Biomedical Journal最新文献

{"title":"Integrating comprehensive genomic profiling in the management of oncology patients: applications and challenges in Taiwan.","authors":"Chen-Yang Huang, Wen-Kuan Huang, Kun-Yun Yeh, John Wen-Cheng Chang, Yung-Chang Lin, Wen-Chi Chou","doi":"10.1016/j.bj.2025.100851","DOIUrl":"https://doi.org/10.1016/j.bj.2025.100851","url":null,"abstract":"<p><p>Comprehensive genomic profiling (CGP) refers to the detailed genomic analysis of cancers for oncology patients. With the rapid development of next-generation sequencing (NGS) technologies, CGP has been widely applied to clinical practice and managing oncology patients. CGP can be performed on the tumor DNA and RNA, as well as non-tumor tissues (e.g., blood, pleural effusion, and ascites). In this article, we review the current evidence supporting the use of CGP in the management of oncology patients, both in real-world practice and the bridging to clinical trials. We also discuss the role of the molecular tumor board on the application of CGP in oncology patients. We provide an overview of the current scheme of CGP reimbursement in Taiwan and the precision oncology branch of the National Biobank Consortium of Taiwan. Finally, we discuss about the potential barriers and challenges of applying CGP in managing oncology patients and the future perspectives of CGP in precision oncology.</p>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":" ","pages":"100851"},"PeriodicalIF":4.1,"publicationDate":"2025-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143787692","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"m6A landscape is more pervasive when Trypanosoma brucei exits the cell cycle","authors":"Lúcia Serra ,&nbsp;Sara Silva Pereira ,&nbsp;Idálio J. Viegas ,&nbsp;Henrique Machado ,&nbsp;Lara López-Escobar ,&nbsp;Luisa M. Figueiredo","doi":"10.1016/j.bj.2024.100728","DOIUrl":"10.1016/j.bj.2024.100728","url":null,"abstract":"<div><div>N6-methyladenosine (m⁶A) is an mRNA modification with important roles in gene expression. In African trypanosomes, this post-transcriptional modification is detected in hundreds of transcripts, and it affects the stability of the variant surface glycoprotein (VSG) transcript in the proliferating blood stream form. However, how the m⁶A landscape varies across the life cycle remains poorly defined. Using full-length, non-fragmented RNA, we immunoprecipitated and sequenced m⁶A-modified transcripts across three life cycle stages of <em>Trypanosoma brucei</em> – slender (proliferative), stumpy (quiescent), and procyclic forms (proliferative). We found that 1037 transcripts are methylated in at least one of these three life cycle stages. While 21% of methylated transcripts are common in the three stages of the life cycle, globally, each stage has a distinct methylome. Interestingly, 47% of methylated transcripts are detected in the quiescent stumpy form only, suggesting a critical role for m⁶A when parasites exit the cell cycle and prepare for transmission by the tsetse fly. In this stage, we found that a significant proportion of methylated transcripts encodes for proteins involved in RNA metabolism, which is consistent with their reduced transcription and translation. Moreover, we found that not all major surface proteins are regulated by m⁶A, as procyclins are not methylated, and that, within the VSG repertoire, not all VSG transcripts are demethylated upon parasite differentiation to procyclic form. This study reveals that the m⁶A regulatory landscape is specific to each life cycle stage, becoming more pervasive as <em>T. brucei</em> exits the cell cycle.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100728"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140847975","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Activating transcription factor 3 is an antitumor gene synergizing with growth differentiation factor 15 to modulate cell growth in human bladder cancer","authors":"Syue-Ting Chen ,&nbsp;Kang-Shuo Chang ,&nbsp;Wei-Yin Lin ,&nbsp;Shu-Yuan Hsu ,&nbsp;Hsin-Ching Sung ,&nbsp;Yu-Hsiang Lin ,&nbsp;Tsui-Hsia Feng ,&nbsp;Chen-Pang Hou ,&nbsp;Horng-Heng Juang","doi":"10.1016/j.bj.2024.100756","DOIUrl":"10.1016/j.bj.2024.100756","url":null,"abstract":"<div><h3>Background</h3><div>The functions of activating transcription factor 3 (ATF3) within the human bladder remain unexplored. This study delves into the expressions, functions, and regulatory mechanisms of ATF3 in human bladder cancer.