This study explored the potential therapeutic targets and mechanisms of Danggui Shaoyao San(DSS) in the prevention and treatment of Alzheimer's disease(AD) through transcriptomics and metabolomics, combined with animal experiments. Fifty male C57BL/6J mice, aged seven weeks, were randomly divided into the following five groups: control, model, positive drug, low-dose DSS, and high-dose DSS groups. After the intervention, the Morris water maze was used to assess learning and memory abilities of mice, and Nissl staining and hematoxylin-eosin(HE) staining were performed to observe pathological changes in the hippocampal tissue. Transcriptomics and metabolomics were employed to sequence brain tissue and identify differential metabolites, analyzing key genes and metabolites related to disease progression. Reverse transcription-quantitative polymerase chain reaction(RT-qPCR) was employed to validate the expression of key genes. The Morris water maze results indicated that DSS significantly improved learning and cognitive function in scopolamine(SCOP)-induced model mice, with the high-dose DSS group showing the best results. Pathological staining showed that DSS effectively reduced hippocampal neuronal damage, increased Nissl body numbers, and reduced nuclear pyknosis and neuronal loss. Transcriptomics identified seven key genes, including neurexin 1(Nrxn1) and sodium voltage-gated channel α subunit 1(Scn1a), and metabolomics revealed 113 differential metabolites, all of which were closely associated with synaptic function, oxidative stress, and metabolic regulation. RT-qPCR experiments confirmed that the expression of these seven key genes was consistent with the transcriptomics results. This study suggests that DSS significantly improves learning and memory in SCOP model mice and alleviates hippocampal neuronal pathological damage. The mechanisms likely involve the modulation of synaptic function, reduction of oxidative stress, and metabolic balance, with these seven key genes serving as important targets for DSS in the treatment of AD.
Animals ; Alzheimer Disease/genetics* ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Mice ; Mice, Inbred C57BL ; Metabolomics ; Transcriptome/drug effects* ; Maze Learning/drug effects* ; Hippocampus/metabolism* ; Humans ; Disease Models, Animal ; Memory/drug effects*

Objective:To explore the effect of horizontal Baduanjin guided by interactive standard reaching theory(ISRT)on cardiac function and quality of life in patients with chronic heart failure(CHF).Methods:This random-ized controlled study enrolled 114 patients diagnosed with CHF in Bozhou People's Hospital between September 2024 and February 2025.Patients were randomly divided into control group(n=57,routine nursing care)and interven-tion group(n=57,horizontal Baduanjin under the guidance of interactive standard theory).Both groups were con-tinuously treated for 3 months.Cardiac function indexes,N-terminal pro-brain natriuretic peptide(NT-proB-NP),self-care ability,quality of life and incidence of major adverse cardiovascular events(MACE)were com-pared between the two groups.Results:Compared to patients in the control group,those in the intervention had significant higher left ventricular ejection fraction(LVEF)[(56.34±0.86)％vs.(46.69±0.91)％],6min walking distance(6MWD)[(477.16±6.62)m vs.(451.39±4.17)m]and the proportion of New York Heart Association(NYHA)class Ⅱ(70.18％vs.35.09％),and significant lower NT-proBNP[(237.76±6.41)pg/ml vs.(377.46±6.85)pg/ml],scores of the European heart failure self-care behavior scale 9(EHFSCB-9)[(12.61±1.81)points vs.(25.51±1.71)points],the physical domain[(3.96±0.91)points vs.(11.05±0.79)points],emotional domain[(4.00±0.85)points vs.(11.95±0.81)points]and other domain[(4.05±0.83)points vs.(13.47±1.15)points]of MLHFQ(P＜0.001 all).Compared to those in the control group,patients in intervention group had significant lower incidence of MACE(7.02％vs.22.81％,P=0.035).Conclusion:Under the guidance of the interactive standard theory,the horizontal Baduanjin exercise may enhance the cardiac function,improve self-care ability and quality of life in patients with chronic heart failure.

