Lawrence Shih-Hsin Wu

Alcohol use disorder (AUD) is a heterogeneous condition. Growing evidence highlights the role of the gut-brain axis in alcohol-related behaviors; however, longitudinal changes in the gut microbiome during pharmacological treatment for AUD remain poorly understood. In this 12-week study, we investigated gut microbiome composition and functional profiles in individuals with AUD undergoing naltrexone treatment. Seventy-two patients meeting DSM-5 criteria for AUD were enrolled. Stool samples were collected at baseline, week 4, and week 12 and analyzed using high-throughput 16 S rRNA gene followed by bioinformatic analyses. Of the 72 enrolled participants, 32 and 23 completed stool sampling at weeks 4 and 12, respectively. Despite significant improvements in drinking behavior over the treatment period, no significant changes in α- or β-diversity were observed, and no distinct clustering of gut microbial communities emerged across timepoints. Notably, the relative abundance of the Eubacterium hallii group increased from baseline and remained elevated through week 12. Several specific bacterial taxa were significantly associated with drinking outcomes and craving severity, particularly abstinence days (or inversely drinking days). At week 12, β-diversity, but not α-diversity, differed significantly between abstinence and non-abstinence groups. Functional enrichment analyses indicated that naltrexone treatment predicted functional reorganization of the gut microbiome based on 16 S inference, characterized by enhanced xenobiotic degradation and remodeling of cofactor-dependent antioxidant metabolism. Naltrexone treatment had limited effects on overall gut microbiota structure. In contrast, AUD patients who achieved sustained abstinence exhibited a distinct gut microbiota profile, suggesting that microbiome functional dynamics may contribute to AUD recovery and reflect treatment response to naltrexone. The absence of a placebo or untreated AUD control group precludes conclusions about the independent effects of naltrexone on gut microbiome changes.

Background :Post-acute sequelae of COVID-19 (PASC) in children present persistent symptoms that impair quality of life. Vitamin D’s immunomodulatory and microbiome-modulating properties suggest its potential to alleviate PASC symptoms. This study evaluated the efficacy of Vitamin D3 supplementation in alleviating symptom severity and to explore associated immunological and microbiome alterations in children with PASC. Methods: In a double-blind randomized controlled clinical trial, 33 children with PASC were assigned to receive 2,000 IU/day of oral vitamin D (Group A, n=21) or placebo (Group B, n=12) for 6 months. Serum 25-hydroxyvitamin D [25(OH)D] levels, symptom severity (Children’s Somatic Symptom Inventory-24, CSSI-24), immune cytokines, and checkpoint proteins were assessed at baseline (M0) and 6 months (M6). Nasal and rectal microbiota were analyzed using 16S rRNA sequencing to evaluate composition and predict functional pathways. Results: Serum 25(OH)D increased significantly in Group A compared with Group B (p < 0.01). Group A demonstrated significant reductions in CSSI-24 scores (p < 0.05), with improvements in neuropsychiatric, respiratory and cardiovascular symptoms. Pro-inflammatory cytokines including IFN-γ (FDR = 0.01) and MIP-1α (FDR = 0.0004) decreased, while checkpoint proteins TIM-3 and HVEM increased (both FDR < 0.05). Nasal microbiota in Group A at M6 showed increased richness and enrichment of Sphingomonas, Paenibacillus, Ralstonia, and Sphingobium, with functional pathways related to xenobiotic metabolism altered after supplementation. Coprobacillus abundance in rectal samples positively correlated with 25(OH)D levels and negatively with IFN-γ (p = 0.02). Pooling participants from both time points across group A indicated that higher 25(OH)D levels were associated with lower symptom scores. Conclusions: Vitamin D3 supplementation improved symptom severity, modulated immune responses, and altered microbiota profiles in children with PASC. These findings support Vitamin D3 as a potential adjunctive therapy for pediatric PASC and highlight host–microbe–immune interactions as therapeutic targets.

