ObjectiveBy establishing a light sleep deprivation （SD） model and using the traditional Chinese medicine （TCM） compound Sancao Anshen prescription for intervention， this study aims to observe one of the effects of Sancao Anshen prescription on the sleep and depressive states of zebrafish in the SD model and explore the mechanism of action of this drug in regulating the cyclic adenosine monophosphate （cAMP）/protein kinase A （PKA） signaling pathway. MethodsA total of 240 six-month-old wild-type AB zebrafish were randomly divided into a blank group， a model group， low-dose， medium-dose， and high-dose groups of Sancao Anshen prescription （0.28， 0.83， 2.48 g·L^-1）， and a melatonin group （0.2 g·L^-1）， with 40 fish in each group. Except for the blank group， all others were exposed to LED lights （150 lux） for three days to construct the sleep deprivation model， and were treated with the corresponding doses of Sancao Anshen decoction and melatonin solution for three days. 24 h movement behavior was used to detect diurnal movement trajectories. A T-shaped maze was employed to detect learning and memory functions， and a new tank experiment was conducted to detect depression-like behaviors in zebrafish. Hematoxylin-eosin （HE） staining was used to observe the morphology of hypothalamic neurons， and transmission electron microscopy was utilized to observe the ultrastructure of hypothalamic cells. Immunohistochemical （IHC） was used to measure the positive expression of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in the hypothalamus， and enzyme-linked immunosorbent assay （ELISA） was employed to determine the content of cAMP， 5-hydroxytryptamine （5-HT）， gamma-aminobutyric acid （GABA）， and glutamic acid （Glu） in brain tissue. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to measure the expression of PKA， cAMP response element-binding （CREB）， and brain-derived neurotrophic factor （BDNF） mRNAs and proteins and their phosphorylation levels （p-PKA， p-CREB） in brain tissue. ResultsCompared with those in the blank group， the resting time and resting count in the model group decreased significantly （P<0.01）， while the gross motor time and gross motor count increased significantly （P<0.01）. The latency time to enter the EC region in each administration group increased significantly （P<0.01）. The exploration time towards the top and the number of times entering the top decreased significantly （P<0.01）， and the incubation period of the first ascent increased significantly （P<0.01）. The number of hypothalamic neurons decreased significantly， and the neurons exhibited irregular shapes， sparse arrangement， and nuclear condensation. Nuclear collapse， nuclear membrane rupture and dissolution， chromatin condensation， mitochondrial swelling and deformity， and plate-like cristae rupture or disappearance were observed. The positive expression of TNF-α and IL-1β in the hypothalamus was significantly decreased （P<0.01）. The content of cAMP， GABA， and 5-HT in brain tissue was significantly downregulated， while the content of Glu was significantly upregulated （P<0.01）. The mRNA of PKA， CREB， and BDNF was significantly downregulated （P<0.01）， and the protein expression of p-PKA， p-PKA/PKA， p-CREB， p-CREB/CREB， and BDNF was significantly decreased （P<0.05， P<0.01）. Compared with those in the model group， the resting time in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group increased （P<0.05， P<0.01）， and the resting count in each administration group significantly increased （P<0.05， P<0.01）. The gross motor time and gross motor count significantly decreased （P<0.05， P<0.01）. The latency time to enter the EC region decreased （P<0.01）， and the exploration time towards the top increased （P<0.01）. The time for the first ascent in the high-dose group of Sancao Anshen prescription and the melatonin group was shortened （P<0.05， P<0.01）， and the number of times entering the top increased （P<0.01）. The morphology of neurons in each administration group improved， with the gap decreased， the nuclear membrane relatively intact， and mitochondrial swelling improved. The positive expression of IL-1β in each administration group significantly decreased （P<0.01）. The positive expression of TNF-α in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group significantly decreased （P<0.01）. The Glu content in the low-dose group of Sancao Anshen prescription decreased， while cAMP and GABA levels increased （P<0.05， P<0.01）. There was no statistically significant difference in 5-HT content. The medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group all showed significant increases in cAMP， 5-HT， and GABA levels （P<0.01）， and a significant decrease in Glu content （P<0.01）. The mRNA of CREB in the low-dose group of Sancao Anshen prescription was significantly upregulated （P<0.01）， while there was no statistically significant difference in the mRNA of PKA and BDNF. The mRNA of PKA， CREB， and BDNF in the medium-dose and high-dose groups of Sancao Anshen prescription and the melatonin group were all significantly upregulated （P<0.01）. The protein expression of p-PKA， p-PKA/PKA， p-CREB， p-CREB/CREB， and BDNF in the high-dose group of Sancao Anshen prescription and the melatonin group increased significantly （P<0.05， P<0.01）， and the protein expression of p-PKA， p-CREB， and BDNF in the medium-dose group of Sancao Anshen prescription increased （P<0.05）. ConclusionThe improvement of sleep and depressive states in zebrafish with the SD model by Sancao Anshen prescription may be related to the inhibition of inflammatory factors such as TNF-α and IL-1β， the reduction in Glu， and the elevation in the content of neurotransmitters such as GABA and 5-HT via the cAMP/PKA signaling pathway.

