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*

Objective To investigate the computed tomography (CT) features of solitary nodular invasive mucinous lung adenocarcinoma (IMA) in stage IA and establish its prediction model. Methods We included 53 lesions of 53 patients with stage-IA IMA and 141 control lesions of 141 patients with invasive non-mucinous lung adenocarcinoma (NIMA) that were confirmed by surgical pathology in our hospital from January 2017 to December 2019. Univariable analysis was used to compare the demographics and CT signs of the two groups. Multivariable logistic regression analysis was performed to determine the main factors influencing solitary nodular IMA. A risk score prediction model was constructed based on the regression coefficients of the main influencing factors. A receiver operating characteristic (ROC) curve was used to assess the performance of the model. Results The univariable analysis showed significant differences between the two groups in age, largest nodule diameter, tumor-lung interface, lobulation, spiculation, air-bronchogram or vacuole sign, vessel abnormalities (P < 0.05). The spiculation sign was different between the two groups, which was longer and softer in the IMA group while shorter and harder in the NIMA group. There was no significant difference in sex, nodule shape, or pleural retraction (P > 0.05), but irregular shapes were slightly more frequent in the IMA group. The multivariable logistic regression analysis showed that obscure tumor-lung interface (odds ratio (OR = 20.930, P < 0.05), air-bronchogram or vacuole sign (OR = 7.126, P < 0.05), spiculation sign (OR = 4.207, P < 0.05), and vessel abnormalities (OR = 0.147, P < 0.05) were the main influencing factors. The prediction model based on those factors’ regression coefficients had an area under the ROC curve of 0.829 (P < 0.05). Conclusion Compared with those with NIMA, patients with solitary nodular IMA in stage IA were older and more likely to have the CT features of obscure tumor-lung interface, air-bronchogram or vacuole sign, and longer and softer spiculation. Based on the regression coefficients of tumor-lung interface, air-bronchogram or vacuole sign, spiculation, and vessel abnormalities, the risk score prediction model showed good predictive performance for solitary nodular IMA.

OBJECTIVE To explore the influence of reusing and recycling times on the quality of ethanol recovered from ethanol deposition solution of aqueous extract of Lonicerae Japonicae Flos. METHODS The ethanol in five consecutive batches of Lonicerae Japonicae Flos water extract ethanol deposition was recovered by vacuum distillation. The impurities in five batches of recovered ethanol and the recovered ethanol when partial coking caused by temperature runaway were qualitatively analyzed by headspace solid phase micro extraction combined with gas chromatography-mass spectrometry(HS-SPME/GC-MS), and the ultraviolet absorbance of each batch of recovered ethanol was also analyzed. RESULTS Styrene was the only impurity in the first and second batch of recovered ethanol; there were five impurities in the third and fourth batch of recovered ethanol; the number of impurities in the fifth batch of recovered ethanol increases to 17. When partial coking was occurred, the exogenous impurities in the recovered ethanol increased dramatically to forty-one, and there were harmful substances such as ninhydrin and naphthalene. The ultraviolet absorbance of the third to fifth batch of recovered ethanol exceed the upper limit stipulated in the second part of Chinese Pharmacopoeia, 2020 Edition. While the ultraviolet absorbance of the recovered ethanol when partial coking takes place exceeds the standard of the pharmacopoeia greatly. CONCLUSION The recovered ethanol should be treated after being reused two times, so that its ultraviolet absorbance level and impurity type are equivalent to that of fresh ethanol before it can be used again. The method provides reference for the quality control and compliance use of recovered ethanol.

Infantile hemangioma is the most common benign tumors in infancy with predictable disease evolution and duration. At present, the basic understanding of the development, proliferation and regression of infantile hemangioma is not sufficient, an ideal lively human infantile hemangioma model which could provide a standard platform for observation of cellular, molecular basis of infantile hemangioma development and drug mechanism is urgently needed. Up to now, a model that completely mimics the biological behavior of infantile hemangiomas has not been successfully established, but the existing research progress has laid a solid foundation for the pathogenesis of hemangioma and clinical targeted therapy. In this paper, the research progress of cell line and animal model of infantile hemangioma and its clinical value in recent years were summarized, as well as the favorable factors that may be used in the construction of infant hemangioma model in the future, providing the latest reference for the selection of animal model of infantile hemangioma.

