This study investigates the pharmacokinetics and metabolic characteristics of three marine-derived piericidins as potential drug leads for kidney disease: piericidin A (PA) and its two glycosides (GPAs), glucopiericidin A (GPA) and 13-hydroxyglucopiericidin A (13-OH-GPA). The research aims to facilitate lead selection and optimization for developing a viable preclinical candidate. Rapid absorption of PA and GPAs in mice was observed, characterized by short half-lives and low bioavailability. Glycosides and hydroxyl groups significantly enhanced the absorption rate (13-OH-GPA > GPA > PA). PA and GPAs exhibited metabolic instability in liver microsomes due to Cytochrome P450 enzymes (CYPs) and uridine diphosphoglucuronosyl transferases (UGTs). Glucuronidation emerged as the primary metabolic pathway, with UGT1A7, UGT1A8, UGT1A9, and UGT1A10 demonstrating high elimination rates (30%-70%) for PA and GPAs. This rapid glucuronidation may contribute to the low bioavailability of GPAs. Despite its low bioavailability (2.69%), 13-OH-GPA showed higher kidney distribution (19.8%) compared to PA (10.0%) and GPA (7.3%), suggesting enhanced biological efficacy in kidney diseases. Modifying the C-13 hydroxyl group appears to be a promising approach to improve bioavailability. In conclusion, this study provides valuable metabolic insights for the development and optimization of marine-derived piericidins as potential drug leads for kidney disease.
Animals ; Male ; Mice ; Aquatic Organisms/chemistry* ; Biological Availability ; Cytochrome P-450 Enzyme System/metabolism* ; Glucuronosyltransferase/metabolism* ; Microsomes, Liver/metabolism* ; Molecular Structure ; Biological Products/pharmacokinetics* ; Pyridines/pharmacokinetics*

Objective:To explore the correlations between the cerebral infarction area and cytokines and immune status in patients with acute ischemic stroke,and to provide the theoretical basis for immunotherapy of the patients with different degrees of cerebral infarction.Methods:Sixty-seven patients with acute ischemic stroke within 72 h of the onset were randomly selected according to the inclusion and exclusion criteria,and were divided into large-area cerebral infarction group(n=34)and non-large-area cerebral infarction group(n=33)on the basis of the biggest infarction area in the sequences of magnetic resonance diffusion-weighted imaging(CDWI).Clinical baseline characteristics such as gender,age,and medical history were collected from the patients in two groups,the serum levels of interleukin(IL)-2,IL-6,IL-10,and IL-17A,tumor necrosis factor-α(TNF-α),and interferon-γ(IFN-γ)were measured using flow cytometry;the absolute values of lymphocytes(LYM#),lymphocyte percentages(LYM％),and neutrophil/lymphocy ratios(NLR)in peripheral blood of the patients caiculated,and the ratios of IFN-γ/IL-4,TNF-α/IL-4,and TNF-α/IL-10 rations were also calculated.The values of National Institutes of Health Stroke Scale(NIHSS)scores of the patients were evaluatd on the basis of the assessment of clinical neurological signs.The correlations of the cerebral infarction area and NIHSS score,cytokines and immune status groups of the patients in two were tested by rank correlation analysis.Results:Compared with non-large-area cerebral infarction group,the serum levels of IL-2,IL-6,IL-10,IL-17A,TNF-α,and IFN-γ as well as the NLR in the peripheral blood of the patients in large-area cerebral infarction group were significantly increased(P＜0.01),while the LYM#,LYM％and TNF-α/IL-4 were significantly decreased(P＜0.01).There was a positive correlation between cerebral infarction area and NIHSS score in the patients in large-area cerebral infarction group(rs=0.521,P＜0.05),and there was a significantly positive correlation between cerebral infarct area and NIHSS score in the patients in non-large-area cerebral infarction group(rs=0.721,P＜0.001).The NIHSS scores were positively correlated with serum IL-6(rs=0.306,P=0.005),IL-4(rs=0.252,P＜0.001),IL-2(rs=0.109,P=0.025),IL-17A(rs=0.405,P＜0.001),and IFN-γ(rs=0.146,P＜0.001)levels in two groups;no correlations were found between NIHSS scores and TNF-α(rs=0.039,P=0.726)and IL-10(rs=0.121,P=0.192)levels.NIHSS scores of the patients in two groups had negative correlatious with the serum level of LYM#(rs=-0.026,P=0.036)and LYM％(rs=-0.008,P=0.002),and had positive correlated with NLR(rs=0.315,P=0.009).Conclusion:The infarction area of the patients with actue cerebral infarction is correlated with the NIHSS score,the inflammatory response,the degree of adaptive immune injury,and the immune status.The have positive correlation with cytokines and immune markers and the overall size of the infarction area.Compared with the patients with non-large-acea cerebral infarction,the immunosuppression of the patients with large-area infarcted areas is more likely to occure.

