ObjectiveThis paper aims to investigate the material basis of the anti-inflammatory efficacy and mechanism of action of Bushen Tongdu prescription （BSTDP）. MethodsThe chemical components of BSTDP and its blood-absorbed components in vivo were systematically identified by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry （UPLC-LIT-Orbitrap-MS）. Network pharmacology was employed to screen blood-absorbed bioactive components and potential targets of this formula. A protein-protein interaction （PPI） network of core targets was constructed to conduct enrichment analysis. Molecular docking was further utilized to verify the binding affinity between key components and targets. The inflammatory model was established and verified in vivo by using a transgenic zebrafish Tg （mpx： GFP）. At three days post-fertilization （3 dpf）， larvae of zebrafish were randomly assigned to blank group， model group， positive drug dexamethasone acetate group （75 μmol·L^-1）， and BSTDP groups with low， medium， and high doses （500， 1 000， and 2 000 mg·L^-1）. The distribution and quantity of neutrophils in the yolk sac region were observed under a fluorescence microscope. The mRNA expression levels of key genes in the toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor kappa-B （NF-κB） signaling pathway and inflammatory factors including interleukin （IL）-1β， IL-6， and tumor necrosis factor-α （TNF-α） were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）. ResultsA total of 120 chemical components were identified in BSTDP， among which 26 original components were confirmed by using serum pharmacochemical methods. A total of 227 common targets linking rheumatoid arthritis （RA） and the blood-absorbed components were screened by network pharmacology. It is suggested that pseudobrucine， vomicine， sinapine， rehmannioside， cinnamyl alcohol glycoside， and methylephedrine exert anti-inflammatory effects by acting on core targets including protein kinase B1 （Akt1）， signal transducer and activator of transcription 3 （STAT3）， tumor necrosis factor （TNF）， TLR4， mitogen-activated protein kinase 14 （MAPK14）， and phosphatidylinositol-4，5-bisphosphate 3-kinase catalytic subunit α （PIK3CA）， thereby modulating multiple signaling pathways such as TLR4 and NF-κB. In vivo verification in zebrafish demonstrates that the maximum tolerable concentration of Bushen Tongdu Formula is 2 000 mg·L^-1. Compared to those in the blank group， zebrafish in the model group showed a significantly higher number of neutrophils in the yolk sac region （P<0.01） and rising mRNA levels of TLR4， MyD88， NF-κB， TNF-α， IL-6， and IL-1β （P<0.01）. Compared to that in the model group， the number of neutrophils was significantly reduced in BSTDP groups with medium and high doses， as well as the dexamethasone acetate group （P<0.05， P<0.01）. There was no statistically significant difference in the low dose group. The mRNA expression levels of TLR4， MyD88， NF-κB， TNF-α， IL-6， and IL-1β were significantly down-regulated （P<0.05， P<0.01）. ConclusionThis paper identifies the material basis of the efficacy of BSTDP， demonstrating that the formula can exert an anti-inflammatory effect through the TLR4/MyD88/NF-κB signaling pathway. The results provide scientific experimental evidence for its further clinical application.

