ObjectiveTo explore the mechanism by which Sini San （SNS） inhibits ferroptosis， alleviates inflammation and myocardial injury， and improves myocardial infarction （MI）. MethodsThe active ingredients of SNS were obtained by searching the Traditional Chinese Medicine System Pharmacology Platform （TCMSP） database， its target sites were predicted using the SwissTargetPrediction Database， and the core components were screened out using the CytoNCA plug-in. The targets of MI and ferroptosis were obtained by using GeneCards， Online Mendelian Inheritance in Man （OMIM） database， DrugBank， Therapeutic Target Database （TTD）， FerrDb database and literature review， respectively. The intersection of these targets of SNS-MI-ferroptosis was plotted as a Venn diagram. The protein-protein interaction （PPI） network was constructed using the STRING database， and the visualization graph was prepared using Cytoscape. The core targets were screened out using the CytoNCA plug-in， and the biological functions were clustered by the MCODE plug-in. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed using the David database. Molecular docking was performed using AutoDock and visualized with PyMOL2.5.2. The Kunming mice were randomly divided into the control group， the model group， the SNS group， and the trimetazidine （TMZ） group. The mice were subcutaneously injected with isoprenaline （ISO， 5 mg·kg^-1·d^-1） to establish an MI model. The drug was continuously intervened for 7 days. The ST-segment changes were recorded by electrocardiogram （ECG）， and the tissue morphology changes were observed by hematoxylin-eosin （HE） staining. Cardiomyocyte ferroptosis was investigated by transmission electron microscopy. Serum creatine kinase （CK）， creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， reduced glutathione （GSH）， and malondialdehyde （MDA） levels were detected by biochemical assay. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of interleukin （IL）-6 and 4-hydroxynonenal （4-HNE）. Immunohistochemical staining was employed to detect IL-6 and phosphorylated signal transducer and transcription activator 3 （p-STAT3） in cardiac tissues. Western blot was used to detect STAT3 and p-STAT3 in cardiac tissues. Real-time PCR was used to detect the levels of IL-6， IL-18， solute carrier family 7 member 11 （SLC7A11）， arachidonic acid 15-lipoxygenase （ALOX15）， and glutathione peroxidase 4 （GPx4） in cardiac tissues. ResultsA total of 121 active ingredients of SNS were obtained， and 58 potential targets of SNS in the treatment of MI by regulating ferroptosis were screened. The three protein modules with a score5 were mainly related to the inflammatory response. The GO function was mainly related to inflammation， and KEGG enrichment analysis showed that SNS mainly regulated ferroptosis- and inflammation- related signaling pathways. Molecular docking indicated that the core component had a higher binding force to the target site. Animal experiments confirmed that SNS reduced the level of p-STAT3 （P0.01）， down-regulated the expression of ALOX15 mRNA （P0.01）， up-regulated the level of serum GSH， and the expressions of SLC7A11 and GPx4 mRNA， reduced MDA and 4-HNE levels （P0.05， P0.01）. Additionally， SNS improved the mitochondrial injury induced by cardiomyocyte ferroptosis， reduced the area of MI， alleviated inflammation and myocardial injury， lowered the levels of serum CK， CK-MB， LDH， IL-6， and the mRNA expression levels of IL-16 and IL-18 （P0.05）， and improved ST segment elevation. ConclusionSNS can reduce ISO-induced STAT3 phosphorylation levels， inhibit ferroptosis in cardiomyocytes， alleviate inflammation and myocardial injury， thereby improving MI.

