1.Integrating Transcriptomics and 3D Organoids to Investigate Mechanism of Periplaneta americana Extract Against Lung Adenocarcinoma
Qiong MA ; Chunxia HUANG ; Jiawei HE ; Yuting BAI ; Xingyue LIU ; Yuxuan XIONG ; Yang ZHONG ; Hengzhou LAI ; Yuling JIANG ; Xueke LI ; Qian WANG ; Yifeng REN ; Xi FU ; Funeng GENG ; Taoqing WU ; Ping XIAO ; Fengming YOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(11):124-132
ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract(PAE) against human-derived lung adenocarcinoma organoids(LUAD-PDOs) and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid(Nor-PDOs) models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8(CCK-8) assay. Experimental groups included the model group(LUAD-PDOs), normal group, model administration group(LUAD-PDOs+PAE), and normal administration group(Nor-PDOs+PAE). Hematoxylin-eosin(HE) staining was used to observe the pathological structures of PDOs, immunohistochemistry(IHC) was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7(CK-7) and Napsin A, TUNEL staining was applied to detect cell apoptosis. RNA sequencing(RNA-Seq) was conducted to identify differentially expressed genes(DEGs), followed by Gene Ontology(GO), Kyoto Encyclopedia of Genes and Genomes(KEGG), and Gene Set Enrichment Analysis(GSEA), alongside protein-protein interaction(PPI) network analysis to screen core mechanisms. Finally, key targets were validated by integrating external database analysis with immunofluorescence(IF). ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures, whereas LUAD-PDOs displayed dense, solid structures. CCK-8 and TUNEL assays revealed that, compared with the model group, PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells, while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention(347 up-regulated and 372 down-regulated)(P<0.05). GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism, including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway(P<0.05). PPI network analysis indicated that breast cancer type 1 susceptibility protein(BRCA1) and checkpoint kinase 1(CHEK1) were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention(P<0.05). Furthermore, survival analysis based on The Cancer Genome Atlas(TCGA) database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD(P<0.05). ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis, its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway, with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.
2.Regulatory effect of astragaloside Ⅰ and calycosin on a mouse model of cholestatic liver fibrosis
Xiaoyu JIANG ; Wei LIU ; Jiamei CHEN ; Ping LIU ; Chunhui LI
Journal of Clinical Hepatology 2026;42(5):1083-1092
ObjectiveTo identify and validate the optimal compatibility dosage of astragaloside Ⅰ (ASⅠ) and calycosin (CY) in the treatment of cholestatic liver fibrosis. MethodsA 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet was used to establish a mouse model of liver fibrosis, and the uniform design method was used to identify the optimal combination ratio of the saponin component ASⅠ and the flavonoid component CY in Astragalus membranaceus. In the uniform design experiment, 80 male C57/BL6J mice were divided into normal group, model group, total astragalosides (TAS) group, groups A — F with a uniform design, and obeticholic acid (OCA) group using a random number table, with 8 mice in each group. The multiple regression analysis was used to establish the optimal regression equation and obtain the potential optimal combination ratio. The in vivo efficacy of the empirically optimal combination identified in the uniform design and the optimal dose combination predicted by the regression equation were compared for validation. A one-way analysis of variance was used for comparison of continuous data between multiple groups; the Levene test was used to determine the homogeneity of variance, and the least significant difference t-test was used for comparison of data with homogeneity of variance between two groups, while the Dunnett T3 test was used for comparison of data with heterogeneity of variance. ResultsIn the uniform design regimen, the JYB combination (3.125 mg/kg ASI+50 mg/kg CY) significantly reduced the serum levels of alanine aminotransferase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), total bile acid (TBA), total bilirubin (TBil), and indirect bilirubin (IBil) in mice with DDC-induced cholestatic liver fibrosis (all P<0.05), and it also reduced hepatic Hyp content (P<0.01), semi-quantified collagen deposition area (P<0.001), and the mRNA expression levels of Acta2, Col1a1, Ck7, Ck19, Adgre1, TLR4, TNF-α, and CCL5 in liver tissue (all P<0.05). The regression equation showed that 50 mg/kg ASⅠ+50 mg/kg CY was the potential optimal combination, which was named as P1 combination. However, subsequent validation experiments showed that P1 combination only significantly improved the serum levels of AST and IBil and hepatic Hyp content in DDC mice (all P<0.05), with no significant impact on hepatic collagen deposition and the mRNA expression levels of Acta2, Ck7, Ck19, Adgre1, and CCL5 (all P>0.05). In contrast, the JYB combination significantly improved the serum levels of ALP, ALT, AST, TBA, TBil, and IBil, hepatic collagen deposition, hepatic Hyp content, and the mRNA expression levels of Acta2, Col1a1, Ck7, Ck19, Adgre1, TLR4, TNF-α, and CCL5 (all P<0.05). ConclusionThis study shows that the JYB combination (3.125 mg/kg ASⅠ+50 mg/kg CY) can significantly alleviate DDC-induced liver fibrosis, with comparable efficacy to total saponins from Astragalus membranaceus, and compared with ASⅠ or CY administered alone, the JYB combination has a significantly better regulatory effect on the serum levels of ALP and TBA.
