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.Treatment Principles and Paradigm of Diabetic Microvascular Complications Responding Specifically to Traditional Chinese Medicine
Anzhu WANG ; Xing HANG ; Lili ZHANG ; Xiaorong ZHU ; Dantao PENG ; Ying FAN ; Min ZHANG ; Wenliang LYU ; Guoliang ZHANG ; Xiai WU ; Jia MI ; Jiaxing TIAN ; Wei ZHANG ; Han WANG ; Yuan XU ; .LI PINGPING ; Zhenyu WANG ; Ying ZHANG ; Dongmei SUN ; Yi HE ; Mei MO ; Xiaoxiao ZHANG ; Linhua ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(5):272-279
To explore the advantages of traditional Chinese medicine (TCM) and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications (DMC), refine key pathophysiological insights and treatment principles, and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine, hosted by the China Association of Chinese Medicine, was held in Beijing, 2024. Experts in TCM, Western medicine, and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis, diagnostic and treatment challenges, and mechanism research related to DMC, ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development, research prioritization, and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM, strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.
3.Molecular Mechanism of Liuwei Dihuangwan Regulating GPNMB Expression and Enhancing Autophagy in Prevention and Treatment of Alzheimer's Disease
Yuxi LIU ; Zhongkang ZHU ; Songnan WANG ; Jiali LIU ; Ye YIN ; Jiarui MIAO ; Shunuo HE ; Danyu ZHAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):1-10
ObjectiveThis study aims to investigate the effect of Liuwei Dihuangwan on the autophagy function in the hippocampus of senescence-accelerated mouse prone 8 (SAMP8) by regulating the expression of glycoprotein non-metastatic melanoma protein B (GPNMB). Furthermore, it is designed to explore the mechanism of the method of tonifying the kidneys and replenishing essence in the treatment of Alzheimer's disease (AD). MethodsIn experiment 1, 24 5-month-old SAMP8 mice were randomly and equally divided into the model group, and the low-, middle- and high-dose(0.59,1.18,2.36 g·kg-1) Liuwei Dihuangwan groups. At the same time, six 5-month-old senescence accelerated mouse resistant 1 (SAMR1) mice were used as the control group. The learning and memory ability was evaluated through novel object recognition experiment. Serum cortisol (Cort), adrenocorticotropic hormone (ACTH) and urine 17-hydroxycorticosteroid (17-OHCS) levels were detected by enzyme-linked immunosorbent assay (ELISA). The ultrastructure of hippocampal neurons was observed by transmission electron microscope (TEM), and the expression levels of hippocampal GPNMB, a disintegrin and metalloproteinase 10 (ADAM10) and autophagy-related proteins were detected by Western blot. In experiment 2, 18 SAMP8 mice were randomly and equally divided into the model group, vector control group (Vector), and GPNMB overexpression group (GPNMBOE). Lentiviral vectors were stereotactically injected into the brain (2 μL per side in the GPNMBOE group). Western blot was used to detect the expression of the above target proteins in the hippocampus; In Experiment 3, 24 SAMP8 mice were randomly and equally divided into the model group, Liuwei Dihuangwan group, Liuwei Dihuangwan+negative control (NC) group, and Liuwei Dihuangwan+GPNMB silencing group (shGPNMB). Before drug treatment, the Liuwei Dihuangwan+NC group and the Liuwei Dihuangwan+shGPNMB group were injected with negative control and GPNMB silencing lentivirus, respectively. Western blot was used to detect the expression of the above target proteins in the hippocampus. ResultsThe novel object discrimination index of mice in the model group was significantly lower than that of mice in the control group (P<0.01). The novel object discrimination index of mice in the medium- and high-dose Liuwei