Exploring the molecular mechanism of Epimedium brevicornu Maxim.in treating breast cancer via network pharmacology and in vitro experiments

Objective:To evaluate the therapeutic effects of Epimedium brevicornu Maxim.(EBM,Yin Yang Huo)on breast cancer using network pharmacology and in vitro validation.It also aimed to explore the novel targets and mechanisms of EBM in the treatment of breast cancer to facilitate the discovery of new drugs and their clinical application.Methods: Network pharmacology was used to identify and screen the components and targets of EBM for breast cancer treatment.Molecular docking was further screened the effective components and targets of EBM.Wound-healing assays and flow cytometry analysis were used to detect the ability of two compounds to intervene in the migration and apoptosis of MDA-MB-231 cells,and their mechanism of action was further explored using western blotting experiments.Results: EBM contained 19 active components.Among them wereβ-anhydroicaritin(Anhy)and isoliquiritigenin(Iso),which were selected for in vitro experiments.Treatment resulted in a dose-dependent suppression of MDA-MB-231 cell viability,with an IC_(50) of 23.73μmol/L for Iso and 21.28μmol/L for Anhy.In the wound healing assay,cells in Anhy and Iso groups exhibited considerable inhibition of migration at 48 h.In flow cytometry analysis,treatment with Iso(20μmol/L)for 96 h resulted in significantly higher levels of both early and late apoptosis in the Iso group than that in the control group(P=.004 and P=.014,respectively).Additionally,both Iso(20μmol/L)and Anhy(10 and 20μmol/L)induced cell necrosis at 96 h.Western blotting revealed that Anhy and Iso increased the expression of Bax and TBK1/NAK.Conclusion: These findings suggested that Anhy and Iso,the two components of EBM,inhibit MDA-MB-231 cell proliferation and migration of and induce their apoptosis,providing substantial support for future studies on breast cancer.

Fang-Xia-Dihuang decoction inhibits breast cancer progression induced by psychological stress via down-regulation of PI3K/AKT and JAK2/STAT3 pathways:An in vivo and a network pharmacology assessment

Background:The development and prognosis of breast cancer are intricately linked to psychological stress.In addition,depression is the most common psychological comorbidity among breast cancer survivors,and reportedly,Fang-Xia-Dihuang decoction(FXDH)can effectively manage depression in such patients.However,its pharmacological and molecular mechanisms remain obscure.Methods:Public databases were used for obtaining active components and related targets.Main active components were further verified by ultra-high-performance liquid chromatography-high-resolution mass spectrometry(UPLC-HRMS).Protein–protein interaction and enrichment analyses were taken to predict potential hub targets and related pathways.Molecule docking was used to understand the interactions between main compounds and hub targets.In addition,an animal model of breast cancer combined with depression was established to evaluate the intervention effect of FXDH and verify the pathways screened by network pharmacology.Results:174 active components of FXDH and 163 intersection targets of FXDH,breast cancer,and depression were identified.Quercetin,methyl ferulate,luteolin,ferulaldehyde,wogonin,and diincarvilone were identified as the principal active components of FXDH.Protein–protein interaction and KEGG enrichment analyses revealed that the phosphoinositide-3-kinase–protein kinase B(PI3K/AKT)and Janus kinase/signal transducer and activator of transcription(JAK2/STAT3)signaling pathways played a crucial role in mediating the efficacy of FXDH for inhibiting breast cancer progression induced by depression.In addition,in vivo experiments revealed that FXDH ameliorated depression-like behavior in mice and inhibited excessive tumor growth in mice with breast cancer and depression.FXDH treatment downregulated the expression of epinephrine,PI3K,AKT,STAT3,and JAK2 compared with the control treatment(p<0.05).Molecular docking verified the relationship between the six primary components of FXDH and the three most important targets,including phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha(PIK3CA),AKT,and STAT3.Conclusion:This study provides a scientific basis to support the clinical application of FXDH for improving depression-like behavior and inhibiting breast cancer progression promoted by chronic stress.The therapeutic effects FXDH may be closely related to the PI3K/AKT and JAK2/STAT3 pathways.This finding helps better understand the regulatory mechanisms underlying the efficacy of FXDH.