</div></div><div><h3>Material and methods</h3><div>Gene expressions were determined by immunoblot, RT-qPCR, and reporter assays. Assays of Ki67, colony formation, Matrigel invasion, and the xenograft animal study were used to assess the cell proliferation, invasion, and tumorigenesis in vitro and in vivo. Silico analysis from TCGA database examined the correlations between GDF15 and ATF3 expressions, clinicopathologic features, and progression-free survival rates.</div></div><div><h3>Results</h3><div>Silico analysis confirmed that ATF3 is an antitumor gene, and the expression positively correlates with GDF15 in bladder cancer tissues. Multivariate analysis revealed that low ATF3/GDF15 but not a single low expression of ATF3 is an independent prognostic factor for progression-free survival of bladder cancer patients. Ectopic overexpression of ATF3 downregulated cell proliferation and invasion in bladder cancer cells in vitro<em>,</em> while ATF3-knockdown reversed these results. Knockdown of ATF3 upregulated EMT markers to enhance cell invasion in vitro and downregulated GDF15, NDRG1, and KAI-1 to elevate tumor growth in vivo. The activation of metformin on ATF3 and GDF15 in bladder cancer cells was blocked by SB431542, a TGFβ receptor inhibitor. ATF3 positively regulated GDF15 expression in bladder cancer cells through a feedback loop.</div></div><div><h3>Conclusions</h3><div>Our results identify that ATF3 is a metformin-upregulated antitumor gene. Results of Silico analysis align with cell-based studies suggesting that low ATF3/GDF15 could be a negative prognostic marker for bladder cancer.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100756"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141466003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pharmaceutical cost savings from the treatment of oncology patients in clinical trials","authors":"Borja Gómez Mediavilla ,&nbsp;Paloma Lanza-León ,&nbsp;Virginia Martínez Callejo ,&nbsp;David Cantarero-Prieto ,&nbsp;María Lanza Postigo ,&nbsp;Matilde Salcedo Lambea ,&nbsp;Yolanda Blanco Mesonero ,&nbsp;María Ochagavia Sufrategui ,&nbsp;Ignacio Durán ,&nbsp;Carmen María Sarabia Cobo","doi":"10.1016/j.bj.2024.100742","DOIUrl":"10.1016/j.bj.2024.100742","url":null,"abstract":"<div><h3>Objective</h3><div>The aim of this study was twofold: to assess the annual pharmaceutical savings associated with the treatment of cancer patients at Marqués de Valdecilla University Hospital and to estimate the cost of innovative antineoplastic therapies that patients receive as experimental treatment, both during clinical trials throughout 2020.</div></div><div><h3>Material and methods</h3><div>An observational and financial analysis of the drug cost related to clinical trials was applied. Direct cost savings to the Regional Health System of Cantabria and the cost of innovative therapies used as an experimental treatment in clinical trials were quantified.</div></div><div><h3>Results</h3><div>This study includes 38 clinical trials with a sample of 101 patients. The clinical trials analyzed provide a total cost savings of €603,350.21 and an average cost saving of €6630.22 per patient. Furthermore, the total investment amounts to €789,892.67, with an average investment of €15,488.09 per patient.</div></div><div><h3>Conclusions</h3><div>Clinical trials are essential for the advancement of science. Furthermore, clinical trials can be a significant source of income for both hospitals and Regional Health Systems, contributing to their financial sustainability.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100742"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140859710","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hirsutella sinensis polysaccharides and Parabacteroides goldsteinii reduce lupus severity in imiquimod-treated mice","authors":"Shih-Hsin Chang ,&nbsp;Yun-Fei Ko ,&nbsp;Jian-Ching Liau ,&nbsp;Cheng-Yeu Wu ,&nbsp;Tsong-Long Hwang ,&nbsp;David M. Ojcius ,&nbsp;John D. Young ,&nbsp;Jan Martel","doi":"10.1016/j.bj.2024.100754","DOIUrl":"10.1016/j.bj.2024.100754","url":null,"abstract":"<div><h3>Background</h3><div>The incidence of autoimmune diseases is increasing in developed countries, possibly due to the modern Western diet and lifestyle. We showed earlier that polysaccharides derived from the medicinal fungus <em>Hirsutella sinensis</em> produced anti-inflammatory, anti-diabetic and anti-obesity effects by modulating the gut microbiota and increasing the abundance of the commensal <em>Parabacteroides goldsteinii</em> in mice fed with a high-fat diet.</div></div><div><h3>Methods</h3><div>We examined the effects of the prebiotics, <em>H. sinensis</em> polysaccharides, and probiotic, <em>P. goldsteinii</em>, in a mouse model of imiquimod-induced systemic lupus erythematosus.