ObjectiveTo explore the mechanism by which Sini San （SNS） inhibits ferroptosis， alleviates inflammation and myocardial injury， and improves myocardial infarction （MI）. MethodsThe active ingredients of SNS were obtained by searching the Traditional Chinese Medicine System Pharmacology Platform （TCMSP） database， its target sites were predicted using the SwissTargetPrediction Database， and the core components were screened out using the CytoNCA plug-in. The targets of MI and ferroptosis were obtained by using GeneCards， Online Mendelian Inheritance in Man （OMIM） database， DrugBank， Therapeutic Target Database （TTD）， FerrDb database and literature review， respectively. The intersection of these targets of SNS-MI-ferroptosis was plotted as a Venn diagram. The protein-protein interaction （PPI） network was constructed using the STRING database， and the visualization graph was prepared using Cytoscape. The core targets were screened out using the CytoNCA plug-in， and the biological functions were clustered by the MCODE plug-in. Gene Ontology （GO） functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed using the David database. Molecular docking was performed using AutoDock and visualized with PyMOL2.5.2. The Kunming mice were randomly divided into the control group， the model group， the SNS group， and the trimetazidine （TMZ） group. The mice were subcutaneously injected with isoprenaline （ISO， 5 mg·kg^-1·d^-1） to establish an MI model. The drug was continuously intervened for 7 days. The ST-segment changes were recorded by electrocardiogram （ECG）， and the tissue morphology changes were observed by hematoxylin-eosin （HE） staining. Cardiomyocyte ferroptosis was investigated by transmission electron microscopy. Serum creatine kinase （CK）， creatine kinase isoenzyme （CK-MB）， lactate dehydrogenase （LDH）， reduced glutathione （GSH）， and malondialdehyde （MDA） levels were detected by biochemical assay. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum levels of interleukin （IL）-6 and 4-hydroxynonenal （4-HNE）. Immunohistochemical staining was employed to detect IL-6 and phosphorylated signal transducer and transcription activator 3 （p-STAT3） in cardiac tissues. Western blot was used to detect STAT3 and p-STAT3 in cardiac tissues. Real-time PCR was used to detect the levels of IL-6， IL-18， solute carrier family 7 member 11 （SLC7A11）， arachidonic acid 15-lipoxygenase （ALOX15）， and glutathione peroxidase 4 （GPx4） in cardiac tissues. ResultsA total of 121 active ingredients of SNS were obtained， and 58 potential targets of SNS in the treatment of MI by regulating ferroptosis were screened. The three protein modules with a score5 were mainly related to the inflammatory response. The GO function was mainly related to inflammation， and KEGG enrichment analysis showed that SNS mainly regulated ferroptosis- and inflammation- related signaling pathways. Molecular docking indicated that the core component had a higher binding force to the target site. Animal experiments confirmed that SNS reduced the level of p-STAT3 （P0.01）， down-regulated the expression of ALOX15 mRNA （P0.01）， up-regulated the level of serum GSH， and the expressions of SLC7A11 and GPx4 mRNA， reduced MDA and 4-HNE levels （P0.05， P0.01）. Additionally， SNS improved the mitochondrial injury induced by cardiomyocyte ferroptosis， reduced the area of MI， alleviated inflammation and myocardial injury， lowered the levels of serum CK， CK-MB， LDH， IL-6， and the mRNA expression levels of IL-16 and IL-18 （P0.05）， and improved ST segment elevation. ConclusionSNS can reduce ISO-induced STAT3 phosphorylation levels， inhibit ferroptosis in cardiomyocytes， alleviate inflammation and myocardial injury， thereby improving MI.