Standardized clinical trial reporting is crucial for ensuring the scientific validity,reproducibility,and clinical translational value of reported results.The Consolidated Standards of Reporting Trials(CONSORT)statement,an internationally recognized guideline for randomized controlled trials(RCTs),has become an important reference standard for writing research papers in medicine since the 2010 version of CONSORT was published.With advancements in scientific research methodologies and the emergence of new forms of clinical trials,the CONSORT working group released an updated version in April 2025,published in journals such as The BMJ.Herein,we provide a systematic interpretation of the core revisions of CONSORT 2025,as well as a comparison with CONSORT 2010 to highlight the key differences.By providing practical,example-based recommendations,we aim to help domestic researchers apply the new guidelines efficiently,thereby improving the quality of clinical trial reports authored by domestic researchers.

A high-quality clinical trial protocol is the cornerstone for ensuring the scientific integrity and ethical compliance of a study.The Standard Protocol Items:Recommendations for Interventional Trials(SPIRIT)has become the international benchmark for developing clinical trial protocols since its release in 2013.To adapt to the developing trends of open science and patient-centered principles,the SPIRIT group completed a comprehensive update in 2025.While retaining its core structure,this updated guideline introduces a new open science module and incorporates several new elements,including patient and public involvement,trial monitoring,and data sharing,alongside substantial revisions of five pre-existing items.In this article,we critically examine the core revisions in SPIRIT 2025 and,through analysis of representative case studies,illustrate the practical application of the new reporting guideline in drafting trial protocols.Our goal is to to provide Chinese researchers with a valuable reference for understanding and implementing this new reporting guideline,thereby enhancing the quality and rigor of clinical trial protocols developed in the country.

Objective To establish a stable in vitro model of CD8+T cell exhaustion.Methods CD8+T cells were isolated and purified from the spleens of ovalbumin-specific CD8+T cell receptor(OT-I)transgenic mice and subjected to chronic antigen stimulation to induce exhaustion in vitro.Flow cytometry was employed to evaluate the expressions of exhaustion markers,secretion of effector cytokines,and transcription factor profiles in CD8+T cells.Exhausted and effector(non-exhausted)CD8+T cells were co-cultured with tumor cells before tumor cell viability was measured to assess the cytotoxic potential of CD8+T cells.Additionally,N-acetyl-L-cysteine(N-AC)was used as a positive control during exhaustion induction to validate the model.Results Chronic stimulation resulted in a significant upregulation of inhibitory receptors,including programmed cell death protein 1(PD-1),T cell immunoglobulin and mucin domain-containing protein 3(TIM-3),T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motifdomain(TIGIT),and lymphocyte activation gene 3(LAG-3).Concurrently,the secretion of key effector cytokines such as tumor necrosis factor-α(TNF-α)and interferon-γ(IFN-γ)was markedly reduced.Exhausted CD8+T cells exhibited diminished cytotoxicity against tumor cells compared to effector CD8+T cells.Notably,treatment with N-AC effectively restored the function of exhausted CD8+T cells and enhanced their anti-tumor activity.Conclusion This study has established an effective in vitro model for CD8+T cell exhaustion.The use of N-AC demonstrates its potential to restore functionality in exhausted CD8+T cells,underscoring the reliability and utility of this model for investigating the anti-tumor potential of exhausted T cells.