</div></div><div><h3>Results</h3><div>The fungal polysaccharides and <em>P. goldsteinii</em> reduced markers of lupus severity, including the increase of spleen weight, proteinuria, and serum levels of anti-DNA auto-antibodies and signal transducer and activator of transcription 4 (STAT4). Moreover, the polysaccharides and <em>P. goldsteinii</em> improved markers of kidney and liver functions such as creatinine, blood urea nitrogen, glomerulus damage and fibrosis, and serum liver enzymes. However, the prebiotics and probiotics did not produce consistent improvements in gut microbiota composition, colonic histology, or expression of tight junction proteins in colon tissues.</div></div><div><h3>Conclusions</h3><div>Our results indicate that <em>H. sinensis</em> polysaccharides and the probiotic <em>P. goldsteinii</em> can reduce lupus markers in imiquimod-treated mice. These prebiotics and probiotics may therefore be added to other interventions conducive of a healthy lifestyle in order to counter autoimmune diseases.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100754"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141431304","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Liver transplantation for advanced hepatocellular carcinoma: Controversy over portal vein tumor thrombosis","authors":"Kun-Ming Chan ,&nbsp;Wei-Chen Lee","doi":"10.1016/j.bj.2024.100757","DOIUrl":"10.1016/j.bj.2024.100757","url":null,"abstract":"<div><div>Liver transplantation (LT) is considered the ideal treatment for hepatocellular carcinoma (HCC) concurrent with underlying cirrhotic liver disease. As well-known, LT for HCC based on the Milan criteria has shown satisfactory outcomes. However, numerous expanded transplantation criteria were proposed to benefit more patients for LT and showed comparable survivals as well. In addition, a modest expansion of transplantation criteria for HCC may be acceptable on the basis of the consensus within the transplantation community. Nonetheless, LT in patients with advanced HCC and portal vein tumor thrombosis (PVTT) recently has received attention and has been reported by many transplantation centers despite being contraindicated. Of those, the LT outcomes in certain HCC patients with PVTT were favorable. Additionally, the advancement of multimodality treatments and the evolution of systemic therapies have emerged as promising therapeutic options for downstaging advanced HCC prior to LT. Somehow, advanced HCC with PVTT could be downstaged to become eligible for LT through these multidisciplinary approaches. Although the available evidence of LT for HCC with PVTT is limited, it is hoped that LT may soon be more widely indicated for these patients. Nevertheless, several unknown factors associated with LT for HCC remain to be explored. Herein, this review aimed to update the developments in LT for patients with advanced HCC.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100757"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141466004","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The role of excess charge mitigation in electromagnetic hygiene: An integrative review","authors":"Isaac A. Jamieson ,&nbsp;J. Nigel B. Bell ,&nbsp;Paul Holdstock","doi":"10.1016/j.bj.2024.100801","DOIUrl":"10.1016/j.bj.2024.100801","url":null,"abstract":"<div><div>The electromagnetic characteristics of many environments have changed significantly in recent decades. This is in large part due to the increased presence of equipment that emits electromagnetic radiation and materials that may often readily gain excess charge. The presence of excess charge can often increase the risk of infection from pathogens and the likelihood of individuals experiencing compromised performance, respiratory problems, and other adverse health issues from increased uptake of particulate matter. It is proposed that adopting improved electromagnetic hygiene measures, including optimized humidity levels, to reduce the presence of inappropriate levels of electric charge can help reduce the likelihood of ill health, infection, and poor performance arising from contaminant inhalation and deposition, plus reduce the likelihood of medical devices and other electronic devices getting damaged and/or having their data compromised. It is suggested that such measures should be more widely adopted within clinical practice guidelines and water, sanitation, and hygiene programs.