OBJECTIVE:Elevated blood pressure increases the risk of cardiovascular diseases.Isometric exercise training has been shown to significantly reduce resting blood pressure,but the factors influencing its effectiveness remain unclear,and specific application guidelines are yet to be established.This study aims to evaluate the impact of isometric exercise training on resting blood pressure through meta-analysis,explore its moderating factors,and provide evidence-based recommendations based on its dose-response relationship.METHODS:Following the PRISMA guidelines,a systematic search was conducted in PubMed,Embase,Cochrane Library,Scopus,and Web of Science databases using keywords"Isometric exercise training,""Systolic blood pressure,"and"Diastolic blood pressure,"covering literature up to September 2024.Randomized controlled trials involving isometric exercise training and resting blood pressure were included.Three independent researchers performed literature screening and data extraction,assessing bias risk and quality grades using the Risk of Bias 2.0 tool and GRADE framework.Main effect pooling,publication bias assessment,subgroup,and regression analysis were conducted using R software(version 4.3.4).RESULTS:A total of 28 articles(comprising 32 randomized controlled trials)involving 977 participants were included.(1)Meta-analysis results indicated that isometric exercise training significantly reduced resting systolic blood pressure(MD=-8.01,95％CI=-9.22 to-6.80,P＜0.01,I2=18.20％,low evidence grade)and diastolic blood pressure(MD=-3.46,95％CI=-4.64 to-2.28,P＜0.01,I2=0％,moderate evidence grade)compared to no exercise.(2)Subgroup analysis results revealed significant influences of gender,health status,exercise modality,frequency,intensity,duration,sets per session,rest duration,and baseline blood pressure on the main effects for both systolic(P＜0.01)and diastolic blood pressure(P＜0.05).(3)Regression analysis results did not show any significant influencing factors,but body mass index(β=-4.11,P=0.091)showed a significant negative trend on the main effect for systolic blood pressure.(4)No significant publication bias was observed in the meta-analysis results(P＞0.05).CONCLUSION:(1)Isometric exercise training significantly lowers systolic(low evidence grade)and diastolic(moderate evidence grade)blood pressure with clinically meaningful thresholds.(2)Participant characteristics(gender,health status,baseline blood pressure,and body mass index)and isometric exercise training protocols(modality,frequency,intensity,duration,cycle,sets per session,and rest duration)influence its antihypertensive effects.(3)The article recommends the optimal blood pressure management prescription:three sessions per week,with four sets per session,each set lasting 2 minutes with a 2-minute rest,at an intensity of 95％HRpeak using isometric wall squat exercises;the intervention period can be adjusted around a 6-week node.Future high-quality research is urgently needed to further validate and support these conclusions.

BACKGROUND:The prevalence of osteoporosis is high in postmenopausal women,but muscle mass,grip strength,and how these factors affect osteoporosis are understudied,and the exact link between them has not been clarified.OBJECTIVE:To investigate the correlation between muscle mass,grip strength,appendicular skeletal muscle index and bone mineral density in postmenopausal women with osteoporosis and to assess the potential values of these indices in predicting and diagnosing postmenopausal osteoporosis.METHODS:Eighty-three postmenopausal women were collected from the outpatient clinic of the Third Affiliated Hospital of Guangzhou University of Chinese Medicine from February 2023 to January 2024.General data were collected.Bone mineral density was detected.T-value,muscle mass of each part,grip strength were recorded.The body mass index and appendicular skeletal muscle index were calculated.The patients were categorized into non-osteoporosis group(n=17)and postmenopausal osteoporosis group(n=66)according to T value and fracture history,and were statistically analyzed accordingly.RESULTS AND CONCLUSION:(1)The body mass,body mass index,bone mineral density of the overall lumbar spine,muscle mass and appendicular skeletal muscle index were higher in the non-osteoporosis group than the osteoporosis group(P＜0.05).(2)Muscle mass was positively correlated with bone mineral density of the overall lumbar spine and individual vertebrae(P＜0.05).(3)Multiple stepwise linear regression analysis showed that body mass and grip strength were linearly and positively correlated with muscle mass;body height and muscle mass were linearly and positively correlated with grip strength,and body mass was linearly and negatively correlated with grip strength.Body mass index was linearly and positively correlated with bone mineral density,and age was linearly and negatively correlated with bone mineral density.(4)Analysis by receiver operating characteristic curve showed that:muscle mass(the area under the curve,sensitivity,specificity and critical value of muscle mass were 0.744,76.50％,74.20％and 36.50 kg,respectively,with P=0.002)and appendicular skeletal muscle index(the area under the curve,sensitivity,specificity and critical value of appendicular skeletal muscle index were 0.739,82.40％,62.10％and 5.81 kg/m2,respectively,and P=0.002)had good predictive value for postmenopausal osteoporosis.To conclude,a reduction in muscle mass and appendicular skeletal muscle index can help to predict the risk of postmenopausal osteoporosis,and the possibility of osteoporosis should be taken into account in postmenopausal women when muscle mass is＜36.50 kg or appendicular skeletal muscle index is＜5.81 kg/m2,in order to prevent the occurrence of postmenopausal osteoporosis.