Objective:To explore the effect of horizontal Baduanjin guided by interactive standard reaching theory(ISRT)on cardiac function and quality of life in patients with chronic heart failure(CHF).Methods:This random-ized controlled study enrolled 114 patients diagnosed with CHF in Bozhou People's Hospital between September 2024 and February 2025.Patients were randomly divided into control group(n=57,routine nursing care)and interven-tion group(n=57,horizontal Baduanjin under the guidance of interactive standard theory).Both groups were con-tinuously treated for 3 months.Cardiac function indexes,N-terminal pro-brain natriuretic peptide(NT-proB-NP),self-care ability,quality of life and incidence of major adverse cardiovascular events(MACE)were com-pared between the two groups.Results:Compared to patients in the control group,those in the intervention had significant higher left ventricular ejection fraction(LVEF)[(56.34±0.86)％vs.(46.69±0.91)％],6min walking distance(6MWD)[(477.16±6.62)m vs.(451.39±4.17)m]and the proportion of New York Heart Association(NYHA)class Ⅱ(70.18％vs.35.09％),and significant lower NT-proBNP[(237.76±6.41)pg/ml vs.(377.46±6.85)pg/ml],scores of the European heart failure self-care behavior scale 9(EHFSCB-9)[(12.61±1.81)points vs.(25.51±1.71)points],the physical domain[(3.96±0.91)points vs.(11.05±0.79)points],emotional domain[(4.00±0.85)points vs.(11.95±0.81)points]and other domain[(4.05±0.83)points vs.(13.47±1.15)points]of MLHFQ(P＜0.001 all).Compared to those in the control group,patients in intervention group had significant lower incidence of MACE(7.02％vs.22.81％,P=0.035).Conclusion:Under the guidance of the interactive standard theory,the horizontal Baduanjin exercise may enhance the cardiac function,improve self-care ability and quality of life in patients with chronic heart failure.

Objective To investigate the improvement effect and mechanism of marmesin on cognitive impairment in Alzheimer's disease(AD)mice.Methods Fifty mice were randomly divided into five groups:blank group,model group,low-and high-dose marmesin groups and donepezil(positive drug)group,with 10 mice in each group.After 21 days of continuous administration,except for the blank group,the mice in other groups were given intraperitoneal injection of scopolamine to establish the AD model.Network pharmacology was used to construct the protein-protein interaction(PPI)network of common targets of marmesin in the treatment of AD,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis were performed to provide further research direction.The cognitive function of AD model mice was evaluated by Morris water maze,open field test and new object recognition test.Nissl staining was used to observe the damage of hippocampal neurons.The levels of acetylcholine(Ach),acetylcholine transferase(ChAT),acetylcholinesterase(AChE),reactive oxygen species(ROS),malondialdehyde(MDA)and catalase(CAT)in hippocampus of mice were detected by kit.The protein expression levels of interleukin 6(IL-6),interleukin 1β(IL-1 β),tumor necrosis factor α(TNF-α),nuclear factor E2-related factor 2(NRF2),silent information regulator homologous protein 3(SIRT3),Kelch-like ECH-associated protein 1(KEAP1),quinone oxidoreductase 1(NQO1)and heme oxygenase 1(HO-1)in hippocampus were detected by Western Blot.Results Compared with the model group,the latency of Morris water maze test was significantly shortened in the high-dose marmesin group,the time of entering the target area in the open field new object test and the movement distance in the central area of the open field were prolonged,the number of neurons in the hippocampal CA1 and CA3 regions was significantly increased,the levels of ChAT and Ach in the hippocampus were significantly increased,AChE level was significantly decreased,CAT level was significantly increased,ROS and MDA levels were significantly decreased,TNF-α expression level was decreased,SIRT3 and HO-1 expression levels were increased,and KE AP1 protein expression level was decreased,the differences being statistically significant(P＜0.05 or P＜0.01 or P＜0.001).Conclusion Marmesin can effectively improve the learning and memory impairment of AD mice,and its mechanism may be related to the activation of NRF2/SIRT3 signaling pathway,thereby alleviating oxidative stress level and neuroinflammation,and repairing cholinergic neuron function.

