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*

To summarize Professor WANG Xixing's clinical experience in treating advanced breast cancer with anxiety and depression from the perspective of shaoyang pivot. It is believed that the core pathogenesis of advanced breast cancer with anxiety and depression lies in the dysfunction of shaoyang pivot （referring to the imbalanced regulatory function of the shaoyang meridian system that governs the transportation and transformation of qi， blood， and body fluids）. This dysfunction can lead to abnormal circulation of qi， blood， and body fluids， as well as the intermingling of phlegm and blood stasis， which further promotes the spread and diffusion of cancer toxin. Meanwhile， it disturbs mental activity， resulting in a condition characterized by stagnation of cancer toxin and concurrent disorders of both the physical body and the spirit. Based on this pathogenesis， the basic therapeutic principles of harmonizing shaoyang， regulating the pivot to calm the spirit， and dissipating masses and resolving toxins are proposed. Clinically， the disease is classified into three syndromes for differentiation and treatment. For shaoyang pivot dysfunction syndrome， treatment should use self-prescribed Chaiqin Hengshu Ningxin Decoction （柴芩衡枢宁神汤）； for sanjiao pivot dysfunction syndrome， treatment should prescribe Chaigui Tongshu Dashen Decoction （柴归通枢达神饮）； for gallbladder function disorder syndrome， treatment should apply Wendan Qishu Shoushen Decoction （温胆启枢守神汤）. Throughout the treatment process， the concept of "simultaneous treatment of cancer and depression" is implemented to smooth the shaoyang pivot， block the vicious cycle where cancer toxin and emotional abnormalities mutually reinforce each other.

Objective To evaluate the protective effect and underlying mechanism of ulinastatin on hepatic ischemia-reperfusion injury.Methods Twenty-four male SD rats were divided into three groups:sham operation group(Sham group),hepatic ischemia-reperfusion injury group(HIRI group)and hepatic ischemia-reperfusion injury+ulinastatin pretreatment group(HIRI+UTI group),with 8 rats in each group.The HIRI rat models were established by occluding hepatic portal vein and hepatic artery for 1 h.In the HIRI+UTI group,the rats were intraperitoneally injected with ulinastatin at 30 min before model establishment,and an equivalent amount of normal saline was given in the Sham and HIRI groups.The rats were sacrificed at 6 h after model establishment.Serum samples were collected to detect alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels.Pathological changes of liver tissues were observed by hematoxylin-eosin(HE)staining.Ultrastructural changes of mitochondria in liver tissues were observed by transmission electron microscopy.The expression of glutathione peroxidase 4(GPX4)was determined by immunofluorescent staining.The contents of malondialdehyde(MDA),glutathione(GSH),Fe,reactive oxygen species(ROS)and GPX4 were detected.The expression levels of GPX4 and acyl-CoA synthetase long-chain family 4(ACSL4)messenger RNA(mRNA)and proteins in liver tissue were measured.Results Compared with the Sham group,serum ALT and AST levels were up-regulated,pathological changes such as congestion,hepatocyte necrosis and abnormal hepatic lobule structure were observed,pathological score was increased,the mitochondria shrank,the membrane density was increased,the mitochondrial crest was damaged or even absent,the contents of ROS,MDA and Fe were elevated,the GSH content was decreased,the fluorescent intensity of GPX4 was weakened,the relative expression levels of ACSL4 mRNA and protein were up-regulated,and the relative expression levels of GPX4 mRNA and protein were down-regulated in the HIRI group(all P＜0.05).Compared with the HIRI group,serum ALT and AST levels were down-regulated,liver tissue injury was alleviated,pathological score was decreased,mitochondrial shrinkage and crest breakage were mitigated,the contents of ROS,MDA and Fe were down-regulated,the GSH content was up-regulated,the fluorescent intensity of GPX4 was enhanced,the relative expression levels of ACSL4 mRNA and protein were down-regulated,and the relative expression levels of GPX4 mRNA and protein were up-regulated in the HIRI+UTI group(all P＜0.05).Conclusions Ulinastatin may alleviate hepatic ischemia-reperfusion injury in rats probably through inhibiting ferroptosis.

