Background Neutrophilic asthma (NA) is a severe, corticosteroid-resistant phenotype associated with neutrophil extracellular traps (NETs) formation and Th17/Treg imbalance. Cold exposure exacerbates NA progression, increasing the global clinical burden. Currently, combination therapy with corticosteroids is considered a viable approach. This study aimed to investigate the therapeutic potential and mechanisms of ginsenoside Rk2 (Rk2) combined with dexamethasone (DEX) against cold-stimulated neutrophilic asthma (CSNA). Methods A corticosteroid-resistant BEAS-2B cell model was established by NTHi stimulation. A CSNA mouse model was established via ovalbumin sensitization, NTHi infection, and cold stimulation, followed by Rk2 and/or DEX treatment. Airway hyperresponsiveness, histopathology, and neutrophil infiltration were measured to assess therapeutic efficacy. Th17/Treg populations were analyzed by flow cytometry. NETs formation was quantified in PMA-stimulated primary mouse neutrophils. A NETs-naïve CD4+ T cell co-culture system was used to examine the causal role of NETs in Th17/Treg imbalance. Transcriptomics and agonist-based gain-of-function experiments were conducted to validate the mechanism of Rk2 combined with DEX. Results Rk2 reversed DEX resistance by rebalancing GRα/GRβ, upregulating HDAC2, and promoting GRα nuclear translocation. Compared with monotherapy, Rk2 combined with DEX ameliorated airway inflammation and restored Th17/Treg homeostasis. Mechanistically, this combination therapy suppressed NETs formation by reducing NETs components, DNA release and ROS generation. NETs drove Th17/Treg imbalance via STAT1/NLRP3 inflammasome axis. Pharmacological activation of either STAT1 or NLRP3 reversed the beneficial effects. Conclusion Our findings demonstrate for the first time that Rk2, a potent natural corticosteroid potentiator, acts in combination with DEX to alleviate CSNA by suppressing the STAT1/NLRP3 inflammasome-NETs-Th17/Treg cascade.

Background/Objectives: Ulcerative colitis is a chronic inflammatory bowel disease marked by a disrupted intestinal barrier and consequent aberrant immune responses. Pseudoginsenoside RT2, an ocotillol-type ginsenoside abundant in Panax herbs, represents a potential therapeutic candidate, yet its anti-ulcerative colitis efficacy and pharmacokinetic profile remain unclear. This study aimed to elucidate RT2's therapeutic potential for ulcerative colitis through a parallel evaluation of pharmacodynamic efficacy and pharmacokinetic properties. Methods: The anti-ulcerative colitis efficacy and in vivo disposition of RT2 were investigated in a trinitrobenzene sulfonic acid-induced rat colitis model. An ultra-performance liquid chromatography-tandem mass spectrometry method was employed to delineate its pharmacokinetic characteristics and quantify its distribution in various tissues following oral administration. Results: Pharmacodynamically, RT2 demonstrated significant efficacy in the UC rat model by repairing the intestinal barrier (by promoting goblet cell regeneration and upregulating tight junction proteins and mucin) and restoring immune homeostasis (by correcting T-helper 17/regulatory T-cell imbalance and reducing pro-inflammatory cytokines while elevating anti-inflammatory cytokines). Pharmacokinetically, RT2 exhibited rapid absorption, slow elimination, and high colonic accumulation, with concentrations in the inflamed colon being significantly higher than those in healthy rats. Furthermore, the biphasic concentration-time profile may account for its prolonged systemic residence time and enhanced local exposure. In summary, through parallel efficacy and pharmacokinetic studies, this work systematically reveals its characteristics as a therapeutic agent that exhibits high colonic accumulation and acts via barrier repair and immunomodulation. Conclusions: These findings provide a theoretical foundation for the development of RT2 as a novel gut-selective drug candidate for UC.

BACKGROUND:Ulcerative colitis (UC) is a chronic, relapsing inflammatory bowel disease characterized by intestinal barrier dysfunction. Pseudoginsenoside RT2, an ocotillol-type ginsenoside widely present in Panax species, possesses unexplored therapeutic potential for UC. PURPOSE:This study investigated the efficacy of RT2 against UC and its mechanisms of intestinal barrier protection by driving epithelial renewal. METHODS:Anti‑UC effects of RT2 were assessed in lipopolysaccharide (LPS)‑stimulated Caco‑2 cells and dextran sulfate sodium (DSS)‑induced mice. Integrated transcriptomic and proteomic analyses were performed to identify key pathways in barrier restoration. Mechanistic validation was conducted using DSS‑induced intestinal organoids (encompassing all epithelial lineages) and LPS‑stimulated IEC-6 cells (non-transformed intestinal epithelial model). Effects on Wnt/β-catenin pathway were assessed with inhibitor ICG-001. RESULTS:In Caco‑2 cells, RT2 restored transepithelial electrical resistance (TEER), regulated cytokines and upregulated tight junction proteins (TJs). In UC mice, RT2 dose‑dependently ameliorated disease activity index (DAI), improved colon length and histopathology, increased anti‑inflammatory cytokines and reduced pro‑inflammatory cytokines. RT2 enhanced TJs and epithelial markers. Multi-omics analysis linked RT2's benefits to Wnt/β-catenin pathway activation. In organoids, RT2 increased budding and Lgr5 expression. In IEC‑6 cells, RT2 restored TEER, promoted proliferation/migration, suppressed apoptosis and upregulated TJs and epithelial markers. Mechanistically, RT2 facilitated Wnt ligand-receptor binding, leading GSK-3β phosphorylation, disassembly of the destruction complex and β-catenin nuclear translocation. All effects were abolished by ICG‑001. CONCLUSIONS:RT2 attenuates UC by activating Wnt/β‑catenin pathway, thereby restoring intestinal barrier integrity via driving epithelial renewal, with efficacy comparable to the positive control drug SASP. RT2 represents a promising natural product candidate for UC treatment.