Dihuangwan groups was significantly higher than that of mice in the model group (P<0.01). Aggregated autolysosomes were observed in the normal hippocampus tissue by TEM. In the model group, mitochondria were dominant, and no typical characteristic autophagosomes were observed. In the low- and medium-dose Liuwei Dihuangwan groups, a small number of autolysosomes and autophagosomes with double-membrane structures were observed. In the high-dose Liuwei Dihuangwan group, the number of autophagosomes and autolysosomes was greater than that in the low- and medium-dose groups. The results of ELISA and Western blot showed that compared with the control group, the levels of serum Cort, ACTH, and urine 17-OHCS in the model group were substantially increased, while the expression of hippocampal ADAM10, Beclin1, and microtubule associated-protein light chain 3-Ⅱ/Ⅰ (LC3 Ⅱ/Ⅰ) was significantly decreased. The expression of GPNMB and ubiquitin binding protein p62 was significantly increased (P<0.05, P<0.01). Compared with the model group, the serum Cort and ACTH levels in the low-, medium-, and high-dose Liuwei Dihuangwan groups were significantly reduced, while only the urine 17-OHCS level in the high-dose group was significantly reduced. The hippocampal GPNMB, ADAM10, Beclin1, and LC3 Ⅱ/Ⅰ expression levels in the medium-, and high-dose groups of Liuwei Dihuangwan were significantly increased compared to the model group, whereas the expression of p62 was significantly reduced (P<0.01). The above indicators showed a progressive trend among the three groups. Compared with the model group, the GPNMBOE group showed a significant increase in GPNMB, ADAM10, Beclin1, LC3 Ⅱ/Ⅰ expression, and a significant decrease in p62 expression (P<0.01). Compared with the model group, the expression of GPNMB, ADAM10, Beclin1, and LC3 Ⅱ/Ⅰ in the hippocampus of the Liuwei Dihuangwan group significantly increased, while the expression of p62 significantly decreased (P<0.01). Compared with the Liuwei Dihuangwan group, the Liuwei Dihuangwan+shGPNMB group showed a significant decrease in GPNMB, ADAM10, Beclin1, LC3 Ⅱ/Ⅰ, and a significant increase in p62 expression (P<0.01). ConclusionLiuwei Dihuangwan can enhance hippocampal autophagy function and improve AD by upregulating GPNMB expression.
4.Genetic differentiation of Anopheles sinensis (Diptera: Culicidae) based on COⅠ and ITS2 sequences
Gang WANG ; Chun-juan SHI ; Dan KONG ; Xuan ZHU ; Ji HE ; Ming-hui ZHAO ; Chong-cai WANG
Acta Parasitologica et Medica Entomologica Sinica 2026;33(2):90-95
Objective Exploring genetic differences among geographic strains of Anopheles sinensis in China. Methods Based on the mitochondrial cytochrome oxidase subunit I(COⅠ )and ribosomal the second internal transcribed spacer(ITS2), genetic distance analyses were conducted on An. sinensis samples collected nationwide. Results Genetic differentiation was observed within An. sinensis geographic strains, primarily reflected in the genetic distances of COⅠ sequences, with some distances between samples exceeding those between closely related species. However, ITS2 sequences showed relatively low levels of differentiation within geographic strains. No significant separation was observed, and gene exchange exhibited a random pattern among geographic strains. Conclusions Significant differences in genetic distance were observed among An. sinensis samples, along with evidence of interspecific continuity among sibling species. For the identification of closely related species using wild-collected Anopheles mosquitoes samples, reliance solely on commonly used barcodes such as COⅠ and ITS2 may be insufficient. Additional molecular markers, including COⅡ and D3 gene fragments, should also be considered. Caution should be exercised when using COⅠ genes to identify cryptic species, especially within Anopheles mosquitoes.
