In vivo最新文献

Background/aim: Nattokinase (NK) is a potent serine protease with various pharmacological properties that include thrombolytic, anti-inflammatory, and antioxidant activities. The aim of this study was to investigate the neuroprotective effects of NK in transient middle cerebral artery occlusion (MCAO)-induced cerebral ischemia-reperfusion injury and determine the mechanisms underlying the effects of NK.

Materials and methods: Rats were administered NK (65 and 130 mg/kg body weight, p.o.) daily for seven days prior to MCAO surgery. The infarct volume, behavioral tests, clotting time, and antioxidant markers in the MCAO model rats were assessed. The involvement of various cellular pathways in the effects of NK was examined.

Results: NK treatment dose-dependently reduced infarct volume and prolonged clotting time in MCAO model rats. Transcription factor activity analysis revealed the involvement of nuclear factor erythroid 2-related factor 2 (Nrf2) activity and enhanced antioxidant enzyme expression following MCAO.

Conclusion: Oral nattokinase confers early neuroprotection after experimental cerebral ischemia through Nrf2-associated antioxidative modulation and a mild, transient anticoagulant effect, supporting its potential as an orally available adjunct in ischemic stroke management.

Background/aim: Brachytherapy (BT) is a standard component of treatment for patients with gynecological cancers, particularly cervical and endometrial cancer. This study aimed to evaluate the potential benefits of local supportive therapies for vaginal toxicity administered during and after vaginal or intracervical BT, within a standardized supportive care framework.

Patients and methods: A retrospective analysis was conducted on 105 patients treated between 2010 and 2016 for endometrial, cervical, or vaginal cancer with primary indication for BT. All patients underwent BT using an Iridium-192 source. Treatment protocols varied by diagnosis: patients with endometrial cancer received 25 Gy in five fractions, while patients with cervical cancer received 20 Gy in 4 fractions and were additionally treated with external beam radiotherapy. Patients were advised to follow a supportive care regimen including local estrogen, topical steroids, and moisturizing agents (e.g., Bepanthen^® ointment). Clinical examinations were performed, and quality of life was assessed using the EORTC QLQ-EN24 questionnaire.

Results: Vaginal toxicity is a well-known complication of radiation therapy. In our cohort, 51% of patients experienced Grade I vaginal late toxicity according to the LENTSOMA scale. No Grade II or higher acute or late toxicities were reported. Adherence with the recommended supportive care was suboptimal; 26% (n=11) of patients did not perform any local supportive therapy despite repeated counseling.

Conclusion: Effective patient education and consistent guidance are crucial for ensuring adherence to supportive care protocols. Properly implemented, local supportive therapy may help mitigate both acute and late toxicity associated with BT. In our collective, a rate of 51% vaginal dryness was observed. However, the specific benefit of adding local vaginal steroid therapy or local estrogen remains inconclusive and requires further investigation.

Background/aim: Brain-to-lung metastasis is clinically rare but represents an underexplored route of cancer dissemination. Understanding this process may reveal novel mechanisms involving cerebrospinal fluid (CSF) and glymphatic clearance pathways. Existing preclinical models fail to replicate this directional spread or allow controlled investigation of central nervous system (CNS)-driven metastasis. This study aimed to develop a reproducible rat model to examine whether tumor cells introduced into the CNS can disseminate to peripheral organs, specifically the lungs.

Materials and methods: We established a brain-to-lung metastatic model in immunocompromised nude rats using intrathecal injection of A549-LUC human lung adenocarcinoma cells. This approach enabled precise tumor placement within the subarachnoid space for controlled modeling of metastatic progression. In vitro bioluminescence assays confirmed robust luciferase activity, peaking at 4 minutes post D-luciferin addition. In vivo imaging using the In Vivo Imaging System (IVIS) was employed to track tumor localization and dissemination over time.

Results: IVIS imaging revealed early tumor localization in the CNS, followed by progressive and asymmetric spread to the lungs, with higher radiance in the right lung. Intracranial tumor detection was limited by poor signal penetration through the skull. Post-mortem hematoxylin and eosin staining confirmed tumor lesions in brain and lung tissues. Physiological monitoring showed initial weight gain followed by decline, and survival analysis indicated a median survival of 36 days, with complete mortality by day 40.

