Successful cloning through somatic cell nuclear transfer (SCNT) faces significant challenges due to epigenetic obstacles. Recent studies have highlighted the roles of H3K4me3 and H3K27me3 as potential contributors to these obstacles. However, the underlying mechanisms remain largely unclear. In this study, we generated genome-wide maps of H3K4me3 and H3K27me3 in mouse pre-implantation NT embryos. Our analysis revealed that aberrantly over-represented broad H3K4me3 domain and H3K27me3 signal lead to increased bivalent marks at gene promoters in NT embryos compared with naturally fertilized (NF) embryos at the 2-cell stage, which may link to relatively low levels of H3K36me3 in NT 2-cell embryos. Notably, the overexpression of Setd2, a H3K36me3 methyltransferase, successfully restored multiple epigenetic marks, including H3K36me3, H3K4me3, and H3K27me3. In addition, it reinstated the expression levels of ZGA-related genes by reestablishing H3K36me3 at gene body regions, which excluded H3K27me3 from bivalent promoters, ultimately improving cloning efficiency. These findings highlight the excessive bivalent state at gene promoters as a potent barrier and emphasize the removal of these barriers as a promising approach for achieving higher cloning efficiency.
Animals ; Mice ; Histone-Lysine N-Methyltransferase/biosynthesis* ; Histones/genetics* ; Nuclear Transfer Techniques ; Female ; Gene Expression Regulation, Developmental ; Promoter Regions, Genetic ; Epigenesis, Genetic ; Embryo, Mammalian/metabolism*

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

This study investigates the clinical differentiation and treatment strategies for bipolar disorder （BD） by analyzing the relationship of its core pathomechanisms including qi stagnation， blood stasis， phlegm turbidity， and heat constraint with bile acid metabolism. The imbalance of "yin in form and yang in function" leads to qi stagnation， blood stasis， phlegm turbidity， and heat constraint， which are critical in the pathogenesis and progression of BD. Bile acids regulate neuroinflammation， neural plasticity， and intestinal flora homeostasis through receptor-mediated pathways. It is believed that the physiological functions of bile acids concretely embody the concept of the "liver is yin in form and yang in function" theory. Clinically， prescriptions such as Sini Powder （四逆散） with the function of venting pathogen and resolving constraint， Wendan Decoction （温胆汤） of drying dampness and resolving phlegm， Longdan Xiegan Decoction （龙胆泻肝汤） of clearing liver and draining fire， and Huanglian Ejiao Decoction （黄连阿胶汤） of nourishing yin and blood can be used to nourish liver yin and restore liver yang function. These strategies may improve BD prognosis by modulating bile acid synthesis and metabolism.

ObjectiveTo analyze the disciplinary knowledge structure and core competency requirements of exercise rehabilitation and health professionals, based on the World Health Organization (WHO) International Classification of Functioning, Disability, and Health (ICF) and the rehabilitation competency framework (RCF), and to identify the key knowledge, skills, and professional qualities required by professionals in the field of health and rehabilitation, thus providing theoretical foundations for the cultivation of high-quality rehabilitation talent. MethodsBased on the WHO Global Competency Framework for Universal Health Coverage, this study adopted the theories and methods of ICF and RCF to systematically analyze the disciplinary knowledge, skills, five core competency domains, and beliefs and values of exercise rehabilitation and health professionals. This analysis identified the competency framework and content required for these professionals, and provided theoretical support for the development of exercise rehabilitation and health educational programs. ResultsFrom the perspective of health services, this study integrated sports into the four aspects of the health service continuum: prevention, intervention, rehabilitation and health promotion. Exercise rehabilitation and wellness was an applied discipline related to health services, with a focus on prevention, rehabilitation and health promotion. It was a multidisciplinary, interdisciplinary and comprehensive field that encompassed knowledge related to physical education and sports sciences, health sciences, rehabilitation sciences, public health, and health management. The ICF, as a scientific framework for understanding and describing health, functioning and disability, provided an important foundation for the construction of this disciplinary system. Based on the WHO global competency framework and RCF, this study constructed a core competency framework for exercise rehabilitation and health professionals. According to the disciplinary knowledge system and professional competency framework, this study established an educational objective system for undergraduate education program of exercise rehabilitation and health. The overall objective was to cultivate, within the framework of comprehensive health service model, interdisciplinary practitioners who possessed health service knowledge and skills, and were equipped with the competence and professional qualities to engage in physical education, sports and health services. Graduates would be able to provide diverse and high-quality physical education, sports and health services to various populations in a multidisciplinary team. Furthermore, they would possess the potential for professional development and be able to enhance their professional practice, learning and research abilities through further education or continuing education. ConclusionThe establishment of the exercise rehabilitation and health major is a response to the growing demand for interdisciplinary professionals in the sports and health service industry. From the perspective of disciplinary development, this field is a result of the interdisciplinary integration of sports science, health science, rehabilitation science, and health management. Based on the conceptual framework of the ICF, this study has constructed a functioning-based exercise rehabilitation and health disciplinary system, integrating the multidisciplinary achievements of physical education and sports sciences, rehabilitation sciences, health sciences, and health promotion. Based on RCF, it has established a core competency framework for exercise rehabilitation and health professionals, providing competency standards for the development of education program of exercise rehabilitation and health.

