In vivo最新文献

Background/aim: The procurement of right lobe grafts while preserving the middle hepatic vein (MHV) tributaries (V5 and V8) just before graft retrieval is technically challenging. Moreover, the safe isolation of the hepatic duct from the Glissonean pedicle is essential in living donor hepatectomy. To date, few studies have reported surgical techniques for preserving the MHV tributaries during right lobe graft procurement.

Patients and methods: This report presents our modified subtraction technique for managing the MHV tributaries and Glissonean pedicle during right lobe graft procurement. First, a subtraction technique for the MHV tributaries was initiated by isolating the right Glissonean pedicle. The lower tip of the tape was placed behind the caudate lobe and passed behind the right hepatic vein. Each end of the tape was then passed behind V5 and V8, followed by dissection of the remaining liver parenchyma between them. Subsequently, a subtraction technique for the Glissonean pedicle was applied to safely isolate the right hepatic duct and hilar plate.

Results: Between September 2011 and May 2025, seven donors underwent right lobe graft procurement using the subtraction technique for the middle hepatic vein tributaries. The mean operative time was 381 min with an estimated blood loss of 217 ml. Using the subtraction technique, all middle hepatic vein tributaries were preserved just before graft retrieval.

Conclusion: This study presents subtraction techniques used during living donor hepatectomy. The technique may facilitate liver parenchyma dissection while preserving MHV tributaries just before graft retrieval and isolating the Glissonean pedicle.

Background aim: N-myristoyltransferase 1 (NMT1), which myristoylates Src, is highly expressed in oral squamous cell carcinoma (OSCC). Although targeted therapies against epidermal growth factor receptor exist, their use is limited by resistance and toxicity, and NMT1-Src interactions remain unexplored. Herein, we aimed to evaluate the role of NMT1-mediated Src myristoylation in the malignant potential of OSCC.

Materials and methods: Myristoylation and the expression of NMT1 in OSCC were assessed using click chemistry and immunocytochemistry. RNA-seq and enrichment analyses were performed to compare OSCC cells with or without NMT1-siRNA treatment. Src activity was determined by measuring Src phosphorylation via western blotting. Expression and binding of NMT1 and Src were analyzed via immunoprecipitation using specific antibodies. HSC-2, HSC-3, WK2, and WK3F derived from human OSCC in vitro were also used to confirm malignancy by siRNA of siNMT1 in OSCC cell lines.

Results: NMT1 and Src expression was detected in all OSCC cell lines. RNA-seq analysis of SAS cells transfected with NMT1-siRNA revealed decreased expression of genes related to cell adhesion and angiogenesis. WK3F cells, which exhibit high malignancy, showed markedly higher NMT1 expression than other cell lines. Immunocytochemistry showed that Src membrane localization was reduced in all of the OSCC cell lines with NMT1 knockdown. Co-immunoprecipitation analysis confirmed that NMT1 was bound to Src during myristoylation.

Conclusion: NMT1 promotes OSCC malignancy by mediating Src myristoylation.

Background/aim: Mesenchymal stem cells (MSCs) can be used for regenerative medicine, particularly in the treatment of neurodegenerative diseases and peripheral nerve injuries. Among these, human placenta-derived mesenchymal stem cells (hPMSCs) possess a high potential for differentiation into various cell types, including those of the neural lineage under appropriate conditions. The primary aim of this study was to isolate and induce the differentiation of hPMSCs into Schwann cell-like cells (SC-like cells) that function to support and play a role in the peripheral nervous system.

Materials and methods: Human placental tissues were isolated, and hPMSCs were cultured and characterized. The isolated hPMSCs were positive for mesenchymal stem cell surface markers CD73, CD90, and CD105. The multipotency of hPMSCs was confirmed by their ability to differentiate into osteocytes, chondrocytes, and adipocytes. To induce Schwann cell differentiation, hPMSCs were cultured in Schwann cell differentiation medium with or without the addition of 5-azacytidine (5-aza). Gene and protein expression analyses were performed to assess Schwann cell-specific markers.