Conclusion: This intrathecal model overcomes limitations of systemic injection techniques by enabling stepwise investigation of brain-to-lung metastasis. While immune interactions are restricted, this reproducible platform supports therapeutic testing - including ultrasound-enhanced glymphatic drug delivery - and provides a foundation for studying rare but clinically significant metastatic routes.

Background/aim: This study aimed to clarify the prognostic and predictive factors of patients with gastric cancer (GC) who are peritoneal lavage cytology positive and negative for other distant metastasis.

Patients and methods: Consecutive patients were selected from the database of Tokyo Metropolitan Cancer and Infectious Diseases Center Komagome Hospital, Department of Gastric Surgery, Tokyo, Japan, according to the following criteria: i) histologically-proven gastric adenocarcinoma; ii) patients who underwent macroscopically curative gastrectomy for gastric cancer as a primary treatment; and iii) a diagnosis of peritoneal washing cytology positive (CY1) disease with no distant metastasis between 2005 and 2025.

Results: A total of 89 patients were eligible for the present study. One-, three- and five-years overall survival (OS) rates of the whole cohort were 68.6%, 41.2%, and 21.3%. Medina survival time was 22.2 months (range=11.3-33.1 months). In the prognostic analysis for OS in the CY positive patients with GC who were negative for other distant metastasis, postoperative chemotherapy status was one of the independent prognostic factors [hazard ratio (HR)=2.228, 95% confidence interval (CI)=1.330-3.734]. One-, three- and five-years OS rates of the patients with postoperative adjuvant chemotherapy were 81.1%, 46.3%, and 33.9%, while that of patients without postoperative chemotherapy were 47.6%, 32.9%, and 4.7% (p=0.001). In addition, the prognostic analysis for OS in the CY positive patients GC who were negative for other distant metastasis and received postoperative chemotherapy, lymph node metastasis status was an independent significant prognostic factor in multivariate analysis (HR=6.312, 95% CI=1.476-26.996, p=0.013).

Conclusion: Postoperative chemotherapy improved the survival of CY positive patients with GC who were negative for other distant metastasis. In addition, lymph node metastasis status was an independent prognostic factor for CY positive patients with GC who were negative for other distant metastasis and received postoperative chemotherapy.

Background/aim: The present study aimed to investigate the association between sleep duration and the risk of newly-onset cancer among middle-aged and elderly adults, and to examine the effect of depression on this association.

Patients and methods: The data from 9,400 participants in the China Health and Retirement Longitudinal Study (CHARLS, 2011-2020) were analyzed. Cox proportional hazards regression models, Kaplan-Meier survival curves, restricted cubic spline (RCS) regression, and threshold-effect analysis were employed to evaluate the relationship between sleep duration and cancer risk. Furthermore, subgroup analyses were conducted to explore potential heterogeneity across populations.

Results: Over a median follow-up period of 9.0 years, 271 participants (2.9%) developed newly-onset cancer. Longer sleep duration was associated with a reduced risk of incident cancer [fully adjusted hazard ratio (HR)=0.65, 95% confidence interval (CI)=0.51-0.84]. RCS regression indicated a nonlinear association between sleep duration and the risk of cancer incidence (nonlinearity p=0.005), with an inflection point at 5.0 h of sleep per night. Specifically, sleeping ≥5 h per day was associated with an 18% lower risk of cancer (HR=0.82, 95% CI=0.74-0.91), whereas shorter sleep duration (<5 h) showed no significant protective effect. This protective effect of sleep was more pronounced among individuals with depression (HR=0.64, 95% CI=0.44-0.92). Subgroup analyses further revealed that the protective effect of sleep was stronger among men, urban residents, individuals without hypertension, and smokers.

Conclusion: Longer sleep duration (≥5 h) is independently associated with a lower risk of newly-onset cancer in middle-aged and elderly adults, particularly in those with depression. These findings highlight the importance of sleep health for cancer prevention.