ObjectiveTo investigate the effects of Huanglian Jiedutang （HLJDT） on cognitive function in mice with ischemic stroke （IS） and to elucidate whether its neuroprotective effects are mediated by inhibition of the nuclear factor-κB （NF-κB） signaling pathway and subsequent suppression of NF-κB-regulated neuronal apoptosis. MethodsAn IS model was established using middle cerebral artery occlusion （MCAO）. Sixty C57BL/6J mice were randomly assigned to five groups （n =12 per group）， i.e.， sham operation， model， HLJDT low-dose （3.9 g·kg^-1·d^-1）， HLJDT high-dose （7.8 g·kg^-1·d^-1）， and Ginkgo biloba extract （GBE， 31.2 mg·kg^-1·d^-1）. Post-operatively， neurological deficit scores （Longa score）， cerebral infarct volume assessed by 2，3，5-triphenyltetrazolium chloride （TTC） staining， and brain water content were evaluated. Learning and memory were assessed using new object recognition （NOR） and fear conditioning （FC） tests. Hippocampal pathology was examined via hematoxylin and eosin （HE） staining. Immunofluorescence detected expression of glial fibrillary acidic protein （GFAP， astrocyte marker）， cellular oncogene Fos （c-Fos， neuronal activation marker）， and glutamate decarboxylase 65 （GAD65）. Western blot measured nuclear factor-κB inhibitor protein α （IκBα）， phosphorylated IκBα （p-IκBα）， NF-κB p65， phosphorylated NF-κB p65 （p-NF-κB p65）， ionic calcium binding adapter molecule 1 （Iba-1）， tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and apoptosis-related proteins， such as cleaved cysteinyl aspartate-specific protease 3 （Caspase-3）， B-cell lymphoma 2 （Bcl-2）， and Bcl-2-associated X protein （Bax）. Real-time quantitative PCR （Real-time PCR） was used to assess mRNA levels of Iba-1， TNF-α， IL-1β， NF-κB p65， cleaved Caspase-3， Bax， and Bcl-2. ResultsCompared with the sham group， the model group exhibited significantly increased neurological deficit scores， brain water content， and cerebral infarct volume （P<0.01）. Hippocampal CA1 neurons were disorganized， showing nuclear pyknosis and karyolysis. NOR exploration time and FC freezing time were significantly reduced （P<0.01）. GFAP and c-Fos expression were increased， while GAD65 expression was decreased （P<0.01）. Cleaved Caspase-3 and Bax were upregulated， Bcl-2 was downregulated， and the Bax/Bcl-2 ratio was elevated （P<0.01）. Expression levels of p-IκBα， p-NF-κB p65， IL-1β， TNF-α， and Iba-1 were significantly increased （P<0.01）. Compared with the model group， HLJDT high-dose， low-dose， and GBE groups showed significant improvements in all parameters （P<0.01）. Among them， the HLJDT high-dose group showed the most pronounced neuronal structural recovery and superior performance in NOR and FC tests （P<0.01）. In this group， GFAP and c-Fos decreased， GAD65 increased （P<0.01）， apoptosis-related protein expression was reversed， and NF-κB signaling and related inflammatory factor expression were suppressed （P<0.01）. ConclusionHLJDT ameliorates cognitive dysfunction in mice after IS， potentially by inhibiting the NF-κB signaling pathway， thereby reducing neuroinflammation and hippocampal neuronal apoptosis.