ObjectiveThis paper aims to investigate the material basis of the anti-inflammatory efficacy and mechanism of action of Bushen Tongdu prescription （BSTDP）. MethodsThe chemical components of BSTDP and its blood-absorbed components in vivo were systematically identified by using ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry （UPLC-LIT-Orbitrap-MS）. Network pharmacology was employed to screen blood-absorbed bioactive components and potential targets of this formula. A protein-protein interaction （PPI） network of core targets was constructed to conduct enrichment analysis. Molecular docking was further utilized to verify the binding affinity between key components and targets. The inflammatory model was established and verified in vivo by using a transgenic zebrafish Tg （mpx： GFP）. At three days post-fertilization （3 dpf）， larvae of zebrafish were randomly assigned to blank group， model group， positive drug dexamethasone acetate group （75 μmol·L^-1）， and BSTDP groups with low， medium， and high doses （500， 1 000， and 2 000 mg·L^-1）. The distribution and quantity of neutrophils in the yolk sac region were observed under a fluorescence microscope. The mRNA expression levels of key genes in the toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor kappa-B （NF-κB） signaling pathway and inflammatory factors including interleukin （IL）-1β， IL-6， and tumor necrosis factor-α （TNF-α） were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）. ResultsA total of 120 chemical components were identified in BSTDP， among which 26 original components were confirmed by using serum pharmacochemical methods. A total of 227 common targets linking rheumatoid arthritis （RA） and the blood-absorbed components were screened by network pharmacology. It is suggested that pseudobrucine， vomicine， sinapine， rehmannioside， cinnamyl alcohol glycoside， and methylephedrine exert anti-inflammatory effects by acting on core targets including protein kinase B1 （Akt1）， signal transducer and activator of transcription 3 （STAT3）， tumor necrosis factor （TNF）， TLR4， mitogen-activated protein kinase 14 （MAPK14）， and phosphatidylinositol-4，5-bisphosphate 3-kinase catalytic subunit α （PIK3CA）， thereby modulating multiple signaling pathways such as TLR4 and NF-κB. In vivo verification in zebrafish demonstrates that the maximum tolerable concentration of Bushen Tongdu Formula is 2 000 mg·L^-1. Compared to those in the blank group， zebrafish in the model group showed a significantly higher number of neutrophils in the yolk sac region （P<0.01） and rising mRNA levels of TLR4， MyD88， NF-κB， TNF-α， IL-6， and IL-1β （P<0.01）. Compared to that in the model group， the number of neutrophils was significantly reduced in BSTDP groups with medium and high doses， as well as the dexamethasone acetate group （P<0.05， P<0.01）. There was no statistically significant difference in the low dose group. The mRNA expression levels of TLR4， MyD88， NF-κB， TNF-α， IL-6， and IL-1β were significantly down-regulated （P<0.05， P<0.01）. ConclusionThis paper identifies the material basis of the efficacy of BSTDP， demonstrating that the formula can exert an anti-inflammatory effect through the TLR4/MyD88/NF-κB signaling pathway. The results provide scientific experimental evidence for its further clinical application.

Sensors have been widely recognized in matrix metalloproteinases(MMP)detection for their high sensitivity,simplicity,speed and easy in vivo and vitro detection.The design thought of existing sensors is based on the structure of MMP-9 molecule or the enzyme activity to output different signals in response to sensing.In this paper,we summarize the current research status and difficulties of MMP-9 sensing detection technology,and further discuss the development prospects of MMP sensor,which will provide a reference for the detection of MMP-9 and other good biological target molecules.

Objective:To evaluate the clinical efficacy and impact on quality of life of the biofeedback (BF) therapy combined with the Wenyang Yiqi Prescription and self-efficacy interventions in patients with functional defecation disorders (FDD).Methods:A prospective, randomized controlled trial design was employed. A total of 128 FDD patients were selected from the Pelvic Floor Center of the Department of Proctology, Suzhou Traditional Chinese Medicine Hospital Affiliated to Nanjing University of Chinese Medicine, between January 2021 and April 2023. These patients were randomly divided into two groups, with 64 cases in each group. Additionally, 41 historical control patients, who had received conventional BF training between June 2015 and January 2018 at the same center, were included for comparison. The historical control group received conventional BF therapy, the intervention group 1 combined BF therapy with self-efficacy interventions, and the intervention group 2 added the Wenyang Yiqi Prescription based on intervention group 1's treatment. All three groups underwent two treatment courses. Constipation symptom scores before and after treatment were performed; self-efficacy was assessed via the Self-Rated Abilities for Health Practices scale (SRAHP); health behaviors were evaluated using the Health-Promoting Lifestyle Profile (HPLP); anxiety and depression were evaluated through Self-Rating Anxiety Scale (SAS) and Self-Rating Depression Scale (SDS); quality of life was measured through SF-36. Clinical efficacy was evaluated based on constipation symptom scores.Results:During the treatment period, seven patients dropped out due to personal reasons, leaving 61 patients in intervention group 1 and 60 patients in intervention group 2 for efficacy analysis.　The total effective rates were 80.32% (49/61) in intervention group 1, 83.33% (50/60) in intervention group 2, and 56.10% (23/41) in the historical control group. The total effective rates of both intervention groups were significantly higher than those in the historical control group ( χ2=7.06, 9.15; P=0.029, 0.010, respectively). After treatment, intervention group 2 showed significantly lower constipation-related straining scores [1 (1, 2) vs. 2 (1, 2), Z=-4.51] compared to intervention group 1 ( P<0.05). Additionally, intervention group 2 had lower scores for straining [1 (1, 2) vs. 2 (1, 2), Z=-3.15] and defecation time [1 (0, 1) vs. 1 (1, 2), Z=-3.13] compared to the historical control group ( P<0.05). Psychological comfort efficacy (19.13 ± 2.51 vs. 16.98 ± 1.86, t=5.36), health responsibility efficacy (23.63 ± 4.69 vs. 22.59 ± 3.05, t=2.06), and overall SRAHP scores (75.98 ± 4.44 vs. 72.33 ± 5.16, t=4.17) were higher in intervention group 2 compared to intervention group 1 ( P<0.05). The HPLP scores (79.33 ± 11.13 vs. 72.80 ± 9.20, t=3.10) of intervention group 2 were higher than those of the historical control group ( P<0.05). Emotional functioning scores (75.98 ± 18.45 vs. 68.92 ± 20.58 and 68.55 ± 18.21, F=20.91) in intervention group 2 were higher than in both intervention group 1 and the historical control group ( P<0.05). Conclusion:The combination of the Wenyang Yiqi Prescription with BF therapy and self-efficacy interventions effectively improves the clinical symptoms and quality of life in FDD patients.

