Advances in Clinical and Experimental Medicine最新文献

Congenital heart disease (CHD) remains the foremost cause of mortality in children under 20 years of age. Given their ease of harvest, robust proliferative capacity and multilineage differentiation potential, stem cells (SCs) have emerged as a promising therapeutic alternative. In particular, mesenchymal stem cells (MSCs) derived from Wharton's jelly in the umbilical cord, i.e., an abundant byproduct of childbirth, especially in lowand middle-income settings, can be induced toward a cardiomyocyte lineage, making them an attractive cell source for cardiac regeneration. Although MSCs can be directed toward a cardiomyocyte lineage using growth factors or chemical cues, most in vitro-differentiated cells remain developmentally immature, with only a small fraction achieving the structural, functional and metabolic maturity required for therapeutic use. In resource-limited laboratories, the primary challenge is to develop a simple, cost-effective protocol that reliably differentiates MSCs into structurally, functionally and metabolically mature cardiomyocytes. This review presents a streamlined, cost-effective in vitro differentiation protocol for umbilical-cord MSCs into cardiomyocytes, designed for laboratories with minimal resources, involving 3 sequential stages. First, induce cardiac mesoderm commitment by treating umbilical cord-derived MSCs (UC-MSCs) with 5-azacytidine (5-Aza) or bone morphogenetic protein (BMP). Next, specify cardiac progenitor cells by adding a Wnt-pathway inhibitor (e.g., IWP-2). Finally, drive cardiomyocyte maturation by supplementing the culture with insulin-like growth factors (IGFs). In laboratories lacking complex bioreactors, seeding the cells onto a simple biocompatible scaffold, such as a collagen or fibrin hydrogel, can further boost differentiation efficiency and promote tissue-like organization.

Background: Septic shock in pediatric intensive care units (PICUs) requires accurate prognostic tools. The Sequential Organ Failure Assessment (SOFA) and the Phoenix Sepsis Score (PSS) are both widely used, yet their comparative effectiveness has not been fully established.

Objectives: To evaluate the prognostic sensitivity of the SOFA and PSS scores in predicting mortality among pediatric patients with septic shock, and to compare their performance across different patient subgroups.

Material and methods: This retrospective study included 110 pediatric patients with septic shock admitted to the PICU of Shanxi Children's Hospital between 2020 and 2024. SOFA and PSS scores were recorded at admission, along with demographic, clinical, and outcome data. Patients with congenital organ abnormalities or severe inherited metabolic disorders were excluded. Predictive accuracy was assessed using correlation analyses and receiver operating characteristic (ROC) curve analysis.

Results: Both SOFA and PSS scores showed moderate correlations with mortality (SOFA: r = 0.57; PSS: r = 0.56), with SOFA demonstrating slightly higher overall predictive accuracy. PSS exhibited greater sensitivity in severe cases. Neurological and respiratory dysfunctions were the strongest predictors of mortality, whereas coagulation parameters had minimal prognostic value. Age-specific analysis revealed that SOFA was more accurate in patients aged 1-3 years and >7 years, while PSS outperformed SOFA in children aged 3-6 years.

Conclusions: Both SOFA and PSS scores are effective tools for predicting mortality in pediatric septic shock. SOFA demonstrated superior overall performance, whereas PSS showed advantages in specific age ranges and disease categories. Using the two scoring systems in combination may support more informed clinical decision-making.

Background: Breast cancer (BC) is a heterogeneous disease classified into 4 molecular subtypes, each with distinct molecular characteristics that influence treatment strategies, clinical outcomes and prognosis. These subtypes are associated with specific changes in cellular metabolism, which may play a crucial role in tumor development and progression.

Objectives: To identify distinctive serum metabolic biomarkers for each molecular BC subtype and to evaluate their associations with estrogen receptor (ER) and human epidermal growth factor 2 (HER2) receptor status, thereby refining molecular classification and informing personalized treatment strategies.

Material and methods: The study utilized the proton nuclear magnetic resonance (1H NMR) metabolomics method to collect serum metabolic profiles from BC patients. Pattern recognition analysis was employed to analyze the metabolic data. Metabolic markers specific to each molecular subtype were selected, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was employed to explore serum metabolic pathway heterogeneity.

