1.A Single-cell Transcriptomic Study of Wenyang Jiedu Granules in Alleviating Influenza Virus Pneumonia by Suppressing Interferon Responses and Apoptotic Pathways in AT1 Cells
Jingyan XIN ; Shengle QIN ; Taoyu CHEN ; Yuntao LIU ; Xiaolu WANG ; Chaofeng LIANG ; Zhongde ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):76-85
ObjectiveTo investigate the molecular mechanisms by which Wenyang Jiedu granules (WYJD) alleviate influenza A virus (IAV)-induced pneumonia based on single-cell transcriptome sequencing. MethodsThirty female BALB/c mice were randomly divided into a blank control group (Control), IAV group, and WYJD low-, medium-, and high-dose groups (WYJD-L, WYJD-M, WYJD-H; 2.925, 5.85, 11.7 g·kg-1, n=6). Except for the Control group, all other groups were intranasally inoculated with IAV subtype H1N1 (A/PR/8/34) to establish an infection model. Two hours after modeling, drug administration was initiated and continued for 5 consecutive days, with daily monitoring of body weight and general condition. On day 6, mice were sacrificed and samples were collected. Lung index was calculated, and histopathological examination of lung tissue was performed. Lung tissues from the Control, IAV, and WYJD-H groups were subjected to single-cell transcriptome sequencing (n=3), focusing on type I alveolar epithelial cells (AT1) to analyze changes in gene expression and signaling pathways. Western blot was used to detect the expression changes of relevant proteins to validate the single-cell sequencing results. ResultsCompared with the Control group, the IAV group exhibited significantly decreased body weight (P<0.05) and significantly increased lung index (P<0.05). Compared with the IAV group, all WYJD-treated groups exhibited significantly increased body weight (P<0.01) and significantly decreased lung index (P<0.01). Single-cell sequencing analysis revealed that WYJD inhibited overactivation of interferon and inflammatory signaling pathways in AT1 cells after IAV infection, including interferon-γ response, interferon-α response, tumor necrosis factor-α/nuclear factor-κB (TNF-α/NF-κB), and interleukin-6/Janus kinase/signal transducer and activator of transcription 3 (IL-6/JAK/STAT3) pathways. Compared with the Control group, the number of AT1 cells in the IAV group showed a decreasing trend. Compared with the IAV group, the WYJD-H group showed an increasing trend, although neither difference was statistically significant. Further analysis of AT1 cell subpopulation gene expression showed that, compared with the Control group, the IAV group exhibited increased expression of pro-apoptotic genes FAS cell surface death receptor (FAS) and cyclin-dependent kinase inhibitor 1A (CDKN1A), a significant increase in tumor protein p53 (Tp53) expression (P<0.05), and significant decreases in expression of the AT1 marker gene advanced glycosylation end-product-specific receptor (AGER) and membrane structural gene caveolin1 (CAV1) (P<0.05, P<0.01). Compared with the IAV group, the WYJD-H group showed significantly decreased expression of FAS, CDKN1A, and Tp53 (P<0.05, P<0.01), and significantly increased expression of AGER and CAV1 (P<0.05, P<0.01). Regarding interferon response-related genes, compared with the Control group, the IAV group showed increased expression of interferon-stimulated gene 15 (ISG15), interferon-induced protein with tetratricopeptide repeats 3 (IFIT3), signal transducer and activator of transcription 2 (STAT2), bone marrow stromal cell antigen 2 (BST2), and C-X-C motif chemokine ligand 10 (CXCL10), with a significant increase in 2′,5′-oligoadenylate synthetase-like protein 1 (OASL1) (P<0.05). Compared with the IAV group, the WYJD-H group showed significantly decreased expression of all the above genes, with highly significant differences for ISG15, IFIT3, STAT2, BST2, and OASL1 (P<0.01), and a significant difference for CXCL10 (P<0.05). Among inflammation-related genes, compared with the Control group, the IAV group showed significantly increased expression of intercellular adhesion molecule 1 (ICAM1), tumor necrosis factor alpha-induced protein 3 (TNFAIP3), keratin 8 (KRT8), tumor necrosis factor receptor superfamily member 1A (TNFRSF1A), and TNFRSF1B (P<0.01), and increased expression of NFKBIA, a negative regulator of NF-κB (P<0.05). Compared with the IAV group, the WYJD-H group showed significantly decreased expression of KRT8 and TNFRSF1B (P<0.05), while ICAM1, NFKBIA, TNFAIP3, and TNFRSF1A showed decreasing trends without statistical significance. Western blot validation showed that, compared with the Control group, protein expression levels of ISG15, FAS, p53, and phosphorylated p65 (p-p65) in lung tissue of the IAV group were significantly increased (P<0.05, P<0.01). Compared with the IAV group, the WYJD-H group showed significantly decreased expression of these proteins (P<0.05, P<0.01). ConclusionWYJD may alleviate viral pneumonia by targeting gene expression in AT1 cells, inhibiting overactivated interferon and inflammatory signaling pathways after IAV infection, and downregulating pro-apoptotic signaling, thereby reducing alveolar epithelial injury.
