1.Allogeneic lung transplantation in miniature pigs and postoperative monitoring
Yaobo ZHAO ; Ullah SALMAN ; Kaiyan BAO ; Hua KUI ; Taiyun WEI ; Hongfang ZHAO ; Xiaoting TAO ; Xinzhong NING ; Yong LIU ; Guimei ZHANG ; He XIAO ; Jiaoxiang WANG ; Chang YANG ; Feiyan ZHU ; Kaixiang XU ; Kun QIAO ; Hongjiang WEI
Organ Transplantation 2026;17(1):95-105
Objective To explore the feasibility and reference value of allogeneic lung transplantation and postoperative monitoring in miniature pigs for lung transplantation research. Methods Two miniature pigs (R1 and R2) underwent left lung allogeneic transplantation. Complement-dependent cytotoxicity tests and blood cross-matching were performed before surgery. The main operative times and partial pressure of arterial oxygen (PaO2) after opening the pulmonary artery were recorded during surgery. Postoperatively, routine blood tests, biochemical blood indicators and inflammatory factors were detected, and pathological examinations of multiple organs were conducted. Results The complement-dependent cytotoxicity test showed that the survival rate of lymphocytes between donors and recipients was 42.5%-47.3%, and no agglutination reaction occurred in the cross-matching. The first warm ischemia times of D1 and D2 were 17 min and 10 min, respectively, and the cold ischemia times were 246 min and 216 min, respectively. Ultimately, R1 and R2 survived for 1.5 h and 104 h, respectively. Postoperatively, in R1, albumin (ALB) and globulin (GLB) decreased, and alanine aminotransferase increased; in R2, ALB, GLB and aspartate aminotransferase all increased. Urea nitrogen and serum creatinine increased in both recipients. Pathological results showed that in R1, the transplanted lung had partial consolidation with inflammatory cell infiltration, and multiple organs were congested and damaged. In R2, the transplanted lung had severe necrosis with fibrosis, and multiple organs had mild to moderate damage. The expression levels of interleukin-1β and interleukin-6 increased in the transplanted lungs. Conclusions The allogeneic lung transplantation model in miniature pigs may systematically evaluate immunological compatibility, intraoperative function and postoperative organ damage. The data obtained may provide technical references for subsequent lung transplantation research.
2.Clinical Advantages of Traditional Chinese Medicine in Treatment of Childhood Simple Obesity: Insights from Expert Consensus
Qi ZHANG ; Yingke LIU ; Xiaoxiao ZHANG ; Guichen NI ; Heyin XIAO ; Junhong WANG ; Liqun WU ; Zhanfeng YAN ; Kundi WANG ; Jiajia CHEN ; Hong ZHENG ; Xinying GAO ; Liya WEI ; Qiang HE ; Qian ZHAO ; Huimin SU ; Zhaolan LIU ; Dafeng LONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):238-245
Childhood simple obesity has become a significant public health issue in China. Modern medicine primarily relies on lifestyle interventions and often suffers from poor long-term compliance, while pharmacological options are limited and associated with potential adverse effects. Traditional Chinese Medicine (TCM) has a long history in the prevention and management of this condition, demonstrating eight distinct advantages, including systematic theoretical foundation, diversified therapeutic approaches, definite therapeutic efficacy, high safety profile, good patient compliance, comprehensive intervention strategies, emphasis on prevention, and stepwise treatment protocols. Additionally, TCM is characterized by six distinctive features: the use of natural medicinal substances, non-invasive external therapies, integration of medicinal dietetics, simple exercise regimens, precise syndrome differentiation, and diverse dosage forms. By combining internal and external treatments, TCM facilitates individualized regimen adjustment and holistic regulation, demonstrating remarkable effects in improving obesity-related metabolic indicators, regulating constitutional imbalance, and promoting healthy behaviors. However, challenges remain, such as inconsistent operational standards, insufficient high-quality clinical evidence, and a gap between basic research and clinical application. Future efforts should focus on accelerating the standardization of TCM diagnosis and treatment, conducting multicenter randomized controlled trials, and fostering interdisciplinary integration, so as to enhance the scientific validity and international recognition of TCM in the prevention and treatment of childhood obesity.