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100801"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142581091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Which outcomes are key to the pre-intervention assessment profile of a child with developmental coordination disorder? A systematic review and meta-analysis","authors":"Bouwien Smits-Engelsman ,&nbsp;Marisja Denysschen ,&nbsp;Jessica Lust ,&nbsp;Dané Coetzee ,&nbsp;Ludvík Valtr ,&nbsp;Marina Schoemaker ,&nbsp;Evi Verbecque","doi":"10.1016/j.bj.2024.100768","DOIUrl":"10.1016/j.bj.2024.100768","url":null,"abstract":"<div><h3>Background</h3><div>Purpose of this study was to determine what key aspects of function should be incorporated to make up a pre-intervention assessment profile of a child with Developmental Coordination Disorder (DCD); more specifically, what aspects of functioning are implicated in DCD and what is their relative impact?</div></div><div><h3>Methods</h3><div>A systematic review and meta-analysis were conducted, for which Pubmed, Web of Science, Scopus and Proquest were searched (last update: April 2023, PROSPERO: CRD42023461619). Case-control studies were included to determine point estimates for performances on field-based tests in different domains of functioning. The risk of bias was assessed, and the level of evidence was estimated. Random-effect meta-analyses were performed to calculate the pooled standardized mean differences for domains of functioning and subgrouping was done for clinically relevant subdomains. Heterogeneity was determined with <em>I</em>^<em>2</em>.</div></div><div><h3>Results</h3><div>121 papers were included for analyses. Data of 5923 children with DCD were included (59.8% boys) and 23 619 Typically Developing (TD) children (45.8% boys). The mean (SD) age of the DCD group was 10.3y (1.2) and 9.3y (1.3) for the TD children. Moderate evidence was found for motor performance, executive functions, sensory processing and perceptions, cognitive functions and sports and leisure activities to be affected in children with DCD.</div></div><div><h3>Conclusion</h3><div>Differences between the two groups varied per domain of functioning. This emphasizes the diversity present within children with DCD and provides a rationale for explaining the heterogeneity in this patient group. Yet, results highlight the potential involvement of all these domains and call for clinicians to be alert not only to examine motor skill difficulties but also other aspects of function. Results indicate the need to develop an individualized pre-intervention multi-dimensional assessment profile for each child with DCD. It also supports the important role that clinicians play in an interdisciplinary team to tackle the difficulties encountered by children with DCD.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100768"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141733499","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Diagnostic accuracy of transthoracic echocardiography for acute type A aortic syndrome: A systematic review and meta-analysis","authors":"Hsin-Tzu Yeh ,&nbsp;Sz-Wei Lu ,&nbsp;Tzu-Heng Cheng ,&nbsp;Jian-Xun Lu ,&nbsp;Chien-Han Hsiao ,&nbsp;Chieh-Ching Yen","doi":"10.1016/j.bj.2024.100747","DOIUrl":"10.1016/j.bj.2024.100747","url":null,"abstract":"<div><h3>Background</h3><div>Transthoracic echocardiography (TTE) is currently recognized as the potential first-line imaging test for patients with suspected acute type A aortic syndrome (AAAS). Direct TTE sign for detecting AAAS is positive if there is an intimal flap separating two aortic lumens or aortic wall thickening seen in the ascending aorta. Indirect TTE sign indicates high-risk features of AAAS, such as aortic root dilatation, pericardial effusion, and aortic regurgitation. Our aim is to summarize the existing clinical evidence regarding the diagnostic accuracy of TTE and to evaluate its potential role in the management of patients with suspected AAAS.</div></div><div><h3>Methods</h3><div>We included prospective or retrospective diagnostic cohort studies, written in any language, that specifically focused on using TTE to diagnose AAAS from databases such as PubMed, EMBASE, MEDLINE, and the Cochrane Library. The pooled sensitivity, specificity, positive likelihood ratio (PLR), negative likelihood ratio (NLR), diagnostic odds ratio (DOR) , and hierarchical summary receiver-operating characteristic (HSROC) curve were calculated for TTE in diagnosing AAAS. We applied Quality Assessment of Diagnostic Accuracy (QUADAS-2) tool and Grading of Recommendations, Assessment, Development and Evaluation (GRADE) quality assessment criteria.