2.Celastrol-loaded ginsenoside Rg3 liposomes boost immunotherapy by remodeling obesity-related immunosuppressive tumor microenvironment in melanoma.
Hongyan ZHANG ; Jingyi HUANG ; Yujie LI ; Wanyu JIN ; Jiale WEI ; Ninghui MA ; Limei SHEN ; Mancang GU ; Chaofeng MU ; Donghang XU ; Yang XIONG
Acta Pharmaceutica Sinica B 2025;15(5):2687-2702
Obesity usually exacerbates the immunosuppressive tumor microenvironment (ITME), hindering CD8+ T cell infiltration and function, which further represents a significant barrier to the efficacy of immunotherapy. Herein, a multifunctional liposomal system (CR-Lip) for encapsulating celastrol (CEL) was utilized to remodel obesity-related ITME and improve cancer immunotherapy, wherein Ginsenoside Rg3 (Rg3) was detected interspersed in the phospholipid bilayer and its glycosyl exposed on the surface of the liposome. CR-Lip had a relatively uniform size (116.5 nm), facilitating favorable tumor tissue accumulation through the interaction between Rg3 and glucose transporter 1 overexpressed in obese tumor cells. Upon reaching the tumor region, CR-Lip was found to induce the immunogenic cell death (ICD) of HFD tumor cells. Notably, the level of PHD3 in HFD tumor cells was effectively boosted by CR-Lip to effectively block metabolic reprogramming and increase the availability of major free fatty acids fuel sources. In vivo, experiments studies revealed that the easy-obtained nano platform stimulated enhanced the production of various cytokines in tumor tissues, DC maturation, CD8+ T-cell infiltration, and synergistic anticancer therapeutic potency with aPD-1 (tumor inhibition rate = 82.1%) towards obesity-related melanoma. Consequently, this study presented an efficacious approach to tumor immunotherapy in obese mice by encompassing tumor eradication, inducing ICD, and reprogramming metabolism. Furthermore, it offered a unique insight into a valuable attempt at the immunotherapy of obesity-associated related tumors.
3.The crucial function of IDO1 in pulmonary fibrosis: From the perspective of mitochondrial fusion in lung fibroblasts and targeted molecular inhibition.
Lei WANG ; Shanchun GE ; Ye ZHANG ; Deqin FENG ; Ting ZHU ; Louqian ZHANG ; Chaofeng ZHANG
Acta Pharmaceutica Sinica B 2025;15(6):3125-3148
The pathogenesis of pulmonary fibrosis (PF) is complex. It is characterized by myofibroblast hyperplasia and deposition of collagen protein. Indoleamine 2,3-dioxygenase 1 (IDO1) is expressed in lung fibroblasts and epithelial cells, but its functions in lung homeostasis and diseases remain elusive. Here, we characterize the critical role of IDO1 in PF patients and bleomycin (BLM)-induced PF mouse models. We find that IDO1 is significantly upregulated in the fibrotic lungs of patients and mice, showing a positive correlation with genes characteristic of fibrosis. Functionally, IDO1 knockout inhibits lung fibroblast proliferation, differentiation, mitochondrial biogenesis, and mitochondrial oxidative phosphorylation. Conversely, IDO1 overexpression and accumulation of kynurenine (Kyn) exacerbate progressive lung fibrosis. Mechanistically, IDO1-deletion activated profound mitochondrial fusion-enhanced potentially the capacity for fatty acid oxidation, along with activation of de novo glycolytic serine/glycine synthesis pathways and mitochondrial one-carbon metabolism. Wedelolactone (WEL), a small molecule IKK inhibitor, is found to strongly bind to IDO1 and effectively protect mice from PF in an IDO1-dependent manner. Collectively, this study characterizes a promotor role for IDO1 in PF and suggests a potential avenue of targeting IDO1 to treat lung diseases.