Objective To compare the efficacy of low-frequency and high-frequency repetitive transcranial magnetic stimulation(rTMS)in patients with tremor-dominant Parkinson's disease.Methods A total of 88 tremor-dominant Parkinson's disease patients who visited Suqian Hospital of Nanjing Drum Tower Hospital Group from January 2021 to June 2024 were randomly assigned to low-frequency rTMS group(treated by 1 Hz stimulation)and high-frequency rTMS group(treated by 5 Hz stimulation),with 44 patients in each group.Treatment was administered once a day,five times a week,for 4 weeks.Another 50 patients were enrolled as the control group.During the rTMS treatment,regular medications were continuously administrated.Before and after treatment,the following assessments were conducted:unified parkinson's disease rating scale part Ⅲ(UPDRS-Ⅲ),Parkinson's disease questionnaire-39(PDQ-39),timed up and go(TUG)test,tremor amplitude,tremor peak frequency,and cortical excitability(resting motion threshold[rMT]and active motion threshold[aMT]).Results After treatment,there were significant differences in the UPDRS-Ⅲ score,TUG time,PDQ-39 score,rMT and aMT values,and tremor amplitude between the two groups(P<0.05),and the high-frequency rTMS group was lower/shorter than the other two group.There was no significant difference in tremor peak frequency between the two groups(P>0.05).Conclusion Both low-frequency and high-frequency rTMS can significantly improve motor function and quality of life in tremor-dominant Parkinson's disease patients.High-frequency rTMS demonstrates better efficacy in improving various symptoms and provides a more effective treatment option for tremor-dominant Parkinson's disease.

Objective:To investigate the clinical, pathological, and molecular biological characteristics of gastric hepatoid adenocarcinoma (HAS) in order to provide reference for clinical treatment.Methods:Thirty-two patients diagnosed with hepatoid adenocarcinoma after radical gastrectomy for gastric cancer at Shanxi Cancer Hospital were included from January 2019 to December 2021. Immunohistochemistry, in situ hybridization, and next-generation sequencing (NGS) methods were used to analyze immune markers and molecular characteristics in the pathological tissues from 32 patients with HAS. Cox regression analysis and Kaplan-Meier method were used to analyze the prognostic factors of overall survival and disease-free survival.Results:Among the 32 patients with HAS, 26 were male, 6 were female; aged 28-77 years, with an median age 62.0 (53.8, 67.2) years. Fifteen cases of HAS were located in the cardia, 10 cases in the antrum, and 7 cases in the body of the stomach. The maximum diameter of the mass was 3-10 cm, and mainly ulcerative in gross. The immunohistochemistry and in situ hybridization results showed that the positive rates of AFP, SALLA4, and Glypican-3 were 68.8% (22/32), 68.8% (22/32), 78.1% (25/32), respectively; Seven patients had microsatellite status of dMMR. Two cases of HER2 gene amplification and 2 cases of EB virus positivity. The NGS results showed that HAS was often accompanied by multiple gene mutations, with 23 cases having ≥ 2 gene mutations and 6 cases having ≥10 gene mutations. The TP53 gene had the highest mutation frequency; 4 cases had genetic structural variations; 28 cases had copy number variation. In addition, there were 7 cases of MSI-H and 9 cases of TMB-H. Follow-up results showed that 12 cases died, 9 cases developed metastasis, and the shortest survival time was 5 months.Conclusions:Gastric HAS is a type of tumor with high invasiveness and poor prognosis. The combined detection of AFP, SALLA4 and Glypican-3 can improve the diagnostic rate of tumors. dMMR/MSI-H and TMB-H patients in HAS are significantly higher than those in ordinary gastric cancer, and the high frequency mutation genes in HAS are often accompanied by multiple potential therapeutic targets. Immunotherapy combined with chemotherapy and targeted therapy are expected to become the treatment direction of HAS.