5.Association of sex hormone levels with all-cause mortality in males: a study based on NHANES
Jiahao WANG ; Shan JIANG ; Yilin ZHAO ; Haolin LI ; He ZHANG ; Di ZHU ; Jinzhi WANG ; Junqi WANG ; Peng GE
Journal of Modern Urology 2026;31(1):33-41
Objective To investigate the association of serum total testosterone(TT),free testosterone(FT),sex hormone-binding globulin(SHBG),and estradiol(E_2)with all-cause mortality rate(AMR)in males and to evaluate their predictive value. Methods Based on data from the National Health and Nutrition Examination Survey(NHANES)cycles(1999—2004 and 2013—2016),the relationships between different sex hormones and AMR was analyzed with Cox proportional hazards regression models. The dose-response relationships between different sex hormones and male AMR were assessed with restricted cubic splines(RCS). The predictive performances of different sex hormones for male AMR were compared with receiver operating characteristic(ROC)curves and the area under the curve(AUC). The survival differences across varying sex hormone levels were evaluated with Kaplan-Meier survival curves and log-rank tests. Results This study involved 3632 adult male participants with a median follow-up of 6.5 years,during which 320 AMR occurred(8.8%). Cox univariate analysis indicated that TT,FT and SHBG were risk factors for AMR(P<0.001). Cox multivariate analysis revealed that FT and SHBG were independent risk factors for AMR(FT:HR=0.90,95%CI:0.86-0.94; SHBG:HR=1.01,95%CI:1.01-1.02). RCS revealed a U-shaped nonlinear relationship between TT,FT and AMR(P<0.001); SHBG showed a positive dose-response relationship with AMR(P<0.001). The AUCs of FT predicting the 5-year,10-year,and 15-year AMR were 0.78(95%CI:0.73-0.82),0.80(95%CI:0.76-0.84),and 0.84(95%CI: 0.80-0.88),respectively,significantly outperforming other indicators. Subgroup analysis revealed that FT was negatively correlated with male AMR,but this association was modulated by age,race,smoking status and diabetes status(P_(interaction)<0.05). Kaplan-Meier survival analysis showed that different levels of TT and FT(Q1-Q4)were significantly positively correlated with survival rate(P<0.001),while different levels of SHBG were significantly negatively correlated with survival rate,with patients in the Q4 group having the highest risk of death(P<0.001). Conclusion Based on US NHANES,studies have shown that the FT and SHBG are independent risk factors for male AMR,and FT demonstrates greater predictive value for male AMR than TT.
6.A panel study on effects of short-term exposure to ambient particulate matter and nitrogen dioxide on serum reproductive hormones in healthy adults
Yan FANG ; Xinghou HE ; Erlu ZHAO ; Bin ZHANG ; Yutong ZHU ; Tong WANG ; Xiaoming SONG ; Hongbing XU ; Wei HUANG
Journal of Environmental and Occupational Medicine 2026;43(6):709-716
Background Air pollution has widespread adverse health effects on human health, but research on its impact on reproductive health and related physiological mechanisms remains insufficient. Objective To investigate the associations between short-term exposure to particulate matter and nitrogen dioxide (NO2) and serum levels of anti-Müllerian hormone (AMH) and testosterone in healthy adults. Methods A total of 152 healthy adults were recruited for a panel study, undergoing four follow-up visits from September 2019 to January 2020. Basic demographic information and serum samples were collected. Reproductive hormone levels were measured by enzyme-linked immunosorbent assay kits. Concurrently, environmental data in the study area were monitored, including fine particulate matter (PM2.5), black carbon (BC), NO2, particle number concentrations in the 5-560 nm size fraction (PNC5_560), and meteorological variables. Linear mixed-effects models were constructed to estimate the effects of short-term exposure to these particulate matter on serum reproductive hormone levels. Results The participants had an average age of (23.9±2.4) years, with 102 women. For each interquartile range (IQR) increment in pollutant moving average concentrations, PM2.5, PNC5_25, PNC100_560, BC, and NO2 were associated with relative decreases in female serum AMH levels by 10.2%-16.9%, 46.2%-58.0%, 16.9%-21.2%, 14.8%-26.5%, and 11.3%-17.6%, respectively. These negative associations were stronger in females with waist-to-hip ratio > 0.76 and physical activity levels ≤ 1217 metabolic equivalents (MET) min per week. Similarly, in males, an IQR increase in the same pollutants was associated with relative decreases in serum free testosterone by 11.1%-17.2%, 35.2%, 10.5%-18.9%, 18.7%-24.2%, and 10.3%-26.0%, respectively. These effects were more pronounce in male with a waist-to-hip ratios > 0.81 and physical activity levels ≤ 1498 MET-min per week. Conclusion Short-term exposure to particulate matter and NO2 may reduce serum levels of AMH and testosterone in healthy adults. Furthermore, these adverse effects appears to be stronger in adults with higher waist-to-hip ratios.