To the Editor: Chemotherapy is one of the mainstay therapies for patients with breast cancer (BC),[1,2] but it is often restricted by dose-limiting toxicities attributed to myelosuppression. Although recombinant human granulocyte colony-stimulating factors (rhG-CSF) are effective for treating and preventing chemotherapy-induced neutropenia, daily administration is inconvenient to patients.[3,4] Telpegfilgrastim (Peijin®, Xiamen Amoytop Biotech Co., Ltd, Xiamen, China) is a Y-shape branched pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF). In this multicenter, randomized, open-label, active drug-controlled, non-inferior, parallel-group, phase 3 study (ClinicalTrials.gov, No. NCT04466137), we evaluated the efficacy and safety of telpegfilgrastim in patients with BC for chemotherapy on the taxotere, adriamycin, and cyclophosphamide (TAC) regimen for the clinical management of neutropenia and febrile neutropenia (FN). The study was approved by the ethics committee of National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College (No. 19-120/1904) with written informed consent obtained from all patients before study entry. Considering the duration of grade 4 neutropenia as 1 day with a standard deviation of 1.5 days in cycle 1 of chemotherapy in telpegfilgrastim (2 mg fixed dose and 33 μg/kg) and control groups, using Power Analysis & Sample Size (PASS) version 15 with a predefined non-inferiority margin of 1 day at a power (1−β) of 0.9 and a one-sided test level of 0.025, the study required minimum sample size of 49 patients in each group. Furthermore, considering the drug exposure and potential dropout, the study enrolled a minimum of 85 patients in each group. Patients were included if they had histopathologically diagnosed BC, were ≥18 years, had a body weight ≥45 kg, a white blood cell count ≥3.5 × 109/L, a platelet count ≥100 × 109/L, an absolute neutrophil count (ANC) ≥1.5 × 109/L, and were suitable for chemotherapy. Patients who had received chemotherapy <2 months before screening, were scheduled for extensive radiotherapy (>25% of the total bone marrow), uncontrolled infections, antibiotics use ≤72 h before screening, or were allergic to rhG-CSF, were excluded. Patients were randomized in a 1:1:1 ratio based on gender (female and male patients with BC), age (≤65 years and >65 years), and history of previous chemotherapy; they received telpegfilgrastim 2 mg or 33 μg/kg or rhG-CSF (Topneuter®, Xiamen Amoytop Biotech Co., Ltd, Xiamen, China)/PEG-rhG-CSF (Xinruibai®, Qilu Pharmaceutical Co., Ltd, Jinan, China) using the Interactive Web Response System. Chemotherapy agents included an intravenous infusion of docetaxel 75 mg/m2 (Jiangsu Aosaikang Pharmaceutical Co. Ltd., Nanjing, China), doxorubicin 50 mg/m2 (Shenzhen Main Luck Pharmaceuticals Inc., Shenzhen, China), and cyclophosphamide 500 mg/m2 (Jiangsu Hengrui Pharmaceuticals Co., Ltd., Lianyungang, China). A 21 day-cycle, which included 4 cycles of chemotherapy, was administered. The treatment group received either 2 mg or 33 μg/kg of telpegfilgrastim subcutaneously on the third day of each chemotherapy cycle (48 ± 12 h). In contrast, the control group received rhG-CSF (Topneuter®) 5 μg/kg once daily subcutaneously until ANC recovered to ≥5.0 × 109/L from the lowest value (usage not exceeding 14 days during cycle 1 of chemotherapy). During cycles 2–4 of chemotherapy, based on the patient's choice, a single injection of PEG-rhG-CSF (Xinruibai®) 6 mg once in each chemotherapy cycle or rhG-CSF (Topneuter®) 5 μg/kg once daily was administered subcutaneously until ANC recovered to ≥5.0 × 109/L from the lowest value (usage not exceeding 14 days during each cycle of chemotherapy). The duration of grade 4 neutropenia in cycle 1 of chemotherapy was considered the primary endpoint, while the duration of grade 4 neutropenia in cycles 2–4 of chemotherapy, the incidence of grade 4 neutropenia across cycles 1–4 of chemotherapy, the duration and incidence of ≥grade 3 neutropenia, the incidence of FN, and a dynamic change in ANC from baseline were considered secondary endpoints. FN is defined as a single oral temperature measurement >101°F (>38.3°C) or a temperature ≥100.4°F (≥38.0°C) sustained for 1 h, with an ANC <0.5 × 109/L or an ANC between 0.5 × 109/L and 1.0 × 109/L that is expected to decrease to <0.5 × 109/L in the next 48 h. Safety was assessed in terms of adverse events (AEs) graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE) version 5.0. Continuous variables were presented as mean ± standard deviation (SD) and median ([minimum value, maximum value]/[Q1, Q3]), whereas categorical variables were expressed as N (%). Using the analysis of variance (ANOVA), differences in the duration of grade 4 neutropenia between the treatment groups were evaluated, and the least-squares mean, and 95% confidence interval (CI) were calculated using the analysis of covariance model (ANCOVA). The non-inferiority margin was set at ± 1 day. Multiple imputation method was used to impute missing data. The chi-squared test was used to analyze the difference in the incidence of neutropenia and FN between the groups. A two-sided test with a significance level of P <0.05 was considered statistically significant. SAS version 9.4 (SAS Institute, Cary, NC, USA) was used to perform all statistical analyses. Between October 16, 2020, and September 1, 2021, 265 patients were enrolled in this study. The patient disposition was presented in Supplementary Figure 1, https://links.lww.com/CM9/C275. The demographic characteristics of patients were shown in Supplementary Table 1, https://links.lww.com/CM9/C275. The mean duration of grade 4 neutropenia during cycle 1 of chemotherapy was 0.58 ± 1.09, 0.74 ± 1.59, and 0.48 ± 0.92 days in the telpegfilgrastim 2 mg group, telpegfilgrastim 33 μg/kg group, and control (rhG-CSF) group, respectively. The least-squares mean difference between the telpegfilgrastim (2 mg and 33 μg/kg) groups and control (rhG-CSF) group was 0.10 (95% CI: −0.19, 0.38; P = 0.51) and 0.22 (95% CI: −0.09, 0.52; P = 0.17), respectively, 95% CI were within the