3.Analysis of soil-borne nematode infection status among rural communities in Yubei, Chongqing
Dan JIANG ; Yong-dong HAO ; Sen-ping YANG ; Xiao-yuan SU ; Hua-jun BAI ; Bo LYU ; Ya-ling RAN ; He-yi GUAN ; Ling HU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):85-89
Objective To analyze the infection status and epidemic trends of soil-borne nematode infections in Yubei, Chongqing City, in 2010,2021, and 2022. Methods The local populations from four survey sites of four towns in 2010 and five sites of five towns in 2021 and 2022 were surveyed regarding their basic information using a unified form. Fecal samples of the participants were collected and tested for soil-borne nematode infections using the modified Kato-Katz thick smear method. Results In 2010, 2 049 participants were surveyed, followed by 1 000 participants in 2021 and 2022. The overall prevalence of parasitic infections declined significantly from 3.86% to 0.20%. In 2010, soil-transmitted nematode included hookworms(3.81%) and roundworms(0.29%). In 2021, the infection rates of roundworms and hookworms were 1.70% and 0.10% respectively. Notably, only Ascaris was identified in 2022(0.20%). The≥60 age group consistently exhibited the highest infection rates across all surveys, followed by the 40-59 age group. The infection rates of males in the three surveys were 3.31%,1.92%, and 0.20% respectively, and those of females were 4.37%, 1.46%, and 0.20% respectively. There was no statistically significant difference in the infection rates between males and females. Educational attainment was inversely associated with infection; in 2010, the highest prevalence was observed among those with primary education or below, whereas in 2021, illiterate or semi-literate individuals showed the highest susceptibility. The occupational distribution of infections in 2010 indicated that retirees (8.33%), farmers(4.86%), and homemakers or unemployed individuals(3.45%)were the most affected. However, in 2021 and 2022, farmers emerged as the predominant occupational group with soil-transmitted nematode infections. Conclusions The infection rate of soil-borne nematodes showed a decreasing trend in Yubei, and the infection species changed from hookworms in 2010 to Ascaris in 2022. Farmers, the elderly, and people with low education levels should continue to be the focus of preventive and control efforts.
4.A new species of Culicoides (Avaritia) (Diptera: Ceratopogonidae) in Heilongjiang Province, China
Ya-yu WANG ; Jiang-fan LI ; Bo-qiao CAI ; Guo-ping LIU
Acta Parasitologica et Medica Entomologica Sinica 2026;33(1):62-65
This study reports a new species of Culicoides(Avaritia)isolated from Xunke County, Heilongjiang Province, China. The new species, Culicoides(Avaritia)dongshanensis Liu et Wang, sp. nov., was identified and illustrated based on female adults. Its diagnostic characteristics are as follows: eyes contiguous over a short distance, with short interfacetal hair, antennal ratio(AR)of 1.20, palpal ratio(PR)of 2.16, wing length of 0.83 mm, mandible with 15 teeth, one pale spot in base cell M1, one pale spot in cell M4, two pale spots in cell A, and two unequal spermathecae. Type specimens were deposited at the Center for Disease Control and Prevention of the Northern Theater Command.