Current status of outcome reporting in randomized controlled trials of traditional Chinese medicine for mammary gland hyperplasia: A systematic review

Objective:To assess outcome indicators in clinical trials and provide a reference for establishing a core outcome set to treat hyperplasia of mammary gland(HMG)with traditional Chinese medicine(TCM).Methods:Eight online databases were searched from their inception to December 31,2022,to assess outcomes reported in randomized controlled trials(RCTs)of HMG treated with TCM.The quality of the included studies was assessed according to the Cochrane Risk of Bias Assessment Tool.All outcomes were extracted,classified,and described.Results:A total of 8249 articles were initially retrieved.Of these,70 articles were eligible and involved 10618 participants with HMG.A total of 17 outcome indicators with a frequency of 271 times were involved and were collected according to six outcome domains.Conclusions:The core outcomes of RCTs of HMG treated with TCM are large and divergent.There are problems in evaluation standards,primary and secondary outcomes,TCM characteristic indicators,long-term prognosis,and standardization of reporting.It is recommended to strengthen the trial design and actively construct the core outcome sets with TCM characteristics for HMG.

Bloodletting puncture and cupping as an adjuvant therapy for breast cancer-related lymphedema in female adults: A non-randomized controlled pragmatic trial

Objective:To evaluate the effectiveness of bloodletting puncture and cupping in relieving breast cancer-related lymphedema.Methods:We conducted a non-randomized controlled study at the Galactophore Department of the 3rd Affiliated Hospital of Beijing University of Chinese Medicine from March 2013 to December 2014.Seventy-five patients undergoing treatment for breast cancer-related lymphedema were divided into two groups in accordance with the patients' treatment choices:the treatment group (n =50) underwent bloodletting puncture and cupping every 5 days for 15 min/session (one session per day) combined with exercise training once a day for 30 minutes;the control group (n =25) underwent the same exercise training alone.Evaluation indexes were measured before and after treatment;these included arm circumference (at the wrist crease,10 cm distal to the wrist crease,the elbow crease,and 10 cm distal to the elbow crease)and visual analogue scale (VAS) score for pain.The safety of the treatment was also evaluated.Results:The treatment period was 50 days for all 75 patients.At the end of the treatment period,the mean reduction in arm circumference was 1.21 (0.65) cm the treatment group,and 0.58 (0.78) cm in the control group.Compared with the control group,the treatment group had a significantly greater reduction in arm circumference (P =.03) and a significantly lower VAS score for pain (P =.01).No patient in either group experienced any adverse events.Conclusion:Bloodletting puncture and cupping effectively reduces arm circumference and relieves upper limb pain in patients with breast cancer-related lymphedema.Our results provide sufficient basis for exploring cupping as long-term therapy for managing breast cancer-related lymphedema.

Accuracy of glomerular filtration rate estimation equations in patients with hematopathy

Renal dysfunction is a common side-effect of chemotherapeutic agents in patients with hematopathy. Although broadly used, glomerular filtration rate(GFR) estimation equations were not fully validated in this specific population. Thus, this study was designed to further assess the accuracy of various GFR equations, including the newly 2012 CKD-EPI equations. Referring to ^(99m)Tc-DTPA clearance method, three Scr-based(MDRD, Peking, and CKD-EPI_(Scr)), three Scys C-based(Steven 1, Steven 2, and CKD-EPI_(Scys C)), and three Scr-Scys C combination based(Ma, Steven 3, and CKD-EPI_(Scr-Scys C)) equations were included. Bias, P_(30), and misclassification rate were applied to compare the applicability of the selected equations. A total of 180 Chinese hematological patients were enrolled.Mean bias, absolute mean bias, P_(30), misclassification rate and Bland-Altman plots of the CKD-EPI_(Scr-Scys C) equation were 7.90 mL/minute/1.73 m^2, 17.77 mL/minute/1.73 m^2, 73.3%, 38% and 79.7 mL/minute/1.73 m^2, respectively.CKD-EPI_(Scr-Scys C) predicted the most precise eGFR both in lymphoma and leukemia subgroups. Additionally, CKDEPI_(Scys C) equation in the rGFR■90 mL/minute/1.73 m^2 subgroup and Steven 2 equation in the rGFR<90 mL/minute/1.73 m^2 subgroup provided more accurate estimates in each subgroup. The CKD-EPI_(Scr-Scys C) equation could be recommended to monitor kidney function in hematopathy patients. The accuracy of GFR equations may be closely related with GFR level and kidney function markers, but not the primary cause of hematopathy.