Objective: To evaluate the diagnostic performance of magnetic resonance spectroscopy (MRS) in discriminating fumarate hydratase-deficient (FH-d) uterine leiomyomas (ULs) from FH-preserved ULs. Materials and Methods: This study consisted of three stages, with independent cohorts recruited for each stage: 1) sample-size estimation was retrospectively performed on UL specimens (diameter ≥3 cm; age, 20–40 years) from our database with immunohistochemistry (IHC) for 2-succinocysteine (2-SC) as the reference, without genetic testing, 2) MRS sequence optimization in confirmed FH germline mutation participants with ultrasound-detected ULs (diameter ≥3 cm), without IHC analysis, and 3) prospective diagnostic test accuracy was evaluated in consecutive participants with ultrasound-detected ULs (diameter ≥3 cm;age, 20–40 years), using IHC for 2-SC for determining the FH status and subsequent genetic testing in those with positive 2-SC results to identify whether FH mutations were germline or somatic in origin. The choline and fumarate peaks in MRS were classified as positive, negative, or technical failure (TF). TFs were analyzed separately and excluded from the primary diagnostic accuracy calculations. T1-, T2-, and diffusion-weighted images were interpreted as hyperintense or hypointense. The enhancement rate and apparent diffusion coefficient were also acquired. Diagnostic performance was compared between MRS and various magnetic resonance imaging (MRI) features. Results: The optimal MRS parameters for the fumarate peak were echo time (TE) = 140 ms and an average of 256. Among the 360 prospective participants, 37 were confirmed to have FH-dULs. MRS showed positive fumarate peaks in 35 of 37 FH-dULs.After excluding six TFs, the positive fumarate peak on MRS showed 94.6% (35/37) sensitivity, 99.7% (316/317) specificity, and 99.2% (351/354) accuracy, all of which were significantly superior to those of other MRI features (P ≤ 0.002). Conclusion A positive fumarate peak on MRS may be a useful imaging biomarker for diagnosing FH-dULs.

Objective: To assess the association of family food environment with dietary and nutrition literacy among junior and senior high school students, providing a basis for nutritional health education and adolescent nutritional improvement. Methods: From January 9 to February 10, 2025, a total of 1 473 junior and senior high school students aged 12-20 were recruited by using a cluster random sampling method combined with convenient sampling from 15 provinces (municipalities and autonomous regions) across China. Data on basic demographic characteristics, home food environment, and dietary and nutrition literacy were collected by using the Food and Nutritional Literacy Questionnaire for Chinese School age Children. Multiple linear regression and Logistic regression models were employed to analyze the association of the home food environment on adolescent dietary and nutrition literacy. Results: The total score of dietary and nutrition literacy among junior and senior high school students was (65.28±9.03). The score in the cognitive domain (67.59±10.56) was higher than that in the skill domain (60.74±9.15), and the score for food preparation skills (56.60±14.28) was the lowest. Students aged 16-20, senior high school students, those for only children, non boarding students, those with urban household registration, those from wealthier families, those whose primary caregivers were parents with higher education levels, and those who previously received nutrition and health education had significantly higher dietary and nutrition literacy scores compared to their respective counterpart groups ( t/F =-8.32, -8.14, 4.76, -2.70, 7.15, 18.29, 10.68, 11.66 , all P <0.05). Multiple linear regression analysis revealed that family food environment was associated with dietary and nutrition literacy, and the model was statistically significant ( R 2=0.20, F=20.54, P <0.05). Lower availability of fruits at home in the past month and lower frequency of family communication regarding dietary and nutrition were associated with lower dietary nutrition literacy scores among junior and senior high school students ( β =-1.35, -1.67, both P <0.01). Among junior and senior high school students, lower frequency of dining out with the family, watching videos during family meals, and being urged by parents to eat more food in the past month were associated with higher scores on dietary nutrition literacy ( β =1.35, 1.13, 0.71, all P <0.01). Logistic regression model results indicated that watching videos during family meals ( OR =1.70) and family communication regarding dietary and nutrition ( OR =0.51) were significantly associated with dietary and nutrition literacy levels (both P <0.05). Conclusions The score of dietary and nutrition literacy among junior and senior high school students is relatively low, and the family food environment is a key factor. It is recommended that nutrition education and improvement initiatives for junior and senior high school students prioritize attention on children and adolescents from households with less favorable food environments.