5.Machine learning-enabled precision transfusion: research progress, challenges, and prospects
Jiang DENG ; Chaojie WANG ; Ning ZHAO ; Liping LYU ; Ping MA ; Ke ZHANG ; Yanyu ZHANG
Chinese Journal of Blood Transfusion 2026;39(7):967-976
Blood transfusion is an important life-support measure in modern medicine. As clinical demand for transfusion continues to rise, blood supply remains under persistent strain, making the efficient utilization of this precious medical resource a critical component of blood management. Accurate prediction of transfusion needs is of great significance for optimizing blood resource allocation and conserving blood products. Traditional transfusion decision-making relies on clinical experience and simplified scoring systems, which are insufficient to meet the demands of precision medicine. Machine learning (ML) technology can integrate multidimensional data—including demographic characteristics, laboratory indicators, surgical information, and dynamic physiological waveforms—to construct high-performance predictive models, offering a new approach to transfusion prediction. This article systematically reviews the application progress of ML in transfusion prediction across clinical scenarios such as trauma, the perioperative period, and obstetrics. In trauma, ML has been applied to early massive transfusion prediction, prehospital transfusion decision-making, and precise prediction in pediatric trauma. In the perioperative field, applications span specialties including traumatic brain injury, orthopedic surgery, cardiovascular and major vascular surgery, and hepatic surgery. In obstetrics, models can effectively predict postpartum hemorrhage and transfusion requirements associated with cesarean section. Studies have shown that algorithms such as random forest, gradient boosting machines, and deep neural networks achieve area under the receiver operating characteristic curve (AUC) values of 0.83-0.98 in massive transfusion prediction, 0.75-0.97 in perioperative scenarios, and 0.80-0.89 in obstetric transfusion prediction—significantly outperforming traditional scoring tools. However, current studies are generally limited by single-center retrospective designs, insufficient external validation, and inadequate reporting standards; limited model interpretability and challenges in clinical integration further constrain practical translation. Future research should focus on constructing multicenter data cohorts, integrating multimodal data, conducting prospective implementation studies, and deeply embedding models within electronic health record systems, thereby advancing the application of ML to optimize clinical transfusion decision-making.
6.Stability of the molluscicidal activity of the fermentation broth of Bacillus indicus F19
Liyun ZHU ; Lei LIN ; Ning XU ; Ping LONG ; Xinran YANG ; Yu ZHOU ; Yanfeng GONG ; Qingwu JIANG ; Yibiao ZHOU
Chinese Journal of Schistosomiasis Control 2026;38(3):304-308
Objective To assess the stability of the molluscicidal activity of the fermentation broth supernatant of Bacillus indicus F19, so as to provide insights into development of novel molluscicides derived from this isolate. Methods Healthy Oncomelania hupensis snails free from Schistosoma japonicum infections and acclimated in the laboratory for 7 days were selected. The fermentation broth supernatant of B. indicus F19 was prepared. For each treatment condition, four experimental groups and one control group were established, with 10 mL of the fermentation broth supernatant each group. O. hupensis snails were exposed to different pH values (3, 6, 9 and 12), temperatures (40, 60, 80 °C and 100 °C), and ultraviolet (UV) exposure durations (1, 2, 3 h and 4 h). Then, the supernatant was diluted with dechlorinated water to concentrations of 50%, 10%, and 5% (v/v), and 10 mL of each dilution solution was added to three separate tubes, with an equal volume of dechlorinated water as a blank control. Twenty O. hupensis snails were transferred to each tube, immersed for 48 h and 72 h, and collected for viability assessment using the shell-tapping method. The mortality of O. hupensis snails was calculated and compared among dilution groups and the blank control group to evaluate the molluscicidal activity of the fermentation broth supernatant of B. indicus F19. Results There was a significant difference in the mortality of O. hupensis snails immersed in 5% fermentation broth supernatants of B. indicus F19 for 48 h at various pH levels (χ2 = 11.556, P < 0.05), and the mortality reduced to 13.33% (8/60) in the experimental groups at a pH value of 12, which was significantly lower than in the blank control group [33.33% (20/60)] (χ2 = 6.708, P < 0.05). There was a significant difference in the mortality of O. hupensis snails immersed in 10% fermentation broth supernatants of B. indicus F19 for 72 h at various pH levels (χ2 = 31.691, P < 0.001), and the mortality reduced to 71.67% (43/60) and 65.00% (39/60) in the experimental