3.SPIN1 promotes the migration and invasion of gastric adenocarcinoma cells by activating the JAK2/STAT3 pathway
XIAO Yao1 ; SUI Wenwen2 ; PAN Yu1 ; ZHAO Yulong3 ; LYU Beibei1
Chinese Journal of Cancer Biotherapy 2026;33(1):37-44
[摘 要] 目的:探究纺锤体蛋白1(SPIN1)促进胃腺癌细胞迁移与侵袭的分子机制。方法: 通过TCGA数据库数据分析胃腺癌组织中SPIN1 mRNA表达与上皮间质转化(EMT)评分、血管生成评分间的相关性。收集2018年8月至2021年11月期间山东第一医科大学附属省立医院手术切除的52例胃腺癌患者的癌组织制成组织芯片,每例均包含胃腺癌组织、对应癌旁组织及淋巴结转移灶,通过免疫组织化学法检测胃腺癌组织中SPIN1与STAT3的蛋白表达水平及相关性。通过Transwell实验研究干扰SPIN1对胃腺癌细胞侵袭与迁移的影响。使用GEPIA2网站分析SPIN1基因与Janus-激酶/信号转导和转录激活因子(JAK/STAT)通路相关因子在胃腺癌中的表达相关性。通过qPCR法、WB法检测干扰SPIN1后JAK/STAT通路相关mRNA和蛋白的表达变化。结果: TCGA数据库数据分析结果显示,SPIN1表达与EMT评分和血管生成评分呈正相关(均P < 0.05)。SPIN1与STAT3在胃腺癌组织和淋巴结转移灶中表达升高(均P < 0.05),在癌旁胃黏膜组织中阴性表达。SPIN1与STAT3的表达显著正相关(P < 0.05)。干扰SPIN1后胃腺癌细胞的迁移、侵袭能力明显降低(P < 0.05或P < 0.01)。GEPIA2网站分析结果显示,SPIN1基因与JAK1、JAK2、STAT1、STAT2及STAT3表达均呈显著正相关(均P < 0.05)。干扰SPIN1后JAK2、STAT3的mRNA水平下降,而JAK1、STAT1、STAT2的mRNA水平变化不明显。WB法实验结果表明,干扰SPIN1后JAK2、STAT3、p-JAK2及p-STAT3的蛋白表达均显著降低(均P < 0.01),过表达SPIN1后JAK2、STAT3、p-JAK2及p-STAT3的蛋白表达均显著升高(均P < 0.01)。结论: SPIN1可通过参与JAK2/STAT3信号通路促进胃腺癌细胞迁移与侵袭。
4.Intervention of Exercise Combined with Shenghui Tang in M1AChR Regulation of Mitochondrial Autophagy Improves Learning and Memory Ability of Alzheimer's Disease Model Rats
Xiaoxue TAO ; Min XIAO ; Kunyang TANG ; Bo WANG ; Chunchun ZHAO ; Rongxiang LIANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):120-130
ObjectiveTo make clear exercise combined with Shenghui Tang interferes in acetylcholine receptor (M1AChR) to improve mitochondrial autophagy and enhance cognition of Alzheimer's disease (AD) model rats through the adenylate activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR) signaling pathway. MethodsForty-eight male SD rats of SPF grade were randomly divided into a blank group, a model group, a Shenghui Tang group (9.3 g·kg-1), an exercise group, an exercise + Shenghui Tang group (9.3 g·kg-1), and a rapamycin group (1.5 mg·kg-1). Except for the blank group, the rat model of AD was constructed by injecting amyloid beta (Aβ1-42) into hippocampus stereotaxically. The exercise group received treadmill exercise for 4 weeks, while the Shenghui Tang group received intragastric administration for 4 weeks, and the exercise + Shenghui Tang group received treadmill exercise and intragastric administration of Shenghui Tang for 4 weeks simultaneously. After the intervention, the Morris water maze test was used to detect the learning and memory ability. Spontaneous behavior was observed in the open field test. The pathological structure of hippocampal neurons was observed by NISSl staining. The expression level of M1AChR in hippocampus was detected by immunohistochemistry (IHC). The autophagy ultrastructure of hippocampal neurons was observed by transmission electron microscopy. The apoptosis rate was evaluated by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). The expression of Beclin1 and microtubule-associated protein light chain 3β (LC3β) was detected by immunofluorescence (IF). The protein expression of M1AChR, AMPK, p-AMPK, mTOR, Beclin1, LC3β, and chelate 1 (SQSTM1/p62) in hippocampus was detected by Western blot. ResultsCompared with the blank group, the model group exhibited significantly increased platform escape latency on the fifth day (P<0.01) and significantly decreased activity distance in the target quadrant and times of crossing the platform (P<0.01). The total movement distance in the open field, the time of movement in the central area, and the average speed obviously decreased (P<0.05). The arrangement of nerve cells in hippocampus CA1 region was dispersed, and the numbers of Nissl bodies and M1AChR positive cells significantly decreased (P<0.01). The expression of TUNEL positive cells was significantly increased (P<0.01). The typical autophagic lysosomal structure decreased. The protein expression of M1AChR, p-AMPK/AMPK, p-mTOR/mTOR, Beclin1, LC3Ⅱ/Ⅰ in hippocampus was significantly decreased (P<0.01), and the protein expression of p62 was significantly increased (P<0.01). Compared with the model group, the exercise + Shenghui Tang group exhibited obviously improved space exploration and positioning navigation ability (P<0.05, P<0.01). The total movement distance in the open field, the time of movement in the central area, and the average speed of movement significantly increased (P<0.01). The number of Nissl bodies significantly increased (P<0.01). The number of M1AChR positive cells in hippocampus was significantly increased (P<0.01). The expression of TUNEL positive cells was significantly decreased (P<0.01). The protein expression of M1AChR, p-AMPK/AMPK, p-mTOR/mTOR, Beclin1, LC3Ⅱ/Ⅰ in hippocampus was significantly increased (P<0.01), while the protein expression of p62 was significantly decreased (P<0.01). Compared with the exercise + Shenghui Tang group, the Shenghui Tang group and the exercise group showed significantly increased platform escape latency on the fifth day (P<0.01) and obviously decreased activity distance in the target quadrant and times of crossing the platform (P<0.05, P<0.01). The total movement distance in the open field, the time of movement in the central area, and the average speed of movement significantly decreased (P<0.01). The number of Nissl bodies and the number of M1AChR positive cells significantly decreased (P<0.01). The expression of TUNEL positive cells was obviously increased (P<0.05). Ultrastructure of the hippocampal region showed decreased autophagy level. The protein expression of M1AChR, p-AMPK/AMPK, p-mTOR/mTOR, LC3Ⅱ/Ⅰ in the hippocampus was obviously decreased in the Shenghui Tang group (P<0.05, P<0.01), while the protein expression of p62 was significantly increased (P<0.01). In the exercise group, the protein expression of M1AChR, p-AMPK/AMPK, Beclin1, and LC3Ⅱ/Ⅰ was obviously decreased (P<0.05, P<0.01), while the protein expression of p-mTOR/mTOR and p62 was significantly increased (P<0.01). ConclusionExercise combined with traditional Chinese medicine can enhance the expression of M1AChR in the hippocampus of AD model rats, induce autophagy through the AMPK/mTOR signaling pathway, and improve the learning and memory ability of AD rats.