4.Research progress on new techniques and methods for identifying active ingredients in traditional Chinese medicine.
Jiaxin ZHANG ; Xinhao ZHU ; Chaofeng ZHANG ; Wangning ZHANG ; Jiangwei TIAN
Chinese Journal of Natural Medicines (English Ed.) 2025;23(10):1153-1170
Recent years have witnessed significant advances in the development of novel techniques and methodologies for identifying active ingredients in traditional Chinese medicine (TCM), substantially advancing research and development efforts. Spectrum-effect correlation analysis, affinity ultrafiltration, high-content screening (HCS) imaging, and cell membrane chromatography (CMC) have emerged as essential tools, effectively linking TCM chemical constituents to their biological effects, thereby enabling efficient active ingredient screening. Additionally, molecular interaction analysis provides deeper insights into TCM-biomolecule interaction mechanisms, enhancing understanding of its therapeutic potential. Computer-aided techniques facilitate TCM active ingredient identification, optimizing the screening process for efficiency and cost-effectiveness. Molecular probe technology, as an emerging methodology, enables precise and rapid screening for novel therapeutic drug discovery. Ongoing technological advancement in this field indicates promising future developments, potentially leading to more effective and targeted TCM-based therapies.
Drugs, Chinese Herbal/chemistry*
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Medicine, Chinese Traditional/methods*
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Humans
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Drug Discovery/methods*
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Animals
5.Research on microscopic histochemical localization of Astragalus membranaceus and its application
Yong HUANG ; Shengchao SHI ; Jian ZHANG ; Chaofeng ZHANG
Journal of China Pharmaceutical University 2025;56(5):566-571
This study compared the chemical composition differences among various root tissues of Astragalus membranaceus through optimization of histochemical localization methods, thereby providing some reference for targeted extraction and enrichment of bioactive components. The chloral hydrate permeabilization technique was enhanced with sodium nitrite-aluminum nitrate-sodium hydroxide reagent for flavonoid staining. Comparative analyses of total saponins, total flavonoids, and astragaloside IV content in xylem, phloem, and periderm tissues were performed using ultraviolet-visible spectroscopy (UV-Vis) and high-performance liquid chromatography (HPLC). By optimizing staining protocols, interference from natural pigmentation was successfully eliminated, thereby enhancing the visualization of tissue-specific chemical distribution. Quantitative analysis revealed that the phloem contained the highest total saponins (6.92%), while the periderm exhibited peak total flavonoids (0.876%) and exclusive enrichment of astragaloside IV (0.850%). The refined histochemical localization method enables precise characterization of phytochemical distribution across root tissues, which can offer guidance for selective extraction and enrichment strategies, and comprehensively promote the utilization and development of Astragalus membranaceus resources.
6.Intraosseous and periosteal arteries of hallux phalanx: a 3D observational study
Zirun XIAO ; Jia CHEN ; Kuo WEI ; Zhenfeng LI ; Chaofeng XING ; Weiguang ZHANG
Chinese Journal of Microsurgery 2025;48(5):547-551
Objective:To explore the feasibility of obtaining three-dimensional (3D) models of intraosseous and periosteal arteries of hallux phalanx using micro-arteriography with micro-CT scan.Methods:From January 2022 to April 2025, the Department of Orthopaedic, the 988th Hospital of the Joint Logistics Support Force of the Chinese PLA conducted a study on 7 fresh-frozen specimens of distal lower limb (right lower limb) from an 85-year-old male, and both lower limbs from an 82-year-old male, a 78-year-old female and a 66-year-old male in the Department of Human Anatomy & Histology and Embryology, Peking University School of Basic Medical Sciences. Red lead oxide powder (Pb 3O 4) was ground and filtered through a 300 mesh, and then mixed with turpentine at ratios of 1 g ∶ 1.5 ml, 1 g ∶ 1.0 ml, and 1 g ∶ 0.5 ml to prepare lead-based contrast agent suspensions. After thawing the specimens at room temperature, the suspensions were injected via the popliteal artery in ascending order of concentration. After injections, the specimens were fixed in 10% methanal for 2 weeks. The proximal and distal phalanges of the hallux, with the surrounding periosteum preserved intact, were then harvested. The harvested specimens were scanned using micro-CT at an ultimate resolution of 12 μm. Subsequently, Mimics Medical software was used to reconstruct 3D models of the intraosseous and periosteal arteries within the phalanges. Results:Periosteal arteries in the proximal phalanx were primarily distributed near the joint region. A consistently large trunk artery entered from plantar side, supplying most of the diaphysis and head. There was a rich periosteal arterial network on both sides of the distal phalanx, which communicates with each other through the arterial arch in the bone groove. However, trunk intraosseous artery could be absent. Intraosseous arteries in the proximal ends of both the proximal and distal phalanges originated from periosteal arteries. These formed an interconnected arteriosomes and coursed parallel to the articular surfaces.Conclusion:The micro-arteriography acquired by micro-CT scan effectively visualizes intraosseous and periosteal arteries and enables the reconstruction and analysis of 3D models of the arteriosomes. The characteristics of arteriolar distribution provide a theoretical basis for osteotomy or internal fixation procedures involving a hallux phalanx.