AIM:To observe the effect and mech-anism of cuttlebone extract regulating vascular en-dothelial growth factor(VEGF)/phosphatidylinosi-tol 3-kinase(PI3K)/protein kinase B(AKT)pathway on refractory wounds in rats.METHODS:Cuttle-bone extract(CE)was obtained by water extraction of cuttlebone.Fifty SD rats were randomly divided into negative Control group,Model group,Cuttle-bone extract low-dose(CE(L))group,Cuttlebone extract high-dose(CE(H))group,and cuttlebone ex-tract high-dose+inhibitor(CE(H)+LY294002)group.After the refractory wound model was successfully established,0.02％furacillin solution or cuttlebone extract solution were applied to the wound area of rats in each group,and the treatment was adminis-tered once a day.After 14 days of treatment for re-fractory wounds,the changes in wound healing,angiogenesis,inflammation and expression of relat-ed regulatory proteins were quantitatively ana-lyzed by measuring skin ulcer wound area,patho-logical sections,immunofluorescence staining,Eli-sa,Western blot,RT-qPCR and other methods.RE-SULTS:Compared with Model group,CE(L)and CE(H)groups can increase the number of epithelial cells and collagen,and promote the healing of re-fractory wound in rats.Serum VEGF,skin tissue mi-crovascular density,P-PI3K,P-AKT,VEGF protein ex-pression and mRNA expression levels of PI3K,Akt,VEGF and eNOS were increased(P＜0.05),while se-rum TNF-α and IL-6 levels were decreased(P＜0.05).LY294002 could partially reverse the repair-ing effect of high dose cuttlebone extract on refrac-tory wound(P＜0.05).CONCLUSION:Cuttlebone ex-tract can regulate the VEGF/PI3K/AKT signaling pathway,inhibit the inflammatory response of re-fractory wounds in rats,induce angiogenesis and promote wound healing.

Objective:To explore the spousal correlations of total cholesterol(TC),total triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),and high-density lipoprotein cholesterol(HDL-C),and to investigate the reasons behind these spousal correlations.Methods:Participants and data were from the baseline survey of family-based cohort studies in Fangshan,Beijing and Tulou,Fujian.The ori-gin of spousal correlations were explored from perspectives of convergence,assortative mating,social ho-mogamy.Pearson's correlation and generalized linear models(GLM)were used to estimate the spousal correlation.Convergence was assessed by Pearson's correlation between the phenotypic differences be-tween couples and the duration of marriage,with GLM used for further validation.Pearson's correlation of genetic risk scores(GRS)and couple-specific Mendelian randomization(MR)were calculated to assess the genetic correlation and possible causal relationships between spouses.Two-independent-sample t-tests were used to compare GRS consistency across subgroups divided by education attainment,couple-specific MR and Q statistics used to test assortative mating in subgroups and intergroup differences.Results:In the study,342 couples(287 couples from Fangshan and 55 couples from Fujian)were included,with the average age of(64.91±8.76)years.Spousal correlations of TC,TG,HDL-C,and LDL-C showed statistically significant associations both before and after adjusting for covariates,with effect sizes of 0.229(95％CI:0.125-0.327),0.257(95％CI:0.155-0.354),0.179(95％CI:0.074-0.280),and 0.181(95％CI:0.076-0.282).For convergence,for each additional year of marriage,ΔTC increased by 0.016 mmol/L(95％CI:0.001-0.033 mmol/L),and ΔLDL-C increased by 0.017 mmol/L(95％CI:0.002-0.031 mmol/L).For assortative mating,GRS correlations and results of couple specific MR didn't show any statistical significance.For social homogamy,no differences in GRS or assortative mating were found between subgroups stratified by education attainment.Conclusion:The blood lipid in participants exhibit spousal phenotypic correlations,however,no effects of convergence,assortative mating or social homogamy were observed.More independent studies with larger sample sizes are warranted to further validate these findings in the future.