Emphasizing symptom efficacy is an important manifestation of the personalized diagnosis and treatment of traditional Chinese medicine （TCM）. However， in current clinical practice of TCM， there are challenges such as diverse symptom expressions， difficulty in standardization， inconsistent evaluation standards for symptom efficacy， lack of universal quantitative methods， and complexity in collecting complete and accurate symptom information. These issues hinder the full and effective utilization of symptom information. Addressing the current research status and existing problems of symptom terminology standardization， quantification and grading of symptom efficacy， and collection of symptom information， this paper proposes methodological strategies for effectively recording and utilizing TCM symptom efficacy information in the era of big data. These strategies include collecting TCM symptom information based on patient reporting， standardizing the evaluation of TCM symptom efficacy from measurement scales and evaluation dimensions， integrating TCM symptom efficacy evaluation into clinical diagnosis and treatment processes， and utilizing artificial intelligence technology to acquire and process TCM symptom efficacy information. TCM symptom efficacy evaluation strategies based on patient perspectives and artificial intelligence technology will help fully explore the value of data elements， promote the objective demonstration of the specific efficacy of TCM， and facilitate the discovery of effective patterns.

To summarize XU Jingshi's clinical experience in treating fever from the perspective of latent pathogen. It is believed that latent pathogen heat accumulation is an important pathogenesis of fever. For latent pathogen in shaoyang and membrane-source， modified Xiaochaihu Decoction （小柴胡汤） plus Qinghao （Artemisiae Annuae Herba） could be used to expel pathogen through membrane and regulate shaoyang； for latent pathogen leading to yin fluids consumption， treatment should clear heat and expel pathogen， and prescribed with Chaihu （Bupleuri Radix）， Huangqin （Scutellaria baicalensis Georgi）， Qinghao （Artemisia carvifolia） combined with Baihu Decoction （白虎汤） or Zhuye Shigao Decoction （竹叶石膏汤） or Qinghao Biejia Powder （青蒿鳖甲散） or Shengmai Decoction （生脉饮） or Siyin Decoction （四阴煎） by syndrome differentiation； for latent pathogen leading to liver wind or reverse transmission to the pericardium， treatment should cool to open pathways， and prescribed with modified Angong Niuhuang Pill （安宫牛黄丸） to clear mind and open the orifices， clear heat and resolve toxins， resolve constraint with aromatics， and expel pathogens to outside； for fever after radiotherapy， chemotherapy or surgery for tumours， prescribed with self-prescribed Fuzheng Anzhong Decoction （扶正安中汤） plus Xiaochaihu Decoction （小柴胡汤） and modified Qinghao Biejia Decoction （青蒿鳖甲汤） to reinforce healthy qi and dispel pathogen， and support the middle jiao.

Objective To evaluate the cost-effectiveness of posaconazole compared with voriconazole in the empiric or diagnostic-driven treatment of invasive mould diseases （IMD） in immunocompromised patients from the perspective of Chinese health system. Methods A decision tree model was constructed based on a phase Ⅲ clinical trial and other publicly available data to evaluate the incremental cost-effectiveness ratio （ICER） of posaconazole versus voriconazole. One-way sensitivity analysis, probability sensitivity analysis, and scenario analysis were conducted. Results The results of base-case analysis indicated that, at a willingness-to-pay （WTP） threshold of 3 times Chinese GDP per capita, posaconazole gained 0.0327 quality-adjusted life years （QALYs） with the cost savings of CNY 1 711.24, demonstrating a clear cost-effectiveness advantage. Sensitivity analysis revealed that the cost of voriconazole and posaconazole significantly influenced the ICER. Under different WTP thresholds, posaconazole consistently maintained its economic advantage over voriconazole. Scenario analysis showed that the incidence of mucormycosis did not impact the results, but a price reduction in voriconazole could change result. Conclusion In the empiric or diagnostic-driven treatment of IMD in immunocompromised patients, posaconazole was both an effective and economically viable choice when compared to voriconazole.

ObjectiveTo observe the effects of electroacupuncture at Zhongwan （CV12） on gastric nociceptive response induced by gastric acid stimulation and explore the underlying mechanisms associated with nuclei of the medullary viscerosensory and visceral motor neurons. MethodsTwenty SD rats were given intragastric administration of 0.5 mol/L diluted hydrochloric acid （0.5 ml/100 g） to induce gastric nociceptive response induction. Eight rats were randomly selected to record the gastric slow wave （GSW） area under the curve， and extracellular discharge frequency of neurons in the nucleus of the solitary tract （NTS） and dorsal motor nucleus of the vagus nerve （DMV） before intragastric administration and at 1， 5， 10， 15， 20， 25， 30， 35， 40， 45， 50， 55， and 60 minutes after intragastric administration. The remaining 12 rats received electroacupuncture intervention at Zhongwan within 5 to 25 minutes after intragastric administration of diluted hydrochloric acid， with a duration of one minute. The GSW area under the curve and extracellular discharge frequency of NTS and DMV neurons were compared between the 1-minute intervals before and after electroacupuncture intervention. ResultsCompared to the baseline before intragastric administration， the area under the curve of GSW significantly increased at 1， 5， 10， 15， 20， and 25 minutes after intragastric administration， and the extracellular discharge frequency of excitatory neurons in the NTS （accounting for 90%， 57/63） significantly increased at 1， 5， 10， 15， 20， 25， 30， 35， and 40 minutes， both reaching peak values at 1 minute after intragastric administration （P＜0.01 or P＜0.05）. The extracellular discharge frequency of inhibitory neurons in the DMV （accounting for 91%， 20/22） showed a non-significant increase at 1 minute after intragastric administration （P＞0.05）， but significantly decreased at other timepoints （P＜0.05）. Compared to the baseline before electroacupuncture intervention， the GSW area under the curve and the extracellular discharge frequency of excitatory neurons in the NTS significantly decreased （P＜0.05）， while the extracellular discharge frequency of inhibitory neurons in the DMV showed no significant difference （P＞0.05）. ConclusionElectroacupuncture at Zhongwan can improve gastric nociceptive response induced by gastric acid stimulation， possibly by reducing the transmission of visceral sensation and decreasing the excitability of NTS neurons in the medulla.