5.Analysis of risk factors and construction of risk prediction model for batroxobin-related severe hypofibrinogenemia
Le CAI ; Yuqing ZHAO ; Jiazhu CUI ; Xiao WEN ; Daihong GUO ; Man ZHU
China Pharmacy 2026;37(4):462-467
OBJECTIVE To investigate the clinical characteristics and risk factors for batroxobin-related severe hypofibrinogenemia (HFIB) and construct a risk prediction model. METHODS A retrospective analysis was conducted on inpatients treated with batroxobin in the First Medical Center of a tertiary hospital from January 1, 2020, to December 31, 2024. Patients were categorized into non-severe HFIB group and severe HFIB group based on the severity of HFIB. Univariate and multivariate Logistic regression analyses were performed to identify the independent influencing factors for batroxobin-related severe HFIB. A nomogram was developed using the “rms” package in R 4.5 software. The predictive performance of the model was evaluated using the receiver operating characteristic curve. Calibration was assessed via the Bootstrap resampling method, and goodness-of-fit was evaluated with the Hosmer-Lemeshow test. RESULTS A total of 1 472 patients were included in this study. Of these, 1 445 developed HFIB, yi elding an incidence of 98.17%. Furthermore, 895 were classified as severe HFIB, accounting for 60.80% of the cohort. Multivariate Logistic regression analysis showed that increased age, high initial dose per 10 kg body weight, use of maintenance dose, and concomitant glucocorticoid use were independent risk factors for batroxobin-related severe HFIB, while high baseline fibrinogen (FIB) level was identified as a protective factor. The model demonstrated an area under the curve of 0.760 (95% CI: 0.735-0.785). The mean absolute error of the calibration curve was 0.006. The P value of the Hosmer-Lemeshow test was 0.609. CONCLUSIONS Batroxobin can rapidly and significantly reduce FIB levels and carries a risk of inducing severe HFIB. Patients with advanced age, high initial dose per 10 kg body weight, use of maintenance dose and concomitant glucocorticoid use had a higher risk of batroxobin-related severe HFIB, while high baseline FIB level had a lower risk of batroxobin-related severe HFIB. The risk prediction model developed based on these factors can be used to predict the likelihood of batroxobin-related severe HFIB.
6.Mechanisms of Tianma Goutengyin in Alleviating Neuronal Injury in Vascular Dementia Model Rats by Inhibiting A1 Astrocyte Activation via Regulating TNF-α/STAT3/α1ACT Signaling Pathway
Xiaoyan WANG ; Min ZHAO ; Feng TIAN ; Min XIAO ; Nan QU ; Fugui LIU ; Chixiao LIU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):56-65
ObjectiveTo investigate the effects of Tianma Goutengyin on the tumor necrosis factor-α (TNF-α)/signal transducer and activator of transcription 3 (STAT3)/α1-antichymotrypsin C-terminal tail fragment (α1ACT) signaling pathway and A1-type astrocytes in a rat model of vascular dementia. MethodsSeventy-two male Sprague-Dawley rats were randomly divided into six groups (n=12 per group): Sham-operated group, model group, Tianma Goutengyin high-, medium-, and low-dose groups (5.13, 10.26, and 20.52 g·kg-1), and a nimodipine group (8.1 mg·kg-1). The vascular dementia model was established by permanent bilateral common carotid artery occlusion, followed by 4 weeks of intervention. Learning and memory ability were evaluated using the novel object recognition test, and behavioral performance was assessed using the forced swimming test. Levels of interleukin-6 (IL-6) and C-C motif chemokine ligand 2 (CCL2) in hippocampal tissue were measured by enzyme-linked immunosorbent assay (ELISA). Hippocampal neuronal morphology was observed by Nissl staining, and apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Immunohistochemistry was used to detect positive expression of brain-derived neurotrophic factor (BDNF), glial fibrillary acidic protein (GFAP), and myelin basic protein (MBP). Western blot analysis was performed to measure the protein expression levels of TNF-α, TNF receptor 1 (TNFR1), phosphorylated STAT3 (p-STAT3), α1ACT, IL-6, complement component 3 (C3), BDNF, S100 calcium-binding protein A10 (S100A10), and GFAP in hippocampal tissue. ResultsCompared with the sham-operated group, the model group showed a significantly reduced relative recognition index in the novel object recognition test (P<0.01), prolonged immobility time and increased immobility frequency in the forced swimming test (P<0.01). Hippocampal IL-6 and CCL2 levels were significantly increased (P<0.01). Nissl staining revealed a marked reduction in neuronal number and loss of Nissl bodies (P<0.01). MBP-positive expression was significantly decreased (P<0.01), apoptosis was significantly increased (P<0.01), BDNF-positive expression was significantly reduced (P<0.05), and GFAP-positive expression was significantly increased (P<0.01). In addition, the protein expression levels of TNF-α, TNFR1, p-STAT3, α1ACT, IL-6, and C3 were significantly elevated (P<0.01), while BDNF and S100A10 expression levels were significantly decreased (P<0.01). Compared with the model group, all Tianma Gouteng yin dose groups exhibited a significant increase in the relative recognition index (P<0.05), shortened immobility time and reduced immobility frequency (P<0.05, P<0.01). IL-6 and CCL2 levels were significantly decreased (P<0.01), neuronal number was significantly increased (P<0.05, P<0.01), and MBP-positive expression was significantly enhanced (P<0.01). Apoptosis was significantly reduced (P<0.01), BDNF-positive expression was significantly increased (P<0.05), and GFAP-positive expression was significantly decreased (P<0.01). Moreover, the protein expression levels of TNF-α, TNFR1, p-STAT3, α1ACT, IL-6, and C3 were significantly decreased (P<0.01), while BDNF and S100A10 protein expression levels were significantly increased (P<0.01). ConclusionTianma Goutengyin may inhibit A1-type astrocyte activation in rats with vascular dementia through the TNF-α/STAT3/α1ACT signaling pathway, thereby reducing neuronal apoptosis and improving learning and memory function.