Results: Distinct metabolic markers were identified for each molecular subtype, demonstrating strong discriminatory power. Additionally, we identified specific serum metabolites whose levels correlate with ER and HER2 expression profiles. The KEGG pathway analysis revealed significant heterogeneity in serum metabolic pathways across different subtypes.

Conclusions: This study demonstrates pronounced metabolic differences across BC subtypes that mirror their distinct molecular profiles and may underlie variations in therapeutic response. These metabolomic insights hold promise for refining tumor classification, improving diagnostic accuracy and guiding more personalized treatment strategies.

Background: The healing rate after treatment in patients with high anal fistula (HAF) remains low. In individuals with HAF, the loose combined cutting seton (LCCS) technique has shown promising effectiveness, demonstrating a high cure rate, low incidence of incontinence and reduced pain levels.

Objectives: To assess the long-term efficacy and safety of LCCS technique in patients with HAF.

Material and methods: The LCCS procedure was conducted in patients with HAF between December 2020 and February 2022. All participants were followed up for 12 months. The primary outcome was fistula healing, while secondary outcomes included fistula recurrence, visual analogue scale (VAS) pain score, severity of fecal incontinence, and quality of life.

Results: A total of 132 patients with HAF were included in the final analysis, with a mean follow-up duration of 17.0 ±3.8 months. At the 12-month follow-up, 130 patients (98.5%) achieved fistula healing. Among them, 103 patients who received primary HAF treatment at our center fully recovered, while 27 of 29 patients previously treated unsuccessfully at other hospitals achieved healing within 12 months, corresponding to a 93.1% success rate. Ninety patients (68.2%) reported no fecal incontinence at follow-up (Wexner Continence Grading Scale (WCGS) score = 0), and 42 patients had a WCGS score of 1. The LCCS procedure was associated with a persistently low risk of postoperative perianal discomfort, with 127 patients (96.2%) scoring 0 and only 5 (3.8%) scoring 1 on the VAS.

Conclusions: The LCCS technique is a safe and effective treatment for patients with HAF.

Background: Plasma ceramides are recognized biomarkers of cardiovascular risk; however, racial and ethnic differences in their levels, as well as their association with cardiovascular health (CVH) among African-American populations, remain insufficiently studied.

Objectives: This study aimed to assess the association between ceramide scores and CVH, as well as atherosclerotic cardiovascular disease (ASCVD) risk, among African-American adults, and to compare ceramide scores between African-American and White adults.

Material and methods: We conducted a secondary analysis of 2 U.S. studies including African-American and White adults. Collected data encompassed demographics, behavioral factors (e.g., diet) and clinical measures (e.g., plasma ceramide levels). Atherosclerotic cardiovascular disease risk was assessed using the American College of Cardiology/American Heart Association (ACC/AHA) 10-year pooled cohort equations, while CVH was evaluated using the American Heart Association (AHA) Life's Essential 8 (LE8) scoring system.

Results: Fifty-eight African-American adults (mean age: 54.6 years; 67.2% women) and 1,103 White adults (mean age: 64.5 years; 52.1% women) were included. Compared with White participants, African-Americans had significantly higher prevalence of obesity, hypertension, diabetes, and hyperlipidemia, but similar ASCVD risk (12.8% vs 12.6%; p = 0.65). No significant associations were observed between ceramide scores and either LE8 or ASCVD risk in African-Americans. Ceramide levels differed by race/ethnicity, with African-Americans showing lower concentrations of 18:0 (0.08 vs 0.10 μmol/L) and 24:1 (0.91 vs 1.17 μmol/L) species compared with White adults (both p < 0.001).

Conclusion: No association was observed between ceramide scores and CVH or ASCVD risk in African-American adults. Despite having a less favorable cardiometabolic profile, African-Americans exhibited lower ceramide levels than White adults. These findings suggest that ceramide scores may not accurately reflect cardiovascular risk in African-American populations.

Background: Human monkeypox is a zoonotic disease with increasing global prevalence. Although several studies have identified its potential risk factors, findings remain inconsistent, highlighting the need for a systematic evaluation.

