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 explore the value of coronary fat attenuation index(FAI)combined with laboratory indicators in predicting the risk of acute coronary syndrome(ACS)in patients with coronary heart disease(CHD).Methods A retrospective analysis was conducted on 454 patients who were diagnosed with CHD in Guangdong Provincial People's Hospital SCAD group(n=233)and an ACS group(n=221).Univariate and multivariate Logistic regression analyses were performed on the FAI values of the main coronary branches[right coronary artery(RCA),left anterior descending branch(LAD),left circumflex branch(LCX)],laboratory indicators,and clinical data,to identify independent risk factors for ACS in CHD patients.Receiver operating characteristic curves were constructed,and area under the curve(AUC)was calculated to evaluate the predictive performance of the independent risk factors and their combinations.Results LAD-FAI,RCA-FAI,and high-sensitivity C-reactive protein(hs-CRP)were independent influencing factors for ACS in CHD patients.The AUC for the prediction of ACS occurrence in CHD patients based on LAD-FAI,RCA-FAI,and elevated hs-CRP values alone were 0.568,0.703,and 0.749,respectively.When these three factors were analyzed in combination,the AUC was 0.815.Conclusion The combined analysis of LAD-FAI,RCA-FAI,and hs-CRP has good predictive performance for assessing the risk of ACS in CHD patients.

Objective To evaluate the potential risk of transmission of angiostrongyliasis by common freshwater snails in Dali Bai Autonomous Prefecture, Yunnan Province, so as to provide insights into local surveillance of angiostrongyliasis. Methods Common freshwater snails were collected from Dali Bai Autonomous Prefecture, Yunnan Province from March to April, 2020, and identified and bred in laboratory. SD rats were infected with third-stage larvae of Angiostrongylus cantonensis that were isolated from commercially available Pomacea canaliculata snails in Dali Bai Autonomous Prefecture, and freshwater snails were infected with the first-stage larvae of A. cantonensis that were isolated from the feces of SD rats 39 days post-infection at room temperature. The developmental process and morphological characteristics of worms in hosts were observed, and the percentages of A. cantonensis infections in different species of freshwater snails were calculated. Then, SD rats were infected with the third-stage larvae of A. cantonensis that were isolated from A. cantonensis-infected freshwater snails, and the larval development and reproduction was observed. Results More than 3 000 freshwater snail samples were collected from farmlands, ditches and wetlands around Erhai Lake in Dali Bai Autonomous Prefecture, and Cipangopaludina chinensis, P. canaliculata, Parafossarulus striatulus, Oncomelania hupensis robertsoni, Galba pervia, Physa acuta, Radix swinhoei, Assiminea spp., Tricula spp. and Bellamya spp. were morphologically identified. A total of 105 commercially available P. canaliculata snails were tested for A. cantonensis infections, and 2 P. canaliculata snails were found to be infected with A. cantonensis, in which the third-stage larvae of A. cantonensis were isolated. Ten species of freshwater snails were artificially infected with the third-stage larvae of A. cantonensis, and all 10 species of freshwater snails were found to be infected with A. cantonensis, with the highest positive rate of A. cantonensis infections in Bellamya spp. (62.3%, 137/204), and the lowest in C. chinensis (35.5%, 11/31). After SD rats were infected with the third-stage larvae of A. cantonensis isolated from different species of freshwater snails, mature adult worms of A. cantonensis were yielded. Conclusions Multiple species of freshwater snails may serve as intermediate hosts of A. cantonensis under laboratory conditions in Dali Bai Autonomous Prefecture of Yunnan Province. Further investigations on natural infection of A. cantonensis in wild snails in Dali Bai Autonomous Prefecture seem justified.

ObjectiveTo establish an analytical method for the determination of resorcinol, ferulic acid, phenethylresorcinol and benzoyl peroxide in freckle whitening cosmetics by high performance liquid chromatography (HPLC), to provide data support for the establishment of cosmetics inspection methods and technical support for the supervision of the cosmetics industry. MethodsThe analytes in samples were extracted by ultrasonic acetonitrile after methanol vortex, and then filtered by centrifugation and microporous filter membrane. Finally, the analytes were separated with a SVEA C4 column (250 mm×4.6 mm, 5 μm). The mobile phase was composed of 0.1% phosphoric acid solution -acetonitrile, and the gradient elution was applied, with a flow rate of 1.0 mL·min^-1. The samples were detected by an ultra-violet detector and quantified by external standard method. ResultsResorcinol, ferulic acid, phenylethylresorcinol and benzoyl peroxide showed good linearity in the experimental range with r>0.999. HPLC was used to investigate the positive spiked recoveries of ferulic acid or phenylethylresorcinol with different matrices. The results showed that the recoveries were all in the range of 87.48% to 101.00%, and the relative standard deviations were all in the range of 3.4% to 4.1%. Furthermore, HPLC also examined the blank matrix spiked with the recoveries ranged from 93.26% to 107.66%, with the relative standard deviation of 0.90% to 2.90%. The limits of detection ranged from 0.000 8% to 0.002%. Among the30 batches of standard freckle whitening cosmetics determined, 6 batches of which were detected with phenethylresorcinol and 1 batch with ferulic acid. ConclusionHPLC is a method with rapidity, simplicity, and high sensitivity, which is suitable for the simultaneous determination of resorcinol, ferulic acid, phenethylresorcinol and benzoyl peroxide in commercially available cosmetics.