7.Effect and Mechanism of Icariin on Improving Spermatogenesis in Exercise-induced Fatigue Model Mice Through Regucalcin
Kunyang TANG ; Min XIAO ; Xiaocui JIANG ; Xiaoxue TAO ; Yue ZOU ; Chunchun ZHAO ; Zhipeng FANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):117-127
ObjectiveThis paper aims to investigate the effects of icariin on spermatogenesis in mice with exercise-induced fatigue and explore the underlying mechanisms. MethodsICR male mice were screened by swimming and randomly divided into normal group, model group, vitamin C group, icariin groups with low, medium, and high doses, and medium-dose icariin+N-nitro-L-arginine methyl ester (L-NAME) group, with 10 mice per group. Except for the normal group, all the other groups underwent weighted swimming training to establish an exercise-induced fatigue model. No gavage was administered during the first two weeks of the weighted training. From week three to four, the icariin groups with low, medium, and high doses received 0.03, 0.06, and 0.12 g·kg-1 icariin via gavage, respectively. The vitamin C group received 0.2 g·kg-1 vitamin C. The L-NAME group received 0.06 g·kg-1 icariin and 0.01 g·kg-1 L-NAME via intraperitoneal injection. The normal and model groups received equivalent physiological saline. After the experiment, body weight and the last exhaustive swimming time were recorded. Blood urea nitrogen (BUN), lactate (LA), lactate dehydrogenase (LDH), malondialdehyde (MDA), testicular testosterone (T), testicular Ca2+/Mg2+-adenosine triphosphatase (ATPase) (micro-assay), and the levels of testicular cyclic guanosine monophosphate (cGMP) were measured by using kits. Sperm CD46 levels were detected by flow cytometry. Testicular seminiferous tubules were observed via hematoxylin-eosin (HE) staining, and the testicular morphometric score (TMS) was used to evaluate the spermatogenic function. Protein expression of regucalcin (RGN, SMP30), cGMP-dependent protein kinase 1 (PKG), and cGMP-dependent protein kinase anchoring protein (GKAP1) was detected by Western blot. Testicular regucalcin expression was examined by immunofluorescence (IF). The epididymal sperm quality of mice was observed under a microscope. Fluorescence-stained sections of stimulated by retinoic acid gene 8 (STRA8), synaptonemal complex protein 3 (SCP3), and transition protein 1(TNP1) in testicular seminiferous tubules were assessed by immunohistochemistry (IHC). ResultsCompared with the normal group, the model group showed decreased body weight and exhaustive swimming time (P<0.01), significantly increased fatigue markers (LA, LDH, and BUN) and lipid peroxidation product MDA (P<0.01), reduced testicular RGN, PKG, GKAP1, testosterone, Ca2+/Mg2+-ATPase, and cGMP levels (P<0.01), decreased sperm motility, sperm count, and TMS scores, and downregulated the expression of STRA8, SCP3, and TNP1. Compared with the model group, the icariin group with high dose exhibited increased exhaustive swimming time (P<0.01), reduced LA, LDH, BUN, and MDA levels (P<0.01), elevated superoxide dismutase (SOD) (P<0.01), upregulated testicular RGN, PKG, GKAP1, testosterone, Ca2+/Mg2+-ATPase, and cGMP levels (P<0.01), improved sperm motility, sperm count, and TMS scores, and enhanced STRA8, SCP3, and TNP1 expression. Compared with the L-NAME group, the icariin group with medium dose showed increased expression of STRA8, SCP3, and TNP1 in the testicular tissue (P<0.01) and elevated cGMP and GKAP1 levels (P<0.01). ConclusionExercise-induced fatigue reduces the expression of RGN and cGMP/PKG/GKAP1 in mice, thereby causing abnormal spermatogenesis and impairing reproductive function in mice. Icariin ameliorates spermatogenic dysfunction in exercise-induced fatigue mice by promoting the expression of RGN and cGMP/PKG/GKAP1, thereby mitigating the damage of exercise-induced fatigue to the reproductive system.