prespecified margin of ±1 day, confirming the non-inferiority of both telpegfilgrastim groups compared to the control (rhG-CSF) group [Supplementary Table 2, https://links.lww.com/CM9/C275]. In cycles 2–4 of chemotherapy, no significant differences in the mean duration of grade 4 neutropenia (P >0.05) between the telpegfilgrastim groups and the control (PEG-rhG-CSF or rhG-CSF) group. No significant differences in the incidence of grade 4 neutropenia across cycles 1–4 of chemotherapy between the telpegfilgrastim groups and the control (rhG-CSF or PEG-rhG-CSF) group. No significant differences in the duration of ≥grade 3 neutropenia, the incidence of grade 3/4 neutropenia, and the FN rates between telpegfilgrastim and control groups across cycles 1–4 of chemotherapy. ANC showed a double-peak pattern [Supplementary Figure 2, https://links.lww.com/CM9/C275], and the stratification analysis based on age or previous chemotherapy is shown in the Supplementary Tables 3 and 4, https://links.lww.com/CM9/C275. All patients in this study experienced at least one AE, with similar incidence, type, and severity across the three groups. Grade ≥3 treatment-emergent adverse events (TEAEs) were 59.6% for the telpegfilgrastim 2 mg group, 70.5% for the telpegfilgrastim 33 μg/kg group, and 62.5% for the control group. TEAEs leading to study discontinuation occurred in 1.1% of patients in each group. Serious adverse events (SAEs) were 5.6% in the telpegfilgrastim 2 mg group, 6.8% in the telpegfilgrastim 33 μg/kg group, and 6.8% in the control group. No study-drug-related deaths were reported [Supplementary Table 5, https://links.lww.com/CM9/C275]. TEAEs related to the study drugs are detailed in Supplementary Table 6, https://links.lww.com/CM9/C275. In a phase 3 study on patients with BC receiving epirubicin and docetaxel, the mean duration of grade 4 neutropenia in cycle 1 was 0.54 days and 0.61 days with 6 mg and 100 μg/kg of mecapegfilgrastim, respectively, and 1.02 days with filgrastim.[5] The duration of grade 4 neutropenia was 1.7 days with pegfilgrastim and 1.8 days with filgrastim, confirming the non-inferiority between the agents.[6] In this study, the duration of grade 4 neutropenia during cycle 1 of chemotherapy was 0.58 days with telpegfilgrastim 2 mg, 0.74 days with telpegfilgrastim 33 μg/kg, and 0.48 days with rhG-CSF. This study also demonstrated the non-inferiority of telpegfilgrastim in the duration of grade 4 neutropenia compared with rhG-CSF in cycle 1 of chemotherapy. The incidence of FN was 4.5% in mecapegfilgrastim 100 μg/kg and 1.82% in the filgrastim group in cycle 1 of chemotherapy in patients with BC. No significant difference was observed between the mecapegfilgrastim and filgrastim groups.[5] In this study, in the telpegfilgrastim 2 mg group, FN only occurred in cycle 1 of chemotherapy, the incidence of FN was 2.2%; the incidence of FN for telpegfilgrastim 33 μg/kg group was 3.4% in cycle 1, 1.2% in cycle 2, and 1.4% in cycle 4 of chemotherapy; and the incidence of FN in the control group was 3.4% in cycle 1 and 1.2% in cycle 2 of chemotherapy. No statistically significant difference was observed in the incidence of FN among the telpegfilgrastim 2 mg, telpegfilgrastim 33 μg/kg, and control groups. The results showed that the efficacy of telpegfilgrastim was equal to that of rhG-CSF and PEG-rhG-CSF. The incidence, type, and severity of AEs were similar among these three groups. The incidence of TEAEs was comparable between the telpegfilgrastim groups and the control group. This study has limitations. It only included Chinese patients with BC for chemotherapy on the TAC regimen, which may not represent all chemotherapy regimens used in clinical practice. There was no long-term follow-up of patients in this study, and only 4 cycles of chemotherapy were observed for safety, potentially introducing bias.[7] In conclusion, this study demonstrated that telpegfilgrastim had a similar efficacy and safety profile compared to rhG-CSF and PEG-rhG-CSF for the prevention of chemotherapy-induced neutropenia. A fixed dose of telpegfilgrastim 2 mg is more convenient and recommended for patients weighing ≥45 kg, while telpegfilgrastim 33 μg/kg may be a better option for patients with low body weight, particularly those weighing <45 kg. Acknowledgments The authors acknowledge Dr. Satya Lavanya Jakki and Dr. Ramandeep Singh (Indegene, Bangalore, India) for medical writing, editorial assistance, which was funded by Fosun Pharma. We also thank Dr. Zucheng Xie and Dr. Xinrui Chen (National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing China) for the editorial assistance. Funding This study was funded by Xiamen Amoytop Biotech Co., LTD and partly supported by the National Science and Technology Major Project for Key New Drug Development (No. 2017ZX09304015). Conflicts of interest The authors declare that there are no conflict of interests. This study was funded by Xiamen Amoytop Biotech Co., LTD, the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Ulcerative colitis (UC) is a multifactorial inflammatory bowel disease associated with intestinal barrier integrity, gut microbiota, and immune homeostasis. According to previous studies, ginsenoside Rk2 (Rk2) demonstrated anti-inflammatory activity in cocultures of Caco-2 and THP-1 cells. This study was the first to demonstrate that Rk2 positively impacted UC by regulating the "barrier-microbiota-immune" axis. Rk2 not only improved inflammatory factor levels and colitis symptoms but also enhanced intestinal epithelial barrier function via increased transepithelial electrical resistance (TEER) values and elevated expressions of tight junction proteins, remodeled gut microbiota via enriching its diversity and composition and increasing short-chain fatty acids (SCFAs) levels, and maintained intestinal homeostasis via restoring Th17/Treg balance. Moreover, the influence of Rk2 on UC might be mediated via the TNFSF14/LTβR/NIK pathway. In summary, Rk2 exerted a beneficial effect on UC by providing multidimensional regulation and was a potential natural therapeutic agent for UC.

