Objective To investigate whether Danshen injection (DSI) can protect human brain organoids from oxidative stress damage by regulating the Nrf2 signaling pathway and its mechanism. Methods A Nrf2 plasmid was constructed, lentivirus was packaged and transfected into human induced pluripotent stem cell (hiPSC) to form a stable cell line. Brain organoids with Nrf2 overexpression (LV-Nrf2 group) and knockdown (sh-Nrf2 group) were constructed using kit. The transfection efficiency was verified by real-time fluorescence quantitative polymerase chain reaction. An oxidative stress injury model was established by inducing hydrogen peroxide (H₂O₂). Intervention was performed either in combination or without DSI pre-treatment. Cell viability was detected by Cell Counting Kit-8 (CCK-8), and the expression levels of Nrf2, malondialdehyde (MDA), interleukin (IL)-1β, sex-determining region Y-box protein 2 (SOX2) and neuronal-specific nuclear protein (NeuN) were detected by Western blot and immunofluorescence staining. Total RNA from human brain organoids was extracted for transcriptome sequencing and analysis. Results LV-Nrf2 group and sh-Nrf2 group brain organoid models were successfully constructed. The CCK-8 results determined that the oxidative stress model was established with 32 mmol/L H₂O₂ intervention conditions. The fluorescence expression of Nrf2 in LV-Nrf2 group was higher than that in control group, and the fluorescence expressions of MDA and IL-1β were lower than those in control group; sh-Nrf2 group showed a damage phenotype of low Nrf2 expression, high MDA and IL-1β expression (all P < 0.05). Compared with H₂O₂ group, the Nrf2 signal in DSI + H₂O₂ group was stronger, and the levels of MDA and IL-1β were lower. Among them, the protective effect of LV-Nrf2 was the most significant. H₂O₂ treatment could down-regulate the protein levels of Nrf2, SOX2 and NeuN, and DSI intervention could effectively reverse the downward trend of these indicators. Sh-Nrf2 could weaken the recovery effect of DSI on SOX2 and NeuN, while LV-Nrf2 could synergistically enhance the regulatory effect of DSI. Conclusions DSI may antagonize oxidative stress damage in human brain organoids through the Nrf2 signaling pathway, and maintaining the stability of the neural cell lineage is its core therapeutic characteristic.

Objective:To translate the Short Self-Regulation Questionnaire (SSRQ) into Chinese, and test its reliability and validity in breast cancer patients.Methods:Using the Brislin translation model, cross-cultural adaptation was conducted by combining expert consultation and a pre-survey on SSRQ. From August to December 2023, 324 breast cancer patients from Department of Breast Surgery of Peking University First Hospital were selected by convenience sampling to test the reliability and validity of SSRQ.Results:A total of 324 questionnaires were distributed and 307 valid questionnaires were recovered, with an effective recovery rate of 94.75% (307/324). The final questionnaire consisted of 30 items. The content validity index at the item level ranged from 0.80 to 1.00, and the content validity index at the scale level was 0.97. The predictive validity result showed a good model fit. The Cronbach's α coefficient for the questionnaire was 0.90, the folded reliability coefficient was 0.85, and the retest reliability coefficient was 0.79.Conclusions:SSRQ has good reliability and validity in breast cancer patients and has value for clinical promotion.