8.Exploring on Processing Mechanism of Enhanced "Invigorating Spleen and Stopping Diarrhea" Effect of Soil-fried Atractylodis Macrocephalae Rhizoma Based on "Microscopic Characterization, Chemical Analysis and Pharmacodynamic Evaluation" Trinity
Guoshun SHAN ; Yuyan XIAO ; Chu YUAN ; Xiuai CHEN ; Qimiao ZHAO ; Xiang LIU ; Hao WU ; Ke ZHANG ; Siqi LIU ; Yongduo YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):182-193
ObjectiveTo analyze the processing mechanism underlying the enhanced effect of invigorating spleen and stopping diarrhea of soil-fried Atractylodis Macrocephalae Rhizoma(AMR) by analyzing the changes of microstructure, chemical composition and anti-ulcerative colitis(UC) activity before and after soil stir-frying. MethodsThe microstructure and elemental composition of AMR before and after soil stir-frying were analyzed by scanning electron microscopy-energy dispersive spectroscopy(SEM-EDS), to investigate the differences in microstructure and the underlying causes. Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) coupled with UNIFI 1.9.2 natural product analysis platform were used to analyze and identify the chemical constituents in raw and soil-fried products, and multivariate statistical methods including principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to explore the differences and sources of chemical constituents between them. A dextran sulfate sodium(DSS)-induced UC mouse model was established. The method of disease activity index(DAI) was used to evaluate the severity of intestinal inflammation. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of colon tissue, enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory factors, Real-time quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to analyze the expressions of key genes and proteins involved in the intestinal mucosal barrier. The 16S rRNA sequencing was used to evaluate the diversity of intestinal flora, headspace gas chromatography-mass spectrometry(HS-GC-MS) was used to explore the levels of short-chain fatty acids(SCFAs) in feces. Base on the above findings, this paper investigated the effects of raw and soil-fried AMR on the biological, chemical, mechanical and immune barriers of model animals, and the differences in pharmacological effects and underlying mechanisms from the perspective of regulating the intestinal mucosal barrier in UC mice. ResultsSEM observation revealed numerous hearth soil particles on the surface of soil-fried AMR, accompanied by bubble-like bulges. At the same time, there were many cracks and folds on the surface of the hearth soil. EDS analysis revealed that the contents of Si, Al, Mg and Ca in soil-fried AMR were significantly higher than those of raw products, and these elements constituted the primary components of hearth soil. UPLC-Q-TOF-MS combined with database comparison was used to identify the chemical constituents of raw and soil-fried AMR. In positive ion mode, a total of 132 components were identified, primarily comprising three categories of terpenoids, polyphenols and amino acids. In negative ion mode, a total of 40 components were identified, primarily polyphenolic and glycoside compounds. Among them, the contents of sesquiterpenes and polyphenolic acids were changed significantly before and after processing. Soil-fried AMR could reduce the DAI score of UC mice, alleviate the shortening of colon length, reduce the levels of pro-inflammatory factors such as interleukin(IL)-17, IL-18, γ-interferon(IFN-γ) and tumor necrosis factor(TNF)-α in serum, increase the levels of anti-inflammatory factors such as secretory immunoglobulin A(sIgA), IL-10, IL-4 and transforming growth factor-β(TGF-β) in serum, increase the expressions of key genes and proteins of intestinal mucosal barrier such as tight junction protein-1(ZO-1), Occludin, Claudin-1 and mucin 2(MUC2) in colonic mucosa, and improve the disorders of intestinal flora diversity and the levels of SCFAs(P<0.05, P<0.01). The raw and stir-fried products of AMR also exhibited the aforementioned effects, but they were weaker than the soil-fried products. Additionally, the auxiliary material hearth soil also had a certain pharmacodynamic effect. ConclusionSoil-fried AMR can enhance the protective effect on intestinal mucosal barrier in UC mice. These changes or heating-induced alterations in the microscopic structure and chemical composition of AMR may be attributed to the dual effects of adsorption of hearth soil.