7.MicroPET/CT-based exploration of the effects of acute sleep deprivation on glucose metabolism and neuroinflammation in rat brain
Mengya DAI ; Zhenyu XIANG ; Yan ZHANG ; Chaofeng LIU ; Jie GAO ; Zhixing QIN ; Hongliang WANG ; Zhifang WU ; Jianguo LI ; Sijin LI
Chinese Journal of Nuclear Medicine and Molecular Imaging 2025;45(9):555-559
Objective:To investigate the effects of acute sleep deprivation (ASD) on hippocampal glucose metabolism and neuroinflammation in rat models.Methods:Twenty SD rats (10 males and 10 females) were divided into four groups (five in each group) by random sampling method: female ASD group, male ASD group, female control group, and male control group. Among them, the ASD group constructed the ASD model. After 72h sleep deprivation, all rats underwent 18F-FDG and N, N-diethyl-2-(2-(4-(2- 18F-fluoroethoxy)phenyl)-5, 7-dimethylpyrazolo[1, 5-a]pyrimidin-3-yl)acetamide ( 18F-DPA-714) microPET/CT brain imaging in 2d to compare the changes of 18F-FDG and 18F-DPA-714 SUV mean in the hippocampus of rats. Brain histopathology, immunohistochemistry and immunofluorescence staining were detected in rats. Independent-sample t test was used to analyze the data. Results:18F-FDG imaging showed the hippocampal SUV mean between ASD group and control group (female: 4.11±0.35 vs 1.89±0.28; male: 3.43±0.47 vs 2.02±0.54) were statistically significant ( t values: 9.65, 3.92, P values: <0.001, 0.002). 18F-DPA-714 imaging showed the hippocampal SUV mean between ASD group and control group (females: 0.28±0.01 vs 0.28±0.02; male: 0.26±0.02 vs 0.31±0.04) were not statistically significant ( t values: -0.18, -2.24, P values: 0.859, 0.056). The 18×10 3 translocator protein (TSPO) immunohistochemistry showed the expression in the hippocampal region of the brain between ASD group and control group (female: 0.19±0.02 vs 0.19±0.01; male: 0.21±0.01 vs 0.20±0.01) were not statistically different ( t values: -0.48, -1.67, P values: 0.651, 0.139). Immunofluorescence staining showed that microglial cytosol in the hippocampal region of the brain decreased after 72h of ASD, and the protrusion points and surrounding branches were significantly reduced. Conclusion:Increased hippocampal glucose metabolism in rats is observed after 72 h of ASD without significant neuroinflammation.