OBJECTIVE:Elevated blood pressure increases the risk of cardiovascular diseases.Isometric exercise training has been shown to significantly reduce resting blood pressure,but the factors influencing its effectiveness remain unclear,and specific application guidelines are yet to be established.This study aims to evaluate the impact of isometric exercise training on resting blood pressure through meta-analysis,explore its moderating factors,and provide evidence-based recommendations based on its dose-response relationship.METHODS:Following the PRISMA guidelines,a systematic search was conducted in PubMed,Embase,Cochrane Library,Scopus,and Web of Science databases using keywords"Isometric exercise training,""Systolic blood pressure,"and"Diastolic blood pressure,"covering literature up to September 2024.Randomized controlled trials involving isometric exercise training and resting blood pressure were included.Three independent researchers performed literature screening and data extraction,assessing bias risk and quality grades using the Risk of Bias 2.0 tool and GRADE framework.Main effect pooling,publication bias assessment,subgroup,and regression analysis were conducted using R software(version 4.3.4).RESULTS:A total of 28 articles(comprising 32 randomized controlled trials)involving 977 participants were included.(1)Meta-analysis results indicated that isometric exercise training significantly reduced resting systolic blood pressure(MD=-8.01,95％CI=-9.22 to-6.80,P＜0.01,I2=18.20％,low evidence grade)and diastolic blood pressure(MD=-3.46,95％CI=-4.64 to-2.28,P＜0.01,I2=0％,moderate evidence grade)compared to no exercise.(2)Subgroup analysis results revealed significant influences of gender,health status,exercise modality,frequency,intensity,duration,sets per session,rest duration,and baseline blood pressure on the main effects for both systolic(P＜0.01)and diastolic blood pressure(P＜0.05).(3)Regression analysis results did not show any significant influencing factors,but body mass index(β=-4.11,P=0.091)showed a significant negative trend on the main effect for systolic blood pressure.(4)No significant publication bias was observed in the meta-analysis results(P＞0.05).CONCLUSION:(1)Isometric exercise training significantly lowers systolic(low evidence grade)and diastolic(moderate evidence grade)blood pressure with clinically meaningful thresholds.(2)Participant characteristics(gender,health status,baseline blood pressure,and body mass index)and isometric exercise training protocols(modality,frequency,intensity,duration,cycle,sets per session,and rest duration)influence its antihypertensive effects.(3)The article recommends the optimal blood pressure management prescription:three sessions per week,with four sets per session,each set lasting 2 minutes with a 2-minute rest,at an intensity of 95％HRpeak using isometric wall squat exercises;the intervention period can be adjusted around a 6-week node.Future high-quality research is urgently needed to further validate and support these conclusions.

BACKGROUND:The prevalence of osteoporosis is high in postmenopausal women,but muscle mass,grip strength,and how these factors affect osteoporosis are understudied,and the exact link between them has not been clarified.OBJECTIVE:To investigate the correlation between muscle mass,grip strength,appendicular skeletal muscle index and bone mineral density in postmenopausal women with osteoporosis and to assess the potential values of these indices in predicting and diagnosing postmenopausal osteoporosis.METHODS:Eighty-three postmenopausal women were collected from the outpatient clinic of the Third Affiliated Hospital of Guangzhou University of Chinese Medicine from February 2023 to January 2024.General data were collected.Bone mineral density was detected.T-value,muscle mass of each part,grip strength were recorded.The body mass index and appendicular skeletal muscle index were calculated.The patients were categorized into non-osteoporosis group(n=17)and postmenopausal osteoporosis group(n=66)according to T value and fracture history,and were statistically analyzed accordingly.RESULTS AND CONCLUSION:(1)The body mass,body mass index,bone mineral density of the overall lumbar spine,muscle mass and appendicular skeletal muscle index were higher in the non-osteoporosis group than the osteoporosis group(P＜0.05).(2)Muscle mass was positively correlated with bone mineral density of the overall lumbar spine and individual vertebrae(P＜0.05).(3)Multiple stepwise linear regression analysis showed that body mass and grip strength were linearly and positively correlated with muscle mass;body height and muscle mass were linearly and positively correlated with grip strength,and body mass was linearly and negatively correlated with grip strength.Body mass index was linearly and positively correlated with bone mineral density,and age was linearly and negatively correlated with bone mineral density.(4)Analysis by receiver operating characteristic curve showed that:muscle mass(the area under the curve,sensitivity,specificity and critical value of muscle mass were 0.744,76.50％,74.20％and 36.50 kg,respectively,with P=0.002)and appendicular skeletal muscle index(the area under the curve,sensitivity,specificity and critical value of appendicular skeletal muscle index were 0.739,82.40％,62.10％and 5.81 kg/m2,respectively,and P=0.002)had good predictive value for postmenopausal osteoporosis.To conclude,a reduction in muscle mass and appendicular skeletal muscle index can help to predict the risk of postmenopausal osteoporosis,and the possibility of osteoporosis should be taken into account in postmenopausal women when muscle mass is＜36.50 kg or appendicular skeletal muscle index is＜5.81 kg/m2,in order to prevent the occurrence of postmenopausal osteoporosis.