9.Effect and Mechanism of Icariin on Improving Spermatogenesis in Exercise-induced Fatigue Model Mice Through Regucalcin
Kunyang TANG ; Min XIAO ; Xiaocui JIANG ; Xiaoxue TAO ; Yue ZOU ; Chunchun ZHAO ; Zhipeng FANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):117-127
ObjectiveThis paper aims to investigate the effects of icariin on spermatogenesis in mice with exercise-induced fatigue and explore the underlying mechanisms. MethodsICR male mice were screened by swimming and randomly divided into normal group, model group, vitamin C group, icariin groups with low, medium, and high doses, and medium-dose icariin+N-nitro-L-arginine methyl ester (L-NAME) group, with 10 mice per group. Except for the normal group, all the other groups underwent weighted swimming training to establish an exercise-induced fatigue model. No gavage was administered during the first two weeks of the weighted training. From week three to four, the icariin groups with low, medium, and high doses received 0.03, 0.06, and 0.12 g·kg-1 icariin via gavage, respectively. The vitamin C group received 0.2 g·kg-1 vitamin C. The L-NAME group received 0.06 g·kg-1 icariin and 0.01 g·kg-1 L-NAME via intraperitoneal injection. The normal and model groups received equivalent physiological saline. After the experiment, body weight and the last exhaustive swimming time were recorded. Blood urea nitrogen (BUN), lactate (LA), lactate dehydrogenase (LDH), malondialdehyde (MDA), testicular testosterone (T), testicular Ca2+/Mg2+-adenosine triphosphatase (ATPase) (micro-assay), and the levels of testicular cyclic guanosine monophosphate (cGMP) were measured by using kits. Sperm CD46 levels were detected by flow cytometry. Testicular seminiferous tubules were observed via hematoxylin-eosin (HE) staining, and the testicular morphometric score (TMS) was used to evaluate the spermatogenic function. Protein expression of regucalcin (RGN, SMP30), cGMP-dependent protein kinase 1 (PKG), and cGMP-dependent protein kinase anchoring protein (GKAP1) was detected by Western blot. Testicular regucalcin expression was examined by immunofluorescence (IF). The epididymal sperm quality of mice was observed under a microscope. Fluorescence-stained sections of stimulated by retinoic acid gene 8 (STRA8), synaptonemal complex protein 3 (SCP3), and transition protein 1(TNP1) in testicular seminiferous tubules were assessed by immunohistochemistry (IHC). ResultsCompared with the normal group, the model group showed decreased body weight and exhaustive swimming time (P<0.01), significantly increased fatigue markers (LA, LDH, and BUN) and lipid peroxidation product MDA (P<0.01), reduced testicular RGN, PKG, GKAP1, testosterone, Ca2+/Mg2+-ATPase, and cGMP levels (P<0.01), decreased sperm motility, sperm count, and TMS scores, and downregulated the expression of STRA8, SCP3, and TNP1. Compared with the model group, the icariin group with high dose exhibited increased exhaustive swimming time (P<0.01), reduced LA, LDH, BUN, and MDA levels (P<0.01), elevated superoxide dismutase (SOD) (P<0.01), upregulated testicular RGN, PKG, GKAP1, testosterone, Ca2+/Mg2+-ATPase, and cGMP levels (P<0.01), improved sperm motility, sperm count, and TMS scores, and enhanced STRA8, SCP3, and TNP1 expression. Compared with the L-NAME group, the icariin group with medium dose showed increased expression of STRA8, SCP3, and TNP1 in the testicular tissue (P<0.01) and elevated cGMP and GKAP1 levels (P<0.01). ConclusionExercise-induced fatigue reduces the expression of RGN and cGMP/PKG/GKAP1 in mice, thereby causing abnormal spermatogenesis and impairing reproductive function in mice. Icariin ameliorates spermatogenic dysfunction in exercise-induced fatigue mice by promoting the expression of RGN and cGMP/PKG/GKAP1, thereby mitigating the damage of exercise-induced fatigue to the reproductive system.