group at pH values of 9 and 12, which was significantly lower than in the blank control group [100.00% (60/60)] (χ2 = 8.571 and 12.110, both P values < 0.01). The mortality of O. hupensis snails both reduced to 23.33% (14/60) following immersion in 5% fermentation broth supernatants of B. indicus F19 heated in a water bath at 80 °C and 100 °C for 48 h, which was lower than that of unheated O. hupensis snails [33.33% (20/60)] and snails heated in a water bath at 40 °C [35.00% (21/60)] and 60 °C [33.33% (20/60)], and there was no significant difference in the mortality of O. hupensis snails among groups (χ2 = 3.898, P > 0.05). The mortality of O. hupensis snails was 68.33% (41/60), 66.67% (40/60), 65.00% (39/60), and 68.33% (41/60) following immersion in fermentation broth supernatants heated in a water bath at 40, 60, 80 °C and 100 °C for 72 h, which was all lower than that of snails in the control group [71.70% (43/60)]; however, there was no significant difference seen in the snail mortality among groups (χ2 = 0.674, P > 0.05). The mortality of O. hupensis snails was 73.33% (44/60), 71.67% (43/60), 71.67% (43/60), and 75.00% (45/60) following 48 h immersion in 10% fermentation broth supernatants of B. indicus F19 irradiated by UV for 1, 2, 3 h and 4 h, 33.33% (20/60), 31.67% (19/60), 35.00% (21/60), and 33.33% (20/60) following 48 h immersion in 10% fermentation broth supernatants of B. indicus F19 irradiated by UV for 1, 2, 3 h and 4 h, and 71.67% (43/60) and 33.33% (20/60) following 48 h immersion in 10% fermentation broth supernatants of B. indicus F19 without UV irradiation, and no significant differences were found among groups (χ2 = 0.229 and 0.150, both P values > 0.05). In addition, the mortality of O. hupensis snails was 76.67% (46/60), 71.67% (43/60), 70.00% (42/60), and 76.67% (46/60) following 72 h immersion in 5% fermentation broth supernatants of B. indicus F19 irradiated by UV for 1, 2, 3 h and 4 h, and 71.67% (43/60) in the control group, and there was no significant difference in the snail mortality among groups (χ2 = 1.193, P > 0.05). Conclusion The molluscicidal action of the fermentation broth supernatants of B. indicus F19 is tolerant to high temperature and UV irradiation; however, the activity is attenuated under alkaline conditions.
7.Activation of α7nAChR optimises thermogenesis in brown adipose tissue of obese mice
Wencheng TIAN ; Sutong WANG ; Hesong FAN ; Yan LI ; Ping JIANG ; Xiao LI
Acta Universitatis Medicinalis Anhui 2026;61(7):1191-1199
ObjectiveTo investigate the mechanism by which activation of α7nicotinic acetylcholine receptors (α7nAChR) optimizes brown adipose tissue (BAT) thermogenesis in obese mice. MethodsObesity was induced in C57BL/6J mice via a high-fat diet. Fifty 8-week-old mice were randomly assigned to five groups: low-fat diet (Control), high-fat diet (HFD), high-fat diet plus a β3-adrenergic receptor agonist (HFD+β3), high-fat diet plus a β3-adrenergic receptor agonist and an α7nAChR selective agonist (HFD+β3+GTS-21), and high-fat diet plus a β3-adrenergic receptor agonist and an α7nAChR selective antagonist (HFD+β3+α-BGT). Haematoxylin and eosin (HE) staining was used to evaluate BAT morphology. Transmission electron microscopy was performed to assess mitochondrial number and lipid droplet morphology in adipocytes. ELISA was used to measure levels of tumour necrosis factor-α (TNF-α), interleukin(IL)-1β,IL-10, transforming growth factor-β (TGF-β), cyclic adenosine monophosphate (cAMP), and norepinephrine (NE). RT-qPCR was conducted to determine mRNA expression levels of vascular endothelial growth factor A (VEGF-A), nitric oxide synthase 2 (NOS2), arginase 1 (Arg1), uncoupling protein 1 (UCP1), PR domain-containing protein 16 (PRDM16), and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) in BAT. Immunohistochemistry was used to detect macrophage markers (CD31, CD86, and CD206). Protein expression levels of UCP1, α7nAChR, nuclear factor-κB p65(NF-κB p65), phosphorylated Janus kinase 2 (p-JAK2), and phosphorylated signal transducer and activator of transcription 3 (p-STAT3) were analyzed by Western blot. ResultsCompared with the Control group, the HFD group showed enlarged lipid droplets and reduced mitochondrial numbers in adipocytes, accompanied by increased TNF-α and IL-1β and decreased IL-10, TGF-β, Arg1, and VEGF-A (all P<0.01). Compared with the HFD+β3 group, the HFD+β3+GTS-21 group showed a smaller lipid droplet area and a higher mitochondrial number, along with reduced levels of pro-inflammatory factors, NOS2, and NF-κB p65, and increased levels of anti-inflammatory factors, Arg1, and phosphorylation of JAK2 and STAT3 (all P<0.01). ConclusionActivation of α7nAChR in combination with a β3-adrenergic receptor agonist effectively enhances thermogenesis, alleviates local adipose tissue hypoxia, and shifts the local inflammatory phenotype toward an M2-like state, thereby reducing inflammation in brown adipose tissue and improving its thermogenic capacity.