Objective:To explore the effects of Ziyu Ointment on granulation tissue,TGF-β and downstream Smad protein in the process of wound healing in rats by constructing a rat model of back trauma.Methods:A rat model of back trauma was constructed,and 100 SD male rats were separated into healthy group,back trauma group,Ziyu Ointment treatment group,petrolatum treatment group,Ziyu Ointment+TGF-β1 activation group,with 20 per group.Wound formation surgery was performed,and wound healing of rats in each group was recorded at 5th,10th,and 15th day after operation;the levels of IL-6,IL-1β and TNF-α in the serum of rats in each group were detected;the granulation tissue formed by wound healing at 15th day was selected for HE and Masson staining,the proportion of blood vessel area and collagen area of granulation tissue of rats in each group were analyzed and calculated;the mRNA expression levels of TGF-β1 and Smad3 in granulation tissues of rats in each group were detected by fluorescence quantitative PCR;the expression levels of TGF-β1,Smad3 and p-Smad3 in granulation tissues of rats in each group were detected by Western blot.Results:The healthy group rats had complete epidermal tissue;the levels of serum IL-6,IL-1β,TNF-α,the expression of TGF-β1 mRNA and protein in the wound granulation tissue,and the phosphorylation of Smad3 in the back trauma group were obviously higher than the healthy group(P<0.05);compared with back trauma group,the wound healing rate,proportion of collagen area,and propor-tion of blood vessel area in Ziyu Ointment treatment group and petrolatum treatment group were obviously increased,the serum IL-6,IL-1β,TNF-α contents,TGF-β1 mRNA and protein expression,and Smad3 phosphorylation were obviously decreased(P<0.05);compared with Ziyu Ointment treatment group,the petrolatum treatment group,Ziyu Ointment+TGF-β1 activation group had obviously lower wound healing rate,collagen area percentage,and blood vessel area percentage,and obviously lower serum IL-6,IL-1β,TNF-α contents,TGF-β1 mRNA and protein expression and Smad3 phosphorylation(P<0.05).Conclusion:Ziyu Ointment can inhibit the expression of related genes and proteins by regulating the TGF-β/Smad signal pathway,thereby alleviating inflammation and promoting wound healing.

Objective To explore the effects of Dushu pills on cognitive ability and mitochondrial dynamics related proteins in mouse model of Alzheimer's disease(AD)based on network pharmacology.Methods The corresponding targets of Dushu pills and its related targets with AD were predicted using TCMSP database.The intersection targets of Dushu pills and AD were obtained by Venny website.The protein interaction network and drug-disease-target network were mapped using String database and Cytoscape software,respectively.GO and KEGG enrichment analysis were performed for intersection targets using David database.The mouse model of AD was established by injecting Aβ25-35 into bilateral ventricles.The learning and memory ability of the mice was detected by behavioral tests,the mitochondrial damage of neurons in the hippocampus was observed by transmission electron microscopy,and the expression of proteins related to mitochondrial dynamics was detected by Western blot.Results A total of 311 intersection targets related to drugs and diseases were screened out from the database.GO and KEGG analysis showed that the relevant targets were concentrated in mitochondria and other components,and concentrated in the pathways of ATP binding and positive regulation of MAPK activity.Compared with the normal group,the learning and memory ability of the model group was decreased(P<0.05),mitochondrial ridges appeared swelling and fracture(P<0.0001),decreased expression of MAPK,Mfn2 and OPA1 proteins in hippocampus(P<0.01),the expression of DRP1 protein increased(P<0.01).Compared with the model group,the learning and memory ability of mice in the medium and high dose groups of Dushu pills was improved(P<0.05),mitochondrial damage was significantly improved(P<0.01),increased expression of MAPK,Mfn2 and OPA1 in the hippocampus(P<0.01),DRP1 protein expression decreased(P<0.01).Conclusion Dushu pills can reduce mitochondrial damage,maintain mitochondrial homeostasis,and improve the cognitive and memory ability of AD mice.