Results: After 14 days of induction, hPMSCs differentiated in Schwann cell differentiation medium showed significant upregulation of Schwann cell-specific genes S100β, P75, GFAP, and PMP22 (p<0.01). Interestingly, the group treated with 5-aza exhibited even higher expression levels of these genes compared to the Schwann cell differentiation medium alone and the control group (p<0.01). Furthermore, protein expression analysis demonstrated that glial fibrillary acidic protein (GFAP) and myelin basic protein (MBP) were highly expressed in the 5-aza-treated condition, confirming enhanced Schwann cell-like differentiation.

Conclusion: This study demonstrates that hPMSCs can be successfully differentiated into Schwann cell-like cells. The addition of 5-aza significantly promoted this differentiation, leading to higher expression of Schwann cell-specific genes and proteins. These findings suggest that 5-aza plays a supportive role in enhancing Schwann cell differentiation from hPMSCs and may provide a promising approach for future clinical applications in peripheral nerve regeneration.

Granulocytes, long considered short-lived effector cells, are increasingly recognized as key modulators of the foreign body response (FBR) to biomaterials and determinants of regenerative outcomes. This review summarizes current evidence on the roles of neutrophils, basophils, and eosinophils in biomaterial-associated inflammation and tissue remodeling. Particular focus is placed on protein adsorption, cytokine release, and downstream effects across diverse biomaterial classes, including bone substitutes, collagen scaffolds, titanium, magnesium, and synthetic polymers. Neutrophils dominate the acute phase through reactive oxygen species, proteases, and neutrophil extracellular traps, which can either support remodeling or drive fibrosis and implant failure. Basophils, though rare, release histamine and Th2 cytokines, enhancing angiogenesis but also contributing to fibrotic encapsulation. Eosinophils are recruited in material-dependent patterns, releasing cytotoxic granules and pro-regenerative mediators, thereby functioning as double-edged regulators of degradation, fibrosis, and vascularization. Overall, granulocytes act as critical, though often overlooked, determinants of biomaterial integration. Incorporating granulocyte biology into biomaterial design-through modulation of surface chemistry, protein adsorption, and degradation kinetics- offers a promising path to guide inflammatory cascades toward constructive remodeling, angiogenesis, and predictable clinical performance.

Hyperglycemia is frequently encountered in critically ill patients and is associated with adverse outcomes, including increased morbidity and mortality. It may be pre-existing due to diabetes mellitus or develop during hospitalization as stress hyperglycemia. This review aims to raise awareness among medical staff regarding the implications of hyperglycemia in critically ill patients, its pathophysiology, monitoring techniques, and current treatment protocols. Therefore, evidence from the international literature is analyzed, with specialized journals serving as reference points, focusing on the impact of glycemic variability, individualized patient management, and emerging technologies such as artificial intelligence-driven biosensors for glucose monitoring. Hyperglycemia is defined as a blood glucose level exceeding 7.0 mmol/l (125 mg/dl) before admission or surpassing 10.0 mmol/l (180 mg/dl) two hours postprandial. Monitoring can be conducted using arterial or capillary blood, continuous glucose monitoring systems, or artificial intelligence-enhanced biosensors. Regardless of their diabetic status, patients present unique clinical features. Those with pre-existing diabetes may experience hypoglycemia, diabetic ketoacidosis, hyperosmolar hyperglycemic state, electrolyte imbalances, and dehydration, necessitating meticulous monitoring and intervention. Special attention must be given to glycemic variability, maintaining levels between 3.8 and 10.0 mmol/l (70-180 mg/dl), and timely interventions to prevent complications. Current treatment protocols prioritize intravenous insulin infusion based on multiple clinical studies on hyperglycemic intensive care unit (ICU) patients. To minimize complications, maintaining glucose levels between 6.1 and 10.0 mmol/l (110-180 mg/dl) is recommended, balancing the risk of hypoglycemia. Hyperglycemia negatively impacts mortality due to systemic damage and the delayed detection of glucose dysregulation. This review underscores the necessity of personalized glycemic control strategies to improve patient outcomes in ICUs.

Background/aim: Maxillary sinus floor elevation is a well-established procedure for increasing bone volume in the posterior maxilla, yet the regenerative outcome depends strongly on the choice of grafting material. This clinical study compared a high-temperature sintered xenograft (Bio-Oss^®) and a low-temperature processed xenograft (Ti-Oss^®) with regard to their regenerative and immunological profiles.