Objectives: To systematically investigate risk factors associated with human monkeypox infections using meta-analysis.

Material and methods: A comprehensive search of PubMed, Scopus, Web of Science, Embase, and The Cochrane Library databases was conducted on all records up to February 19, 2024. Eligible studies assessing risk factors for monkeypox were included. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated, and heterogeneity was evaluated using I2 statistics.

Results: Of the 1,844 articles identified, 9 studies met the inclusion criteria after screening, no publication bias was identified, and the meta-analysis results showed strong robustness. Human immunodeficiency virus (HIV) infection significantly increased monkeypox risk (OR = 2.21, 95% CI: 1.13-4.34, p = 0.02, I2 = 93%). Concurrent sexually transmitted infections (STIs) were also a significant risk factor (OR = 1.84, 95% CI: 1.46-2.33), as was body mass index (BMI) higher than 30 kg/m2 (OR = 1.18, 95% CI: 0.19-7.53, p = 0.86), lower economic status (OR = 0.33, 95% CI: 0.01-9.36, p = 0.52), education level (OR = 0.74, 95% CI: 0.30-1.79, p = 0.50), or men who have sex with men (MSM) status (OR = 1.22, 95% CI: 0.84-1.75, p = 0.29).

Conclusion: HIV infection and concurrent STIs significantly increase monkeypox risk, underscoring the need for targeted prevention, including screening and risk reduction strategies in vulnerable populations, particularly MSM.

Background: Late diagnosis and chemotherapy resistance, particularly to 5-fluorouracil (5-FU), contribute to the low survival rate in cholangiocarcinoma (CCA) patients. Identifying relevant genes and pathways, as well as novel targeted molecules, is crucial to overcoming 5-FU resistance and improving treatment outcomes for CCA patients.

Objectives: This study aimed to determine the potential molecules associated with 5-FU resistance in CCA cells.

Material and methods: Transcriptomic datasets from 4 stable 5-FU-resistant cell lines and their corresponding parental lines were retrieved from the Gene Expression Omnibus. A series of bioinformatics analyses were conducted to identify key genes upregulated in 5-FU-resistant cells compared to their parental counterparts. The expression levels of candidate genes identified through bioinformatics analysis were validated in CCA tissues and cell lines.

Results: Differential gene expression, protein-protein interaction, and Hub genes analysis revealed 8 genes that were significantly upregulated in 5-FU resistance cells compared to their parental cells. Six of the 8 genes, including TCP1, RPS6, RPS29, HSPA5, RPS15A, and NOTCH1, were upregulated in patient CCA tissues. Using real-time PCR, only the expression levels of NOTCH1 and TCP1 were significantly higher in the 5-FU insensitive CCA cell lines, KKU-213A and KKU-213B, than that of the 5-FU sensitive CCA cell line, KKU-055. A similar result was observed in stable 5-FU-resistant cell lines (KKU-213A-FR and KKU-213B-FR) compared to their parental cells.

Conclusions: The bioinformatic analysis and PCR results revealed that NOTCH1 and TCP1 might be associated with 5-FU resistance and serve as potential molecular targets to enhance 5-FU sensitivity in CCA cells.

Background: Hyperlipidemia is a major risk factor for cardiovascular diseases and is associated with complications such as atherosclerosis and tendon injury. Though atorvastatin reduces cholesterol, genetic variants (CYP2D6-4, SULT1A1, CYP2C192) affect its response. These genetic variations influence atorvastatin metabolism, thereby affecting its therapeutic effectiveness.

Objectives: To advance personalized therapeutic drug monitoring and improve lipid profile management, this study aims to develop a robust and LC-MS/MS method for quantifying atorvastatin levels in human plasma. Additionally, to investigate the influence of genetic polymorphisms - particularly CYP2D6-4-on plasma concentrations of atorvastatin in patients with hyperlipidemia.

Material and methods: Ethical approval for the study was obtained from the appropriate institutional review boards, and written informed consent was obtained from all participants. Atorvastatin was measured using LC-MS/MS. PCR-based methods were used for genotyping. Statistical analyses were performed to evaluate relationships between plasma atorvastatin levels and genetic variants.