Objective To construct the respiratory rehabilitation calisthenics for school-age children with asthma,and evaluate its effects,in order to provide a guidance for the scientific and effective implementation of respiratory rehabilitation in clinical work.Methods By convenience sampling method,the school-age children with asthma treated in the respiratory outpatient department of a tertiary specialized children's hospital in Chongqing from De-cember 2023 to February 2024 were selected.The children were randomly divided into an experimental group and the control group(25 of each group)by a random number table.Both groups were given routine asthma drug in-halation treatment and health education.Additionally,the experimental group received respiratory rehabilitation calis-thenics training and the control group received conventional aerobic exercise training.After 3-month intervention,the effects of adherence to respiratory rehabilitation training,lung functions,exercise capacity,inspiratory muscle strength and occurrence of adverse events were evaluated between 2 groups.Results 24 children of each group completed the study.After 3-month intervention,the adherence to respiratory rehabilitation training,exercise capacity and in-spiratory muscle strength were improved in the experimental group compared with those in the control group,and all differences were statistically significant(P＜0.05).There were no adverse events in both groups.Conclusion The respiratory rehabilitation calisthenics for school-age children with asthma constructed in this study was scientif-ic,feasible and safe,and could improve exercise capacity and inspiratory muscle strength for children with asthma.

Objective:To investigate the influences of asiaticoside(AS)on inflammatory response and oxidative stress in rats with mesangial proliferative glomerulonephritis(MsPGN)by regulating the rat sarcoma(Ras)/extracellular signal regulated kinase(ERK)/mitogen activated protein kinase(MAPK)signaling pathway.Methods:Sixty SPF SD rats were randomly divided into sham operation group(Sham group),MsPGN model group(Model group),low-dose AS group(AS-L group,40 mg/kg),high-dose AS group(AS-H group,80 mg/kg),high-dose AS+Ras/ERK/MAPK signaling pathway activator group(AS-H+anisomycin group,80 mg/kg AS+10 mg/kg anisomycin),with 12 rats in each group.Rat model of MsPGN was established by modified serum immunization.The 24 h urine protein content of rats in each group was detected by Coomassie brilliant blue method.HE staining and PSA staining were used to observe the pathological changes of renal tissue of rats in each group.TUNEL staining was used to detect apoptosis of rat glomerular cells in each group.Levels of serum creatinine(Scr),superoxide dismutase(SOD),blood urea nitrogen(BUN),malondialdehyde(MDA),and levels of IL-18,IL-1β,TNF-α,SOD and MDA in renal tissues were measured by ELISA.Protein levels of Ras,p-ERK1/2,ERK1/2,MAPK and p-MAPK in renal tissues of rats in each group were detected by Western blot.Results:①AS alleviated inflammatory reaction and oxidative stress injury in MsPGN rats.Compared with Sham group,24 h urine protein content,glomerular cell apoptosis rate,serum Scr,BUN,MDA levels,renal tissue IL-18,IL-1β,TNF-α,MDA levels in model group were significantly increased(P<0.05),while SOD level in serum and renal tissue was significantly decreased(P<0.05).HE staining showed that the glomerulus of rats had severe mesangial hyperplasia,a large amount of matrix deposition,and the basement membrane was significantly thickened.Compared with model group,24 h urine protein content,glomerular cell apoptosis rate,serum Scr,BUN,MDA levels,renal tissue IL-18,IL-1β,TNF-α,MDA levels in AS-H group rats were significantly decreased(P<0.05),while SOD level in serum and renal tissue was significantly increased(P<0.05),HE staining showed alleviated mesangial hyperplasia injury.②AS downregulated the Ras/ERK/MAPK signaling pathway.Compared with Sham group,expression of Ras protein and phosphorylation level of ERK1/2 and MAPK in Model group were significantly increased(P<0.05);compared with Model group,expression of Ras protein,phosphory-lation level of ERK1/2 and MAPK in AS-H group were significantly decreased(P<0.05),and anisomycin reduced the inhibitory effect of AS on inflammatory response and oxidative stress injury in MsPGN rats(P<0.05).Conclusion:AS may reduce the inflammatory re-action and oxidative stress injury in MsPGN rats by down-regulating Ras/ERK/MAPK signaling pathway.

