Black tea consumption shown an association with the prevalence of metabolic syndrome (Mets) negatively. Theabrownin (TB) composes high content in black tea, and interferes tea quality of infusion color and flavor. To explore the functions of Ninghong black tea TB in attenuating Mets, and illustrate the beneficial potentials in health promotion, mice fed a HFD were intervened with TB (150 and 300 mg/kg) for 11 weeks. Biochemical indexes were determined. Multi-omics analysis also was conducted. TB decreased adipogenesis-and lipogenesisrelated gene expression, reduced adipocyte expansion and lipid accumulation in white adipose tissue, and prevented HFD-induced obesity. TB reduced lipogenesis-related gene expression, enhanced lipolysis-and fatty acid oxidation-related gene expression, and alleviated hepatic lipid accumulation and liver steatosis in mice. TB exposure to mice down-regulated the mRNA levels of the inflammatory genes of TNF alpha, IL-6, Mip1a, Mcp1, and Socs3. While, TB increased glucose and insulin tolerance via enhancing PI3K/Akt/Gsk-3 beta signaling, improved glucose consumption, and reduced PEPCK, G6Pase, PGC1 alpha, FoxO1 gene expression and glucose production, by reducing FoxO1-PGC1 alpha activities and activating AMPK signaling. Metabolomics analysis reported that TB alleviated lipid biosynthesis and arachidonic acid metabolism, which attenuated HFD-induced inflammation. The intermediates entering the gluconeogenesis pathway were reduced in TB-intervened mice, confirming the inhibitory effects on hepatic gluconeogenesis. Overall, Ninghong black tea TB prevented the development of obesity, decreased lipid accumulation via inhibiting lipid biosynthesis and promoting fatty acid oxidation. TB increased glucose and insulin tolerance, attenuated hepatic gluconeogenesis and hyperglycemia, alleviated HFDinduced inflammation, eventually improved Mets in obese mice.

OBJECTIVES:Metabolic disorders in newborns can cause significant morbidity and mortality if not diagnosed and managed early. In this study, we developed and validated a reliable LC-MS/MS method to simultaneously quantify methylmalonic acid, methylcitric acid, malonic acid, ethylmalonic acid, and total homocysteine associated with metabolic disorders. DESIGN AND METHODS:According to Clinical Laboratory Standards Institute (CLSI) guidelines, various analytical performances (i.e., precision, linearity, accuracy and reference intervals) were rigorously evaluated. Meanwhile, sample preparation, chromatographic separation and mass spectrometric detection and incorporating stable isotope-labeled internal standards were optimized. Furthermore, calibrators and quality controls were developed to ensure standardization and traceability. RESULTS:The coefficient of variations for precision were less than 10.0 %. Meanwhile, the results showed high accuracy (recoveries: 94.57 %-109.60 %) and robust linearity (R2 > 0.9935) over clinically relevant ranges. The limits of detection and limits of quantification for all analytes were established, enabling sensitive detection of pathological elevations. Reference intervals for pediatric populations (ages 0-1 month and 2 months to 18 years) were determined, providing essential baselines for clinical interpretation. CONCLUSIONS:The newly developed method demonstrated good analytical performance and offers a standardized, high-throughput solution for clinical laboratories to improve early diagnosis and personalized management of metabolic diseases.

