type I IFN (IFN-I) responses in macrophages. Mechanistically, GABPA directly binds to the Sting1 promoter, thereby transcriptionally sustaining basal STING expression. Upon viral infection, activated IκB kinase epsilon (IKKε) phosphorylates GABPA at S447, which further enhances its transcriptional activity and amplifies the STING-IFN-I signaling cascade. Notably, Gabpa deficiency protects mice from DSS-induced colitis, and this protective effect is abolished upon Sting1 deletion. Strikingly, patients with inflammatory bowel disease (IBD) or systemic lupus erythematosus (SLE) exhibit elevated GABPA expression that positively correlates with STING hyperactivation and enhanced inflammatory signatures. Moreover, a GABPA-blocking peptide effectively suppresses aberrant IFN-β production in peripheral blood mononuclear cells (PBMCs) from SLE patients, highlighting its therapeutic potential for interferonopathies.
Due to the late stage at which most gastric cancer patients are diagnosed, the prognosis is often poor. Current treatments have limited efficacy for advanced cases. This study delves into the involvement of circARID1A and the mechanisms behind its role in gastric cancer progression. GES-1 cell line and GC cell lines HGC-27, AGS, MKN45, and SNU-16 were cultured and subjected to various assays. RNA interference and overexpression techniques were used to modulate circARID1A and miR-147a levels in vitro, while xenograft tumor models in BALB/c nude mice were used to assess tumor growth in vivo. Cellular growth, migration, and invasion were assessed using CCK8, colony formation, and Transwell assays. Dual-luciferase reporter assays, qPCR and Western blot were utilized to investigate the interactions between circARID1A, miR-147a, and SNAI2. CircARID1A promotes GC cell proliferation, migration, and invasion by regulating the miR-147a/SNAI2 axis. In vivo, knockdown of circARID1A inhibits tumor growth in subcutaneous xenograft models.
Journal of the International Society of Sports NutritionRobyn Aitkenhead, Mark Waldron, Gillian E Conway, Katy Horner, Shane M Heffernan
BACKGROUND: Exercise-induced gastrointestinal symptoms (GIS) are commonly reported, with either Likert-type scales or visual analog scales (VAS) used to quantify the incidence and magnitude of symptoms. However, these measurement tools lack consistency in their design, reporting, and key aspects such as the validity of these tools is unclear. This study aimed to systematically review and critically discuss the various Likert-type and VAS scales used to assess GIS during exercise. METHODS: A search conducted in August 2025, in accordance with PRISMA guidelines, identified full-text articles evaluating GIS during exercise or sport. RESULTS: In total, 126 studies were included, representing 50 different scales and 13 scale lengths. The most frequently used measures were 11-point Likert-type scales and VAS (46%). Only one study assessed the reliability of an 11-point exercise-specific scale, and only one study used a validated retrospective questionnaire developed for assessing chronic GIS in runners. Most studies (92%) did not justify their scale selection, often adapting tools from non-exercise contexts or using custom-designed measures. CONCLUSION: The current review highlights substantial heterogeneity in GIS measurement during exercise and a paucity of evidence on validity and reliability. In addition to identifying current methodological limitations, the current review provides practical recommendations to support improved GIS assessment and reporting in exercise settings. The development of a standardized, fully validated exercise-specific GIS scale remains a priority, as current tools range from those with limited (partial) psychometric evaluation to those with no formal validation evidence, thereby limiting consistency, reliability, and comparability across studies and in practice.
Human vaccines & immunotherapeuticsMaheen Humayun, Lina S Sy, Emily Rayens, Lei Qian, Jun Wu, Bradley K Ackerson, Yi Luo, Yanjun Cheng, Avanish R Patel, Zendi Solano, Britta Amundsen, Justine De…
Individuals with inflammatory bowel disease (IBD) are at increased risk of herpes zoster (HZ). Using a matched cohort design, we evaluated the vaccine effectiveness (VE) of recombinant zoster vaccine (RZV) against HZ among IBD patients aged ≥18 years. Within the Kaiser Permanente Southern California population, individuals who received 2 RZV doses were matched up to 1:3 to RZV-unvaccinated individuals. Individuals were followed from 31 days after the receipt of the 2nd RZV dose until the occurrence of a censoring event. VE was calculated using Cox proportional hazards regression. A self-controlled case series analysis was used to compare the rate of flares in the 30‑day risk window following vaccination with the comparison window among RZV recipients using conditional Poisson regression. The adjusted VE of 2 RZV doses (≥4 weeks apart) against HZ was 65.4% (95% confidence interval [CI]: 40.6%-79.9%) in all IBD patients, 62.7% (17.2%-83.2%) in ulcerative colitis patients, and 70.2% (34.5%-86.4%) in Crohn's disease patients. The VE of 2 doses administered 4 weeks-6 months apart in IBD patients was 62.9% (34.3%-79.1%). The rate ratio for IBD flares was 0.80 (95% CI: 0.58-1.12). In conclusion, RZV provided protection against HZ in IBD patients with no increased risk of IBD flares observed.