Patients and methods: Eight patients underwent split-mouth sinus augmentation with both materials, and biopsies were retrieved at 6 months after implant placement. Histological, histomorphometrical, and immunohistochemical analyses were performed to assess bone formation, material resorption, and tissue compatibility. Immunohistochemistry was applied to evaluate the local immune response, focusing on macrophage polarization and multinucleated giant cell activity. Quantitative histomorphometry determined the relative areas of newly formed bone, residual graft material, and connective tissue.

Results: Histopathological and histomorphometrical analyses demonstrated comparable levels of new bone formation in both groups, confirming reliable osteoconduction. Immunohistochemical evaluation revealed tartrate-resistant acid phosphatase isoform 5a (TRAP5A) expression in multinucleated giant cells adherent to both materials. Interestingly, Bio-Oss^® induced a higher proportion of anti-inflammatory (CD163⁺) macrophages, whereas Ti-Oss^® triggered a significantly greater number of pro-inflammatory multinucleated giant cells (CD11c⁺).

Conclusion: These findings indicate that although both xenogeneic substitutes (Bio-Oss^® and Ti-Oss^®) achieve bone regeneration, they elicit distinct immune responses, which may influence long-term remodeling and graft integration. Consideration of osteoimmunological properties is therefore essential when selecting biomaterials for clinical sinus augmentation.

Background/aim: Ramucirumab, a monoclonal antibody targeting vascular endothelial growth factor receptor-2 (VEGFR-2), has been shown to prolong survival in patients with advanced gastric cancer when combined with paclitaxel as second-line chemotherapy in the RAINBOW study. The most common adverse events include hypertension, proteinuria, and hemorrhage, reflecting its anti-angiogenic activity. Although the VEGF signaling pathway is strongly implicated in the pathogenesis of hemangiomas, the development of new hemangiomas during VEGFR-2 inhibition is rarely reported.

Case report: A 38-year-old woman with stage IV gastric adenocarcinoma presented with peritoneal dissemination and lymph node metastasis. She initially received FOLFOX plus immune checkpoint inhibitor therapy, achieving a partial response before disease progression. Second-line treatment with paclitaxel and ramucirumab was initiated. During therapy, chest computed tomography revealed a newly developed contrast-enhancing nodule in the left breast parenchyma, raising suspicion of metastasis or primary breast carcinoma. Breast ultrasonography confirmed a hypervascular lesion, and ultrasound-guided biopsy demonstrated numerous capillary-sized vessels lined by bland endothelial cells, consistent with capillary hemangioma. Owing to deterioration of her general condition, chemotherapy was temporarily discontinued. Despite subsequent third-line FOLFIRI and fourth-line trifluridine/tipiracil treatment, the patient eventually died of disease progression. Notably, follow-up chest CT scans showed gradual reduction and eventual spontaneous disappearance of the breast hemangioma, independent of systemic therapy.

Conclusion: This is a rare case of a breast parenchymal capillary hemangioma arising during ramucirumab therapy for advanced gastric cancer, which regressed spontaneously. Clinicians should recognize that atypical vascular lesions may develop paradoxically during VEGFR-2 blockade and confirm histological diagnosis to avoid misinterpretation as metastasis or new primary malignancy.

Background/aim: Hereditary hemorrhagic telangiectasia (HHT) is a rare disease with an incidence of 1:5,000. HHT is inherited in an autosomal dominant manner and is associated with vascular malformations. It particularly affects the genes ENG (HHT1) and ACVRL1 (HHT2). Clinically, patients typically exhibit pronounced recurrent epistaxis. The aim of this study was to evaluate if ACVRL1 knockdown in HMEC-1 endothelial cells could induce a HHT2-like phenotype which could be deployed as a model for HHT2.

Materials and methods: The human immortalized endothelial cell line HMEC-1 was used for the experiments. RNAi knockdown was performed using a pool of four siRNAs targeting the ACVRL1 gene. The gene knockdown was verified using RT-qPCR and Western blot analysis. The effects of ACVRL1 knockdown on angiogenesis were compared to a non-target (NT) small RNA control using a tube formation assay. The expression of 84 endothelia-associated genes was analyzed with RT-qPCR.