Background: The clinical features and risk factors of post‑acute sequelae of SARS‑CoV‑2 infection (PASC) in children remain incomplete. This study aims to clarify the manifestations and identify predictors of PASC in pediatric patients previously hospitalized for COVID‑19. Methods: We conducted this retrospective cohort study at the China Medical University Children’s Hospital in Taiwan, including 143 children aged 3-18 hospitalized for the acute COVID-19 during the Omicron pandemic from April 2022 to July 2023. PASC was assessed through phone calls, online survey, and in-person follow-ups. Data encompassing demographics, clinical presentations, laboratory results, and acute-phase treatment were collected and compared between children with and without PASC. Each child was assigned total clinical scores based on the acute-phase symptoms and treatment. We evaluated the association of the total clinical scores with PASC among study children. The receiver operating characteristic (ROC) analysis was used to determine the optimal cutoff for total clinical scores, which best differentiated occurrence of PASC. Results: Among the 143 discharged children, 35.7% (n = 51) developed PASC. Children with PASC tended to be older (mean age: 7.6 vs. 6.3 years, p = 0.05) and greater body weight (29.1 vs. 23.7 kg, p = 0.05). PASC children exhibited higher reverse transcriptase-polymerase chain reaction (RT-PCR) cycle threshold (Ct) values (17.1 vs. 14.2, p = 0.04) during the acute phase than those without PASC. There were no significant disparities in other laboratory results, length of intensive care unit (ICU) stays, bacterial coinfections, oxygen supplementation, and medication usage between two groups. Notably, compared to the control group, the total clinical scores were significantly higher in the PASC group (37.9 vs. 31.5, p = 0.02), with an optimal cutoff value of 37.7 yielding an area under the curve (AUC) of 0.70. Conclusions: This study identified a significant proportion of children developing PASC following hospitalization for acute COVID-19. Older age, greater body weight, and larger RT-PCR Ct values during the acute phase emerged as potential risk factors for developing PASC. Moreover, the clinical scoring system constructed in this study may help to predict the risk of developing PASC after discharge.

Post-acute sequelae of SARS-CoV-2 infection (PASC) is characterized by persistent symptoms following SARS-CoV-2 infection. Children with PASC are at risk of developing cardiac complications. Echocardiography has been instrumental in identifying cardiac abnormalities. This study applies deep learning to enhance the detection and understanding of echocardiographic changes in children with PASC. A case–control study was conducted at a pediatric tertiary center in central Taiwan. Children under 18 years who tested positive for SARS-CoV-2 and experienced symptoms for longer than 4 weeks were recruited between July 1, 2022, and July 31, 2023, during the Omicron variant surge. Echocardiographic data were also collected from a control group, consisting of children who presented with similar symptoms and received medical care in the same pediatric tertiary center in 2018. Children with congenital or structural heart disease, inflammatory conditions, or arrhythmias were excluded. Echocardiographic images were analyzed using a ResNet-50-based deep learning model to identify cardiac abnormalities. A total of 270 children with PASC and 400 age-matched control children were included. Standard echocardiographic parameters, including EF, FS, chamber dimensions, and valvular assessment, did not reveal abnormalities in the PASC group. The deep learning model achieved an accuracy of 96.6%, sensitivity of 96.7%, specificity of 96.2%, and balanced accuracy of 96.4%. AI-assisted echocardiographic analysis demonstrated high performance in distinguishing cardiac function between PASC and controls. Deep learning models enhance the detection of subtle cardiac changes in children with PASC. Although the deep learning model demonstrated high performance in distinguishing PASC from controls, the clinical significance of these subtle image-based differences remains uncertain and requires further evaluation in large-scale studies with long-term follow-up.

Background: Allergen sensitization patterns vary across regions; however, few studies have directly compared pediatric sensitization profiles between Japan and Taiwan. Objective: This study aimed to compare allergen sensitization patterns in children from Japan and Taiwan using the multiple antigen simultaneous test (MAST) and latent class analysis (LCA). Methods: A total of 217 Japanese and 166 Taiwanese children (aged 2–12 years) were enrolled. Based on responses to the International Study on Asthma and Allergy in Childhood questionnaire, participants were classified as either healthy or as having allergic rhinitis (AR), bronchial asthma (BA), or atopic dermatitis. Serum samples were analyzed using a commercial MAST kit available in Japan, and 20 allergens shared with the panel used in Taiwan were selected for analysis. Sensitization was defined as a lumicount value ≥2.78 (equivalent to class 2 or higher). LCA was conducted on the combined dataset, and the optimal number of latent classes was determined using the Akaike Information Criterion. Results: LCA identified 4 latent classes: low-sensitized (LS), house dust mite–dominant (HDM-D), inhalant-sensitized (IH-S), and poly-sensitized (PS). The HDM-D class was more prevalent in Taiwan (50%) than in Japan (6.9%). In contrast, the IH-S and PS classes were more common in Japan (36.4% vs. 3% and 8.8% vs. 2.4%, respectively). AR and BA were primarily associated with HDM-D in Taiwan and with IH-S in Japan, which includes HDM and Japanese cedar pollen. HDM and crab sensitization were strongly correlated in Taiwan. Conclusion: MAST combined with LCA reveals distinct regional sensitization profiles, suggesting divergent pathways in allergic disease development.