Background:In recent years, the increasing number of elderly individuals has highlighted frailty as a significant public health issue. Although the potential health benefits of bilirubin in adults are recognized, studies on the link between bilirubin levels and frailty are sparse. This study explores the association between serum total bilirubin (STB) and frailty in individuals aged 45-85 who participated in the National Health and Nutrition Examination Survey (NHANES) from 2015 to 2020. Materials and methods:STB levels were measured using the Jendrassik-Grof method. Frailty was evaluated using a frailty index that included 49 deficits across seven domains. Survey weighted logistic regression analyses and Restricted Cubic Spline (RCS) techniques were used to examine the relationship between STB and frailty. Subgroup analyses were performed to confirm the consistency of the observed association. Results:The study comprised a cohort of 8,603 individuals aged between 45 and 85 years, of whom 54.7% were female, and 3,037 were identified as frail. In models that were fully adjusted, each unit increase in STB was associated with a 5% reduction in the risk of frailty. Participants in the second and third tertiles of STB exhibited statistically significant lower odds of frailty compared to those in the lowest tertile, with odds ratios (ORs) of 0.75 and 0.59, respectively. RCS analysis revealed an L-shaped correlation between STB levels and frailty, exhibiting statistically significant non-linearity (P = 0.0075), with an inflection point at 17.1 μmol/L of STB (the upper limit of normal). Below this threshold, a negative correlation is evident, whereas a weak positive correlation is observed for values exceeding 17.1 μmol/L. Further subgroup analysis within the physiological range of bilirubin suggests that the negative association between STB and frailty is more pronounced in individuals younger than 60 years. Conclusion:This study reveals a negative relationship between STB levels and frailty among middle-aged and elderly individuals, suggesting that elevated bilirubin concentrations within the physiological range may reduce the risk of frailty.

AIM To identify the pathogenic gene variant in a family with lacrimo-auriculo-dento-digital syndrome [LADD (MIM 149730)] showing congenital lacrimal duct dysplasia as the main clinical manifestation and lay the foundation for future research on the pathogenic gene. METHODS Ophthalmological examinations, including slit-lamp biomicroscopy and lacrimal duct probing, and computed tomography dacryocystography (CT-DCG) were performed for all participants. The family pedigree was drawn, genetic features were analyzed, and the genomic DNA of the subjects was extracted. Pathogenic genes were screened via whole exome sequencing (WES) and confirmed using Sanger sequencing. RESULTS Six patients belonged to this three-generation family, and their clinical manifestations included congenital nasolacrimal duct obstruction, congenital absence of lacrimal puncta and canaliculi, lacrimal fistulae, and limb deformities. This pattern indicates autosomal dominant inheritance. Diagnosis was based on the clinical characteristics of LADD syndrome, which presented in all the patients in this family. A novel frameshift mutation in the FGF10 gene (NM_004465.1), c.234dupC (p.Trp79Leus*15), was identified in all patients via WES. The variant was confirmed by Sanger sequencing and classified as a "pathogenic mutation" according to the American College of Medical Genetics and Genomics (ACMG) variant interpretation guidelines. CONCLUSION A novel frameshift mutation in the FGF10 gene is found in all patients. This finding helps this family with LADD syndrome receiving a more accurate clinical diagnosis and genetic counseling by extending the mutation range of the FGF10 gene.

OBJECTIVE To observe the change of receptor binding domain（RBD） neutralizing antibody in health care workers infected with severe acute respiratory syndrome coronavirus 2（SARS-CoV-2） and understand the immune response of high-risk population so as to provide evidence for follow-up. METHODS From Dec 1, 2022 to Dec 25, 2022, totally 385 health care workers who were infected with SARS-CoV-2 were recruited from a general hospital. The epidemiological data and clinical symptoms were collected, the expressions of SARS-CoV-2 specific antibodies [ S-IgM, S-IgG and RBD neutralizing antibody（ gold immunochromatography assay） in peripheral blood were detected, and the expression trends were analyzed by JPR model. RESULTS JPR model analysis showed that the expression levels of S-IgM, S-IgG and RBD neutralizing antibody were remarkably elevated after symptoms emerged for 4 and 6 days, the expression level of S-IgM began to decline after symptoms emerged for 8 days. The multivariate analysis based on generalized linear mixed model indicated that there was correlation between gender, fever and RBD neutralizing antibody, the expression level of RBD neutralizing antibody declined with the increase of age; vaccination could increase the expression of RBD neutralizing antibody. CONCLUSION The SARS-CoV-2 specific antibodies S-IgM, S-IgG and RBD neutralizing antibody are elevated remarkably after the typical symptoms emerge for 4-6 days, RBD neutralizing antibody maintains a certain level for a long time period. The RBD neutralizing antibody shows a downward trend with the increase of age.