Objective:To investigate the influences of asiaticoside(AS)on inflammatory response and oxidative stress in rats with mesangial proliferative glomerulonephritis(MsPGN)by regulating the rat sarcoma(Ras)/extracellular signal regulated kinase(ERK)/mitogen activated protein kinase(MAPK)signaling pathway.Methods:Sixty SPF SD rats were randomly divided into sham operation group(Sham group),MsPGN model group(Model group),low-dose AS group(AS-L group,40 mg/kg),high-dose AS group(AS-H group,80 mg/kg),high-dose AS+Ras/ERK/MAPK signaling pathway activator group(AS-H+anisomycin group,80 mg/kg AS+10 mg/kg anisomycin),with 12 rats in each group.Rat model of MsPGN was established by modified serum immunization.The 24 h urine protein content of rats in each group was detected by Coomassie brilliant blue method.HE staining and PSA staining were used to observe the pathological changes of renal tissue of rats in each group.TUNEL staining was used to detect apoptosis of rat glomerular cells in each group.Levels of serum creatinine(Scr),superoxide dismutase(SOD),blood urea nitrogen(BUN),malondialdehyde(MDA),and levels of IL-18,IL-1β,TNF-α,SOD and MDA in renal tissues were measured by ELISA.Protein levels of Ras,p-ERK1/2,ERK1/2,MAPK and p-MAPK in renal tissues of rats in each group were detected by Western blot.Results:①AS alleviated inflammatory reaction and oxidative stress injury in MsPGN rats.Compared with Sham group,24 h urine protein content,glomerular cell apoptosis rate,serum Scr,BUN,MDA levels,renal tissue IL-18,IL-1β,TNF-α,MDA levels in model group were significantly increased(P<0.05),while SOD level in serum and renal tissue was significantly decreased(P<0.05).HE staining showed that the glomerulus of rats had severe mesangial hyperplasia,a large amount of matrix deposition,and the basement membrane was significantly thickened.Compared with model group,24 h urine protein content,glomerular cell apoptosis rate,serum Scr,BUN,MDA levels,renal tissue IL-18,IL-1β,TNF-α,MDA levels in AS-H group rats were significantly decreased(P<0.05),while SOD level in serum and renal tissue was significantly increased(P<0.05),HE staining showed alleviated mesangial hyperplasia injury.②AS downregulated the Ras/ERK/MAPK signaling pathway.Compared with Sham group,expression of Ras protein and phosphorylation level of ERK1/2 and MAPK in Model group were significantly increased(P<0.05);compared with Model group,expression of Ras protein,phosphory-lation level of ERK1/2 and MAPK in AS-H group were significantly decreased(P<0.05),and anisomycin reduced the inhibitory effect of AS on inflammatory response and oxidative stress injury in MsPGN rats(P<0.05).Conclusion:AS may reduce the inflammatory re-action and oxidative stress injury in MsPGN rats by down-regulating Ras/ERK/MAPK signaling pathway.