Human vaccines & immunotherapeuticsYangjun Gao, Wenqi Zhao, Qiuli Wang, Min Liu, Wenxia Wang, Fei Su, Suxia Yang, Xuefeng Shen, Wenxuan Fan, Yuexiang Zhang, Xiaoling Zhang, Yunyi du, Hui Wang, X…
Radiofrequency hyperthermia (RFH) increases local tumor temperature and may enhance sensitivity to systemic therapies. We evaluated the association between adding RFH to first-line immune checkpoint inhibitor (ICI) plus chemotherapy and outcomes in patients with HER2-negative advanced gastric/gastroesophageal junction (GEJ) adenocarcinoma. This study adopted a single-center, retrospective, observational design. Patients treated between January 2021 and January 2024 were retrospectively enrolled and grouped by receipt of RFH (hyperthermia vs. non-hyperthermia). The primary endpoint was overall survival (OS); secondary endpoints were progression-free survival (PFS), objective response rate (ORR), disease control rate (DCR), and safety. A 1:1 propensity score matching (PSM) analysis was performed. A total of 212 patients were included: 119 in the hyperthermia group and 93 in the non-hyperthermia group. In the overall cohort, RFH was associated with longer median PFS (10.17 vs. 7.06 months; HR = 0.70; p = .011) and median OS (19.31 vs. 14.98 months; HR = 0.71; p = .018). ORR did not differ significantly between groups (21.0% vs. 21.5%; p = .930), whereas DCR was higher with RFH (96.6% vs. 86.0%; p = .005). After PSM, 168 patients were matched; balance improved, but residual imbalance remained. Matched-cohort estimates remained directionally similar for PFS (9.72 vs. 7.09 months; HR = 0.67; p = .013) and OS (19.02 vs. 15.12 months; HR = 0.67; p = .015). Recorded grade ≥3 immune-related adverse events occurred in 5.0% and 4.3% of patients, respectively. RFH-related events were mainly transient local skin erythema and induration, and no RFH-related serious adverse events were recorded. RFH added to ICIs and chemotherapy was associated with longer PFS and OS, requiring confirmation in prospective randomized studies.
PulmonologyRaffaella Manzo, Giuseppe Fiorentino, Anna Michela Gaeta, Antonietta Coppola, Valentina Di Spirito, Maurizia Lanza, Angela Mirizzi, Ilaria Ferrarotti, Nuccia G…
BACKGROUND: :Alpha-1 antitrypsin deficiency (AATD) is a rare genetic disorder mainly affecting the lungs and liver, but its psychological, functional, and relational burden remains insufficiently characterizedcharacterised. OBJECTIVE: To describe emotional distress, health-related quality of life, interpersonal and sexual difficulties, and daily functional limitations in individuals with AATD. METHODS: We conducted a cross-sectional online survey in Italy between October and December 2025 among individuals with AATD. Socio-demographic, clinical, and psychosocial data were collected using an ad hoc questionnaire, the Hospital Anxiety and Depression Scale (HADS), and the 12-item Short Form Health Survey (SF-12). HADS subscales were reconstructed from item-level responses, applying reverse scoring where appropriate. RESULTS: The study included 130 participants (mean age 56.3 ± 13.5 years; 53.8% female). Pulmonary involvement was the most frequent manifestation (68.5%). Disease-related limitations were reported in daily functioning by 57.7% and in social participation by 53.1%. Clinically significant symptoms were present in 46.2% for anxiety and 17.7% for depression. The SF-12-derived ad hoc unweighted physical and mental descriptive composites were 56.68 ± 27.47 and 54.53 ± 26.91. Women reported higher anxiety and poorer mental-composite scores, while participants receiving augmentation therapy showed worse depressive and perceived-health burden. Sexual-life changes were reported by 37/96 partnered participants (38.5%). Fifteen of 128 respondents (11.7%; two preferred not to answer) reported current psychological support. In exploratory covariate-adjusted models, functional and perceived-health correlates remained associated with clinically significant anxiety. CONCLUSIONS: In this cross-sectional survey, respondents with AATD reported substantial emotional, functional, and relational burden, while few were receiving psychological support. The self-selected sample and absence of a non-AATD comparison group preclude causal or disease-specific inference; the findings support evaluation of brief psychological screening and defined referral pathways in multidisciplinary AATD care.
