In vivo最新文献

Background/aim: Imeglimin, a novel oral antidiabetic agent, was approved in 2021 for the treatment of type 2 diabetes mellitus (T2DM). Phase III clinical trials demonstrated its safety and efficacy in managing T2DM. However, its safety profile in patients with heart failure has not been thoroughly evaluated in real-world clinical settings.

Patients and methods: We analyzed cases of patients with heart failure (stage B or higher) who were newly prescribed imeglimin, based on electronic medical records from June 2022 to June 2024. Baseline clinical data at the initiation of imeglimin therapy were collected, and cardiovascular events, adverse effects (e.g., lactic acidosis), and blood test results, including glycated hemoglobin A1c (HbA1c), were assessed as of July 2024.

Results: A total of 21 patients met the inclusion criteria. HbA1c levels significantly decreased after an average of 312.1±205.8 days of imeglimin therapy (baseline vs. on therapy: 8.2±1.0% vs. 7.5±0.7%, p=0.001). Alanine aminotransferase levels were also significantly reduced (baseline vs. on therapy: 30.9±23.8 IU/l vs. 22.0±12.3 IU/l, p=0.022). No adverse drug reactions were observed during the treatment period. Major adverse cardiovascular events occurred in three patients (14%), although a clear association with imeglimin remains uncertain.

Conclusion: Imeglimin demonstrated safety and efficacy in T2DM in patients with coexisting heart failure.

Background/aim: Vaccine refusal or delay remains a significant public health concern, leading to lower vaccination rates and increasing the risk of preventable diseases.

Patients and methods: The study included 404 mothers and 413 children, assessing vaccination coverage and conducting telephone interviews with mothers who declined vaccines to understand their reasons.

Results: Children of mothers who supported vaccination were more likely to be fully immunized compared to those with hesitant mothers. Among the incompletely vaccinated or unvaccinated children, 73.08% had mothers from the hesitant group (GNV). However, 90.05% of hesitant mothers still vaccinated their children with all recommended vaccines, while 9.95% maintained their refusal. Only 3.22% of the total sample, all from the GNV group, refused vaccination entirely. The primary reasons for refusal included fear of side effects, lack of trust in vaccines or the healthcare system, negative vaccination experiences, and influence from media or social platforms.

Conclusion: While vaccination behaviors may improve as a child grows, a significant proportion of hesitant parents continue to exist across different population groups, contributing to suboptimal vaccination coverage rates. The consistent implementation of unified, nationwide strategies aimed at increasing trust in vaccines and the vaccination process is essential for achieving protective vaccination rates.

Background/aim: Immune checkpoint blockade has achieved great success as a targeted immunotherapy for solid cancers. However, small molecules that inhibit programmed death 1/programmed death ligand 1 (PD-1/PD-L1) binding are still being developed and have several advantages, such as high bioavailability. Previously, we reported a novel PD-1/PD-L1-inhibiting small compound, SCL-1, which showed potent antitumor effects on PD-L1⁺ tumors. These effects were dependent on CD8⁺ T-cell infiltration and PD-L1 expression on tumors. The present study investigated the in vivo antitumor activity of SCL-1 in various mouse syngeneic tumor models.

Materials and methods: Twelve syngeneic mice models of tumors, such as colon, breast, bladder, kidney, pancreatic, non-small cell lung cancers, melanoma, and lymphomas, were used for in vivo experiments. Tumor mutation burden (TMB) was analyzed by whole exome sequencing (WES) using reference DNA from mouse blood. The proportion of CD8⁺ T-cells infiltrating tumors before and after treatment was assessed using flow cytometry and immunohistochemistry (IHC).

Results: SCL-1 had a markedly greater antitumor effect (11 sensitive tumors and 1 resistant tumor among the 12 tumor types) than the anti-mouse PD-1 antibody (8 sensitive tumors and 4 resistant tumors). In addition, the tumor growth inhibition rate (%) was more closely associated with TMB in the SCL-1 group than in the anti-PD-1 antibody group. Furthermore, in vivo experiments using PD-L1 gene knockout and lymphocyte-depletion technologies demonstrated that the antitumor activity of SCL-1 was dependent on CD8⁺ T-cell infiltration and PD-L1 expression in tumors.

Conclusion: SCL-1 has great potential as an oral immunotherapy that targets immune checkpoint molecules in cancer treatment.

Background/aim: Lymphangioleiomyomatosis (LAM) belongs to the perivascular epithelioid cell tumor (PEComa) family. The relationship between LAM and tuberous sclerosis complex (TSC) is of particular concern in a subset of women with clinically occult LAM involving the pelvic lymph nodes. This study aimed to investigate the clinicopathological features of incidental nodal LAM detected during the surgical staging of gynecological tumors.