</div></div><div><h3>Results</h3><div>Ten studies (2886 patients) were included in the meta-analysis. The pooled sensitivity and specificity of direct TTE signs were 58% (95% CI, 38–76%) and 94% (95% CI, 89–97%). For any TTE signs, the pooled sensitivity and specificity were 91% (95% CI, 85–94%) and 74% (95% CI, 61–84%). The diagnostic accuracy of direct TTE signs was significantly higher than that of any TTE signs, as measured by the area under the HSROC curve [0.95 (95% CI, 0.92–0.96) vs. 0.87 (95% CI, 0.84–0.90)] in four studies.</div></div><div><h3>Conclusions</h3><div>Our study suggests that TTE could serve as the initial imaging test for patients with suspected AAAS. Given its high specificity, the presence of direct TTE signs may indicate AAAS, whereas the absence of any TTE signs, combined with low clinical suspicion, could suggest a lower likelihood of AAAS.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100747"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140910761","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The establishment of a molecular diagnostic platform for mitochondrial diseases: From conventional to next-generation sequencing","authors":"Ni-Chin Tsai ,&nbsp;Chai-Wai Liou ,&nbsp;Yin-Hua Cheng ,&nbsp;Hao-Ting Lien ,&nbsp;Tzu-Ling Lin ,&nbsp;Tsu-Kung Lin ,&nbsp;Min-Yu Lan ,&nbsp;Pi-Lien Hung ,&nbsp;Tzu-Jou Wang ,&nbsp;Chen-Hao Lee ,&nbsp;Yi-Chih Liang ,&nbsp;Kuo-Chung Lan","doi":"10.1016/j.bj.2024.100770","DOIUrl":"10.1016/j.bj.2024.100770","url":null,"abstract":"<div><h3>Background</h3><div>The aim of this study was to create a molecular diagnostic platform and establish a diagnostic pipeline for patients highly suspected of mitochondrial disorders. The effectiveness of three methods, namely, traditional restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR), Sanger sequencing for hotspot detection and whole mitochondrial DNA (mtDNA), and third-generation (Nanopore) whole mtDNA sequencing, will be compared in diagnosing patients with suspected primary mitochondrial diseases (PMDs). The strengths and limitations of different methods are also discussed.</div></div><div><h3>Material and methods</h3><div>A single-center prospective cohort study was conducted to validate the diagnostic pipeline for suspected mitochondrial diseases. In the first stage, a PCR-based method with five sets of primers was used to screen for eight hotspots (m.3243A &gt; G, m.3460G &gt; A, m.8344A &gt; G, m.8993T &gt; G, m.9185T &gt; C, m.11778G &gt; A, m.13513G &gt; A, and m.4977deletion) using either RFLP or direct Sanger sequencing. Sanger sequencing was also used to confirm the RFLP-positive samples. In the second stage, for samples with negative screening results for the eight hotspots, mitochondrial whole-genome sequencing was performed using Sanger sequencing or third-generation nanopore sequencing.</div></div><div><h3>Results</h3><div>Between June 2020 and May 2023, 30 patients from ages 0 to 63 with clinically suspected mitochondrial disease were enrolled. The positive yield for the diagnosis of PMDs was 8/30 = 26.7%, and the sensitivity of the heteroplasmy level for the RFLP-based method was approximately 5%. The remaining 22 patients who tested negative at the first stage were tested using Sanger sequencing or the third-generation sequencing Nanopore, and all tested negative for pathological mtDNA mutations. Compared to the Sanger sequencing method, the results of RFLP-PCR were compromised by the limitations of incomplete RFLP enzyme digestion. For whole-genome sequencing of mtDNA, Sanger sequencing, instead of nanopore sequencing, is preferred at our institution because of its cost-effectiveness.</div></div><div><h3>Conclusions</h3><div>In our highly selective cohort, most tested positive in the first stage of the 8 hot spots screen. Sanger sequencing is a conventional and accurate method for mitochondrial disease screening, at least for the most common hot spots in the region. The results revealed that Sanger sequencing is an accurate method with the benefit of being more cost-effective. This integral platform of molecular diagnosis bears the advantages of being relatively low cost and having a shorter reporting time, facilitating crucial identification of patients with clinical evidence of such disorders. This diagnostic flowchart has also been translated into routine clinical use in the tertiary hospital.</div></div>","PeriodicalId":8934,"journal":{"name":"Biomedical Journal","volume":"48 2","pages":"Article 100770"},"PeriodicalIF":4.1,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141756991","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}