Background: The postacute sequelae of coronavirus disease 2019 (PASC), also known as pediatric long coronavirus disease (COVID), can significantly affect the quality of life (QoL) of affected children. Currently, there are no standardized screening tools to identify high-risk children. The Kinder Lebensqualität fragebogen (KINDL) is a psychometric method for measuring QoL in children.Purpose: This study used the KINDL questionnaire and Children's Somatic Symptom Inventory-24 (CSSI-24) to evaluate symptom burden and establish a screening threshold for pediatric PASC.Methods: We performed a cross-sectional study of children diagnosed with PASC defined as the presence of at least one persistent symptom lasting more than 4 weeks after a confirmed COVID-19 infection. Symptoms were assessed using a structured checklist developed by our team. The correlation between the KINDL and CSSI-24 scores was analyzed, and receiver operating characteristic (ROC) curve analysis was used to determine the optimal KINDL cutoff for identifying high-risk cases. QoL scores were also compared with those of historical cohorts to evaluate the impact of the PASC.Results: We included 84 participants: 46 children (mean age, 8.74±1.77 years; 41.3% girls) and 38 adolescents (mean age, 14.50±1.56 years; 44.7% girls). KINDL and CSSI-24 scores showed a significant negative correlation (r=-0.44, P<0.001), suggesting that increased somatic symptoms were associated with a reduced QoL. The ROC analysis identified a KINDL cutoff score of 74.75 (area under the curve, 0.82) for detecting high-risk children. Compared to historical cohorts, children with PASC had QoL scores comparable to child-reported norms from 2010 but lower than parent-reported norms from the same year.Conclusion: KINDL and CSSI-24 are practical tools for screening for pediatric PASC in outpatient settings. A KINDL cutoff of 74.75 may help clinicians identify children with PASC who require early intervention. Further studies in larger and more diverse populations are required to validate these findings.

Background Maternal and newborn vitamin D levels are closely linked, but the role of genetic factors is not fully understood. This study investigates how maternal vitamin D levels and variations in vitamin D metabolism genes influence cord blood vitamin D concentrations. Methods A total of 456 mother–newborn pairs were included. Maternal blood was collected during the second trimester (24–28 weeks), and cord blood at birth. Genetic polymorphisms in vitamin D receptor (VDR), vitamin D binding protein (VDBP), and cytochrome P450 27B1 (CYP27B1) genes were genotyped. Linear regression and other statistical methods assessed associations with cord blood vitamin D levels. Results Maternal and newborn vitamin D levels showed a linear correlation (R² = 0.3854, p < 0.001). Maternal CYP27B1 rs10877012 and the G allele count in mother–newborn pairs were significantly associated with cord blood levels (p = 0.0009). The newborn-to-maternal vitamin D ratio, reflecting transfer efficiency, was influenced by VDBP variants (maternal rs4588, p = 0.0203; newborn rs7041, p = 0.0171). Conclusion Maternal vitamin D status is the key driver of cord blood vitamin D levels, with genetic variants in CYP27B1 and VDBP playing additional roles in determining both the levels and the efficiency of vitamin D transfer from mother to newborn. Impact Demonstrates a strong linear correlation between maternal and newborn vitamin D levels. Identifies a significant association between maternal CYP27B1 polymorphism (rs10877012) and cord blood vitamin D concentration. Shows that VDBP genetic variants influence the efficiency of vitamin D transfer from mother to fetus. Supports the development of personalized vitamin D supplementation strategies for pregnant women to optimize neonatal outcomes.

Chronic obstructive pulmonary disease (COPD) involves persistent inflammation and dysregulated lipid metabolism, with foamy macrophages playing a central role in disease progression. Exosomes—vesicles transporting microRNAs (miRNAs)—mediate intercellular communication, but their contribution to foamy macrophage-driven COPD remains unclear. This study investigates the role of exosomal miRNAs, particularly let-7, in modulating lipid metabolism and inflammation in foamy macrophages. Bone marrow-derived macrophages (BMDMs) were treated with oxidized low-density lipoprotein (oxLDL) and lipopolysaccharide (LPS) to induce foamy macrophage formation. Exosomal miRNA profiles were analyzed, and the function of let-7c-3p was assessed via transfection. Foamy macrophages released significantly more exosomes (392.7 × 10⁷ particles) than controls (284.9–302.5 × 10⁷), without differences in exosome size or molecular content. The miRNA sequencing and qRT-PCR confirmed downregulation of exosomal let-7c-3p in foamy macrophages, correlating with increased RNF8 and decreased RXR expression—markers of disrupted PPAR/RXR signaling. Pathway analysis implicated let-7c-3p in regulating PPAR/RXR, WNT/β-catenin, and pulmonary fibrosis pathways. Transfection with let-7 mimics reduced lipid accumulation (52% to 19%), suppressed RNF8, restored RXR, and lowered IL-6 and TNF-α levels, indicating strong anti-inflammatory and lipid-modulating effects. Loss of exosomal let-7c-3p aggravates lipid dysregulation and inflammation in COPD by impairing PPAR/RXR signaling. Restoring let-7 expression reverses these effects, highlighting its potential as a diagnostic biomarker and therapeutic target.