Patients and methods: During the study period of 10 years, we identified 17 patients with pelvic nodal LAM that was incidentally detected during surgery for gynecological neoplastic lesions. We conducted immunostaining to assess the diagnostic utility of a panel of PEComa markers.

Results: Two of the 17 patients (11.8%) were diagnosed with TSC before surgery without any pulmonary symptoms. During the follow-up, both patients developed pulmonary and extrapulmonary LAMs. All affected nodes were multiple and unilateral in the pelvic region. The mean nodal size was 5.4 mm, and the mean proportion of the area involved in the LAM was 34.1%. In two patients with TSC, the largest affected node measured 19.3 mm and 7.6 mm, respectively, and the proportion of the area replaced by LAM was 99% and 90%, respectively. The most frequently expressed markers were human melanoma black 45 and cathepsin K, which showed 100% positivity in all the examined cases.

Conclusion: While most small nodal LAMs incidentally discovered during surgery have insignificant prognostic value, larger nodal LAMs occupying most of the nodal parenchyma at reproductive age should raise awareness of pulmonary and extrapulmonary LAMs as well as TSC.

Background/aim: Clear cell renal cell carcinoma (ccRCC) represents the most common type of renal cancer. When resectable, nephrectomy is the only radical treatment for ccRCC, however metastasis is already present at 30% of the patient population. Although great progress has been made in the field of targeted therapy with the emergence of immune checkpoint inhibitors (ICIs) the cure of metastatic ccRCC (mccRCC) remains far from achieved. Additionally, the need of biomarkers capable of predicting their therapeutic efficacy with the aim to ameliorate patient treatment and management is crucial. This study aimed to investigate potential changes in the expression of 3 cuproptosis-related lncRNAs, FOXD2-AS1, MINCR, LINC02154, in the blood of mccRCC patients that receive ICI-based treatments and whether these changes could be used to distinguish patients with clinical benefit (CB) from patients with progressive disease (PD).

Materials and methods: Peripheral blood from 31 mccRCC patients was obtained, prior to administration of ICI-immunotherapy. Using the RECIST criteria patients were subdivided into CB and PD groups. The fold change of FOXD2-AS1, MINCR, LINC02154 was evaluated using qRT-PCR.

Results: The tested lncRNAs showed an increase in peripheral blood with the most notable up-regulation in FOXD2-AS1 and LINC02154 (fold change of 3.7 and 3.8 respectively), followed by MINCR, (fold change of 2.6) in the PD patient group.

Conclusion: Cuproptosis-related lncRNAs FOXD2-AS1, MINCR, LINC02154 show promise for the prediction of patient response (CB vs. PD) in ICI-based therapeutic schemes in patients with mccRCC.

Background/aim: Breast cancer stem cells (BCSCs) are a subpopulation of tumor cells that play a role in therapeutic resistance. In this study, we demonstrated that sertaconazole, an antifungal agent, displayed a potent inhibition on cancer stem cells (CSCs) and investigated the mechanism of action involved in its anti-BCSC effect.

Materials and methods: The effect of sertaconazole on BCSCs was investigated using a mammosphere formation assay, a colony formation assay, and a cell migration assay. In addition, CD44^high/CD24^low and ALDEFLOR analyses, an apoptosis assay, quantitative real-time PCR, western blotting, an electrophoretic mobility shift assay, and a cytokine profiling assay were performed.

Results: Sertaconazole inhibited cell proliferation, colony formation, cell migration, mammosphere formation, and mammosphere proliferation. It also induced apoptosis of breast cancer cells. It decreased the subpopulation of CD44^high/CD24^low and aldehyde dehydrogenase-expressing cells. It also reduced the DNA binding of Stat3 and nuclear protein expression levels of phosphorylated Stat3. Furthermore, it reduced the IL-8 mRNA levels of the mammosphere.

Conclusion: Sertaconazole can inhibit the Stat3 and IL-8 signaling pathways and induce CSC death. Thus, sertaconazole might be a potential inhibitor of BCSCs.

Background/aim: Low levels of vitamin D are a widespread global issue. This study aimed to determine the optimal vitamin D3 supplementation dose for healthy young adults by comparing the effectiveness of gradually increasing cholecalciferol doses over two years.

Patients and methods: Thirty-five volunteers participated in a two-season pilot study conducted from October to April to avoid sunlight-induced vitamin D3 synthesis. The participants used oil-based drops of cholecalciferol, increasing their dose from 1,000 to 2,000, 4,000, and then 8,000 IU daily for 60 days with a 30-day break.