Background/aim: Osteosarcoma is the most common malignant bone tumor in pediatric and young adult patients. Osteosarcoma is also refractory to immune checkpoint inhibitors (ICIs). It has been recently demonstrated that methionine restriction (MR) increases the response to ICIs in melanoma and colon cancer. The present study aimed to determine whether MR alone can be an immunotherapeutic for osteosarcoma.

Materials and methods: K7M2 murine osteosarcoma cells and 143B human osteosarcoma cells were used for the present study. Cell viability and the half-maximal effective concentration (EC₅₀) of methionine for K7M2 and 143B were determined with the WST-8 cell-viability reagent. Western immunoblotting was used to compare programmed cell death receptor ligand 1 (PD-L1) expression in K7M2 and 143B cells treated with and without MR. K7M2 cells were subcutaneously implanted in immunocompetent BALB/c mice and T-cell-deficient nude (nu/nu) mice to determine the efficacy of an MR diet on tumor growth and enhancing CD8-positive T-cell tumor infiltration in BALB/c mice. Tumor-infiltrating lymphocytes in the tumor of BALB/c mice were determined with immunohistochemistry.

Results: The EC₅₀ values of methionine for K7M2 and 143B were 14.18 μM and 20.85 μM, respectively. Both cell lines had a strong dependence on methionine at the concentration range of 4 to 32 μM. MR using methionine-depleted medium in vitro decreased PD-L1 expression in 143B and K7M2, compared to untreated control cells (p<0.05, respectively). The MR diet significantly suppressed the growth of K7M2 tumors in immunocompetent BALB/c mice (p<0.05), but not in T-cell-deficient nu/nu mice. The MR diet enhanced CD8-positive T-cell infiltration in the K7M2 tumor growing in BALB/c mice (p<0.05).

Conclusion: MR alone is a potential immunotherapeutic for osteosarcoma. The present results suggest MR is a T-cell stimulant and not a cause of T-cell exhaustion.

Background/aim: Gastric emptying (GE) scintigraphy is a noninvasive technique that is useful for assessing gastric accommodation and emptying. Further, it can facilitate dynamic acquisition, which provides continuous images over time. Hence, we considered the quantitative evaluation of residual gastric motility function using dynamic images in post-gastrectomy patients. The current study aimed to establish a protocol for dynamic GE scintigraphy, a novel method that can quantitatively assess residual gastric motility function in patients who underwent gastrectomy.

Patients and methods: A preliminary analysis of four healthy volunteers who underwent dynamic GE scintigraphy examinations was performed. The following factors were considered during the development of the examination protocols: direction of acquisition, start time for acquisition, frame time, acquisition time, and end of the acquisition period. A clinical analysis of 14 postoperative patients who underwent dynamic GE scintigraphy examinations was performed. The examination protocol used in the clinical study was based on the results of the preliminary study. Four indices, including T_peak, T_1/2, lag phase, and gastric peristaltic frequency, were calculated.

Results: The T_peak was 7.9±3.8 min (mean±standard deviation), the T_1/2 reached 52.3±19.2 min, and the lag phase was 4.5±2.5 min. The peristaltic frequency was almost equal in all cases, and there were no significant differences between the major and minor axes.

Conclusion: The novel dynamic GE scintigraphy protocol was established to quantitatively assess residual gastric motility in postgastrectomy patients.

Background/aim: Immune checkpoint inhibitors are widely used in treating metastatic urothelial carcinoma and metastatic renal cell carcinoma. However, the incidence, timing, and clinical impact of immune-related adverse events in real-world practice remain unclear. This study aimed to investigate their onset patterns and outcomes.

Patients and methods: We retrospectively analyzed 210 patients with metastatic urothelial carcinoma (n=127) or metastatic renal cell carcinoma (n=83) treated with immune checkpoint inhibitors between 2017 and 2023 at a single academic institution. Events were graded using Common Terminology Criteria for Adverse Events version 5.0. Associations with progression-free and overall survival were assessed. We also evaluated cumulative incidence, corticosteroid or immunosuppressant use, and outcomes after rechallenge.