Results: The LC-MS/MS method demonstrated excellent linearity, accuracy, precision, and stability, for the quantification of atorvastatin in human plasma. Higher atorvastatin concentrations were tied to CYP2D6-4. Furthermore, the study validated the analytical method for consistent and reliable measurement of atorvastatin levels in clinical samples.

Conclusions: This study successfully developed and validated a straightforward and reliable LC-MS/MS method for quantifying atorvastatin levels in human plasma. Significant CYP2D64 - atorvastatin links highlight the value of pharmacogenetic dosing. Integrating pharmacogenetics - especially in the Jordanian population - may enhance the safety, efficacy, and individualization of atorvastatin therapy.

Background: Acute cholecystitis (AC) is a common biliary disorder, most often caused by gallstones obstructing the cystic duct and leading to gallbladder inflammation.

Objectives: This study aimed to compare the therapeutic efficacy and complication rates of laparoscopic cholecystectomy (LC) performed using the Calot's triangle approach vs traditional LC techniques in the treatment of AC.

Material and methods: A retrospective analysis was conducted on 120 patients diagnosed with AC, with 60 patients undergoing LC using the Calot's triangle approach (study group) and 60 patients treated with traditional LC techniques (control group). Surgical parameters, including operation time, intraoperative hemorrhage, postoperative recovery times, and 30-day postoperative complications were recorded. Intraoperative adhesion formation was evaluated through direct visualization and graded based on severity. Postoperative pain was assessed using the visual analogue scale (VAS).

Results: There was no statistically significant difference in the baseline characteristics between the 2 groups, confirming their comparability. The study group (Calot's triangle approach) demonstrated significantly shorter average operation time, postoperative exhaust time, and diet recovery time compared to the control group. Additionally, patients in the study group had significantly lower intraoperative bleeding, lower VAS pain scores at 24 h and 72 h postoperatively, and a lower overall complication rate compared to the control group (p < 0.05).

Conclusions: The LC Calot's triangle approach demonstrated shorter operation times and lower rates of certain complications compared with traditional LC techniques. However, the absence of statistically significant differences in some key outcomes highlights the need for further research to fully evaluate its clinical advantages and long-term benefits.

Background: The precise causal relationship between circulating inflammatory factors and sepsis has not yet been fully elucidated.

Objectives: To identify biomarkers that enable earlier and more accurate diagnosis of sepsis.

Material and methods: The causal relationships between 41 circulating inflammatory factors, C-reactive protein (CRP), and procalcitonin (PCT) with sepsis and 28-day sepsis-related mortality were evaluated using two-sample bidirectional Mendelian randomization (MR) analyses.

Results: This study revealed negative causal associations between genetically predicted circulating inflammatory factors-interleukin-6 (IL-6) (odds ratio [OR] = 0.923; 95% confidence interval [CI], 0.854-0.998; p = 0.044), RANTES (OR = 0.926; 95% CI, 0.862-0.994; p = 0.033), and macrophage inflammatory protein-1β (MIP1β) (OR = 0.957; 95% CI, 0.919-0.996; p = 0.032) - and the risk of sepsis. Furthermore, positive causal relationships were observed between CRP and sepsis (OR = 1.140; 95% CI: 1.055-1.232; p = 0.001), as well as between CRP and 28-day sepsis-related mortality (OR = 1.241; 95% CI: 1.034-1.489; p = 0.020). Platelet-derived growth factor-BB (PDGF-BB) levels were also elevated in sepsis (OR = 1.136; 95% CI: 1.003-1.286; p = 0.044). Mediation analysis indicated that CRP mediated the effects of IL-6 and RANTES on sepsis, accounting for 25.87% (OR = 0.980; 95% CI: 0.961-0.998) and 2.04% (OR = 1.002; 95% CI: 0.991-1.012) of the total effect, respectively. The robustness of these associations was confirmed through leave-one-out sensitivity analysis and funnel plots.

Conclusions: This study enhances our understanding of the mechanisms underlying sepsis and its associated mortality, and underscores the therapeutic potential of targeting inflammatory factors in its management.