7.Chinese clinical expert consensus on multi-arm robotic-assisted uniportal pulmonary resection
Deping ZHAO ; Xuefei HU ; Chenlu YANG ; Ye NING ; Yuming ZHU ; Lei JIANG ; Wenxin HE ; Chang CHEN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(09):1370-1374
Multi-arm robotic-assisted uniportal pulmonary resection is an emerging minimally invasive technique that enables anatomical pulmonary resection through a single incision using a multi-arm robotic platform. This consensus was initiated by Shanghai Pulmonary Hospital and developed through a multidisciplinary national expert panel. Using evidence-based methodology and expert survey, it provides a comprehensive overview of the clinical application of this technique, including its definition, indications and contraindications, technical considerations, and learning curve. As the first expert consensus focused on multi-arm robotic-assisted uniportal pulmonary resection, this consensus highlights the feasibility and developmental potential of the technique, which integrates existing robotic systems with uniportal minimally invasive principles. The consensus also offers theoretical and practical guidance for the standardized and safe implementation of the technique.
8.Mechanism of Shengxian Yixin Granule (升陷益心颗粒) in the Treatment of Heart Failure with Preserved Ejection Fraction Based on Macrophage-Mediated Inflammatory Response
Zhengwei DONG ; Rui YU ; Qiaozhi LI ; Huan ZHAO ; Jingjing WEI ; Genhao FAN ; Xinlu WANG ; He WANG ; Yongxia WANG ; Mingjun ZHU
Journal of Traditional Chinese Medicine 2026;67(19):2086-2094
ObjectiveTo investigate the effect and mechanism of Shengxian Yixin Granule (升陷益心颗粒, SYG) in treating heart failure with preserved ejection fraction (HFpEF). MethodsExperiment 1: Forty male C57BL/6 mice were randomly divided into control group, model group, low-dose [25 mg/(kg·d)], medium-dose [50 mg/(kg·d)], and high-dose [(75 mg/(kg·d)] SYG groups, with 8 mice per group. Except for the control group, HFpEF was induced by a high-fat diet combined with Nω-nitro-L-arginine methyl ester in drinking water for 16 weeks. After 10 weeks of modeling, treatment groups received corresponding doses of SYG by gavage, while the control group and model group received equal volume of normal saline, for 6 weeks. After 6 weeks of administration, the body weight, blood pressure including systolic blood pressure (SBP) and diastolic blood pressure (DBP), and heart weight-to-tibia length ratio (HW/TL) were recorded. Echocardiography was used to evaluate left ventricular ejection fraction (LVEF) and the ratio of early to late diastolic transmitral flow velocity (E/A). Plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) and C-reactive protein (CRP) levels were measured by ELISA. The expression level of brain natriuretic peptide (BNP) in heart tissue was detected by qRT-PCR. Blood lipid levels, including low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), and total cholesterol (TC), were determined by a biochemical analyzer. Splenic proportions of M1 macrophages, M2 macrophages, T helper 17 (Th17) cells, and regulatory T (Treg) cells were analyzed by flow cytometry. Experiment 2: RAW 264.7 cells were stimulated with lipopolysaccharide (LPS) to establish an M1 macrophage polarization model, and then treated with SYG-containing serum for 24 h. The cells were divided into four groups, including blank serum group, high-dose SYG-containing serum group, LPS group, and LPS plus high-dose SYG-containing serum group. Combined with asignal transducer and activator of transcription 3 (Stat3) inhibitor for 24 h, the cells were divided into a lipopolysaccharide group, a high-dose SYG-containing serum group, an inhibitor group, and high-dose SYG-containing serum plus inhibitor group. The expression levels of inducible nitric oxide synthase (iNOS) and interleukin-1β (IL-1β) in each group were detected by