8.Construction of Mouse Models of Psoriasis-like Lesions Induced by Cold Exposure Combined with Imiquimod and Evaluation of Therapeutic Efficacy of Kaixuan Jiedu Core Prescription
Meiqi SUN ; Xue XIAO ; Jiarong WU ; Jiaqi LI ; Ningxin ZHANG ; Mengyao JIANG ; Huan LIU ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(19):69-78
ObjectiveTo establish the mouse models of psoriasis-like lesions induced by continuous cold exposure or intermittent cold exposure combined with imiquimod (IMQ), and to evaluate the interventional effects of Kaixuan Jiedu core prescription (KXJD) on the two models. MethodsMale C57BL/6J mice were selected and classified into two experimental batches. The first batch of 36 mice was randomized into a room temperature group, a continuous cold exposure (10 ℃/24 h) group, and an intermittent cold exposure (10 ℃/6 h) group. Each group was further divided into a normal subgroup and a model subgroup (topical application of IMQ to induce skin lesions), with 6 mice in each subgroup, for modeling and evaluation. The second batch of 54 mice, with 6 in each group, were subjected to the same temperature grouping with an additional KXJD (30.42 g·kg-1, continuous gavage for 5 days) group. Comprehensive evaluation of model characteristics and KXJD efficacy was conducted through Psoriasis Area and Severity Index (PASI) scoring, skin temperature measurement by infrared thermography, histopathological observation by hematoxylin-eosin (HE) staining, detection of vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule 1 (CD31) by immunohistochemistry, detection of Claudin-1 and Occludin by immunofluorescence assay, determination of serum levels of tumor necrosis factor-α (TNF-α) and interleukin (IL)-10 by enzyme-linked immunosorbent assay (ELISA), and quantification of mRNA levels of IL-17A, IL-23, IL-6, and chemokine ligand 20 (CCL20) in skin lesions by quantitative Real-time polymerase chain reaction (Real-time PCR). ResultsModel mice in all temperature groups exhibited typical psoriasis-like skin lesions. Compared with the normal groups, the model groups showed increased PASI scores, decreased skin temperatures (P<0.05), obvious epidermal thickening, parakeratosis, and dermal inflammatory cell infiltration, as well as elevated mRNA levels of IL-17A, IL-23, IL-6, and CCL20 (P<0.05). Cold exposure further aggravated psoriasis. The total PASI score of the intermittent cold exposure model group was higher than that of the room temperature model group (P<0.05). The serum IL-10 did not show a compensatory elevation, and the blood vessels presented a characteristic of elevated CD31 expression (P<0.05) without a synchronous increase in VEGF. The continuous cold exposure model group exhibited more significant dermal capillary tortuosity and dilation, with the highest mRNA levels of IL-17A, IL-23, IL-6, and CCL20 among all groups. Compared with the respective model groups, KXJD intervention alleviated skin lesions, reduced epidermal thickness and inflammatory cell infiltration, and increased skin temperature, with the temperature increase being particularly significant in the intermittent cold exposure+KXJD group (P<0.05). Furthermore, KXJD down-regulated the expression of VEGF and CD31, restored the expression of Claudin-1 and Occludin, decreased the mRNA levels of IL-17A and IL-23 (P<0.05), and reduced the serum TNF-α level. ConclusionThis study successfully established compound psoriasis-like mouse models induced by cold exposure combined with IMQ. It confirms that cold aggravates the severity of psoriasis by exacerbating the closure of Xuanfu (sweat pores), microcirculation disorders, and immune imbalance. Moreover, different cold exposure patterns have distinct mechanism differences. Continuous cold exposure focuses on enhancing the inflammatory response via the IL-23/IL-17 axis and angiogenesis, simulating chronic aggravation under a long-term cold environment. Intermittent cold exposure tends to impair immune regulation and induce microvascular endothelial stress, corresponding to acute exacerbations caused by sudden temperature drops. KXJD can effectively alleviate psoriasis-like skin lesions under cold conditions by unblocking Xuanfu, regulating vasomotor function, and correcting abnormal immune-inflammatory responses.