Journal of investigative surgery : the official journal of the Academy of Surgical ResearchHong-Yu Xu, Wei-Zhen Tang, Tai-Hang Liu, Fei Han, Qin-Yu Cai, Hao-Wen Chen, Yi-Fan Zhao, Wang Qi Rui Liu, He-Zhe Hua, Jun Ding, Peng Guo, Zhi-Yong Xiang
BACKGROUND: Complications after pancreaticoduodenectomy are common and may be life-threatening. Management of the common bile duct stump may affect biliary pressure, drainage, and postoperative outcomes. This study seeks to compare open versus ligated bile duct stump management during pancreaticoduodenectomy. METHODS: This retrospective cohort included 125 patients who underwent pancreaticoduodenectomy between 2019 and 2025. Surgical outcomes and postoperative recovery were compared between the open-stump and ligation groups. Multivariable logistic regression identified associations between stump management and postoperative complications; biochemical indicators were also assessed. RESULTS: After adjustment, the opening group had significantly lower risks of pancreatic fistula (aOR = 0.18, 95%CI: 0.05 - 0.68, P = 0.011) and intra-abdominal and gastrointestinal bleeding (aOR = 0.09, 95%CI: 0.01 - 0.77, P = 0.028) versus the ligation group. The opening group also demonstrated a shorter surgical duration (315 minutes vs. 420 minutes, P < 0.001), reduced postoperative hospitalization (24 days vs. 30 days, P = 0.002), and lower costs (109,375 RMB vs. 137,518 RMB, P < 0.001). No significant changes were seen in liver function or coagulation markers. CONCLUSION: Keeping the common bile duct open markedly enhances perioperative management during pancreaticoduodenectomy, thus serving as a significant reference for refining surgical protocols.
Annals of medicineKeijo Viiri, Chen Chongtham, Markku Mäki
BACKGROUND: Celiac disease (CeD) is an autoimmune enteropathy in which diagnosis and disease monitoring rely on serology and small-intestinal histology. However, conventional histological assessment is semi-quantitative, subject to interobserver variability, and relatively insensitive to subtle mucosal changes, particularly in treated patients or during early disease. Recent advances in transcriptomic technologies have enabled detailed characterization of intestinal mucosal gene expression, revealing disease-specific molecular signatures. This review aims to synthesize current evidence on molecular histomorphometry and to evaluate the emerging role of transcriptomic profiling in improving diagnosis, disease monitoring, and therapeutic trial endpoints in CeD. In addition, we propose practical guidelines and a methodological pipeline for implementing transcriptomic analyses in this context. DISCUSSION: Transcriptomic analyses consistently identify characteristic epithelial and immune activation signatures in active CeD, including interferon-driven pathways, epithelial stress responses, and loss of mature enterocyte differentiation. Importantly, persistent molecular abnormalities are often detectable in patients adhering to a gluten-free diet despite histological normalization, highlighting limitations of conventional endpoints. Molecular histomorphometry integrates gene expression data with morphologic features to provide continuous, quantitative measures of mucosal injury. Regression-based molecular indices demonstrate improved sensitivity, reproducibility, and dynamic range compared with traditional histology. In clinical trials, these approaches have potential to enable detection of treatment effects with smaller sample sizes and shorter study durations, and they have shown particular promise as endpoints in gluten challenge models and studies of therapies such as transglutaminase 2 inhibitors. We further outline a practical analytical pipeline, including study design considerations, data processing, model construction, and reporting standards. CONCLUSIONS: Molecular histomorphometry and transcriptomic profiling represent a significant advancement in the assessment of CeD, offering quantitative, qualitative and highly sensitive measures of mucosal pathology. Coupled with clear methodological guidance, these approaches have the potential to complement or, in selected contexts, surpass traditional histological evaluation in both clinical practice and drug development. Continued validation, harmonization, and standardization will be essential for broader implementation and regulatory acceptance.
Pegylated interferon-α (PEG-IFNα) exhibits suboptimal efficacy in achieving a functional cure in patients with chronic hepatitis B (CHB), partially due to impaired hepatitis B virus (HBV)-specific T-cell function. We conducted a pilot study (ClinicalTrials.gov: NCT06457477) to explore the efficacy, safety, and immunological changes associated with a response-guided sequential strategy of anti-PD-1 antibody (αPD-1) and PEG-IFNα. Ten virologically suppressed CHB patients receiving nucleos(t)ide analogues (NAs), all of whom had previously received PEG-IFNα and had HBsAg levels ≤200 IU/mL at enrolment, were included. The patients continued background NA therapy and received αPD-1 (sintilimab) for up to 24 weeks; PEG-IFNα was added at week 12 if HBsAg decline was <1 log10 IU/mL. HBV-specific T-cell responses targeting surface, core, and polymerase peptide pools were quantified, and plasma inflammatory proteins were longitudinally assessed. Nine patients completed the study, and six achieved HBsAg loss: three during NAs plus αPD-1 treatment (Group A) and three after PEG-IFNα add-on (Group B). All six maintained HBsAg negativity 24 weeks after treatment discontinuation. Most adverse events were grade 1-2. Increases in HBV-specific IL-2 and IFN-γ responses were observed in several patients with HBsAg loss, whereas such responses remained limited in nonresponders (Group C). Among 12 prespecified cytokines, IL-10 showed an early longitudinal difference, with higher levels in Group A at week 3. Exploratory analyses showed that week-3 IL-10 was positively associated with early ALT elevation and subsequent HBsAg decline. These findings support further evaluation of response-guided sequential immunotherapy strategies in larger, controlled, and well-characterized cohorts.Trial registration: ClinicalTrials.gov identifier: NCT06457477..