Objective:To study the relationship between human leukocyte antigen (HLA) Ⅱ and nuclear factor κB (NF-κB) gene polymorphisms and susceptibility to community-acquired pneumonia in children and the therapeutic effect of amoxicillin clavulanate potassium.Methods:A total of 120 children with community-acquired pneumonia admitted to the Baoding Second Central Hospital from February 2018 to January 2021 were selected as the study objects (observation group). According to the effect of amoxicillin and clavulanate potassium treatment, they were divided into effective group and ineffective group, and 120 healthy children in the same period were selected as the control group. The polymorphisms of HLAⅡ and NF-κB gene were compared between the two groups, and the correlation between the polymorphisms of HLAⅡ and NF-κB gene and the susceptibility of children to community-acquired pneumonia and the therapeutic effect of amoxicillin clavulanate potassium was analyzed.Results:The gene frequency of HLA-DQA1*0201 [86(71.67%) vs 40(33.33%)], HLA-DQA1*0301 [72(60.00%) vs 20(16.67%)] of the observation group was significantly higher than that of the control group, the gene frequency of HLA-DQA1*0401 [30(25.00%) vs 96(80.00%)] was significantly lower than that of control group (all P<0.05). The gene frequency of HLA-DRB1*07 [92(76.67%) vs 48(40.00%)], HLA-DRB1*15 [72(60.00%) vs 40(33.33%)], HLA-DRB1*16 [96(80.00%) vs 76(63.33%)] of the observation group was significantly higher than that of the control group, and the gene frequency of HLA-DRB1*11 [36(30.00%) vs 104(86.67%)] was significantly lower than that of control group (all P<0.05). The expression of 94ins/delATTG*II in the observation group was significantly higher than that in the control group [40(33.33%) vs 24(20.00%), P<0.05]. The gene frequency of HLA-DQA1*0201 [12(38.71%) vs 74(83.15%)], HLA-DQA1*0301 [9(29.03%) vs 63(70.79%)] in the ineffective group was significantly lower than that of the effective group, the gene frequency of HLA-DQA1*0401 [18(58.06%) vs 12(13.48%)] was significantly higher than that of the effective group (all P<0.05). The gene frequency of HLA-DRB1*07 [29(93.55%) vs 63(70.79%)], HLA-DRB1*15 [29(93.55%) vs 43(48.31%)], HLA-DRB1*16 [29(93.55%) vs 67(75.28%)] of the ineffective group was significantly higher than that of the effective group, and the gene frequency of HLA-DRB1*11 [17(54.84%) vs 19(21.35%)] was significantly higher than that of the effective group (all P<0.05). The expression of 94ins/delATTG*II gene in the ineffective group was significantly higher than that in the effective group [15(48.39%) vs 25(28.09%), P<0.05]. Conclusions:Polymorphisms of HLAⅡ and NF-κB genes are associated with susceptibility to community-acquired pneumonia and efficacy of amoxicillin and clavulanate potassium in children.