Background/Aims: Ubiquitination is widely involved in the progression of hepatocellular carcinoma (HCC) by regulating various cellular processes. However, systematic strategies for screening core ubiquitin-related genes, clarifying their functions and mechanisms, and ultimately developing potential therapeutics for patients with HCC are still lacking. Methods: Cox and LASSO regression analyses were performed to construct a ubiquitin-related gene prediction model for HCC. Loss- and gain-of-function studies, transcriptomic and metabolomics analysis were used to explore the function and mechanism of UBE2S on HCC cell glycolysis and growth. Results: Based on 1,423 ubiquitin-related genes, a four-gene signature was successfully constructed to evaluate the prognosis of patients with HCC. UBE2S was identified in this signature with the potential to predict the survival of patients with HCC. E2F2 transcriptionally upregulated UBE2S expression by directly binding to its promoter. UBE2S positively regulated glycolysis in a HIF-1α-dependent manner, thus promoting the proliferation of HCC cells. Mechanistically, UBE2S enhanced K11-linkage polyubiquitination at lysine residues 171 and 196 of VHL independent of E3 ligase, thereby indirectly stabilizing HIF-1α protein levels by mediating the degradation of VHL by the proteasome. In particular, the combination of cephalomannine, a small molecule compound that inhibits the expression of UBE2S, and PX-478, an inhibitor of HIF-1α, significantly improved the anti-tumor efficacy. Conclusions UBE2S is identified as a key biomarker in HCC among the thousands of ubiquitin-related genes and promotes glycolysis by E3 enzyme-independent ubiquitination, thus serving as a therapeutic target for the treatment of HCC.

OBJECTIVE: To observe the therapeutic effect of electroacupuncture at acupoints of liver meridian in patients with diminished ovarian reserve (DOR) of liver depression. METHODS: A total of 62 patients with DOR of liver depression were randomly divided into an electroacupuncture group (31 cases, 1 case discontinued) and a western medication group (31 cases, 1 case was eliminated). Electroacupuncture was applied at bilateral Taichong (LR 3), Ligou (LR 5), Ququan (LR 8), Jimai (LR 12) in the electroacupuncture group, with continuous wave, in frequency of 2 Hz and current of 0.5-1.0 mA, 30 min each time, once every other day, 3 times a week. Femoston was taken orally in the western medication group, oral estradiol tablets were taken for the first 14 days, followed by oral estradiol/progesterone complex tablets for the rest 14 days, 1 tablet a day. Both groups were treated for 3 consecutive menstrual cycles. Before and after treatment, the scores of TCM syndrome, self-rating anxiety scale (SAS) and self-rating depression scale (SDS) were observed, serum levels of follicle stimulating hormone (FSH) and anti-Müllerian hormone (AMH) were detected, and antral follicle count (AFC), peak systolic velocity (PSV) and resistance index (RI) of ovarian artery were measured by color Doppler ultrasound in the two groups, and the clinical efficacy was evaluated after treatment. RESULTS: After treatment, the scores of primary symptom and secondary symptom, as well as the total scores of TCM syndrome were decreased compared with those before treatment (P<0.01), the scores of SAS and SDS, as well as the serum FSH levels and RI of ovarian artery were decreased compared with those before treatment (P<0.01), while the serum AMH levels, AFC and PSV of ovarian artery were increased compared with those before treatment (P<0.05, P<0.01) in the two groups. After treatment, in the electroacupuncture group, the primary symptom score of TCM syndrome was higher than that in the western medication group (P<0.01), the secondary symptom score of TCM syndrome and the scores of SAS and SDS were lower than those in the western medication group (P<0.05, P<0.01). The total effective rate was 70.0% (21/30) in the electroacupuncture group and 73.3% (22/30) in the western medication group respectively, there was no significant difference in the total effective rate between the two groups (P>0.05). CONCLUSION Electroacupuncture at acupoints of liver meridian can effectively improve the clinical symptoms, anxiety and depression, regulate the serum sex hormone levels, increase AFC and improve ovarian blood supply in DOR patients of liver depression.
Humans ; Female ; Electroacupuncture ; Adult ; Acupuncture Points ; Meridians ; Ovarian Reserve ; Young Adult ; Liver Diseases/physiopathology* ; Liver/metabolism* ; Ovary/physiopathology* ; Treatment Outcome ; Depression/therapy*

