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*

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.

Self-organized blastoids from extended pluripotent stem (EPS) cells possess enormous potential for investigating postimplantation embryo development and related diseases. However, the limited ability of postimplantation development of EPS-blastoids hinders its further application. In this study, single-cell transcriptomic analysis indicated that the "trophectoderm (TE)-like structure" of EPS-blastoids was primarily composed of primitive endoderm (PrE)-related cells instead of TE-related cells. We further identified PrE-like cells in EPS cell culture that contribute to the blastoid formation with TE-like structure. Inhibition of PrE cell differentiation by inhibiting MEK signaling or knockout of Gata6 in EPS cells markedly suppressed EPS-blastoid formation. Furthermore, we demonstrated that blastocyst-like structures reconstituted by combining the EPS-derived bilineage embryo-like structure (BLES) with either tetraploid embryos or tetraploid TE cells could implant normally and develop into live fetuses. In summary, our study reveals that TE improvement is critical for constructing a functional embryo using stem cells in vitro.
Pregnancy ; Female ; Animals ; Mice ; Tetraploidy ; Blastocyst ; Embryo, Mammalian ; Cell Differentiation ; Embryonic Development

Chemically defined medium is widely used for culturing mouse embryonic stem cells (mESCs), in which N2B27 works as a substitution for serum, and GSK3β and MEK inhibitors (2i) help to promote ground-state pluripotency. However, recent studies suggested that MEKi might cause irreversible defects that compromise the developmental potential of mESCs. Here, we demonstrated the deficient bone morphogenetic protein (BMP) signal in the chemically defined condition is one of the main causes for the impaired pluripotency. Mechanistically, activating the BMP signal pathway by BMP4 could safeguard the chromosomal integrity and proliferation capacity of mESCs through regulating downstream targets Ube2s and Chmp4b. More importantly, BMP4 promotes a distinct in vivo developmental potential and a long-term pluripotency preservation. Besides, the pluripotent improvements driven by BMP4 are superior to those by attenuating MEK suppression. Taken together, our study shows appropriate activation of BMP signal is essential for regulating functional pluripotency and reveals that BMP4 should be applied in the serum-free culture system.
Animals ; Bone Morphogenetic Protein 4/metabolism* ; Cell Differentiation ; Chromosomal Instability ; Endosomal Sorting Complexes Required for Transport ; Mice ; Mitogen-Activated Protein Kinase Kinases/metabolism* ; Mouse Embryonic Stem Cells/cytology* ; Pluripotent Stem Cells/cytology* ; Signal Transduction ; Ubiquitin-Conjugating Enzymes

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Objective:To explore the effect of nurse-led management model in adherence of patients with obstructive sleep apnea hypopnea syndrome (OSAHS) who underwent non-invasive mechanical ventilation.Methods:92 patients with severe OSAHS diagnosed by polysomnography(PSG) and treated with non-invasive mechanical ventilation in Tianjin medical university general hospital from September 2019 to August 2020 were enrolled. The patients were divided into intervention group (45 cases) and control group (47 cases) by random number table. Then, basing on routine treatment and subsequent visit, the patients in intervention group received regular telephone follow-up interviews within one week of ventilator purchasing, and the interview was based on a standard telephone follow-up manuscript designed by Duffy and lasted for six months. The patients in control group received routine nursing treatment and regular subsequent visit, and were not followed up by telephone, but were encouraged to have telephone consultation. The Epworth sleepiness scale(ESS) scores of the two groups before and after the intervention and the compliance of non-invasive mechanical ventilation of the two groups after the intervention were compared.Results:After the intervention, the ESS scores of the two groups were lower than those before the intervention ( P<0.001), and the ESS scores of the intervention group were lower than those of the control group ( P<0.001). After the intervention, the compliance of noninvasive mechanical ventilation in the intervention group was better than that in the control group ( P<0.001). Conclusions:The nurse-led management model in combination with telephone follow-up can improve the sleepiness of patients and the adherence of using non-invasive mechanical.

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.

Objective:To explore the influence of multidimensional integration and 360° support on the function of family and marriage, and quality of life in women patients with systemic lupus erythematosus.Methods:Totally 196 patients with systemic lupus erythematosus from Department of Rheumatology and Immunology, The Second Affiliated Hospital of the Air Force Medical University from August 2016 to November 2017 were included. According to random number table method, these patients were divided into observation group and control group as 98 cases each. Conventional care and hospital discharge were used for control group. On the basis of this, multidimensional integration and 360° support were used for patients of observation group. The function of family and marriage, quality of life in patients were assessed before and after 3 months of the intervention. The treatment adherence was evaluated in 3 months and 6 months after intervention.Results:Before intervention, the marriage family function score, marital satisfaction, conflict resolution methods and the relationship with friends and family, husband and wife exchange scores of the observation group were (2.3 ± 0.5), (24.6 ± 6.1), (25.7 ± 7.1), (28.2 ± 6.9), (28.8 ± 6.9) points, respectively. Three months after intervention, these scores were (2.5 ± 0.7), (31.6 ± 5.0), (31.7 ± 5.3), (28.1 ± 6.8), (29.0 ± 7.1) points, respectively. There was statistically significant difference between before and after the intervention ( t values were -2.371 - 8.631, P < 0.01). These scores of control group before the intervention were (2.3 ± 0.6), (24.5 ± 6.2), (25.2 ± 7.2), (32.5 ± 6.0), (33.9 ± 6.3) points, respectively. Three months after intervention, these scores were (2.3 ± 0.4), (24.5 ± 6.2), (26.1 ± 6.9), (29.1± 4.8), (28.5 ± 7.2) points. Significant differeces were found between before and after the intervention in control group ( t values were -3.878-6.323, P < 0.05 or 0.01). There was statistically significant difference between the two groups after the intervention ( t values were 2.675-8.631, P<0.01). As for observation group, planning (62.8 ± 27.2 vs. 75.5 ± 25.4) and intimate relationship (62.8 ± 25.2 vs. 78.2± 24.9) in quality of life were obviously difference before and after 3 months of intervention ( t values were 3.050, 3.639, P < 0.01). As for control group, planning (62.5 ± 27.6 vs. 65.7 ± 24.9) and intimate relationship (65.8 ±25.2 vs. 63.5 ± 23.8) in quality of life were obviously difference before and after 3 months of intervention ( t values were 2.375, 3.132, P < 0.01). There was statistically significant difference between the two groups after the intervention ( t values were 3.050, 3.639, P < 0.01). The treatment adherence of observation group was significant better than control group. After 6 months intervention, the treatment adherece of observation group was 83.67% (82/98), while the treatment adherece of control group was 44.89% (44/98), significant differences were found btween the two groups ( χ2 value was 0.511, P < 0.01). Conclusion:Multidimensional integration and 360° support obviously improved function of family and marriage, improved the understanding of disease, and self-management ability of patients. Therefore, it can increase the treatment adherence and improve quality of life in SLE patients.

Breast cancer is a malignant tumor originating from breast epithelial tissue. Ferroptosis is a novel type of programmed cell death which differs from apoptosis and necrosis. Research found that the accumulation of lipid peroxides in cells, a crucial process of ferroptosis, can be induced by multiple mechanisms. The ferroptosis regulation is closely related to the occurrence and development of breast cancer, and it induced by drugs is a potential and valuable research direction in breast cancer.