Results: Tube formation ability was significantly affected by ACVRL1 knockdown. In particular, the parameters total tube length, total segment length, total master segment length, total branching length, total mesh area and branching interval were significantly increased whereas total isolated branch length, number of master junctions, number of branches and the number of isolated segments decreased after ACVRL1 knockdown. Significant changes in the expression of angiogenesis related genes were detected by qPCR analysis and discussed.

Conclusion: Knockdown of ACVRL1 in HMEC-1 endothelial cells leads to pathological angiogenesis with enhanced tube formation capacity, which is similar to the angiogenesis in vivo in patients with HHT2. The presented HMEC-1 cell-based system therefore has the potential to be deployed as an in vitro model for HHT2 studies.

Background/aim: Obesity represents a significant global health challenge and is closely linked to the prevalence of metabolic syndromes and liver disorders. In Taiwan, the Ministry of Health and Welfare has instituted specific evaluation protocols for functional foods to mitigate body fat accumulation, underscoring mechanisms beyond caloric restriction. The present study sought to assess the anti-adipogenic and metabolic effects of pterostilbene (PTS) using a rat model of high fat diet (HFD)-induced obesity, in compliance with Taiwan's regulatory standards for health claims about "difficult-to-form body fat".

Materials and methods: Male Sprague-Dawley (SD) rats were fed a HFD for six months to induce obesity. Subsequently, the SD rats were administered PTS orally at dosages of 30 or 50 mg/kg/day for 45 days. This study evaluated various parameters, including body weight, food intake, feed efficiency, body fat percentage, liver weight, and serum biochemical markers.

Results: Forty-five day 30 and 50 mg/kg PTS oral treatment significantly reduced body weight gain, food intake, and feed efficiency (all p<0.05). Remarkably, over 70% of the rats exhibited reduction in body fat exceeding 0.02%, meeting the established regulatory efficacy standards. Furthermore, notable improvements were observed in the aspartate aminotransferase (AST), alanine aminotransferase (ALT), triglycerides (TG), total cholesterol (TC), and glucose (GLU) levels (all p<0.05).

Conclusion: PTS is a promising natural compound for the formulation of health foods aimed at enhancing fat metabolism and providing liver protection. The observed effects are consistent with physiological expectations and aligned with the regulatory efficacy criteria established in Taiwan.

Background/aim: Rhupus syndrome, an overlap of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), is a rare condition characterized by heterogeneous clinical manifestations and difficult management. Conventional therapies, including glucocorticoids, disease-modifying anti-rheumatic drugs (DMARDs), immunosuppressants, and biologics, are often limited by drug intolerance and steroid-related complications. Molecular hydrogen (H₂) has emerged as a potential adjuvant therapy due to its antioxidant and immunomodulatory properties. This report aimed to evaluate the clinical efficacy of H₂ as an adjunct to rituximab in a patient with refractory Rhupus and multiple drug intolerances.

Case report: We report a 47-year-old male with a 27-year history of rheumatoid arthritis, who subsequently developed systemic lupus erythematosus, fulfilling criteria for Rhupus syndrome. His disease course was complicated by multiple drug intolerances, including adverse reactions to methotrexate, sulfasalazine, hydroxychloroquine, azathioprine, leflunomide, and rituximab at standard dosing. Despite reduced-dose rituximab and mycophenolic acid, disease activity persisted and lupus nephritis was confirmed. In October 2023, oral molecular hydrogen capsule therapy was initiated as adjuvant treatment, resulting in significant clinical improvement, which was paralleled by characteristic changes in T- and B-cell subsets, normalization of anti-dsDNA antibody, erythrocyte sedimentation rate (ESR), and complement levels, and successful discontinuation of prednisone. Hydrogen therapy was well tolerated without major adverse effects. The patient developed avascular necrosis from prior corticosteroid use but recovered well after hip arthroplasty in January 2025. Disease control was sustained with mycophenolic acid and continued hydrogen therapy.

Conclusion: This case suggests molecular hydrogen therapy as a potential adjuvant for refractory Rhupus with multiple drug intolerances, showing immune modulation, reduced inflammation, steroid withdrawal, and sustained control with low-dose rituximab. Further studies are needed to confirm its efficacy and standardize its use.