Objective:To explore the application effect of early nursing intervention based on NNN-link in social disorders of children with autism spectrum disorders (ASD) .Methods:A total of 126 hospitalized children with ASD in the Rehabilitation Center of Henan Children's Hospital from January to May 2023 were selected as research subjects by convenience sampling. They were randomly divided into an intervention group ( n=63) and a control group ( n=63) using a random number table. The control group received routine care, while the intervention group received early nursing intervention based on NNN-link in addition to routine care. The Social Disorder-related Nursing Diagnosis Outcome Scales and the Childhood Autism Rating Scale (CARS) scores were compared between the two groups before and after intervention. Results:After the early nursing intervention, the scores for family atmosphere, participation in leisure activities, participation in play, social skills, and social participation were higher in the intervention group compared to the control group, the differences were statistically significant ( P<0.05). The CARS scores of the intervention group were also lower than those of the control group, the differences were statistically significant ( P<0.05) . Conclusions:Early nursing intervention based on NNN-link can effectively improve the social disorder conditions of children with ASD, reduce their autism symptoms, and promote recovery.

Objective To investigate the effect of sigma-1 receptor(sig-1R)inhibition on apoptosis in the dorsal root ganglion(DRG)of rats with sciatic nerve chronic constriction injury(CCI)-mediated neuropathic pain.Methods Rats undergoing intrathecal intubation were randomly divided into three groups with 12 in each:sham group,model group(CCI modeling one week after intrathecal intubation)and sig-1R inhibitor group(BD1047 group)that was injected intrathecally on the fourth,fifth,and sixth days after the CCI operation.The mechanical withdrawal threshold(MWT)of rats was detected on the day before surgery and then first,third,fifth and sev-enth days after surgery.The expression of sig-1R,Bcl-2 and Bax was detected by Western blot and immunofluo-rescence in DRG cell;TUNEL staining microscopy was used to observe the apoptosis of DRG cells.The changes of endoplasmic reticulum and mitochondria were observed by transmission electron microscopy of DRG cells.Results Compared with the sham group,the model group showed a decreased MWT at all time points after surgery,up-regulation of DRG cell apoptosis,up-regulation of sig-1R and Bax,down-regulation of Bcl-2,swelling of en-doplasmic reticulum,disruption of mitochondrial membrane and reduction of mitochondrial cristae in the DRG cell endoplasmic reticulum after surgery(P＜0.05).BD1047 group showed elevated MWT at the fifth and seventh postoperative days,down-regulation of DRG cell apoptosis,down-regulated expression of sig-1R and Box,up-regulated expression of Bcl-2 and slightly damaged endoplasmic reticulum as well as mitochondria of DRG cells compared with model group(P＜0.05).Conclusions Inhibition of sig-1R up-regulates mechanical withdrawal threshold and reduces DRG cell apoptosis in CCI rats.

Objective:To assess the feasibility and efficacy of hypofractionated radiotherapy with simultaneous integrated boost (HFRT-SIB) for patients with non-osteogenic spinal metastases who were not suitable for stereotactic body radiotherapy (SBRT).Methods:A totoal of 26 spinal metastatic lesions in 24 patients with non-osteogenic spinal metastases admitted to Beijing Luhe Hospital affiliated to Capital Medical University from March 2020 to April 2023 were retrospectively analyzed. In our institution, a higher-dose fractionated regimen for spinal metastases was introduced, at a dose of 3 Gy/f ×10 times (5 times a week, once a day) to the clinical target volume (CTV), and simultaneous integrated boost (SIB) of 4 Gy/f ×10 times (5 times a week, once a day) to the gross tumor volume (GTV). The primary endpoint was the overall relief (OR) rates of pain at 12 and 24 weeks post-radiotherapy, while secondary endpoints included the pain response time and duration of pain relief, the incidence of adverse events post-radiotherapy, and the 1-year overall survival (OS) rate. The differences before and after treatment were analyzed by Wilcoxon signed-rank test. OS was assessed using Kaplan-Meier method.Results:The median follow-up was 11.7 (8.0, 17.0) months. At 12 weeks post-radiotherapy, 92% (22/24) of patients achieved OR, and 81% (17/21) obtained OR at 24 weeks post-radiotherapy. The median pain response time was 4 (3, 5) d. At the last follow-up, 42% (10/24) of patients maintained pain relief for over 1 year, with a median duration of pain relief up to 11.0 (6.0, 16.9) months. The 1-year OS rate was 66.3%, and no patients experienced grade 3 or higher acute or chronic adverse events.Conclusion:HFRT-SIB can effectively alleviate pain, and achieve sustained pain control, and does not lead to severe adverse effects in patients with non-osteogenic spinal metastases.