Objective:To investigate the current status of treatment delay and analyze its influencing factors in patients with breast cancer-related lymphedema.Methods:Using convenience sampling, 218 patients with breast cancer-related lymphedema from The First Affiliated Hospital of Zhengzhou University between April 2024 and January 2025 were enrolled. The General Information Questionnaire, Lymphedema Self-Management Support Scale for Breast Cancer Survivors (LSMS-BCs), Brief Illness Perception Questionnaire for Breast Cancer-related Lymphedema (BIPQ-BCRL), Perceived Barriers to Health Care-Seeking Decision-Chinese (PBHSD-C), and Health Literacy Scale for Chronic Patients (HLSCP) were used to conduct a cross-sectional survey. Logistic regression identified predictors of treatment delay, with model fit assessed by the Hosmer-Lemeshow test. Discriminative ability was evaluated using receiver operating characteristic (ROC) curve analysis.Results:The study included 218 female BCRL patients, aged (58.31 ± 10.54) years. Among 218 patients, 76 experienced treatment delay, the incidence of treatment delay was 34.8% (76/218). Independent risk factors included junior high school education or below, no regular arm circumference measurement, low self-management support scores, low illness perception scores, high perceived barriers to healthcare-seeking scores, and low health literacy scores (Wald χ2 values were 7.75-15.15, all P<0.05). The Hosmer-Lemeshow test indicated good model fit ( χ2=6.21, P>0.05). The combined predictive model demonstrated significantly better discrimination than individual factors, the area under the curve of ROC was 0.846 ( P<0.01). Conclusions:The incidence of treatment delay is relatively high among breast cancer-related lymphedema patients. Nursing staff should pay special attention to patients with a junior high school education or below, no regular arm circumference measurement, low LSMS-BCs scores, low BIPQ-BCRL scores, high PBHSD-C scores and low HLSCP scores, implement timely interventions to reduce treatment delay in lymphedema patients.

Objective·To explore the potential role of SUMO-specific protease 1(SENP1)in ferroptosis.Methods·The Cancer Genome Atlas(TCGA)database was used to analyze the correlation between the expression levels of SENP1 and the ferroptosis-related genes,acyl-CoA synthetase long chain family member 4(ACSL4)and glutathione peroxidase 4(GPX4).Ferroptosis in human fibrosarcoma HT1080 cells,murine fibrosarcoma MCA-205 cells,and human embryonic kidney 293T cells was induced by RAS-selective lethal 3(RSL3).Quantitative real-time PCR(RT-qPCR)and Western blotting were used to detect the expression of SENP1.In 293T cells,immunoprecipitation-mass spectrometry was used to investigate the interacting proteins of SENP1 in the process of ferroptosis.The Flag-SENP1 plasmid was transiently transfected into 293T cells,and the overexpression efficiency of SENP1,along with the expression levels of ferroptosis-related genes ACSL4 and GPX4,was assessed by RT-qPCR and Western blotting.Results·TCGA database analysis showed that the expression of SENP1 was positively correlated with ACSL4 and negatively correlated with GPX4 in most tumor tissues.RT-qPCR and Western blotting showed that the expression level of SENP1 was significantly down-regulated in RSL3-treated HT1080,MCA-205,and 293T cells.Immunoprecipitation-mass spectrometry showed that SENP1 enriched SUMO molecules in the process of ferroptosis.Western blotting showed that the level of ACSL4 protein increased after SENP1 overexpression,and there was no significant change in the level of GPX4 protein.RT-qPCR showed that after SENP1 overexpression,there was no significant change in the mRNA levels of ACSL4 and GPX4.Conclusion·SENP1 gene expression is downregulated during ferroptosis,and may regulate the stability of ferroptosis-related protein ACSL4.

Objective:To establish a detection method for fatty acid composition and methoxylaniline value in pharmaceutical excipient castor oil.Methods:The detection of fatty acid composition involves pre-test of the sam-ple using the methanol sodium methylation method,followed by direct injection analysis using gas chromatography,and finally calculating the content of each fatty acid composition using the area normalization method with correction factors.The detection of methoxylaniline value is achieved by dissolving and diluting the sample with isooctane:isopropanol(8∶2,volume ratio),reacting with 4-methoxylaniline,measuring the absorbance at a wavelength of 350 nm,and calculating the methoxylaniline value.Results:Under the composition of fatty acids,the 8 kinds of fatty acids have good separation degree.The methyl ricinoleate and methyl stearate have good linear relationship in the range of 0.1-5.0 mg·mL-1.The repeatabilityand intermediate precision(RSD)ofthe determination results of each fatty acid in the sample are all less than 5%.It is also found that only α-type of linolenic acid is present in castor oil,without γ-type.In the determination of methoxyaniline value,the blank solvent does not interfere with the determination of the sample,and the repeatability RSD is 3.6%.Conclusion:The detection method for fatty acid composition and meth-oxylaniline value established in this article is accurate and reliable,and can be used for the detection of fatty acid composition and methoxylaniline value in pharmaceutical excipient castor oil.