Objective:To analyze the blood differential metabolites of patients with intrahepatic cholestasis (IHC) by liquid chromatography-mass spectrometry metabolomics technology so as to find potential metabolic target.Method:Serum samples were collected from thirty patients with intrahepatic cholestasis and thirty healthy individuals after metabolomics analysis. The differential metabolites were initially screened based on the multiple differences and significance. KEGG enrichment analysis was performed on the differential metabolites to determine the candidate targets. The potential clinical application value of these characteristic metabolites was analyzed using the receiver operating characteristic curve.Result:A total of thirty patients with intrahepatic cholestasis and thirty healthy adults were included. The age difference between the two groups was not statistically significant ( P>0.05). The clinical condition was consistent with the statistically significant differences in liver biochemical indicators, blood routine, coagulation, and inflammatory indicators between the two groups ( P<0.05). Furthermore, a blood metabolomics screening analysis revealed 99 differentially expressed metabolites associated with intrahepatic cholestasis. Of these, 15 showed statistically significant differences. Glucose, lipid, and energy metabolisms were the various primary types of differential metabolites involved. The receiver operating characteristic curve>0.9 included the following twelve kinds of metabolites: 1H-indole-3-carboxaldehyde, 6-hydroxy-1H-indole-3-acetamide, phenylalanyl tryptophan, 1-methylguanosine, 2-ethoxy-5-methylpyrazine, p-hydroxybenzaldehyde, 5-(2-chlorophenyl)-3,4-dihydro-2H-pyrrole, methylthioadenosine, alanylisoleucine, anabsinthin, N-acetyl-DL-histidine monohydrate, N-methylnicotinamide, and others. The fifteen metabolites that were previously identified and calculated according to the differential quantitative value of the metabolite corresponding ratio exhibited fold-changes in the upregulated and downregulated potential biomarkers (phenylalanine tryptophan, phenylalanine, 5'-methylthioadenosine, anabsinthin, and N-methylnicotinamide) in combination with the area under the receiver operating characteristic curve>0.9. Conclusion:Phenylalanyl tryptophan, phenylalanylalanine, 5'-methylthioadenosine, anabsinthin, and N-methylnicotinamide may serve as potential metabolic markers to distinguish patients with cholestasis from healthy controls. N-methylnicotinamide, among them, is of great importance as a potential marker.

Objective To investigate the regulatory effect of miRNA-933 on the apoptosis and proliferation of human hepatic stellate cell line LX-2 and its mechanism.Methods Firstly,with human liver tissue for research,gene microarray technology was used to detect the differentially expressed genes in liver tissue between liver cirrhosis/chronic hepatitis B tissue and normal liver tissue,among which the significantly differentially expressed miRNAs were identified,and thus miRNA-933 was determined as the research object.Then,with the human hepatic stellate cell line LX-2 for research,miRNA-933 mimic and inhibitor(miRNA-933 siRNA)were used to construct the LX-2 models of overexpression and knockdown,and the cells transfected with mimic-NC(overexpression)or siRNA-NC(knockdown)were established as the negative control group.Quantitative real-time PCR and Western blot were used to measure the expression levels of miRNA-933 and activation biomarkers;techniques such as cell proliferation assay and flow cytometry were used to investigate the effect and mechanism of miRNA-933 on cell apoptosis,proliferation,and activation.The independent-samples t test was used for comparison of continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups,and Bonferroni correction was also performed.Results A total of 18 significantly differentially expressed miRNAs were obtained based on the results of gene microarray,among which miRNA-933 was significantly downregulated(P<0.05).After LX-2 cells were transfected with miRNA-933 mimic or siRNA,compared with the negative control group,miRNA-933 siRNA significantly downregulated the expression of miRNA-933(P=0.000 7),while miRNA-933 mimic significantly upregulated the expression of miRNA-933(P=0.000 3).Western blot and quantitative real-time PCR showed that miRNA-933 siRNA significantly upregulated the expression of collagen Ⅰ and α-SMA(P<0.001),while miRNA-933 mimic significantly inhibited the expression of collagen Ⅰ and α-SMA(P<0.05).Flow cytometry showed that compared with the negative control group,miRNA-933 siRNA significantly downregulated the apoptosis rate of LX-2 cells(P=0.031 9),and miRNA-933 mimic significantly upregulated the apoptosis rate of LX-2 cells(P=0.005 5).Western blot showed that compared with the negative control group,miRNA-933 siRNA could inhibit the expression of Caspase-3(P=0.006 7)and poly(ADP-ribose)polymerase-1(PARP-1)(P=0.003 0)and upregulate the expression of B-cell lymphoma-2(Bcl-2)in LX-2 cells(P=0.002 0),while miRNA-933 mimic could significantly upregulate the expression of Caspase-3(P=0.011 8)and PARP-1(P=0.049 5)and downregulated the expression of Bcl-2(P=0.002 1).Cell proliferation assay showed that compared with the negative control group,miRNA-933 siRNA could promote the proliferation of LX-2 cells(P=0.011 5),while on the contrary,miRNA-933 mimic could inhibit the proliferation of LX-2 cells(P=0.001 2).Western blot and quantitative real-time PCR showed that miRNA-933 siRNA significantly inhibited the expression of Kruppel-like factor 6(KLF6)and downregulated the expression of activating transcription factor 4(ATF4),activating transcription factor 3(ATF3),and C/EBP homologous protein(CHOP),while miRNA-933 mimic promoted the expression of the above proteins(all P<0.05).Conclusion This study shows that miRNA-933 may promote cell apoptosis and inhibit cell activation and proliferation by promoting the activation of the KLF6/ATF4/ATF3/CHOP/Bcl-2 signal axis in LX-2 cells.