Objective:To study the relationship between human leukocyte antigen (HLA) Ⅱ and nuclear factor κB (NF-κB) gene polymorphisms and susceptibility to community-acquired pneumonia in children and the therapeutic effect of amoxicillin clavulanate potassium.Methods:A total of 120 children with community-acquired pneumonia admitted to the Baoding Second Central Hospital from February 2018 to January 2021 were selected as the study objects (observation group). According to the effect of amoxicillin and clavulanate potassium treatment, they were divided into effective group and ineffective group, and 120 healthy children in the same period were selected as the control group. The polymorphisms of HLAⅡ and NF-κB gene were compared between the two groups, and the correlation between the polymorphisms of HLAⅡ and NF-κB gene and the susceptibility of children to community-acquired pneumonia and the therapeutic effect of amoxicillin clavulanate potassium was analyzed.Results:The gene frequency of HLA-DQA1*0201 [86(71.67%) vs 40(33.33%)], HLA-DQA1*0301 [72(60.00%) vs 20(16.67%)] of the observation group was significantly higher than that of the control group, the gene frequency of HLA-DQA1*0401 [30(25.00%) vs 96(80.00%)] was significantly lower than that of control group (all P<0.05). The gene frequency of HLA-DRB1*07 [92(76.67%) vs 48(40.00%)], HLA-DRB1*15 [72(60.00%) vs 40(33.33%)], HLA-DRB1*16 [96(80.00%) vs 76(63.33%)] of the observation group was significantly higher than that of the control group, and the gene frequency of HLA-DRB1*11 [36(30.00%) vs 104(86.67%)] was significantly lower than that of control group (all P<0.05). The expression of 94ins/delATTG*II in the observation group was significantly higher than that in the control group [40(33.33%) vs 24(20.00%), P<0.05]. The gene frequency of HLA-DQA1*0201 [12(38.71%) vs 74(83.15%)], HLA-DQA1*0301 [9(29.03%) vs 63(70.79%)] in the ineffective group was significantly lower than that of the effective group, the gene frequency of HLA-DQA1*0401 [18(58.06%) vs 12(13.48%)] was significantly higher than that of the effective group (all P<0.05). The gene frequency of HLA-DRB1*07 [29(93.55%) vs 63(70.79%)], HLA-DRB1*15 [29(93.55%) vs 43(48.31%)], HLA-DRB1*16 [29(93.55%) vs 67(75.28%)] of the ineffective group was significantly higher than that of the effective group, and the gene frequency of HLA-DRB1*11 [17(54.84%) vs 19(21.35%)] was significantly higher than that of the effective group (all P<0.05). The expression of 94ins/delATTG*II gene in the ineffective group was significantly higher than that in the effective group [15(48.39%) vs 25(28.09%), P<0.05]. Conclusions:Polymorphisms of HLAⅡ and NF-κB genes are associated with susceptibility to community-acquired pneumonia and efficacy of amoxicillin and clavulanate potassium in children.

Plant-derived bioactive peptides have become a research hotspot in the fields of food and medicine due to their high source safety,easy absorption and utilization by the human body,and potential edible and medicinal value.Bioactive peptides can be classified into antihypertensive,hypoglycemic,hypolipidemic,anticancer,antioxidant,antimicrobial,and anti-inflammatory peptides according to their functions.Among these,antihypertensive,hypoglycemic,and hypolipidemic peptides are collectively referred to as cardiovascular active peptides,which can be used for the treatment and prevention of cardiovascular diseases and have an important role in the development of modem biomedicine.This review focused on the preparation methods,separation,purification,and identification techniques of bioactive peptides,as well as their mechanisms of action and applications in regulating cardiovascular diseases,aiming to provide a reference for further development and application of plant-derived cardiovascular peptides.

Plant-derived bioactive peptides have become a research hotspot in the fields of food and medicine due to their high source safety,easy absorption and utilization by the human body,and potential edible and medicinal value.Bioactive peptides can be classified into antihypertensive,hypoglycemic,hypolipidemic,anticancer,antioxidant,antimicrobial,and anti-inflammatory peptides according to their functions.Among these,antihypertensive,hypoglycemic,and hypolipidemic peptides are collectively referred to as cardiovascular active peptides,which can be used for the treatment and prevention of cardiovascular diseases and have an important role in the development of modem biomedicine.This review focused on the preparation methods,separation,purification,and identification techniques of bioactive peptides,as well as their mechanisms of action and applications in regulating cardiovascular diseases,aiming to provide a reference for further development and application of plant-derived cardiovascular peptides.