10.Exploring on Processing Mechanism of Enhanced "Invigorating Spleen and Stopping Diarrhea" Effect of Soil-fried Atractylodis Macrocephalae Rhizoma Based on "Microscopic Characterization, Chemical Analysis and Pharmacodynamic Evaluation" Trinity
Guoshun SHAN ; Yuyan XIAO ; Chu YUAN ; Xiuai CHEN ; Qimiao ZHAO ; Xiang LIU ; Hao WU ; Ke ZHANG ; Siqi LIU ; Yongduo YU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):182-193
ObjectiveTo analyze the processing mechanism underlying the enhanced effect of invigorating spleen and stopping diarrhea of soil-fried Atractylodis Macrocephalae Rhizoma(AMR) by analyzing the changes of microstructure, chemical composition and anti-ulcerative colitis(UC) activity before and after soil stir-frying. MethodsThe microstructure and elemental composition of AMR before and after soil stir-frying were analyzed by scanning electron microscopy-energy dispersive spectroscopy(SEM-EDS), to investigate the differences in microstructure and the underlying causes. Ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) coupled with UNIFI 1.9.2 natural product analysis platform were used to analyze and identify the chemical constituents in raw and soil-fried products, and multivariate statistical methods including principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to explore the differences and sources of chemical constituents between them. A dextran sulfate sodium(DSS)-induced UC mouse model was established. The method of disease activity index(DAI) was used to evaluate the severity of intestinal inflammation. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of colon tissue, enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory factors, Real-time quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to analyze the expressions of key genes and proteins involved in the intestinal mucosal barrier. The 16S rRNA sequencing was used to evaluate the diversity of intestinal flora, headspace gas chromatography-mass spectrometry(HS-GC-MS) was used to explore the levels of short-chain fatty acids(SCFAs) in feces. Base on the above findings, this paper investigated the effects of raw and soil-fried AMR on the biological, chemical, mechanical and immune barriers of model animals, and the differences in pharmacological effects and underlying mechanisms from the perspective of regulating the intestinal mucosal barrier in UC mice. ResultsSEM observation revealed numerous hearth soil particles on the surface of soil-fried AMR, accompanied by bubble-like bulges. At the same time, there were many cracks and folds on the surface of the hearth soil. EDS analysis revealed that the contents of Si, Al, Mg and Ca in soil-fried AMR were significantly higher than those of raw products, and these elements constituted the primary components of hearth soil. UPLC-Q-TOF-MS combined with database comparison was used to identify the chemical constituents of raw and soil-fried AMR. In positive ion mode, a total of 132 components were identified, primarily comprising three categories of terpenoids, polyphenols and amino acids. In negative ion mode, a total of 40 components were identified, primarily polyphenolic and glycoside compounds. Among them, the contents of sesquiterpenes and polyphenolic acids were changed significantly before and after processing. Soil-fried AMR could reduce the DAI score of UC mice, alleviate the shortening of colon length, reduce the levels of pro-inflammatory factors such as interleukin(IL)-17, IL-18, γ-interferon(IFN-γ) and tumor necrosis factor(TNF)-α in serum, increase the levels of anti-inflammatory factors such as secretory immunoglobulin A(sIgA), IL-10, IL-4 and transforming growth factor-β(TGF-β) in serum, increase the expressions of key genes and proteins of intestinal mucosal barrier such as tight junction protein-1(ZO-1), Occludin, Claudin-1 and mucin 2(MUC2) in colonic mucosa, and improve the disorders of intestinal flora diversity and the levels of SCFAs(P<0.05, P<0.01). The raw and stir-fried products of AMR also exhibited the aforementioned effects, but they were weaker than the soil-fried products. Additionally, the auxiliary material hearth soil also had a certain pharmacodynamic effect. ConclusionSoil-fried AMR can enhance the protective effect on intestinal mucosal barrier in UC mice. These changes or heating-induced alterations in the microscopic structure and chemical composition of AMR may be attributed to the dual effects of adsorption of hearth soil.

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