The application of mesenchymal stem cell (MSC)-derived extracellular vesicles (EVs) holds significant promise in anti-aging cosmetics, regenerative medicine, and drug delivery systems. However, their limited production efficiency remains a critical barrier to advancing related therapies and pharmaceutical applications. In this study, a library of triterpene saponins was screened, leading to the re-discovery of an oleanane-type triterpene saponin Lucyoside H (1), along with its structural analogs, Chikusetsusaponins IVa (2), IV (3), and V (4), which were found to increase the production of EV from human adipose-derived mesenchymal stem cells (ADMSCs) at a concentration ranging from 10 to 100 µM. A comparative analysis of the chemical structures and activities of all evaluated compounds, along with oleanolic acid (5), revealed that (i) disubstitution of glycosyl on C-3 and C-28 of oleanane aglycone was crucial to the promoting effect; (ii) 3-O-β-D-glucuronopyranosyl substitution achieved the highest efficiency in the promoting effect and alteration of this group attenuated the activity. These results highlight the potential for developing a natural product-based approach to increase EV production by MSCs.

ETHNOPHARMACOLOGICAL RELEVANCE:Panax ginseng C.A. Meyer has been used traditionally for treating gastrointestinal disorders. Oleanolic acid 28-O-β-D- glucopyranoside (OAG), a major pentacyclic triterpenoid in ginseng, has demonstrated the antioxidant and the anti-ulcerative colitis effects in our previous studies. However, whether OAG intervenes UC by alleviating oxidative stress injury through regulating ferroptosis remains unclear. AIM OF THE STUDY:To explore the underlying mechanism of OAG regulating ferroptosis to alleviate UC. MATERIALS AND METHODS:A TNBS-induced UC rat model, which closely mimics the immunological characteristics of human UC, was established to evaluate the therapeutic efficacy of OAG by analyzing the disease activity index (DAI), colonic macroscopic damage index (CMDI), biochemical indicators, Th17/Treg cell ratio and IL-17 levels, pathomorphological alterations, tight junction proteins (TJs), intestinal epithelial ultrastructure. Subsequently, an integrated multi-omics analysis was conducted to elucidate whether ferroptosis pathways underlie OAG's efficacy against UC and to screen for key candidate genes and proteins, which were further validated using quantitative PCR and western blotting. Finally, molecular docking and the cellular thermal shift assay were performed to confirm the direct binding of OAG to key ferroptosis-related proteins in an RSL3-induced ferroptosis model of Caco-2 cells. RESULTS:OAG treatment significantly ameliorated UC symptoms, attenuated oxidative stress and inflammatory cell infiltration, reduced Th17/Treg cell ratio and IL-17 levels, and enhanced intestinal epithelial barrier integrity. Ferroptosis inhibition was identified as the crucial mechanism mediating OAG's therapeutic effects, with direct interactions between OAG and key ferroptosis-related proteins (GPX4, NRF2, HO-1, and x-CT) executing its protective effects against UC. CONCLUSIONS:This study demonstrates that OAG effectively ameliorates TNBS- induced UC by modulating Nrf2/x-CT/GPX4-mediated ferroptosis pathway, furthering our understanding of its molecular mechanism and supporting its therapeutic potential for UC treatment.