Objective:To explore the effects of Ziyu Ointment on granulation tissue,TGF-β and downstream Smad protein in the process of wound healing in rats by constructing a rat model of back trauma.Methods:A rat model of back trauma was constructed,and 100 SD male rats were separated into healthy group,back trauma group,Ziyu Ointment treatment group,petrolatum treatment group,Ziyu Ointment+TGF-β1 activation group,with 20 per group.Wound formation surgery was performed,and wound healing of rats in each group was recorded at 5th,10th,and 15th day after operation;the levels of IL-6,IL-1β and TNF-α in the serum of rats in each group were detected;the granulation tissue formed by wound healing at 15th day was selected for HE and Masson staining,the proportion of blood vessel area and collagen area of granulation tissue of rats in each group were analyzed and calculated;the mRNA expression levels of TGF-β1 and Smad3 in granulation tissues of rats in each group were detected by fluorescence quantitative PCR;the expression levels of TGF-β1,Smad3 and p-Smad3 in granulation tissues of rats in each group were detected by Western blot.Results:The healthy group rats had complete epidermal tissue;the levels of serum IL-6,IL-1β,TNF-α,the expression of TGF-β1 mRNA and protein in the wound granulation tissue,and the phosphorylation of Smad3 in the back trauma group were obviously higher than the healthy group(P<0.05);compared with back trauma group,the wound healing rate,proportion of collagen area,and propor-tion of blood vessel area in Ziyu Ointment treatment group and petrolatum treatment group were obviously increased,the serum IL-6,IL-1β,TNF-α contents,TGF-β1 mRNA and protein expression,and Smad3 phosphorylation were obviously decreased(P<0.05);compared with Ziyu Ointment treatment group,the petrolatum treatment group,Ziyu Ointment+TGF-β1 activation group had obviously lower wound healing rate,collagen area percentage,and blood vessel area percentage,and obviously lower serum IL-6,IL-1β,TNF-α contents,TGF-β1 mRNA and protein expression and Smad3 phosphorylation(P<0.05).Conclusion:Ziyu Ointment can inhibit the expression of related genes and proteins by regulating the TGF-β/Smad signal pathway,thereby alleviating inflammation and promoting wound healing.

Objective To explore the effects of Dushu pills on cognitive ability and mitochondrial dynamics related proteins in mouse model of Alzheimer's disease(AD)based on network pharmacology.Methods The corresponding targets of Dushu pills and its related targets with AD were predicted using TCMSP database.The intersection targets of Dushu pills and AD were obtained by Venny website.The protein interaction network and drug-disease-target network were mapped using String database and Cytoscape software,respectively.GO and KEGG enrichment analysis were performed for intersection targets using David database.The mouse model of AD was established by injecting Aβ25-35 into bilateral ventricles.The learning and memory ability of the mice was detected by behavioral tests,the mitochondrial damage of neurons in the hippocampus was observed by transmission electron microscopy,and the expression of proteins related to mitochondrial dynamics was detected by Western blot.Results A total of 311 intersection targets related to drugs and diseases were screened out from the database.GO and KEGG analysis showed that the relevant targets were concentrated in mitochondria and other components,and concentrated in the pathways of ATP binding and positive regulation of MAPK activity.Compared with the normal group,the learning and memory ability of the model group was decreased(P<0.05),mitochondrial ridges appeared swelling and fracture(P<0.0001),decreased expression of MAPK,Mfn2 and OPA1 proteins in hippocampus(P<0.01),the expression of DRP1 protein increased(P<0.01).Compared with the model group,the learning and memory ability of mice in the medium and high dose groups of Dushu pills was improved(P<0.05),mitochondrial damage was significantly improved(P<0.01),increased expression of MAPK,Mfn2 and OPA1 in the hippocampus(P<0.01),DRP1 protein expression decreased(P<0.01).Conclusion Dushu pills can reduce mitochondrial damage,maintain mitochondrial homeostasis,and improve the cognitive and memory ability of AD mice.

Objective:To translate the Short Self-Regulation Questionnaire (SSRQ) into Chinese, and test its reliability and validity in breast cancer patients.Methods:Using the Brislin translation model, cross-cultural adaptation was conducted by combining expert consultation and a pre-survey on SSRQ. From August to December 2023, 324 breast cancer patients from Department of Breast Surgery of Peking University First Hospital were selected by convenience sampling to test the reliability and validity of SSRQ.Results:A total of 324 questionnaires were distributed and 307 valid questionnaires were recovered, with an effective recovery rate of 94.75% (307/324). The final questionnaire consisted of 30 items. The content validity index at the item level ranged from 0.80 to 1.00, and the content validity index at the scale level was 0.97. The predictive validity result showed a good model fit. The Cronbach's α coefficient for the questionnaire was 0.90, the folded reliability coefficient was 0.85, and the retest reliability coefficient was 0.79.Conclusions:SSRQ has good reliability and validity in breast cancer patients and has value for clinical promotion.