ObjectiveTo investigate the mechanism of modified Si Junzitang and Shashen Maidong Tang ［Yiqi Yangyin Jiedu prescription （YQYYJD）］ in enhancing the sensitivity of cisplatin in epidermal growth factor receptor tyrosine kinase inhibitor （EGFR-TKI）-resistant lung adenocarcinoma cells based on aerobic glycolysis. MethodsThe effects of different concentrations of YQYYJD （0， 2， 3， 4， 5， 6， 7， 8 g·L^-1） and cisplatin （0， 3， 6， 9， 12， 15， 18， 21， 24， 27 mg·L^-1） on the proliferation and activity of PC9/GR cells were detected by the cell counting kit-8 （CCK-8） assay after 24 hours of intervention. The half-maximal inhibitory concentration （IC₅₀） for PC9/GR cells was calculated to determine the concentrations used in subsequent experiments. PC9/GR cells were divided into blank group （complete medium）， YQYYJD group （5 g·L^-1）， cisplatin group （12 mg·L^-1）， and combined group （YQYYJD 5 g·L^-1 + cisplatin 12 mg·L^-1）. After 24 hours of intervention， cell viability was measured using CCK-8 assay. Cell proliferation was assessed by colony formation assay， and cell migration was evaluated by scratch and Transwell assays. Glucose consumption， lactate production， and adenosine triphosphate （ATP） levels were measured by colorimetric assays. The expression levels of glycolysis-related proteins， including hexokinase 2 （HK2）， phosphofructokinase P （PFKP）， pyruvate kinase M2 （PKM2）， lactate dehydrogenase A （LDHA）， glucose transporter 1 （GLUT1）， and monocarboxylate transporter 4 （MCT4）， were determined by Western blot. ResultsBoth YQYYJD and cisplatin inhibited the viability of PC9/GR cells in a concentration-dependent manner. The IC₅₀ of PC9/GR cells for YQYYJD and cisplatin were 5.15 g·L^-1 and 12.91 mg·L^-1， respectively. In terms of cell proliferation， compared with the blank group， the cell survival rate and the number of colonies formed in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group showed a further significant reduction in cell survival rate and colony formation （P<0.01）. In terms of cell migration， compared with the blank group， the cell migration rate and the number of cells passing through the Transwell membrane in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group exhibited a further significant reduction in cell migration rate and the number of cells passing through the Transwell membrane （P<0.01）. In terms of glycolysis， compared with the blank group， glucose consumption， lactate production， and ATP levels in the YQYYJD group， cisplatin group， and combined group were significantly decreased （P<0.01）. Compared with the YQYYJD and cisplatin groups， the combined group showed a further significant reduction in glucose consumption， lactate production， and ATP levels （P<0.05）. Compared with the blank group， the protein expression levels of HK2， PFKP， PKM2， and LDHA in the YQYYJD， cisplatin， and combined groups were significantly decreased （P<0.01）. The combined group showed a further significant reduction in the expression levels of these proteins compared with the YQYYJD and cisplatin groups （P<0.01）. No significant differences were observed in the protein expression levels of GLUT1 and MCT4 among the groups. ConclusionYQYYJD can synergistically inhibit the proliferation and migration of PC9/GR cells and enhance their sensitivity to cisplatin. The mechanism may be related to the downregulation of the expression of glycolysis-related rate-limiting enzymes， including HK2， PFKP， PKM2， and LDHA， thereby inhibiting glycolysis.

