Human naïve pluripotent stem cells (PSCs) hold great promise for embryonic development studies. Existing induction and culture strategies for these cells, heavily dependent on MEK inhibitors, lead to widespread DNA hypomethylation, aberrant imprinting loss, and genomic instability during extended culture. Here, employing high-content analysis alongside a bifluorescence reporter system indicative of human naïve pluripotency, we screened over 1,600 chemicals and identified seven promising candidates. From these, we developed four optimized media-LAY, LADY, LUDY, and LKPY-that effectively induce and sustain PSCs in the naïve state. Notably, cells reset or cultured in these media, especially in the LAY system, demonstrate improved genome-wide DNA methylation status closely resembling that of pre-implantation counterparts, with partially restored imprinting and significantly enhanced genomic stability. Overall, our study contributes advancements to naïve pluripotency induction and long-term maintenance, providing insights for further applications of naïve PSCs.
Humans ; DNA Methylation/drug effects* ; Genomic Instability ; Pluripotent Stem Cells/metabolism* ; Cell Culture Techniques/methods* ; Cells, Cultured

AIM To study the chemical constituents from the leaves of Citrus reticulata Blanco and their anti-inflammatory activities.METHODS The 85%ethanol extract from the leaves of C.reticulata was isolated and purified by silica gel,D101 macroporous resin,MCI,ODS and Sephadex LH-20,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The Griess method was used to determine their inhibitory activities on lipopolysaccharide-induced NO production in macrophages RAW 264.7 cells.The mice foot swelling inflammation model induced by carrageenan was established,and the levels of IL-1β,TNF-α were detected.RESULTS Twelve compounds were isolated and identified as nobiletin(1),tangeretin(2),5-demethylinoblitin(3),5,4'-dihydroxy-6,7,8,3'-tetramethoxy flavone(4),5-hydroxy-7,8,3',4'-trimethoxyflavanone(5),3,5,6,7,8,3',4'-heptamethoxyflavanone(6),hesperetin(7),5-hydroxy-6,7,3',4'-tetramethoxyphenone(8),β-balsam alcohol(9),stigmaster-5-en-3β-alcohol(10),p-hydroxybenzaldehyde(11),vanillin(12).Compounds 1,4,6,7,10 and 12 had strong inhibitory activites on NO release in LPS-induced RAW 264.7 cells,and the IC50 values were(25.21±2.10),(37.77±0.50),(38.19±1.58),(21.89±1.73),(43.81±1.18),(47.98±2.55),(41.23±1.11),(43.80±1.43)μmol/mL,respectively.Compounds 2-3 reduced IL-1β and TNF-α levels(P＜0.05,P＜0.01).CONCLUSION Compounds 6-7,9 are isolated from this plant for the first time.Compounds 1-4,8 exhibit strong in vitro anti-inflammatory activities,and compounds 2-3 exhibit significant in vivo anti-inflammatory activities.

Objective This study aimed to clarify the anti-influenza virus activity of Bingyanqing(BYQ),as well as to explore the mechanism of BYQ in treating influenza through network pharmacology and experimental verification.Methods The impact of BYQ on mortality,lung index,and viral load in an influenza mouse model was detected.We collected the ingredients and targets of BYQ formula by searching databases including TCMSP,Swiss Target Prediction and consulting the literature.The"ingredients-common target"network for anti-influenza effect of BYQ was constructed using Cytoscape software.The protein-protein interaction(PPI)network was constructed using the STRING database and Cytoscape software,and the core targets were screened.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed for common targets.The effect of BYQ on reducing influenza-induced oxidative damage,the expression of antioxidant and pro-oxidant enzyme,P65 phosphorylation and nuclear factor E2-related factor 2(Nrf2)nuclear translocation in vivo were investigated.Results BYQ significantly reduced mortality,lung index,pulmonary viral load and lung injury in a mouse model of influenza.We obtained one hundred and ninety-three of BYQ active compounds,which corresponded to three hundred and thirty-eight targets.There are 180 influenza-related targets among them.Nine targets,including IκBα kinase α(CHUK),IκBα kinase γ(IKBKG),NF-κB p65(RELA),tumor necrosis factor(TNF)and interleukin 6(IL6),were identified as potential core targets.GO analysis indicated that BYQ is involved in several biological functions,including antibacterial and antioxidant stress responses.KEGG analysis revealed the involvement of several viral and immune-related pathways for BYQ in treating influenza,including herpes simplex virus,influenza A virus,TNFα and toll-like receptor pathways.In vivo studies showed that high-dose BYQ significantly reduced pulmonary malondialdehyde(MDA)levels(P＜0.01)and increased total superoxide dismutase(SOD)activity(P＜0.001)in a mouse mode of influenza compared to oseltamivir phosphate.The treatment group with the combination of BYQ&oseltamivir phosphate had lower levels of NADPH oxidase 2(NOX2)and 4(NOX4)(P＜0.001),and higher levels of catalase(CAT)and glutathione peroxidase 4(GPX4)(P＜0.001)in the lungs than oseltamivir phosphate group.The combined treatment group showed more significant Nrf2 nuclear translocation(P＜0.05)than the oseltamivir phosphate group.However,there was no significant difference in P65 phosphorylation levels between the combination treatment group and the oseltamivir phosphate group(P＜0.05),but P65 phosphorylation levels in both groups were lower than in the model group(P＜0.05).Conclusion BYQ exhibits significant anti-influenza virus activity,manifests a dual effect by inhibiting the synthesis of pro-oxidant enzymes and promoting the antioxidant system,thereby alleviates the oxidative stress damage caused by influenza.