Objective To investigate the effect and mechanism of hypericin on osteoporosis(OP)in rats with chronic obstructive pulmonary disease(COPD).Methods COPD combined with OP rat model was established by cigarette combined with bacteria.The rats were randomly divided into control group,model group(COPD combined with OP model was constructed),experimental-L group(50 mg·kg-1 hypericin was given by intragastric administration after constructing COPD combined with OP model),experimental-H group(100 mg·kg-1 hypericin was given intragastric administration after constructing COPD combined with OP model),positive group(subcutaneous injection of 16 U·kg-1 salmon calcitonin after constructing COPD combined with OP model);each group was given 12 rats for 90 days.The lung function of rats was detected by pulmonary function apparatus;bone mineral density(BMD)was detected by micro-computed tomography(CT);serum bone metabolism and inflammatory factors were detected by enzyme-linked immunosorbent assay(ELISA);Western blot assay was used to detect the relevant indicators of the pathway.Results The levels of forced vital capacity(FVC)in control group,model group,experimental-H group and positive group were(10.42±1.40),(4.10±0.60),(6.75±0.37),(4.18±0.33)mL,respectively;BMD levels were(0.31±0.04),(0.12±0.02),(0.28±0.03),(0.29±0.04)g·mm-3,respectively;bone alkaline phosphatase(BALP)levels were(200.04±20.03),(80.80±6.00),(148.16±14.23),(173.97±23.55)U·L1,respectively;interleukin-1β(IL-1β)levels were(122.60±8.70),(695.59±74.84),(422.41±44.86),(527.90±39.36)pg·mL-1,respectively;phosphorylated p38 mitogen-activated protein kinase(p-P38)protein expression levels were 0.99±0.11,0.36±0.05,0.79±0.08,0.36±0.04,respectively.Compared with the control group,the above indexes in the model group had statistical significance(all P＜0.05);the above indexes in experimental-H group were significantly different from those in model group(all P＜0.05).Conclusion Hypericin can inhibit inflammatory response,improve bone metabolism and biomechanics.

Aim To study the effects of high-fat diet on testicular germ cell apoptosis in mice through endoplasmic reticulum stress. Methods C57BL/6J male mice were assigned into normal group and high-fat diet group randomly, with six mice in each group. The mice in normal group or high-fat diet group were fed with regular or high-fat diet continuously for five months. The mice were weighed, anesthetized, and euthanized to collect testicular and epididymal tissue for analysis. The testicular tissue was weighed and their indices were calculated. Epididymal tissue was collected for semen analysis. The morphological alterations of testicular tissue were observed using hematoxylin-eosin ( HE ) staining. The apoptosis of germ cells was detected by TUNEL staining and the apoptotic indices were calculated. The expression levels of apoptosis and endoplasmic reticulum stress-related proteins in testicular tissue were detected by Western blot. The protein expression and localization of GRP78 in testicular tissue were further detected by immunofluorescence. Results The results showed that compared to the normal group, the high-fat diet group had a significant increase in body weight, a significant decrease in testicular index, sperm concentration, and sperm vability, loose arrangement of germ cells, significant thinning of the seminiferous epithelium, no significant change in the diameter of seminiferous tubules, a significant increase in germ cell apoptosis , with an increased apoptosis index, and significant increase in expression of Bax and cleaved-caspase-12,and a significant decrease in Bcl-2 protein expression. The expression levels of GRP78 , p-IREl, XBP1, and ATF6a proteins were significantly up-regulated, while p-PERK, p-eIF2a, ATF4 protein expression showed no significant changes. Immunofluorescence results further showed a significant increase in the expression of GRP78 protein in the testicular tissue,with no significant changes in the expression location. Conclusions High-fat diet can induce the apoptosis of mouse testicular germ cells, and the mechanism may be related to the activation of endoplasmic reticulum stress IRE1 and ATF6 signaling pathway.

COVID-19 has been prevalent for three years. The virulence of SARS-CoV-2 is weaken as it mutates continuously. However, elderly patients, especially those with underlying diseases, are still at high risk of developing severe infections. With the continuous study of the molecular structure and pathogenic mechanism of SARS-CoV-2, antiviral drugs for COVID-19 have been successively marketed, and these anti-SARS-CoV-2 drugs can effectively reduce the severe rate and mortality of elderly patients. This article reviews the mechanism, clinical medication regimens, drug interactions and adverse reactions of five small molecule antiviral drugs currently approved for marketing in China, so as to provide advice for the clinical rational use of anti-SARS-CoV-2 in the elderly.