ObjectivePneumonia is an infectious inflammation of the alveoli， distal airway， and interstitium caused by bacterial， viral， and other pathogens. Maxing Shigantang， originated from Treatise On Cold Damage Diseases， is a classic prescription for treating pneumonia， with significant clinical efficacy. However， its treatment mechanism is still elusive. MethodIn that paper， the transcriptome-based multi-scale network pharmacology was used to reveal the overall pharmacological mechanism of Maxing Shigantang in treating pneumonia from six scales of tissue， cell， pathological process， biological process， signaling pathway， and target. ResultAt the tissue level， Maxing Shigantang mainly acted on the focal tissue of pneumonia-lung and the main inflammatory immune tissues-blood and spleen. Analysis of cell， pathological process and biological process suggested that Maxing Shigantang could treat pneumonia by reversing inflammatory and immune functions and improving cardiopulmonary and vascular injury caused by pneumonia. Analysis of signaling pathway and target showed that Maxing Shigantang regulated inflammatory immune response pathways such as "coronavirus disease-COVID-19" and "Toll-like receptor signaling pathway"， and related targets such as "MAPKAPK3" and "NRG1". ConclusionThis paper， from molecular to tissue levels， indicated Maxing Shigantang treated pneumonia mainly by regulating inflammatory immune response and improving cardiopulmonary and vascular injury.

Lacunar stroke (LS) is one of the subtypes of small cerebral vascular disease. Recent studies have shown that LS has a high risk of cognitive impairment, which imposes a heavy burden on patients, their families and society. However, at present, there is a lack of comprehensive discussion on the related factors, pathogenic mechanism, clinical features, prevention and treatment of cognitive impairment in LS patients. Therefore, this article reviews the relationship between LS and cognitive impairment, in order to provide clinical basis for early prevention and rehabilitation plan, and improve long-term follow-up awareness and comprehensive management ability of cognitive function in LS patients.

Objective:To investigate the efficacy of Kechuanning combined with western medicine on acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and its effects on serum amyloid A, interleukin 1β and procalcitonin levels. Methods:A total of 104 patients with AECOPD who received treatment in Yongkang Hospital of Traditional Chinese Medicine from January 2019 to December 2020 were included in this study. They were randomly assigned to receive either symptomatic treatment with western medicine alone ( n = 52, control group) or symptomatic treatment with western medicine combined with Kechuanning ( n = 52, observation group). Therapeutic effects, latency to clinical symptom relief, pre- and post-treatment pulmonary function, serum inflammatory factor levels, and blood gas analysis indexes were compared between the two groups. Results:Total response rate in the observation group was significantly higher than that in the control group [86.54% (45/52) vs. 67.31%(35/52), χ2 = 4.99, P < 0.05]. Latency to rale disappearance, latency to cough disappearance, length of hospital stay in the observation group were (8.25 ± 1.38) days, (10.05 ± 1.53) days, and (12.65 ± 2.28) days, which were significantly shorter than those in the control group [(9.41 ± 1.46) days, (12.19 ± 1.61) days, (14.36 ± 2.14) days, t = 4.16, 6.98, 3.61, all P < 0.05]. After treatment, forced vital capacity (FVC), forced expiratory volume in the first second (FEV 1), and FEV 1/FVC value in the observation group were (1.88 ± 0.5) L, (64.13 ± 5.72)%pred, (59.43 ± 5.57)%, respectively, which were significantly higher than those in the control group [(1.65 ± 0.51) L, (60.22 ± 5.60)% pred, (54.16 ± 5.19)%, t = 2.17, 3.52, 4.99, all P < 0.05]. Arterial partial pressure of oxygen (PaO 2) and blood oxygen saturation (SpO 2) in the observation group were (9.18 ± 0.89) kPa and (96.26 ± 2.13)%, respectively, which were significantly higher than those in the control group [(8.74 ± 0.76) kPa, (94.07 ± 2.08)%, t = 2.71, 5.305, both P < 0.05]. Partial pressure of carbon dioxide (PaCO 2) in the observation group was significantly lower than that in the control group [(7.32 ± 0.27) kPa vs. (7.63 ± 0.32) kPa, t = 5.34, P < 0.05]. Serum amyloid protein, interleukin-1β and procalcitonin levels in the observation group were (43.84 ± 6.15) mg/L, (3.24 ± 0.51) μg/L, (1.55 ± 0.37) ng/L, respectively, which were significantly lower than those in the control group [(55.26 ± 3.46) mg/L, (4.19 ± 0.56) μg/L, (2.03 ± 0.46) ng/L, t = 9.23, 9.04, 5.86, all P < 0.05]. Conclusion:Kechuanning as an adjuvant therapy for AECOPD can greatly improve lung function and hypoxia, alleviate clinical symptoms, reduce inflammatory reactions, and have a definite clinical effect. The study is innovative and scientific and is worthy of clinical reference.