ObjectiveTo investigate the clinical efficacy and safety of state-target formula Angelica and Safflower spot removal essence in the treatment of melasma. MethodsA randomized，double-blind，placebo-controlled parallel-group trial was conducted，selecting patients with Qi stagnation and blood stasis type of melasma who visited the dermatology outpatient department of Beijing Hospital of Integrated Traditional Chinese and Western Medicine from September 2022 to February 2023. In accordance with the double-blind principle，a random number table was generated to randomly divide the patients into an experimental group （50 cases） and a control group （50 cases）. The experimental group applied the Angelica and Safflower spot removal essence topically，while the control group applied a placebo topically，with continuous treatment for 8 weeks and a follow-up of 1 month. Melasma Area and Severity Index （MASI），Physician Global Assessment （PGA），patient self-assessment scoring，VISIA detection and scoring were performed before medication and on the first day after stopping medication，and all adverse events were recorded. ResultsIn a study involving 100 patients，87 cases were included in the analysis. The experimental group consisted of 45 cases，with 5 dropouts，and the control group had 42 cases，with 8 dropouts. The baseline data of the patients in both groups were consistent，with no statistically significant differences. After treatment，the total effective rate of the experimental group was 75.56% （34/45），significantly higher than the control group's 2.38% （1/42），and the difference was statistically significant （χ²=48.38，P<0.01）. Compared with before treatment，after treatment，the MASI score，VISIA image spot，brown spot，and red area score of the experimental group patients were significantly reduced （P<0.05，P<0.01），while there was no statistically significant difference in the control group patients. Compared with the control group after treatment，the experimental group showed more significant improvement （P<0.05，P<0.01）. VISIA image analysis showed that after 8 weeks of treatment，the experimental group showed significant improvement in surface spots，deep spots，and red areas，while the control group remained unchanged or showed an increasing trend. 42.22% （19/45） of the experimental group had a PGA score of 3 or below，while 88.89% （40/45） had a PGA score of 4 or below. 80.95% （34/42） of the control group had a PGA score of 5 or above，and the difference between the two groups was statistically significant （χ²=38.26，P<0.01）. There was no statistically significant difference in the recurrence rate and adverse reaction rate within one month between the experimental group and the control group. ConclusionThe state-target formula Angelica and Safflower spot removal essence is clinically effective and safe in the treatment of melasma.

Objective To systematically identify the key genes regulating the exit from na?ve pluripotency in embryonic stem cells(ESCs)in order to provide novel targets and theoretical insights into the mechanisms for pluripotency transition and early cell fate determination.Methods Nanog-green fluorescent protein(Nanog-GFP)reporter-labeled ESCs were infected with a genome-wide Brie knockout library,and further cultured under leukemia inhibitory factor/serum(LIF/S)conditions for 14 d.Flow cytometry was used to sort Nanog-GFP?(na?ve-state)and Nanog-GFP-(primed state)cell populations,followed by genomic DNA extraction and high-throughput sequencing.Model-based Analysis of Genome-wide CRISPR/Cas9 Knockout(MAGeCK)was applied to identify differential genes between GFP?/Input,GFP?/Input,and GFP?/GFP? groups.Metascape and Gene Set Enrichment Analysis(GSEA)were conducted for functional enrichment analysis.Then the obtained candidate genes were employed to construct knockout models,and their roles were assessed through cell morphology observation,Nanog-positive rate detection,colony formation assays,and pluripotency gene expression analysis.Results The GFP?/Input screening revealed 2 921 negatively regulated genes(mainly enriched in basic life processes,such as RNA metabolism and cell cycle)and 1 393 positively regulated genes(enriched in the processes of nervous system development,carbohydrate metabolism,and vascular system development).In the GFP?