Humans ; Parkinson Disease/diagnosis* ; Machine Learning

Parkinson's disease (PD) is a common neurodegenerative disease in middle-aged and elderly people. In particular, increasing evidence has showed that astrocyte-mediated neuroinflammation is involved in the pathogenesis of PD. As a precious traditional Chinese medicine, bear bile powder (BBP) has a long history of use in clinical practice. It has numerous activities, such as clearing heat, calming the liver wind and anti-inflammation, and also exhibits good therapeutic effect on convulsive epilepsy. However, whether BBP can prevent the development of PD has not been elucidated. Hence, this study was designed to explore the effect and mechanism of BBP on suppressing astrocyte-mediated neuroinflammation in a mouse model of PD. PD-like behavior was induced in the mice by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) (30 mg·kg^-1) for five days, followed by BBP (50, 100, and 200 mg·kg^-1) treatment daily for ten days. LPS stimulated rat C6 astrocytic cells were used as a cell model of neuroinflammation. THe results indicated that BBP treatment significantly ameliorated dyskinesia, increased the levels of tyrosine hydroxylase (TH) and inhibited astrocyte hyperactivation in the substantia nigra (SN) of PD mice. Furthermore, BBP decreased the protein levels of glial fibrillary acidic protein (GFAP), cyclooxygenase 2 (COX2) and inducible nitric oxide synthase (iNOS), and up-regulated the protein levels of takeda G protein-coupled receptor 5 (TGR5) in the SN. Moreover, BBP significantly activated TGR5 in a dose-dependent manner, and decreased the protein levels of GFAP, iNOS and COX2, as well as the mRNA levels of GFAP, iNOS, COX2, interleukin (IL) -1β, IL-6 and tumor necrosis factor-α (TNF-α) in LPS-stimulated C6 cells. Notably, BBP suppressed the phosphorylation of protein kinase B (AKT), inhibitor of NF-κB (IκBα) and nuclear factor-κB (NF-κB) proteins in vivo and in vitro. We also observed that TGR5 inhibitor triamterene attenuated the anti-neuroinflammatory effect of BBP on LPS-stimulated C6 cells. Taken together, BBP alleviates the progression of PD mice by suppressing astrocyte-mediated inflammation via TGR5.
Humans ; Mice ; Rats ; Animals ; Aged ; Middle Aged ; Parkinson Disease/pathology* ; Astrocytes/pathology* ; Powders/therapeutic use* ; Ursidae/metabolism* ; NF-kappa B/metabolism* ; Neuroinflammatory Diseases ; Neurodegenerative Diseases/metabolism* ; Cyclooxygenase 2/metabolism* ; Lipopolysaccharides/pharmacology* ; Bile ; Mice, Inbred C57BL ; Microglia ; Disease Models, Animal

Cartilage has poor self-recovery because of its characteristics of no blood vessels and high extracellular matrix. In clinical treatment, physical therapy or drug therapy is usually used for mild cartilage defects, and surgical treatment is needed for severe ones. In recent years, cartilage tissue engineering technology provides a new way for the treatment of cartilage defects. Compared with the traditional surgical treatment, cartilage tissue engineering technology has the advantages of small wound and good recovery. The application of microcarrier technology in the design of tissue engineering scaffolds further expands the function of scaffolds and promotes cartilage regeneration. This review summarized the main preparation methods and development of microcarrier technology in recent years. Subsequently, the properties and specific application scenarios of microcarriers with different materials and functions were introduced according to the materials and functions of microcarriers used in cartilage repair. Based on our research on osteochondral integrated layered scaffolds, we proposed an idea of optimizing the performance of layered scaffolds through microcarriers, which is expected to prepare bionic scaffolds that are more suitable for the structural characteristics of natural cartilage.
Cartilage ; Extracellular Matrix/chemistry* ; Technology ; Tissue Engineering/methods* ; Tissue Scaffolds/chemistry*

Objective? To explore the effects of IDEAS visits model on anxiety in ear, nose and throat (ENT) patients before day surgery. Methods? Totally 200 ENT patients before day surgery were selected using convenient sampling from July 2017 to June 2018. 5 cases fell off before the end of study. Totally 97 patients admitted between July and December 2017 were finally included into the control group, while 98 patients admitted between January and June 2018 were finally included into the intervention group. Patients in the control group received conventional visits before surgery, whereas patients in the intervention group received visits according to IDEAS model. Blood pressure, pulse rate and Visual Analogue Score (VAS) on pain were compared between the two groups before and after intervention. Amsterdam Preoperative Anxiety and Information Scale (APAIS) was used to evaluate the patients' anxiety. Results? There was no statistically significant difference in blood pressure and pulse rate between the two groups before intervention (P＞ 0.05); blood pressure and pulse rate were lower in the intervention group than in the control group, and the VAS of the intervention group was lower than that of the control group after intervention (P＜ 0.05). There was no statistically significant difference in APAIS scores between the two groups before intervention (P＞0.05); while the APAIS score of the intervention group was lower than that of the control group (P＜0.05). Conclusions? IDEAS visits model can reduce the preoperative anxiety in ENT patients before day surgery.