Background Uropathogenic Escherichia coli (UPEC) constitutes a primary pathogen responsible for urinary tract infections (UTIs), characterized predominantly by colonization initiated through adhesion, persistence via biofilm formation, and invasion facilitated by lipid rafts. Cynaroside (Cyn), a natural flavonoid, emerges as a potential anti-virulence compound; nonetheless, the involvement of Caveolin-1 (CAV1) in UPEC adhesion and invasion processes has yet to be fully elucidated. Purpose To investigate the effects and mechanisms of Cyn on UPEC adhesion and invasion in bladder epithelial cells and in a rat UTI model. Methods We evaluated Cyn across bacterial, human bladder epithelial (5637), and rat UTI models using standard microbiological assays, imaging, and transcriptomics with quantitative PCR validation. Adhesion/invasion, barrier function, and cytokines were measured in 5637 cells, with CAV1 dependence tested by small interfering RNA (siRNA) and rescue experiments. Oral efficacy was assessed by urinary colony-forming units (CFU), inflammatory indices, histopathology, and bladder tissue protein expression. Results No direct bacteriostasis was observed, yet UPEC motility and biofilm formation declined, fimbrial/flagellar ultrastructure was disrupted, and virulence genes (fimH, papG, fliC) were downregulated. In 5637 cells, Cyn decreased adhesion and invasion, restored barrier function, and lowered IL-6 and TNF-α; these effects were attenuated in the CAV1 knockdown condition, but in the rescue experiment, reconstitution of CAV1-Mut partially restored these effects. In rats, oral Cyn reduced urinary CFU, improved inflammatory indices and bladder pathology, and decreased adhesion-related proteins (Uroplakin Ia, Uroplakin Ib, Uroplakin III, integrin alpha-3, integrin beta-1, and CAV1). Conclusions Cyn functions as an anti-virulence natural product that mitigates UPEC pathogenesis by inhibiting bacterial virulence programs and modulating CAV1-mediated epithelial processes, supporting its potential for UTI management.

[Objective] To establish an androgenic alopecia model in C57BL/6 mice, observe the improvement effect of Black Ginseng extract on alopecia, and explore its mechanism using network pharmacology and molecular docking methods. [Methods] An androgenic alopecia model was established by subcutaneous injection of testosterone propionate in mice. Different doses of Black Ginseng extract groups were set up and compared with Minoxidil. The improvement effect of Black Ginseng extract on androgenic alopecia was evaluated based on hair growth, dermal thickness, hair follicle morphology and count, serum hormone levels, and skin tissue VEGF and TGF-β content. Based on previous studies, the chemical constituents of Black Ginseng extract were identified. Swiss Target Prediction and Symmap databases were used to obtain targets of Black Ginseng’s chemical constituents, combined with alopecia-related targets from Malacard, DisGeNET, and Genecards databases to construct a "Black Ginseng-chemical composition-intersection target-alopecia" interaction network, screening key chemical components and targets, followed by molecular docking analysis. [Results] Mice in the Black Ginseng extract groups showed better hair growth than the model group, with increased dermal thickness, decreased serum testosterone and dihydrotestosterone levels, increased estradiol levels, increased VEGF expression, and decreased TGF-β expression in skin, all in a dose-dependent manner. Key chemical components Deoxyoleanolic acid, Ginsenoside Rb1, Ginsenoside Rg3, Ginsenoside Rh4, and Campesterol, as well as key targets INS, AR, VEGFA, PPARG, LEP, and CASP3, were identified. Molecular docking results showed stable binding between key components and targets. [Conclusion] Black Ginseng extract improves androgen-induced alopecia in mice, likely by regulating targets such as AR and pathways like MAPK. This provides a scientific basis for further research and development of Black Ginseng.