Chinese herbal medicine for subacute thyroiditis:a systematic review of randomized controlled trials

OBJECTIVE:To evaluate the effectiveness and safety of Chinese herbal medicine(CHM) in the treatmentofsubacutethyroiditis.METHODS: Randomized controlled trials found in PubMed, Cochrane Library, and three Chinese databases were selected. RevMan 5.2 software was used to analyze the data with relative risk or mean difference, expressed with 95% of confidence interval. The quality of trials was assessed and gradedthe quality of evidence with GRADE profiler software.RESULTS: Twenty one studies were included. CHM was superior to Western Medicine(include prednisone and NSAIDs) in abating fever, relieving thyroid pain, recovering blood sedimentation, improving thyroid function, and preventing hypothyroidism(P<0.05), while no statistical differences were found in eliminating goiter and reducing relapse rate.CHM plusWestern Medicine were superior toWestern Medicine in abating fever, relieving thyroid pain, eliminating goiter, and reducing relapse rate,while no statistical differences(P>0.05) were found in recovering blood sedimentation and improving thyroid function. The incidence of adverse reactions in treatment group was lower than that in control group(relative risk was 0.12 and 95% confidence interval was 0.03-0.51). The methodological quality of trials is generally poor with a high risk of bias.CONCLUSION: CHM(particularly CHM combined withWestern Medicine) used to treat subacute thyroiditis may improve clinical symptoms and signs,reduce relapse rate, and alleviate the side effects of hormones. Due to poor methodological quality of included trials, further more high-quality studies are warranted to confirm the effectiveness and safety of CHM.

Effectiveness of external Sanjierupi Gao on mastalgia caused by mammary gland hyperplasia: a placebo controlled trial

OBJECTIVE:To evaluate the curative effect of external application of the Chinese drug,Sanjierupi Gao,on mastalgia caused by mammary gland hyperplasia.METHODS:This randomized,double-blinded,and placebo controlled study enrolled 260 patients with mammary gland hyperplasia from five hospitals.Patients were randomly and equally divided in-to a Sanjierupi Gao treatment group and a placebo control group.An adhesive plaster was applied to the most painful area on either breast for 7 h a day.Treatment lasted for two menstrual cycles without application during menstruation.Mastalgia was used as the main index of curative effect.The change before and after treatment in days of mastalgia,the time to alleviate pain,pain extent,and severe pain scores were observed.RESULTS:Compared to the control group,the treatment group had significantly fewer days of mastalgia(P<0.01),a significantly lower severe pain score(P<0.01),and significantly less subjective pain and tenderness(P<0.05 and P<0.01,respectively).Three days before the follow-up visit,the pain score in the treatment group was significantly lower than that in the control group(P<0.05).A non-parametric test was used to compare the time to alleviate mastalgia between the two groups and found no statistical difference(Z=0.313,P=0.754).CONCLUSION:Application of Sanjierupi Gao can decrease mastalgia duration in patients with mammary gland hyperplasia during menstruation and alleviate the extent of mastalgia.The time to alleviate pain is psychologically influenced.

Fluorescence imaging mitochondrial copper(Ⅱ) via photocontrollable fluorogenic probe in live cells

Monitoring mitochondrial derived copper（Ⅱ） in live cells is highly demanded, but accurately detecting is unmet due to the interference with cytoplasmic copper（Ⅱ）. Herein, we have reported the design,synthesis and characterization of photocontrollable fluorogenic probe, MCu-3, which is equipped with a photo-labile group（nitrobenzyl group） and mitochondria targeting unit（triphenylphosphonium salt）.This novel probe showed an intense fluorescence enhancement in response to copper（Ⅱ） without interference from other metal cations in the biological condition（p H 6–9）. The detection limit is 1.7 ×10^（-7） mol/L in HEPES buffer. The confocal fluorescence imaging results demonstrated MCu-3 can visualize mitochondrial copper（Ⅱ） in live mammalian cells. The clear advantage of our photocontrollable method is successful to avoid the influence of cytoplasmic copper（Ⅱ） during mitochondria specific detection.