Background: The aim of the present study was to compare the difference in drug resistance between CSCs and their original hepatocellular carcinoma (HCC) cells by analyzing the expression of genes and activity of signaling pathways associated with drug resistance, in an attempt to identify specific molecules involved in sorafenib resistance.Methods: Cell cycle and apoptosis analyses were performed with flow cytometry. The sensitivity of the cell lines to chemotherapeutic drugs was measured using the cell counting Kit-8. RNA-Seq is used to identify differentially expressed genes between Human cancer cell lines and CSCs. RT-PCR and Immunofluorescence analysis are used to analyze candidate genes that were associated with sorafenib resistance of liver CSCs. Finally, the signal transduction pathways associated with sorafenib resistance were detected by western blottingResults: CSC cell lines are more resistant to sorafenib than their original HCC cells. After Sorafenib treatment, the number of G1-phase liver cancer cells increased significantly, indicating that sorafenib can stop the cell cycle. The MAPK pathway is susceptible to sorafenib treatment in liver cancer cells, and the expression of ERK1/2 is decreased as a response to increasing concentrations of sorafenib administration. Among genes associated with sorafenib resistance, PDGFRB is specifically downregulated in liver CSCs. Moreover, the MAPK pathway inhibitor U0126 combined with sorafenib can effectively inhibit the viability of liver CSCs and increase their sensitivity to sorafenib.Conclusions: These findings may serve as a theoretical basis for developing liver CSC-targeted therapies for sorafenib-resistant HCC to improve the clinical outcome of patients with advanced HCC.

Supplementary Figure S1. Typical images of spiked Hep3B cells after negative enrichment for CTCs.

目的 探讨导管消融术(CA)治疗肥厚型心肌病(HCM)合并心房纤颤(AF)的长期结果、临床疗效及安全性,总结复发性心律失常的预测因素.方法 检索2013-01至2022-01纳入Pubmed、CNKI、NOS等电子数据库检索符合标准的18篇HCM合并AF患者进行CA术后的文献,利用Revman软件进行Meta分析,总结CA成功率、不良反应及并发症,评估CA对HCM伴AF临床疗效、长期结果及安全性.结果 HCM合并AF患者826例经CA治疗,停止口服药物后随访,Meta分析示,临床观察转复率69.98％,95％CI:1.99(1.72,2.30),P<0.00001.围手术期及术后并发症共64例,95％CI为0.09(0.08,0.10),P<0.00001.在HCM合并AF患者中行CA转复成功率高,能有效转复为窦性心律,部分患者在口服降心率药物的协助下能维持窦性心律.CA术后预测复发/成功的协变量为左心房直径(LAD)、左室射血分数(LVEF).结论 CA治疗HCM合并AF转复率高,临床效果显著,并发症少,病死率低,安全性好,LAD和左房容积指数是CA术后AF复发的最重要因素.

OBJECTIVE:To investigate the factors affecting phenotypes in the patients of methylmalonic acidemia combined with homocysteinemia cblC type with MMACHC c.609G>A homologous variant. METHODS:A retrospective study on the clinical manifestations, complications, treatment, and outcome in 164 patients of cblC type with MMACHC c.609G>A homologous variant was conducted. The patients were diagnosed by biochemical and genetic analysis from January 1998 to December 2020. RESULTS:Among the 164 patients, 2 cases were prenatally diagnosed and began treatment after birth. They are 3 and 12 years old with normal physical and mental development. Twenty-one cases were diagnosed by newborn screening. Among them, 15 cases had with normal development. They were treated from the age of two weeks at the asymptomatic period. Six cases began treatment aged 1 to 3 months after onset. Their development was delayed. One hundred and forty-one cases were clinically diagnosed. Their onset age ranges from a few minutes after birth to 6 years old. 110 cases had early-onset (78.0%). 31 cases had late-onset (22.0%). Five of them died. 24 patients lost to follow-up. Of the 141 clinically diagnosed patients, 130 (92.2%) with psychomotor retardation, 69 (48.9%) with epilepsy, 39 (27.7%) with anemia, 30 (21.3%) had visual impairment, 27 (19.1%) had hydrocephalus, 26 (18.4%) had feeding difficulties, 7 (5.0%) with liver damage, and 5 (3.5%) with metabolic syndrome. The frequency of hydrocephalus and seizures was significantly higher in the early-onset group. The urinary methylmalonic acid increased significantly in the patients with epilepsy. During the long-term follow-up, the level of plasma total homocysteine in the seizure-uncontrolled group was significantly higher than that in the seizure-controlled group, the difference had a statistical significance (P<0.05). CONCLUSION:Most of the patients with MMACHC c.609G>A homozygous variant had early-onset disease, with a high mortality and disability rate. If not treated in time, it will lead to neurological damage, resulting in epilepsy, mental retardation, hydrocephalus, and multiple organ damage. Pre-symptomatic diagnosis and treatment are crucial to prevent irreversible neurological damage. Neonatal screening and prenatal diagnosis are important to improve the outcome of the patients.