Results: Immune-related adverse events occurred in 78 patients (37.1%), more frequently in metastatic renal cell carcinoma (47.0%) than in metastatic urothelial carcinoma (30.9%). Overall, grade ≥3 events occurred in 22.0% of patients, with higher incidence in renal cell carcinoma (27.7%) than in urothelial carcinoma (17.3%). In metastatic urothelial carcinoma, their presence was linked to longer survival; no difference was seen in metastatic renal cell carcinoma. Most events developed within six months; only 3.4% occurred after one year. High-dose corticosteroids were administered in 13.8% of patients, and 72.4% of them successfully completed tapering. Immunosuppressants were used in 1.4%. Eleven patients underwent rechallenge, of whom two experienced recurrence.

Conclusion: Immune-related adverse events showed distinct incidence and prognostic relevance between urothelial and renal cell carcinoma. These findings underscore the need for treatment-specific monitoring and provide practical insights into managing toxicities and decisions regarding immunotherapy rechallenge.

Background/aim: Biologic enhancement of bone grafts with enamel matrix derivative (EMD) may improve regenerative outcomes by modulating the host tissue response. This study evaluated the in vivo tissue reaction to a biphasic bone substitute (Maxresorb^®), used alone or in combination with EMD, in a subcutaneous mouse model.

Materials and methods: Forty BALB/c mice received subcutaneous implants of Maxresorb^®, EMD, Maxresorb^® + EMD, or underwent sham surgery. Tissue samples were harvested at 15- and 30-days post-implantation. Histological and immunohistochemical analyses were performed to assess inflammation, neovascularization, fibrosis, and the presence of macrophages and biomaterial-associated multinucleated giant cells (BMGCs). Statistical analysis was conducted using ANOVA (p≤0.05).

Results: The Maxresorb^® + EMD group showed higher inflammatory scores, increased numbers of macrophages and BMGCs, and enhanced neovascularization compared with the other groups, particularly at 30 days. Despite this elevated cellular activity, all test conditions were classified as non-irritant or slightly irritant according to ISO 10993-6.

Conclusion: The combination of EMD and Maxresorb^® enhanced early tissue responses without adverse effects. These findings support the biocompatibility of this combination and suggest potential regenerative benefits through biologically driven material functionalization.

Background/aim: Decompressive craniectomy (DC) is a life-saving intervention performed in patients with severe neurological deterioration; however, early postoperative mortality remains high and reliable biochemical predictors are not well defined. This study aimed to identify early clinical and perioperative biochemical predictors of 30-day mortality in patients undergoing DC.

Patients and methods: This single-center retrospective cohort study included 102 patients who underwent DC between 2019 and 2024. Demographic, clinical, radiological, and biochemical parameters including sodium, potassium, hematocrit, international normalized ratio (INR), C-reactive protein (CRP), and neutrophil to lymphocyte ratio (NLR) were analyzed. The primary outcome was 30-day mortality. Univariable analyses were conducted using Chi-square and t-tests. Multivariable logistic regression models were constructed using a hierarchical approach to evaluate independent predictors. Model performance was assessed using likelihood ratio Chi-square, Hosmer-Lemeshow goodness-of-fit statistics, and area under the receiver operating characteristic curve (AUC).

Results: Thirty-day mortality occurred in 70.6% of patients. In the multivariable model, preoperative INR >1.20 (OR=25.60; 95% CI=1.89-345.82; p=0.015) and discharge NLR >3.13 (OR=5.19; 95% CI=1.01-26.66; p=0.048) were identified as strong independent biochemical predictors of mortality. Conversely, preoperative GCS 8-13 was independently associated with improved short-term survival (OR=0.16; 95% CI=0.03-0.89; p=0.036). The predictive accuracy of the model significantly improved after adding biochemical variables, with Pseudo R² increasing from 0.115 to 0.252 and an AUC of 0.826, indicating good discrimination and calibration (Hosmer-Lemeshow p=0.765).

Conclusion: Preoperative INR and discharge NLR are strong independent predictors of early mortality after decompressive craniectomy, while preoperative GCS remains a key indicator of improved survival. These routinely available biochemical markers may enhance postoperative risk stratification and inform clinical decision-making in critically ill neurosurgical patients.