Ethnopharmacological relevance Yinhua Miyanling Tablet (YMT), a traditional Chinese medicine consisting of 10 herbs, has been widely used clinically to treat urinary tract infections (UTIs), however, its therapeutic mechanism is not fully understood. Aim of the study To investigate the mechanism of YMT in treating UTIs through network pharmacology, multi-omics and experimental validation. Materials and methods Clinically, blood and urine samples from YMT-treated UTI patients were collected for transcriptomic and metabolomic analyses. Computationally, compounds that are related to YMT were obtained from the databases, relevant targets were identified, and UTI-related targets were analyzed to determine the core signaling pathways. Subsequently, an integrated approach combining multi-omics and network pharmacology assisted in identifying the key pathways underlying therapeutic effects of YMT on UTI. Finally, a mouse model of UTI was established using uropathogenic Escherichia coli (UPEC), and the therapeutic mechanism of YMT on UTI was validated by ELISA, qRT-PCR and Western blotting. Results After taking YMT, patients showed reduced levels of urinary bacteria, white blood cells, and serum inflammatory factors (CRP, IL-6 and TNF-α). Multi-omics analysis combined with network pharmacology demonstrated that YMT significantly inhibited the TLR/MAPK/NFκB signaling pathway. In vivo experiments confirmed that YMT attenuated UPEC-induced pathological changes in bladder structural, reduced the expression of bladder proteins (TLR4, MyD88, p-p38 MAPK and p-p65 NFκB), increased protein expression of IκB-α, and attenuated the release of inflammatory factors (TNF-α, IL-6 and IL-1β) in mice. Conclusion YMT is effective in treating UTI by down-regulating the TLR4/p38MAPK/p65NFκB pathway, thereby providing a scientific basis for its clinical application.

This review covers the structures of diterpenoids, including chain (72), monocyclic (9), labdane-type (67), clerodane-type (127) abietane-type (716), (ent)-kaurane-type (89), grayanane-type (331), ingenane-type (55), tigliane-type (154), daphnane-type (237), and aconitine-type diterpene alkaloids (265) with rich biological activities reported in 2013–2023. And the drugs in clinical use or under clinical investigation of diterpenoids and leading compounds were summarized.

Superoxide dismutase (SOD) is a vital antioxidant enzyme that exerts antioxidative and anti-inflammatory effects on the host. In this study, a novel thermostable SOD of Alicyclobacillus sp. (AliSOD) from a hot spring was overexpressed in Escherichia coli, and enzymatic properties were identified. Mn2+ plays a decisive role in enzyme activity, indicating that AliSOD is MnSOD. Specifically, AliSOD was determined to be dimeric with a subunit molecular mass of 23.0 kDa, and the specific activity was confirmed to be as high as 24990.8 U·mg-1. AliSOD demonstrated exceptional thermal stability, broad pH stability, and resistance to urea, exhibiting minimal loss of activity at 70 °C and remarkable tolerance in an alkaline environment. Moreover, AliSOD significantly alleviated oxidative stress in diquat-injured cells (P < 0.01). It also increased intracellular SOD expression and activated the Nrf2 protein downstream of the Keap1-Nrf2 signaling pathway (P < 0.05). Overall, AliSOD exhibits excellent thermostability and specific activity, indicating potential applications in the pharmaceutical, food, and animal feed industries.

Steroid-resistant asthma (SRA), resisting glucocorticoids such as dexamethasone (DEX), is a bottleneck in the treatment of asthma. It is characterized by a predominantly neutrophilic inflammatory subtype and is prone to developing into severe refractory asthma and fatal asthma. Currently, there is a lack of universally effective treatments for SRA. Moreover, since cold stimulation does increase the risk of asthma development and exacerbate asthma symptoms, the treatment of cold-stimulated SRA (CSRA) will face greater challenges. To find effective new methods to ameliorate CSRA, this study established a CSRA mouse model of allergic airway inflammation mimicking human asthma for the first time and evaluated the alleviating effects of 80% ethanol extract of mountain-cultivated ginseng (MCG) based on multi-omics analysis. The results indicate that cold stimulation indeed exacerbated the SRA-related symptoms in mice; the DEX individual treatment did not show a satisfactory effect; while the combination treatment of DEX and MCG could dose-dependently significantly enhance the lung function; reduce neutrophil aggregation; decrease the levels of LPS, IFN-γ, IL-1β, CXCL8, and IL-17; increase the level of IL-10; alleviate the inflammatory infiltration; and decrease the mucus secretion and the expression of MUC5AC. Moreover, the combination of DEX and high-dose (200 mg/kg) MCG could significantly increase the levels of tight junction proteins (TJs), regulate the disordered intestinal flora, increase the content of short-chain fatty acids (SCFAs), and regulate the abnormal gene profile and metabolic profile. Multi-omics integrated analysis showed that 7 gut microbes, 34 genes, 6 metabolites, and the involved 15 metabolic/signaling pathways were closely related to the pharmacological effects of combination therapy. In conclusion, integrated multi-omics profiling highlighted the benefits of MCG for CSRA mice by modulating the interactions of microbiota, genes, and metabolites. MCG shows great potential as a functional food in the adjuvant treatment of CSRA.