9.Kaixuan Jiedu Core Prescription Alleviates Restraint Stress-exacerbated Imiquimod-induced Psoriasis-like Skin Lesions via Regulation of HPA Axis and Neuroinflammation
Ningxin ZHANG ; Haoruo YANG ; Jiaqi LI ; Mengyao JIANG ; Jiarong WU ; Xinqian LIU ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(19):79-88
ObjectiveTo investigate the ameliorative effect and potential mechanisms of Kaixuan Jiedu core prescription (KXJD) on imiquimod (IMQ)-combined restraint stress-induced psoriasis-like skin lesions in mice. MethodsA total of 36 C57BL/6J mice were randomly assigned to six groups (n=6 per group): Normal group, restraint stress blank group, model group, restraint stress model group, KXJD group (30.42 g·kg-1), and methotrexate group (0.001 g·kg-1). Psoriasis-like lesions were induced by topical IMQ application combined with restraint stress. After 5 days of intervention, the severity of skin lesions was evaluated using the Psoriasis Area and Severity Index (PASI). The following parameters were assessed: Hypothalamic-pituitary-adrenal (HPA) axis-related hormones, including adrenocorticotropic hormone (ACTH), corticotropin-releasing hormone (CRH), and corticosterone (CORT), central neuronal markers, including neuronal nuclei antigen (NeuN), microtubule-associated protein 2 (MAP2), and beta Ⅲ-tubulin (β-Ⅲ tubulin), microglial activation markers, including ionized calcium-binding adapter molecule-1 (Iba-1) and cluster of differentiation 11b (CD11b), and skin nerve fiber markers, including protein gene product 9.5 (PGP9.5) and serotonin (5-HT), as well as the expression levels of inflammatory cytokines tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β). ResultsCompared with the normal group, both the model group and the restraint stress model group exhibited more severe psoriasis-like skin lesions, with significantly elevated serum levels of ACTH, CRH, and CORT (P<0.01). The expression of glucocorticoid receptor (GR) and corticotropin-releasing hormone receptor 1 (CRHR1) in the skin was significantly downregulated. The expression of NeuN, MAP2, β-Ⅲ tubulin, Iba-1, and CD11b in the hypothalamus and hippocampus was significantly upregulated, suggesting abnormal changes in neuronal number or structure and aberrant activation of glial cells (P<0.05, P<0.01). In addition, the expression of PGP9.5 and 5-HT in the skin, as well as the mRNA levels of TNF-α, IL-6, and IL-1β were all markedly elevated (P<0.05, P<0.01). Compared with the model group, the restraint stress model group showed further exacerbation in most parameters. Compared with the restraint stress model group, the KXJD group exhibited significantly reduced PASI scores (P<0.01), significantly decreased serum CRH, ACTH, and CORT levels (P<0.05, P<0.01), upregulated skin GR and CRHR1 expression, inhibited abnormal neuronal changes and microglial activation in the hypothalamus and hippocampus, attenuated skin nerve fiber hyperplasia and amine signaling molecule release, and significantly reduced pro-inflammatory cytokine mRNA expression (P<0.05, P<0.01). ConclusionKaixuan Jiedu Core prescription can effectively ameliorate IMQ combined with restraint stress-induced psoriasis-like skin lesions in mice by modulating HPA axis function, inhibiting central neuropathological changes, and attenuating skin nerve fiber hyperplasia, amine signaling molecule (5‑HT) release and local inflammatory responses.