白血病是一种造血干细胞异常增殖与发育的进行性恶性疾病,其治疗的选择性十分有限,而微小残留病(MRD)已成为白血病复发的直接原因,也是患者生存的重要预后因素,因此MRD的检测有利于对白血病患者进行疾病状态评估、疗效判定、指导治疗及预测复发.临床广泛使用流式细胞术对MRD进行检测,但其检测能力较低,而新兴技术的快速发展则有望改善白血病患者的MRD检测,如质谱分析及第二代测序技术可以同时检测多个基因,对MRD的检测均具有较高的敏感性.本文对质谱分析及第二代测序技术检测白血病MRD的相关优点与局限性进行综述.

目的 探讨串联质谱法在新生儿原发性肉碱吸收障碍(CUD)中的应用.方法 使用串联质谱分析技术检测2015-2017年在该院进行新生儿游离肉碱及酰基肉碱标本10.9万份,并对二次召回游离肉碱(C0)依然降低的患儿肉碱转运蛋白基因进一步测序确诊.结果 串联法初筛发现了124例疑似原发肉碱吸收障碍的患儿,通过测序确诊3例原发肉碱吸收障碍的患儿均检测到SLC22A5基因突变,1例纯合突变,2例杂合突变.患儿血游离肉碱水平均低于参考值,且不同种类的酰基肉碱水平均有下降.结论 串联质谱分析技术对新生儿原发性肉碱吸收障碍的诊断有重要的提示意义,联合基因测序技术对新生儿遗传代谢病的早期诊断、治疗及预后意义重大.

Cerebral creatine deficiency syndromes (CCDSs) are a group of rare mendelian disorders mainly characterized by intellectual disability, movement anomaly, behavior disorder and seizures. SLC6A8, GAMT, and GATM are known genes responsible for CCDS. In this study, seven pediatric patients with developmental delay were recruited and submitted to a series of clinical evaluation, laboratory testing, and genetic analysis. The clinical manifestations and core biochemical indications of each child were basically consistent with the diagnosis of CCDS. Genetic diagnosis determined that all patients were positive for SLC6A8 or GAMT variation. A total of 12 variants were identified in this cohort, including six novel ones. The frequency of these variants, the Revel scores and the conservatism of the affected amino acids support their pathogenicity. Our findings expanded the mutation spectrum of CCDS disorders, and provided solid evidence for the counseling to affected families.

The metabolism of plasticizing phthalates may correlate with an increased risk of benign prostatic hyperplasia in humans. Diethylphthalate (DEHP) and its metabolites interfere with sex hormone function, causing inflammation and oxidative stress at low doses. The effects may contribute to the development of benign prostate hyperplasia.