OBJECTIVE: To observe the effects of electroacupuncture (EA) at "Shenting" (GV24) and "Baihui" (GV20) on mitochondrial autophagy in hippocampal neurons and silent information regulator sirtuin 1 (Sirt1)/forkhead box O3 (FOXO3)/PTEN-inducible kinase 1 (PINK1)/Parkin pathway in rats with learning-memory impairment after cerebral ischemia reperfusion injury. METHODS: A total of 35 male SD rats were randomly divided into a sham operation group (9 rats) and a modeling group (26 rats). In the modeling group, middle cerebral artery occlusion method was used to establish the middle cerebral artery ischemia-reperfusion (MCAO/R) model, and 18 rats of successful modeling were randomly divided into a model group and an EA group, 9 rats in each one. EA was applied at "Shenting" (GV24) and "Baihui" (GV20) in the EA group, 30 min a time, once a day for 14 days. After modeling and on 7th and 14th days of intervention, neurologic deficit score was observed; the learning-memory ability was detected by Morris water maze test; the morphology of neurons in CA1 area of hippocampus was detected by Nissl staining; the mitochondrial morphology was observed by transmission electron microscopy; the protein expression of Beclin-1, microtubule-associated protein 1 light chain 3B (LC3B), P62, Sitrt1, FOXO3, PINK1 and Parkin was detected by Western blot. RESULTS: After modeling, the neurologic deficit scores in the model group and the EA group were higher than that in the sham operation group (P<0.001); on 7th and 14th days of intervention, the neurologic deficit scores in the model group were higher than those in the sham operation group (P<0.001), the neurologic deficit scores in the EA group were lower than those in the model group (P<0.05, P<0.01). After modeling, the escape latency in the model group and the EA group was prolonged compared with that in the sham operation group (P<0.001); on 9th-13th days of intervention, the escape latency in the model group was prolonged compared with that in the sham operation group (P<0.001), the escape latency in the EA group was shortened compared with that in the model group (P<0.05, P<0.01, P<0.001). The number of crossing plateau in the model group was less than that in the sham operation group (P<0.001); the number of crossing plateau in the EA group was more than that in the model group (P<0.05). In the model group, in CA1 area of hippocampus, the number of neurons was less, with sparse arrangement, nuclear fixation, deep cytoplasmic staining, and reduction of Nissl substance; the morphology of mitochondrion was swollen, membrane structure was fragmented, and autophagic lysosomes were formed. Compared with the model group, in the EA group, in CA1 area of hippocampus, the number of neurons was increased, the number of cells of abnormal morphology was decreased, and the number of Nissl substance was increased; the morphology of mitochondrion was more intact and the number of autophagic lysosomes was increased. Compared with the sham operation group, in the model group, the protein expression of Beclin-1, FOXO3, PINK1, Parkin and the LC3BⅡ/Ⅰ ratio in hippocampus were increased (P<0.01, P<0.001), while the protein expression of P62 was decreased (P<0.05). Compared with the model group, in the EA group, the protein expression of Beclin-1, Sirt1, FOXO3, PINK1, Parkin and the LC3BⅡ/Ⅰratio in hippocampus were increased (P<0.001, P<0.01), while the protein expression of P62 was decreased (P<0.001). CONCLUSION EA at "Shenting" (GV24) and "Baihui" (GV20) can relieve the symptoms of neurological deficits and improve the learning-memory ability in MCAO/R rats, its mechanism may relate to the modulation of Sirt1/FOXO3/PINK1/Parkin pathway and the enhancement of mitochondrial autophagy.
Animals ; Electroacupuncture ; Male ; Rats, Sprague-Dawley ; Rats ; Forkhead Box Protein O3/genetics* ; Reperfusion Injury/metabolism* ; Ubiquitin-Protein Ligases/genetics* ; Brain Ischemia/complications* ; Mitochondria/genetics* ; Autophagy ; Protein Kinases/genetics* ; Sirtuin 1/genetics* ; Humans ; Memory Disorders/psychology* ; Signal Transduction