Vaginal birth after cesarean (VBAC) can reduce the cesarean section rate and has many benefits for both mother and infant. Decision-making aids can promote communication between doctors and patients regarding childbirth methods and increase pregnant women's informed consent and decision-making participation in VBAC, thereby supporting pregnant women in making choices and improving the quality of childbirth decisions. This article summarizes the content of VBAC decision-making aids, evaluation of different VBAC decision-making aids, application of VBAC decision-making aids, and obstacles to the application of VBAC decision-making aids in China to provide reference for the development and application of domestic childbirth decision-making aids.

Objective:To conduct systematic review of mother-infant attachment measurement tools based on Consensus-based Standards for the Selection of Health Measurement Instruments (COSMIN) guidelines.Methods:The researches on mother-infant attachment measurement tools in PubMed, Scopus, Embase, Web of Science, China Biology Medicine disc, China National Knowledge Infrastructure, WanFang Data and VIP was searched by computer, and the search period was from establishment of the databases to October 30, 2023. Two reviewers trained in evidence-based methodology independently screened the literature, extracted and summarized the data, and systematically evaluated the attributes of the measurement tools using the COSMIN guideline bias risk list and good measurement attribute standards.Results:A total of 35 studies were included, including seven maternal-infant attachment measurement tools. Among them, the content validity quality of the Maternal-fetal Attachment Tool was sufficient (evidence quality was advanced), the structural validity quality was uncertain (evidence quality was intermediate), the internal consistency quality was sufficient (evidence quality was advanced) and the hypothesis testing quality was sufficient (evidence quality was advanced), which was recommended at level A.Conclusions:This study systematically evaluates seven measurement tools for maternal-infant attachment, among which the Maternal-fetal Attachment Tool is class A tool and is recommended for use.

This study investigated the epidemiological and genotypic characteristics of group A rotavirus(RVA)in children under 5 years of age with diarrhea at a viral diarrhea surveillance sentinel hospital in Guangdong Province from 2021 to 2022,to provide a basis for RVA prevention and control.A total of 1 858 fecal samples from children under 5 years of age with diarrhea in 2021-2022 were collected.Rotavirus antigen was detected with ELISA,and positive samples were further sequenced and categorized.Among the 1 858 samples,156 were RVA positive.The positivity rates in boys and girls were 8.76％and 7.87％,respectively.Significant differences were observed in the detection rates of RVA infection among age groups.The rate of RVA infection increased gradually from December to April of the following year.In 2021,the main endemic strains of RVA in the Guangdong region were of the G9P[8]subtype,and the rare G8P[8]subtype increased in China.In 2022,the G8P[8]subtype surpassed the G9P[8]subtype for the first time.In cases of diarrhea in infants younger than 5 years in Guangdong Province from 2021 to 2022,the RVA genotypes were diverse,the G9P[8]genotype significantly decreased,and the G8P[8]subtype became a dominant genotype.Continuous RVA genotype monitoring remains necessary to assess the risk of RVA-related disea-ses.