Results: Supplementing with 1,000 IU/day raised vitamin D levels to the recommended range (above 75 nmol/l), but levels dropped below this range after a 30-day break. A dose of 2,000 IU/day maintained vitamin D levels within the recommended range, even after the break. Increasing the dose to 4,000 IU/day produced a rapid rise, though levels dropped more significantly after stopping supplementation. With 8,000 IU/day, both the rise and subsequent decline in vitamin D levels were more pronounced.

Conclusion: Effective vitamin D supplementation in healthy young adults can be achieved with a daily dose of 2,000 IU during winter. However, 4,000 IU/day was more effective for maintaining levels above 100 nmol/l, supporting broader health benefits. Regular monitoring of [25(OH)D], calcium, and phosphorus levels is essential.

Background/aim: Lactate is a physiologically active substance secreted by skeletal muscle that has been suggested to stimulate muscle mass gain. However, the molecular mechanism for lactate-associated muscle hypertrophy remains unclear. The purpose of the present study was to investigate whether oral administration of lactate increases muscle mass under different loading conditions.

Materials and methods: Male C57BL/6J mice were divided into 1) control, 2) lactate, 3) unloading, 4) unloading with lactate, 5) reloading after unloading, and 6) reloading after unloading with lactate groups. Mice in the unloading and reloading after unloading groups were subjected to hindlimb suspension (HS) for two weeks and 2-week ambulation recovery after HS, respectively. Mice of the lactate groups were orally administered sodium lactate five days per week. The changes in muscle mass (muscle weight and protein content) and intracellular signals in fast plantaris and slow soleus muscles were evaluated.

Results: Oral administration of lactate increased the muscle mass and suppressed 5'AMP-activated protein kinase (AMPK) phosphorylation in both plantaris and soleus muscles under normal weight-bearing and unloading conditions. However, during reloading after unloading, lactate administration increased muscle mass and suppressed AMPK phosphorylation in the plantaris muscle, but not in the soleus muscle.

Conclusion: Lactate administration is an effective countermeasure for unloading-associated skeletal muscle atrophy. This anabolic effect of lactate on skeletal muscle mass may differ depending on muscle types.

Background/aim: The incidence and characteristics of pediatric thrombotic events have become increasingly recognized, due to the enhanced utilization of advanced diagnostic techniques. Pediatric thrombosis remains less frequent than in adults, often manifesting in those with underlying congenital or acquired risk factors. This study aimed to establish epidemiological data on pediatric thrombotic events in Bihor County, Romania, highlighting the challenges of diagnosis in smaller medical centers and proposing a relevant diagnostic and treatment algorithm.

Patients and methods: This retrospective study, conducted over 22 years at the Emergency County Clinical Hospital Bihor, identified 39 pediatric patients diagnosed with thrombotic events using electronic medical records.

Results: Most patients (82.1%) were diagnosed between 2013 and 2024, with a slight male predominance. The age distribution shows two peak clusters: newborns up to one year and adolescents. The majority of cases (53.8%) were venous thromboembolism, followed by arterial thromboembolism at 41%, while 5.1% involved both types. Cerebral thrombosis was the most common, followed by lower and upper limb events. Inherited thrombophilia factors were found in all patients tested, with antithrombin, protein S, and protein C deficiencies identified. Malignancy was the most frequently acquired risk factor, and PAI-1 4G/5G was the most common genetic variant detected among inherited factors.

Conclusion: This study highlights the significant rise in pediatric thromboembolism recognition over the past two decades; however, underdiagnosis remains an issue. Improved awareness among healthcare professionals is crucial, particularly for unprovoked thrombosis cases where a thorough thrombophilia panel and the involvement of a multidisciplinary team may be necessary.

Background/aim: Transient ischaemic attack (TIA) is characterised by a temporary neurological dysfunction resulting from focal ischaemia in the brain, spinal cord or retina without acute infarction. These episodes typically last less than 24 hours and are significant predictors of subsequent ischaemic strokes. Hypertension is a major risk factor for cerebrovascular events, and primary aldosteronism (PA) is recognised as a common cause of secondary hypertension. This case report presents a male patient with secondary hypertension due to Conn Syndrome, a form of PA, who experienced a TIA manifesting as left leg weakness, underscoring the heightened stroke risk associated with secondary hypertension.

Case report: A 78-year-old male with secondary hypertension caused by Conn Syndrome presented with an episode of left leg weakness that resolved within 24 hours. After ruling out other potential causes such as metabolic disturbances, infections, and structural brain lesions, he was diagnosed with TIA and treated with dual antiplatelet therapy. A carotid ultrasound revealed significant stenosis, leading to a referral for carotid endarterectomy. Long-term management included clopidogrel monotherapy and optimising hypertension control.

Conclusion: This case highlights the increased stroke risk in patients with Conn Syndrome-related hypertension, emphasising the importance of early recognition and optimising hypertension management in patients with secondary hypertension to prevent future cerebrovascular events.