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.

【Objective】 To evaluate the safety and efficacy of balloon-expandable vertebral body stenting （VBS） in treating elderly patients with osteoporotic vertebral compression fractures. 【Methods】 Thirty-eight elderly patients with osteoporotic vertebral compression fractures were randomly divided into two groups according to the parity of their hospital admission numbers to receive VBS and traditional percutaneous kyphoplasty （PKP）， respectively. The two groups were compared regarding changes in the intensity of pain， functional score， imaging parameters related to vertebral compression， and sagittal Cobb angle as well as the incidence of bone cement leakage after treatment， to evaluate the safety and efficacy of VBS. 【Results】 All the patients underwent the operations smoothly， which were performed with the bilateral percutaneous puncture technique under local anesthesia by the same surgeon， and were followed up for more than half a year. Both VBS group and PKP group showed significant improvement in the visual analogue scale score， the Oswestry disability index， the height of the fractured vertebral body， and the Cobb angle at 3 days， 1 month， 3 months and 6 months after treatment （all P<0.05）. There were significant differences in those indicators between the two groups （all P<0.05）. 【Conclusion】 VBS is an effective treatment approach for elderly patients with osteoporotic vertebral compression fractures. It can effectively recover fractured vertebral body height， relieve patients’ pain， and have fewer complications such as bone cement leakage.

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Objective:To understand the situation of death reports of HIV/AIDS patients in China from 2011 to 2019, including the situation of reporting delay and the risk factors of reporting delay, and to provide scientific basis for formulating relevant policies.Methods:The Joinpoint regression model was used to describe the changing trend of the proportion of HIV/AIDS patients who were reported within 6 months after death, and the logistic regression model was used to analyze the risk factors of delayed-reporting of HIV/AIDS deaths.Results:From 2011 to 2019, 241, 019 cases of HIV/AIDS patients were reported dead in China, and 237, 282 cases were included in the study. The proportion of reporting within 6 months after death declined during 2011-2018 (APC=-0.57%, p=0.047). Logistic regression model showed that ethnic minorities ( OR=1.34, 95% CI: 1.29-1.39), domestic workers and unemployed workers ( OR=1.52, 95% CI: 1.48~1.61), high school education or above ( OR=1.15, 95% CI: 1.09~1.20), patients who were over 45 years old when they were diagnosed ( OR=1.16, 95% CI: 1.12~1.21), duration of diagnosis was more than six months ( OR=1.63, 95% CI: 1.57~1.70), infection route was heterosexual transmission ( OR=1.21, 95% CI: 1.13~1.30) or injection drug ( OR=1.26, 95% CI: 1.17~1.36) and the inconsistency between the permanent residence registration place and the reporting place ( OR=1.08, 95% CI: 1.04~1.13) are more prone to delayed-reporting, in the cases of divorced or widowed ( OR=0.87, 95% CI: 0.84~0.91) patients, those who were receiving national free antiviral treatment ( OR=0.78, 95% CI: 0.76~0.81), and the death place was located in medical institutions ( OR=0.90, 95% CI: 0.86~0.94) and no loss to follow-up at the last follow-up before death ( OR=0.25, 95% CI: 0.24~0.26) deaths were more likely to be reported in time. Conclusions:From 2011 to 2019, 91.63% of the dead HIV/AIDS patients in China can be reported within 6 months, and the proportion of the dead HIV/AIDS patients reported within 6 months from 2011 to 2018 shows a slightly downward trend. Ethnic minorities, those who are engaged in domestic work and unemployed, high school or above education, patients who were over 45 years old when they were diagnosed, duration of diagnosis was more than six months, infection route was heterosexual transmission or injection drug and the inconsistency between the permanent residence registration place and the reporting place are the risk factors for delayed-reporting, while divorce or widowhood, receiving antiviral treatment, dying in medical institutions, no loss to follow-up at the last follow-up visit before death are the protective factors for delayed-reporting.