Objective:To assess the efficacy of non-pharmaceutical intervention on improving sleep quality in pregnant women by network Meta-analysis.Methods:Randomized controlled clinical trials on non-pharmaceutical intervention to improve sleep quality in pregnant women were searched on Cochrane Library, PubMed, Web of Science, Embase, China Biology Medicine disc, CNKI and Wanfang Database by computer, and the search time limit was from the establishment of databases to December 2022. Two researchers independently screened the literature, extracted data and evaluated the risk of bias in the included studies. Stata 14.0 software was used to conduct a network meta-analysis.Results:Finally, a total of 26 randomized controlled trial were included, including 2 363 patients and 7 non-pharmaceutical intervention measures. The best probability ranking results of network meta-analysis showed that in terms of total score evaluation of Pittsburgh Sleep Quality Index, relaxation intervention, traditional Chinese medicine treatment, psychological intervention, exercise intervention, music therapy, sleep health education and routine nursing were ranked from high to low. In terms of dimensional evaluation of Pittsburgh Sleep Quality Index, relaxation intervention, traditional Chinese medicine treatment, psychological intervention and music therapy had better effects.Conclusions:Current evidence suggests that relaxation intervention, traditional Chinese medicine treatment, psychological intervention and music therapy may be more effective interventions in improving the sleep quality of pregnant women.

Objective:To explore the advantage of Fuzheng Huayu capsule in patients with hepatitis B cirrhosis based on neutrophil/lymphocyte ratio (NLR) risk stratification in reducing the incidence of hepatocellular carcinoma (HCC).Methods:916 cases diagnosed with hepatitis B cirrhosis and followed up for five years from January 2011 to January 2016 at Beijing Ditan Hospital Affiliated with Capital Medical University were included, and clinical data were collected. Patients were divided into a combination group and an antiviral group according to whether they were treated with anti-fibrosis for≥6 months. The antiviral group was treated with entecavir or tenofovir disoproxil, while the combination group was treated with Fuzheng Huayu capsules based on the antiviral therapy. The incidence of HCC was compared between the two groups of patients within five years. The advantaged groups treated with Fuzheng Huayu capsule were explored based on NLR risk stratification. The independent sample t-test and Mann-Whitney U test were used to compare measurement data between two groups. Categorical variable data were compared using either the χ2 test or Fisher's exact probability method. The incidence of HCC in the two groups of patients was analyzed through the Kalplan-Merier curve and compared using the log-rank method. Results:There were 299 (32.6%) and 617 (67.4%) cases in the combined group and the antiviral group, respectively. A total of 154 (16.8%) patients developed HCC during the follow-up period. The five-year cumulative incidence of HCC in the combination group was lower than that in the antiviral group (10.7% vs. 19.8%, χ2 = 11.848, P = 0.000 4). Patients with baseline NLR>3 had an increased risk of HCC. According to NLR risk stratification, there were 191 cases in the low-risk group (NLR<1.4), 462 cases in the medium-risk group (NLR1.4 ~ 3.0), and 263 cases in the high-risk group (NLR>3). Among medium to high-risk patients, the incidence of HCC was significantly reduced in the combination group (11.5% vs. 19.4%, χ2 = 4.519, P = 0.029; 13.2% vs. 26.2%, χ2 = 5.258, P = 0.019), while there was no statistically significant difference in the incidence of HCC among the low-risk group ( P = 0.38). Conclusion:Compared with antiviral treatment alone, Fuzheng Huayu capsules combined with antiviral treatment can better reduce the five-year HCC incidence rate in patients with hepatitis B cirrhosis. Medium-and high-risk patients with NLR stratification are the most advantageous population to be treated with Fuzheng Huayu capsules.

The "student-centered" flipped classroom teaching model can improve students' academic achievement, improve cognition, and cultivate innovation ability. However, it has obvious deficiencies in the integrity and systematization of knowledge as well as education. The traditional teaching concept based on " teacher-centered" has the unique advantages of systematization of knowledge learning and education. Therefore, we integrated the merits of these two different teaching models and introduced the semi-flipped classroom teaching model into the Biochemistry teaching of 2020 stomatology, pharmacy and preventive medicine majors in Cheeloo Medical College, Shandong University, compared with the traditional teaching of 2019 same majors. The data on the improvement of students ' academic achievement and self-cognition were analyzed. The results showed that the students' achievements of the semi-flipped classroom teaching model were better than those of the traditional teaching (P<0. 01). The students' self-cognitions were significantly improved after the implementation of semi-flipped classroom teaching (P<0. 01 or P<0. 05). This study provides a reference for the related teaching and research work in medical colleges.