Background: Due to its anti-oxidative effects, bilirubin may protect against a spectrum of diseases. However, the role of bilirubin in patients with benign prostatic hyperplasia (BPH) is poorly explored. This study aimed to investigate the cross-sectional associations between serum indirect bilirubin (IBIL) and prostate volume (PV) in patients with BPH. Methods: The medical records of 722 BPH patients were retrospectively analyzed. Body mass index (BMI) was calculated as body weight (kg)/height (m)(2). PV was obtained as height (cm) x width (cm) x length (cm) x pi/6. Other biochemical indexes were measured by the automatic biochemical analyzer. A univariable linear regression analysis was performed to detect confounders. The IBIL-PV relationship was examined using unadjusted and covariate-adjusted regression models. Furthermore, a segmented linear regression was conducted to analyze the linear trend of IBIL levels and PV. Finally, the sensitivity analysis was stratified by BMI and low-density lipoprotein cholesterol (LDL-C) cutoffs. Results: In this study, the mean age of the patients was 68 (range, 43- 93) years. By univariable line regression test, we observed that PV was positively correlated with age, BMI, and LDL-C (beta=0.113, 0.096, and 0.135, respectively). IBIL was negatively associated with PV in full adjusted model in men age <= 75 years (beta=-1.01; 95% CI: -1.81, -0.22; P=0.01). A statistically significant inverse trend was observed between serum IBIL intervals and PV in patients aged <= 75 years (adjusted for age, BMI, and LDL-C, P for trend >= 0.015). In sensitivity analysis, significantly negative IBIL-PV relationship only existed in men with normal BMI (adjusted beta=-1.328; 95% CI: -2.467, -0.190; P=0.022), overweight men (adjusted beta=-1.296; 95% CI: -2.519, -0.074; P=0.038), and men with normal LDL-C level (adjusted beta=-1.017; 95% CI: -1.869, -0.164; P=0.019). Conclusions: IBIL is negatively associated with PV in the non-obese population <= 75 years with normal LDL-C. These results suggest that higher serum IBIL possibly provides a degree of protection to BPH by mitigating oxidative stress (OS) related to aging and lipid peroxidation. Nevertheless, these preliminary findings from a single-center, retrospective study have limitations and need to be confirmed by future studies.

目的 分析直出式绿激光前列腺剜除术(Green LEP)治疗良性前列腺增生(BPH)的学习曲线.方法 对2021年1月至2022年4月期间,由解放军总医院泌尿外科1名泌尿科医生行Green LEP手术的72例BPH患者进行回顾性分析.根据手术时间先后分为A、B、C 3组,每组24例.比较3组患者年龄、前列腺体积、总前列腺特异抗原(tPSA)等基线特征,以及剜除时间、血红蛋白(Hb)变化、术后近期并发症等手术相关指标,并予评估术者的学习曲线.结果 3组患者除年龄以外的基线特征差异无统计学意义,相比A组,B、C两组的手术剜除时间短[36.92(24.50,42.59)min vs 50.87(36.38,79.47)min;25.98(19.88,36.37)min vs 50.87(36.38,79.47)min,P<0.05],前列腺组织剜除效率高[0.90±0.42mL/min vs 0.56±0.32mL/min;0.94±0.39mL/min vs 0.56±0.32mL/min P<0.05].24例手术后,操作者熟练掌握Green LEP手术.3组患者术后3个月IPSS评分和QoL评分与术前相比有显著改善(P<0.05).结论 采用Green LEP术治疗BPH安全、有效,其学习曲线约为24例.

Background: The coronavirus disease (COVID-19) pandemic has presented challenges in the care of patients with chronic dis-eases. We identified the challenges faced by Chinese patients with Duchenne muscular dystrophy (DMD) during the pandemic. Methods: An online cross-sectional survey was conducted between March 27 and June 30, 2021. Results: Of the 2105 valid questionnaire responses, 2,056 patients were from non-lockdown areas. In these areas, 42.8% reduced outside daily activities, 49.4% reduced rehabilitation service use, 39.7% postponed regular follow-ups, and 40.8% reported accelerated motor function decline. These figures generally increased for patients from lockdown areas-67.3% reduced outside daily activities, 44.9% reduced rehabilitation service use, 79.6% postponed regular follow-ups, and 55.1% reported accelerated motor function decline. Ambulation loss was most commonly reported in September and March before 2020; however, this trend was absent in 2020. Regarding the informed prices of disease-modifying drugs in Europe and the United States, 86.7% could afford a maximum of one-twentieth of the prices, 8.0% could afford one-tenth of the prices, and only 0.6% of the patients could afford the full prices. Conclusions: Implementation of standardized care for DMD in China is consistent with global practices, and the COVID-19 pandemic has affected the care of patients with chronic diseases worldwide, particularly in lockdown areas. Telemedicine is an effective model for providing healthcare to such patients. Healthcare workers should assist patients and establish more robust chronic disease management systems. Collaboration between governmental and non-governmental entities could address the cost of disease -modifying drugs in China and other developing countries. (C) 2022 The Japanese Society of Child Neurology Published by Elsevier B.V. All rights reserved.