Acute kidney injury (AKI) is a common, multicause clinical condition that, if ignored, often progresses to chronic kidney disease (CKD) and end-stage kidney disease, with a mortality rate of 40-50%. However, there is a lack of universal treatment for AKI. Inflammation is the basic pathological change of early kidney injury, and inflammation can exacerbate AKI. Macrophages are the primary immune cells involved in the inflammatory microenvironment of kidney disease. Therefore, regulating the function of macrophages is a crucial breakthrough for the AKI intervention. Our team chemically modified pyxinol, an ocotillol-type ginsenoside, to prepare PJ16 with higher solubility and bioavailability. In vitro, using a model of macrophages stimulated by LPS, it was found that PJ16 could regulate macrophage function, including inhibiting the secretion of inflammatory factors, promoting phagocytosis, inhibiting M1 macrophages, and promoting M1 transition to the M2c macrophage. Further investigation revealed that PJ16 may shield renal tubular epithelial cells (HK-2) damaged by LPS in vitro. Based on this, PJ16 was validated in the animal model of unilateral ureteral obstruction, which showed that it improves renal function and inhibits renal tissue fibrosis by decreasing inflammatory responses, reducing macrophage inflammatory infiltration, and preferentially upregulating M2c macrophages. In conclusion, our study is the first to show that PJ16 resists AKI and fibrosis by mechanistically regulating macrophage function by modulating the phenotypic transition from M1 to M2 macrophages, mainly M2c macrophages.

The purpose of this study was to explore the therapeutic effect of the oral administration of pseudo-ginsenoside RT4 (RT4) on ulcerative colitis (UC), and to determine the rate of absorption and distribution of RT4 in mice with UC. Balb/c mice were induced using dextran sulfate sodium salts (DSS) to establish the UC model, and 10, 20, or 40 mg/kg of RT4 was subsequently administered via gavage. The clinical symptoms, inflammatory response, intestinal barrier, content of total short-chain fatty acids (SCFAs), and gut microbiota were investigated. Caco-2 cells were induced to establish the epithelial barrier damage model using LPS, and an intervention was performed using 4, 8, and 16 µg/mL of RT4. The inflammatory factors, transient electrical resistance (TEER), and tight-junction protein expression were determined. Finally, pharmacokinetic and tissue distribution studies following the intragastric administration of RT4 in UC mice were performed. According to the results in mice, RT4 decreased the disease activity index (DAI) score, restored the colon length, reduced the levels of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1β), and boosted the levels of immunosuppressive cytokine IL-10, increased the content of SCFAs, improved the colonic histopathology, maintained the ultrastructure of colonic mucosal epithelial cells, and corrected disturbances in the intestinal microbiota. Based on the results in caco-2 cells, RT4 reduced the levels of TNF-α, IL-6, and IL-1β; protected integrity of monolayers; and increased tight-junction protein expression. Additionally, the main pharmacokinetic parameters (Cmax, Tmax, t1/2, Vd, CL, AUC) were obtained, the absolute bioavailability was calculated as 18.90% ± 2.70%, and the main distribution tissues were the small intestine and colon. In conclusion, RT4, with the features of slow elimination and directional distribution, could alleviate UC by inhibiting inflammatory factors, repairing the intestinal mucosal barrier, boosting the dominant intestinal microflora, and modulating the expression of SCFAs.

Ulcerative colitis (UC), an incurable and recurrent inflammatory bowel disease, presents a significant threat to health and highlights the need for novel therapeutic strategies. Oleanolic acid 28-O-β-D-glucopyranoside (OAG) is a naturally occurring pentacyclic triterpenoid found in ginseng. In this study, we demonstrated that OAG exhibited remarkable anti-UC activity in LPS-induced Caco-2 cells and DSS-induced model mice. First, OAG alleviated the symptoms of UC by mitigating weight loss, reducing the DAI score, and increasing colon length. Second, the inflammatory response was inhibited after OAG intervention, evidenced decreases in the spleen coefficient, cytokine levels, and inflammatory cell infiltration in colon tissue. Thirdly, OAG also enhanced intestinal epithelial barrier function, as evidenced by elevated TEER values, increased expression of tight junction proteins, diminished bacterial translocation, and maintained intact ultrastructure of colonic mucosal cells. Notably, compared with 5-aminosalicylic acid, OAG demonstrated superior efficacy in enhancing mucosal barrier function. Fourth, OAG increased microbial diversity, promoted the abundance of beneficial bacteria, reduced the abundance of harmful bacteria, and rebalanced the gut microbiome. Finally, the PI3K-AKT and MAPK signaling pathways were identified as crucial mechanisms underlying the therapeutic effects of OAG against UC through multi-omics. In summary, we identified OAG as a novel therapeutic agent against UC, demonstrating anti-inflammatory, barrier-preserving, and gut microbiota-modulating effects, highlighting its promising potential as a candidate UC drug.