8.Clinical efficacy analysis of autologous rib cartilage reconstruction for finger hemiarticular defects
Zhiyu HU ; Chaofeng XING ; Tao YANG ; Jia CHEN ; Zirun XIAO ; Xiazhi LIU ; Li SONG ; Beibei CHENG ; Yingjie XIONG ; Guangchao ZHANG ; Songfeng YANG
Journal of Clinical Surgery 2025;33(6):641-645
Objective Exploring the clinical efficacy of using autologous rib cartilage grafting to reconstruct finger hemiarticular defects.Methods From August 2022 to February 2024,for 6 patients with hemiarticular surface defects in the metacarpophalangeal joints and interphalangeal joints of 8 fingers,costal cartilage was used for joint remodeling and transplantation to reconstruct the hemiarticular surface defects of the fingers.All 8 joint transplants in 6 patients were two-stage surgeries.In the first stage,antibiotic bone cement was used to fill the space-occupying lesions,and in the second stage,costal cartilage joint remodeling was performed to reconstruct the finger joint defects.Postoperative follow-up and assessment of fracture healing according to Paley fracture healing scoring criteria.Outpatient and WeChat follow-up,upper limb function is evaluated according to the upper limb functional assessment standards of the Chinese Medical Association Hand Surgery Society.Record VAS pain score.Results In this group,there were a total of 6 patients with 8 cases of hemiarticular defects.Among them,2 patients had two joint surgical sites,while the remaining 4 patients had a single joint surgical site.There were 2 cases of metacarpophalangeal joint head defects,2 cases of proximal articular surface defects,3 cases of proximal articular head defects,and 1 case of thumb proximal articular head defect.After surgery,8 out of 6 patients'hand wounds healed successfully.All patients were followed up for 6-12 months postoperatively,with an average of 9 months.The VAS pain score(affected finger)for the last follow-up was 0-2 points,with an average of 0.6 points.According to Paley's scoring criteria,all 6 patients had excellent fracture healing.According to the evaluation criteria for upper limb functional assessment of the Chinese Medical Association Hand Surgery Society,3 cases were rated as excellent,3 cases were rated as good,and 2 cases were rated as fair.Conclusion For patients with half joint defects on one side of the finger,rib rib cartilage was used for joint reconstruction,which significantly improved the joint shape and function at the defect site,and reduced joint pain scores.
9.The application progress of music intervention in nursing of patients with psychiatric disorders
Haijiao LI ; Dongshuai WEI ; Ruiyan WANG ; Rongxun LIU ; Chaofeng ZHANG ; Jie JIA ; Yange WEI
Chinese Journal of Nursing 2025;60(6):764-768
Music intervention is an innovative nursing method that integrates music,medicine,and psychology.It is safe,effective,and easy for implemention.It has demonstrated positive effects in alleviating psychotic symptoms,stabilizing emotions,and improving sleep,and it is gradually being applied in psychiatric disorders.This article provides a review of the theoretical basis and current research status of music intervention,and offers a perspective on future applications,with the aim of providing a reference for the application of music intervention in the rehabilitation nursing of mental illnesses.
10.Survival Analysis of Biliary Tract Carcinoma Patients Receiving Integrated Traditional Chinese and Western Medicine Treatment:A Retrospective Cohort Study
Junke XIAO ; Chaofeng ZHOU ; Sai WANG ; Yu ZHANG ; Jiaojiao GUO ; Shangzhi YANG ; Xiaoyan MU ; Zhizhong GUO
Herald of Medicine 2025;44(8):1236-1241
Objective To explore the impact of integrated traditional Chinese and Western medicine on the overall survival(OS)of patients with biliary tract carcinoma(BTC),and to analyze the related factors affecting the OS of patients with BTC,so as to provide accurate prognosis assessment for clinical diagnosis and treatment.Methods The medical records of 198 patients with BTC in Henan Provincial Hospital of Traditional Chinese Medicine from February 2018 to October 2023 were retrospectively analyzed.The Cox proportional hazards model was utilized to analyze the independent prognostic factors affecting the OS of BTC patients.Survival curves were constructed using the Kaplan-Meier method,and the Log-rank test was employed to compare median OS differences among various groups.Results A total of 116 patients with BTC were included.There were 59 cases(50.86%)in the Western medicine group,with a median OS of(14±2.77)months,and the 1-,2-and 3-year OS rates were 81.36%,61.02%and 46.11%,respectively.There were 57 cases(49.14%)in the integrated traditional Chinese and Western medicine group,with a median OS of(29±3.91)months,and the 1-,2-and 3-year OS rates were 82.32%,67.64%and 58.00%,respectively.There was a statistically significant difference in survival time between the two groups(P=0.038).Univariate analysis indicated that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy and integrated traditional Chinese and Western medicine were significantly correlated with OS in BTC patients(all P<0.05).Multivariate analysis revealed that age,clinical stage,surgical treatment,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine had significant effects on the OS of patients with BTC(all P<0.05).Conclusion The median OS of the traditional Chinese and Western medicine groups was significantly longer than that of the Western medicine group,and the long-term OS rate of the patients was significantly better than that of the Western medicine group.Age,clinical stage,surgery,chemotherapy,targeted therapy,local therapy,and integrated traditional Chinese and Western medicine are independent prognostic factors affecting OS in patients with BTC.

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