/Input screening,2 765 negatively regulated genes(enriched in RNA metabolism,cell cycle,and other fundamental processes)and 1 303 positively regulated genes(enriched in neural development,cell survival,and endothelial migration)were identified.The GFP?/GFP? comparison identified 1 001 negatively regulated genes[involved in stress response and inhibition of mitogen-activated protein kinase(MAPK)signaling]and 983 positively regulated genes[related to fibroblast growth factor/extracellular signal-regulated kinase(FGF/ERK)signaling pathway and glucose metabolism).These genes,were not only known pluripotency regulators(e.g.,Nanog,Nr5a2,Klf2,Klf4)and exit-associated genes(e.g.,Gata6,Grb2,Zeb1,Fgfr1),but also some novel candidates(e.g.,Dmrt1,Rxra,Zbtb14 and Tmem41b).Functional validation showed that transient knockout of Dmrt1,Tmem41b,and Hic2 significantly increased the proportion of Nanog? cells(P<0.01),suggesting their role in suppressing ground-state exit.ESCs with stable Dmrt1 knockout exhibited a more na?ve-state phenotype,presenting compact,dome-shaped colonies,with increased ratio of undifferentiated colonies(P<0.01),up-regulation of ground-state markers(Nanog,Nr5a2,Dppa3,P<0.01),and down-regulation of primed-state markers(Fgf5,Lefty1,Dnmt3b,P<0.01).Rescue experiments for Dmrt1 expression reversed these above phenotypes.Conclusion A candidate gene set regulating exit from na?ve pluripotency in ESC is screened out and identified with genome-wide CRISPR.Our findings implicate Dmrt1 plays a critical role in promoting the exit.

Objective To observe the therapeutic effect and mechanism of external application with Shilian Powder combined with oral administration of Shumai Capsules for the treatment of rats with diabetic foot ulcer(DFU).Methods Eight male rats with successfully modeled foot ulcer(DF)were used as the control group.While 24 male rats with successfully modeled DFU were randomly divided into DFU group,Shumai Capsules group and Shilian Powder combined with Shumai Capsules group,with eight rats in each group.After the corresponding interventions,we determined the wound healing rate,histopathological changes of wound,levels of inflammatory factors such as interleukin(IL)-6,IL-1β and vascular endothelial growth factor(VEGF)in serum,levels of Apelin and Relaxin,and protein expressions of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT)and VEGF in wound tissue,as well as mRNA expressions of PI3K,AKT,Relaxin and Apelin.Results Compared with the control group,the DFU group showed a significant decrease in wound healing rate,VEGF level in serum and wound,wound Relaxin level,protein and mRNA levels of wound AKT(P<0.05),and a significant increase in serum IL-6 and IL-1β levels,wound Apelin level,wound PI3K protein and mRNA levels(P<0.05),and the reduced wound granulation tissue and formation of new capillaries and increased inflammatory cell infiltration were seen under the microscope.Compared with the DFU group,the wound healing rate,VEGF level in serum and wound,wound Relaxin and Apelin levels,protein and mRNA levels of wound PI3K and AKT in the Shumai Capsules group and Shilian Powder combined with Shumai Capsules group were significantly increased(P<0.05),and serum IL-6 and IL-1β levels were significantly decreased(P<0.05),and the increased wound granulation tissue and formation of new capillaries and reduced inflammatory cell infiltration were seen under the microscope.Compared with the Shumai Capsules group,the wound healing rate,wound VEGF level,wound Relaxin and Apelin levels,protein and mRNA levels of wound PI3K and AKT in Shilian Powder combined with Shumai Capsules group were significantly increased(P<0.05),and the serum IL-6 and IL-1β levels were significantly decreased(P<0.05),and the increased wound granulation tissue and formation of new capillaries and reduced inflammatory cell infiltration were seen under the microscope.Conclusion External application with Shilian Powder combined with oral administration of Shumai Capsules can promote the wound healing in rats with DFU,its mechanism is related to the activation of PI3K/AKT/Relaxin/Apelin signaling pathway.