目的 研究在体能训练过程中尿液肌红蛋白增高对学员尿液代谢水平的变化影响.方法 随机选取112人序贯进行队列动作训练、40km(6h)和55 km(11h)徒步训练,分别在静息状态、队列动作训练后、6h 40km徒步运动后和11h 55km徒步运动后收集尿液.现场即时测定尿液肌红蛋白水平,并将剩余尿液分装冻存,用于尿液有机酸检测.其中尿液肌红蛋白>1 ng/mL被分为肌红蛋白升高组,尿液肌红蛋白<1 ng/mL被分为正常组.结果 在训练过程中,正常组各指标均较静息状态有显著变化(P<0.05),而肌红蛋白升高组琥珀酸、2-酮戊二酸、苹果酸、顺乌头酸和柠檬酸无显著变化,其余指标均较静息状态差异有统计学意义(P<0.05).与正常组相比,在6h 40km徒步运动后,肌红蛋白升高组丙酮酸、2-酮戊二酸、3-羟基丁酸、2-羟基异戊酸、2-甲-3羟基丁酸水平降低(P<0.05),在11h 55km运动后,2-酮戊二酸和戊烯二酸低于正常组(P<0.05).肌红蛋白升高组在6h 40km和11h 55km运动后乳酸水平增高,棕榈酸水平较低,差异无统计学意义(P>0.05).结论 在运动过程中,肌红蛋白升高组较正常组无氧酵解产生更多乳酸,有氧氧化和脂肪酸代谢较弱.密切监测尿液肌红蛋白及尿液代谢水平的变化有助于揭示机体在运动中的肌肉微损伤情况及变化过程,指导科学训练,减少运动损伤的发生.

目的 为评价SARS-CoV-2感染后机体免疫应答水平,构建一种用于人血清、血浆和全血中的新型冠状病毒SARS-CoV-2中和抗体的定量检测方法.方法 利用双抗原夹心法建立S-RBD中和抗体结合方法,通过荧光免疫层析法进行检测,建立SARS-CoV-2中和抗体定量检测方法,展开性能评估、优化和样本验证.双抗原夹心法试剂能够检测到所有亚型的中和抗体结合时间分辨荧光技术实现高灵敏度检测,此外不受内源性免疫球蛋白和异嗜性抗体影响,特异性高.结果 通过400例健康人群检测定义参考范围0～10U/L;性能试验结果显示最低检测限为6.5U/L,弱阳性和中阳性样品的重复性分别为12.4％(13.1±1.62U/L)和10.5％(57.1±6.02U/L),与常见8种呼吸道病毒抗体无交叉反应.检测结果显示,新冠疫苗接种2周后能检测到中和抗体,14～21d时水平在118.37±20.21U/L;新冠感染轻症患者中和抗体在感染7d后显著升高,1个月内能维持较高水平(均值高于500U/L).结论 本研究开发的荧光免疫层析试剂条灵敏度高,可以即时定量检测机体内中和抗体水平,可用于初步评价疫苗接种保护效果和保护持久性,以及感染后机体的免疫应答效果和持续性.

Background: The purpose of the study was to investigate the levels of amino acids and acylcarnitines in newborns of the Tibet Autonomous Region for the first time and to provide an experimental basis for the diagnosis of genetic metabolic diseases. Methods: We detected concentrations of 43 kinds of amino acids, acylcarnitines and succinylacetone in the dried blood spots of 18482 newborns using liquid chromatography tandem mass spectrometry and diagnose the case by gene sequencing. We compared the indexes between Tibet and our lab, where most data come from an inland area and Han Chinese people. Then we compared amino acid and acylcarnitine levels of seven regions in Tibet and explored their impact factors. Results: We described the levels of amino acids and acylcarnitines in Tibet newborns using 95% confidence intervals. The distribution of amino acid and acylcarnitines were different in Tibet. Conclusion: This study has contributed to filling in the blanks of Tibet newborn screening, which should be considered in the newborn metabolic disease screening in this area.