Infantile hemangioma is the most common benign tumors in infancy with predictable disease evolution and duration. At present, the basic understanding of the development, proliferation and regression of infantile hemangioma is not sufficient, an ideal lively human infantile hemangioma model which could provide a standard platform for observation of cellular, molecular basis of infantile hemangioma development and drug mechanism is urgently needed. Up to now, a model that completely mimics the biological behavior of infantile hemangiomas has not been successfully established, but the existing research progress has laid a solid foundation for the pathogenesis of hemangioma and clinical targeted therapy. In this paper, the research progress of cell line and animal model of infantile hemangioma and its clinical value in recent years were summarized, as well as the favorable factors that may be used in the construction of infant hemangioma model in the future, providing the latest reference for the selection of animal model of infantile hemangioma.

To explore the role of progesterone in the pathogenesis and development of hemangioma in nude mice.
 Methods: The hemangioma model was established. Progesterone was injected intramuscularly at different doses (0.2, 0.4, and 0.8 mg/d) for one week. Camellia oil (0.4 mL/d) was injected intramuscularly as control. The size of hemangioma was observed dynamically. The subcutaneous implants were harvested on the 14th and 28th days. The expression of vascular endothelial growth factor in the tumor tissues were evaluated using immunohistochemistry and microvessel density (MVD) was counted under the microscope.
 Results: On the 14th day, the expression of vascular endothelial growth factor (P<0.01 and P<0.05, respectively) was higher, the volume of tumors (All P<0.01) and MVD (All P<0.01) were greater in the high-dose progesterone group than those in the control and low-dose progesterone group. On the 28th day, there was no significant difference in the volume of tumors among 4 groups (P>0.05). The expression of vascular endothelial growth factor (P<0.01) was lower, and MVD (All P<0.05) were less in the middle-dose and high-dose progesterone group than those in the control and low-dose progesterone group.
 Conclusion: Progesterone promotes angiogenesis via upregulation of vascular endothelial growth factor expression, promotion of hemangiomas proliferation, suggesting that excessive progesterone supplementation may be one of the initial factors for early hemangioma formation.
Animals ; Cell Line, Tumor ; Hemangioma ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; Progesterone ; Vascular Endothelial Growth Factor A

This article summarizes the pharmacological treatment of infantile hemangioma in soft tissue.We aim to review the literature and summarize the mechanisms,methods,efficiency and adverse reactions of beta blockers,glucocorticoids,bleomycin and imiquimott,and to analyze the rational options.Beta blockers,as oral and external use,have high efficiency and few side effects,and can be used as a first -line medication for infantile hemangiomas.Imiquimod external use and topical glucocorticoid and bleomycin injection have good efficacy and potential adverse reactions.They can be used as an alternative treatment for infantile hemangioma.

Objective:To determine drug dose and usage of timolol maleate eye drops in the treatment of superficial infantile hemangioma.Methods:A total of 250 superficial hemangioma infants were recruited and assigned into 5 groups (n=50 for each group):an external application group and 4 exterior coating groups (2,4,6,8 times per day).We evaluated the therapeutic effect of different methods for drug application (external application or exterior coating) and the frequency for drug administration on superficial infantile hemangioma.Results:The external application group (twice a day and 0.5 hour per time) showed better effect than that in the exterior coating group with twice a day (P＜0.001).The difference in therapeutic effect between the exterior coating group with 6 times a day and exterior coating group with twice a day or with 3 times a day was significant (P＜0.001).The differences in drug efficacy were not found among the exterior coating group with 6 times a day,the exterior coating group with 8 times a day,or the external application group with twice a day (All P＞0.05).Conclusion:Drug dose may affect the therapeutic effect oftimolol maleate eye drops in superficial hemangioma infants,and exterior coating with 6 times a day may achieve the best curative effect.

Skin sensitization (allergic contact dermatitis, ACD), is a serious condition caused by small reactive molecules and is characterized by a delayed-type hypersensitivity .Animal tests were usually used in the evaluation of sensitizing potential of chemical substances in the past .However , there is an increasing interest from the public for reducing and ultimately replacing animal tests .The European Union (EU) has posed a ban on animal testing of cosmetic ingredients that includes skin sensitization since 2013.Therefore, alternative in vitro tests are the main tendency in chemical substances and cosmetic sensitizing potential research in the future .In this study, different kinds of in vitro test methods that were adopted by OECD or on research (LLNA, DPRA, KeratinoSens TM, h-CLAT) were reviewed through recent years literature , comprehensive introduction and evaluation were made to obtain reliable hazard and potency information on potential skin sensitizers .