10.Kaixuan Jiedu Core Prescription Alleviates Psoriatic Skin Lesions in Mice by Modulating Cold-sensitive TRPM8 Neuron-derived Signaling
Xue XIAO ; Bin YANG ; Meiqi SUN ; Haoruo YANG ; Ningxin ZHANG ; Jiaqi LI ; Huan LIU ; Mengyao JIANG ; Yuanyao SHE ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(19):89-101
ObjectiveTo investigate the ameliorative effects of Kaixuan Jiedu core prescription (KXJD) on skin lesions in psoriasis-like mouse models under cold environment exposure, and to analyze its influences on transient receptor potential (TRP) channels and related neuroimmune regulatory factors. MethodsThirty-six C57BL/6J mice were randomized into 6 groups, with 6 mice in each group. Two feeding conditions were set: Normal temperature and cold [simulating a cold environment at (10±0.5) ℃, for 6 h daily]. Mice were induced to develop psoriasis-like lesions by applying imiquimod externally. The model mice were allocated into model groups and KXJD (30.42 g·kg-1, continuous gavage for 5 days) groups. Normal mice were used as the control group. Specifically, mice were allocated into normal temperature, normal temperature model, normal temperature+KXJD, cold exposure control, cold exposure model, and cold exposure+KXJD groups. The pathological changes in skin lesions were observed by hematoxylin-eosin (HE) staining. The expression of cluster of differentiation (CD) 3+ T lymphocytes, CD11c+ dendritic cells (DCs), phosphorylated extracellular signal-regulated kinase (p-ERK), and substance P (SP) were detected by immunofluorescence assay. The protein level of transient receptor potential cation channel subfamily M member 8 (TRPM8) in the skin tissue was determined by Western blot. The expression of TRPM8, transient receptor potential cation channel subfamily V member 1 (TRPV1), transient receptor potential cation channel subfamily A member 1 (TRPA1), and transient receptor potential cation channel subfamily V member 2 (TRPV2) at the protein and mRNA levels was determined by immunohistochemistry and Real-time PCR, respectively. The levels of calcitonin gene-related peptide (CGRP) and neuropeptide Y (NPY) in the serum were analyzed by enzyme-linked immunosorbent assay (ELISA). The enrichment analysis of differentially expressed genes (DEGs) and TRP pathway network construction were conducted based on the GEO database. The co-expression of TRPM8 and CGRP in the skin lesions was verified by immunofluorescence double labeling. ResultsBoth the normal temperature and cold exposure model groups showed typical psoriasis-like skin lesions. Compared with the normal temperature and cold exposure control groups, the model groups had excessive epidermal keratinization, thickened spinous layer, and inflammatory infiltration in the dermis, with increased pathological scores (P<0.05), increased infiltration of CD3+ and CD11c+ cells and expression of p-ERK and SP, upregulated mRNA levels of TRPM8, TRPA1, and TRPV2, downregulated mRNA level of TRPV1 (P<0.05), and reduced content of CGRP and increased content of NPY in the serum. Compared with the normal temperature and cold exposure model groups, KXJD reduced the pathological manifestations and pathological scores of psoriasis-like skin lesions (P<0.05), and inhibited the infiltration of CD3+ and CD11c+ cells and the expression of p-ERK and SP. Gene enrichment analysis suggested that the DEGs of psoriasis were significantly enriched in the interleukin (IL)-17 signaling pathway and TRP channel inflammatory regulation. Compared with the normal temperature and cold exposure model groups, KXJD reversed the abnormal mRNA levels of genes related to the TRP channel subfamilies (P<0.05), increased the CGRP level, and decreased the NPY level. Immunofluorescence double labeling further confirmed that compared with the model groups, KXJD down-regulated the co-expression of TRPM8 and CGRP in the skin lesions. ConclusionKXJD may ameliorate psoriasis-like skin lesions by downregulating the overexpressed cold-sensitive receptor TRPM8 in skin lesions and correcting the disorder of neuropeptide (such as SP and CGRP) release mediated by it, thereby inhibiting the IL-23/helper T cell 17 (Th17) core inflammatory pathway, suppressing the infiltration of inflammatory cells and the activation of the ERK signaling pathway, and regulating the Xuanfu (sweat pore)-TRPM8-neuroimmune response axis.


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