Background: Yinhua Miyanling tablets (YMT), comprising 10 Chinese medicinal compounds, is a proprietary Chinese medicine used in the clinical treatment of urinary tract infections. Medicinal compounds, extracts, or certain monomeric components in YMT all show good effect on ulcerative colitis (UC). However, no evidence supporting YMT as a whole prescription for UC treatment is available. Purpose: To evaluate the anti-UC activity of YMT and elucidate the underlying mechanisms. The objective of the study was to provide evidence for the add-on development of YMT to treat UC. Methods: First, YMT's protective effect on the intestinal barrier was evaluated using a lipopolysaccharide (LPS)induced Caco-2 intestinal injury model. Second, the UC mouse model was established using dextran sodium sulfate (DSS) to determine YMT's influence on symptoms, inflammatory factors, intestinal barrier, and histopathological changes in the colon. Third, an integrated method combining metabolomics and network pharmacology was employed to screen core targets and key metabolic pathways with crucial roles in YMT's therapeutic effect on UC. Molecular docking was employed to identify the key targets with high affinity. Finally, western blotting was performed to validate the mechanism of YMT action against UC. Results: YMT enhanced the transepithelial electrical resistance value and improved the expression of proteins of the tight junctions dose-dependently in LPS-induced Caco-2 cells. UC mice treated with YMT exhibited alleviated pathological lesions of the colon tissue in the in vivo pharmacodynamic experiments. The colonic lengths tended to be normal, and the levels of inflammatory factors (TNF-alpha, IL-6, and iNOS) along with those of the core enzymes (MPO, MDA, and SOD) improved. YMT effectively ameliorated DSS-induced colonic mucosal injury; pathological changes along with ultrastructure damage were significantly alleviated (evidenced by a relatively intact colon tissue, recovery of epithelial damage, repaired gland, reduced infiltration of inflammatory cells and epithelial cells arranged closely with dense microvilli). Seven key targets (IL-6, TNF-alpha, MPO, COX-2, HK2, TPH, and CYP1A2) and four key metabolic pathways (arachidonic acid metabolism, linoleate metabolism, glycolysis, and gluconeogenesis and tyrosine biosynthesis) were identified to play vital roles in the treatment on UC using YMT. Conclusions: YMT exerts beneficial therapeutic effects on UC by regulating multiple endogenous metabolites, targets, and metabolic pathways, suggestive of its potential novel application in UC treatment.

Background: (-)-Syringaresinol (SYR), a natural lignan with significant antioxidant and anti-inflammatory activities, possesses various pharmacological benefits including cardio-protective, antibacterial, anticancer, and anti-aging effects. It was shown that the effectiveness of (+)-syringaresinol diglucoside on the ulcerative colitis (UC) was attributed to the active metabolite (+)-syringaresinol (the enantiomor of SYR). However, the efficacy of SYR against UC remains unclear, and the associated molecular mechanism has not been revealed yet Purpose: This study aimed to assess the protective effect of SYR in UC and its underlying mechanism Study design and methods: We examined SYR's protective impact on the intestinal epithelial barrier and its ability to inhibit inflammatory responses in both a lipopolysaccharide (LPS)-induced Caco-2 cell model and a dextran sodium sulfate (DSS)-induced UC mouse model. We also explored the potential signaling pathways regulated by SYR using transcriptome analysis and western blot assay Results: In Caco-2 cells, SYR significantly increased trans-epithelial electrical resistance, reduced tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL -6), interferon-gamma (IFN-gamma), and cyclooxygenase-2 (COX -2) levels, and enhanced cellular tight junction protein expression and distribution. In mice with UC, oral treatment with SYR (10, 20, 40 mg.kg(-1)) dose-dependently increased body weight, colon length, and expression of tight junction proteins, decreased disease activity index score, spleen coefficient, cytokine serum levels, bacterial translocation, and intestinal damage, and also preserved the ultrastructure of colonic mucosal cells. Transcriptomics indicated that the anti-UC effect of SYR is mediated via the PI3K-Akt/MAPK/Wnt signaling pathway. Conclusion: In summary, SYR effectively mitigated the development of UC by enhancing the intestinal epithelial barrier function and attenuating the inflammatory response. The plant-derived product SYR might be a potentially effective therapeutical agent against UC.

Saponins, the major bioactive components of Panax ginseng C. A. Mey. (Renshen in Chinese), are gradually emerging as research hotspots owing to the possession of various pharmacological activities. This review updates the ginsenosides list from P. ginseng and the steam-processed ginseng (red ginseng and black ginseng) up to 271 by June of 2024, encompassing 241 saponins from different parts of P. ginseng (roots, stems, leaves, flowers, berries, and seeds), 103 from red ginseng, and 65 from black ginseng, respectively. Among 271 saponins, there are a total of 249 (1−249) dammarane type (with a−z subtypes) tetracyclic triterpene saponins reported from each part of P. ginseng and steam-processed ginseng, two (250−251) lanostane type tetracyclic triterpene saponins identified from red ginseng, 18 (252−269) oleanane type pentacyclic triterpenoid saponins discovered from each part of P. ginseng and steam-processed ginseng, and two (270−271) ursane type pentacyclic triterpenoid saponins reported from red ginseng. Overall, this review expounds on the chemical diversity of ginsenosides in multiple aspects, such as chemical structure, spatial distribution and subtype comparison, processed products, and transformation. This facilitates more in-depth research on ginsenosides and contributes to the future development of ginseng.