qRT-PCR. The protein levels of iNOS, IL-1β, Stat3, and phosphorylated Stat3 (p-Stat3) were detected by Western Blotting. ResultsExperiment 1: Compared to the control group, the model group showed significantly decreased E/A ratio, and increased cardiac BNP expression, plasma NT-proBNP,HW/TL ratio, body weight, TC, LDL-C, SBP, and DBP, as well as shortened exhaustion distance and exhaustion time (P<0.05 or P<0.01). Compared to the model group, the E/A ratio increased in the low-, medium-, and high-dose SYG groups; cardiac BNP expression decreased in the low- and high-dose groups, and plasma NT-proBNP levels decreased in the high-dose group; the HW/TL ratio and body weight decreased, while exhaustion distance and exhaustion time increased in the medium- and high-dose groups; TG levels decreased in the low- and medium-dose groups, and LDL-C levels decreased in the medium- and high-dose groups; SBP and DBP decreased in the low-, medium-, and high-dose groups (P<0.05 or P<0.01). Compared to the low-dose SYG group, body weight decreased, and both exhaustion distance and exhaustion time increased in the medium- and high-dose groups (P<0.05 or P<0.01). Compared to the medium-dose SYG group, the exhaustion time increased in the high-dose SYG group (P<0.01). Experiment 2: In the LPS‑induced M1 macrophage polarization model, compared to the blank serum group, the LPS group showed significantly increased mRNA and protein expression of iNOS and IL‑1β, as well as the p‑Stat3/Stat3 ratio (P<0.01). Compared to the LPS group, the high‑dose SYG-containing serum plus LPS group showed significantly decreased mRNA and protein expression of iNOS and IL‑1β, as well as the p‑Stat3/Stat3 ratio (P<0.05 or P<0.01). Compared to the LPS group, the inhibitor group also showed significantly reduced these parameters (P<0.05 or P<0.01). ConclusionSYG can improve cardiac function, ventricular remodeling, exercise capacity, and lipid metabolism in HFpEF mice, while inhibiting inflammatory responses. Its mechanism may be related to the regulation of the Stat3 signaling pathway and inhibition of M1 macrophage polarization.
9.Mechanism of Shengxian Yixin Granule (升陷益心颗粒) in the Treatment of Heart Failure with Preserved Ejection Fraction Based on Macrophage-Mediated Inflammatory Response
Zhengwei DONG ; Rui YU ; Qiaozhi LI ; Huan ZHAO ; Jingjing WEI ; Genhao FAN ; Xinlu WANG ; He WANG ; Yongxia WANG ; Mingjun ZHU
Journal of Traditional Chinese Medicine 2026;67(19):2086-2094
ObjectiveTo investigate the effect and mechanism of Shengxian Yixin Granule (升陷益心颗粒, SYG) in treating heart failure with preserved ejection fraction (HFpEF). MethodsExperiment 1: Forty male C57BL/6 mice were randomly divided into control group, model group, low-dose [25 mg/(kg·d)], medium-dose [50 mg/(kg·d)], and high-dose [(75 mg/(kg·d)] SYG groups, with 8 mice per group. Except for the control group, HFpEF was induced by a high-fat diet combined with Nω-nitro-L-arginine methyl ester in drinking water for 16 weeks. After 10 weeks of modeling, treatment groups received corresponding doses of SYG by gavage, while the control group and model group received equal volume of normal saline, for 6 weeks. After 6 weeks of administration, the body weight, blood pressure including systolic blood pressure (SBP) and diastolic blood pressure (DBP), and heart weight-to-tibia length ratio (HW/TL) were recorded. Echocardiography was used to evaluate left ventricular ejection fraction (LVEF) and the ratio of early to late diastolic transmitral flow velocity (E/A). Plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) and C-reactive protein (CRP) levels were measured by ELISA. The expression level of brain natriuretic peptide (BNP) in heart tissue was detected by qRT-PCR. Blood lipid levels, including low-density lipoprotein cholesterol (LDL-C), triglycerides (TG), and total cholesterol (TC), were determined by a biochemical analyzer. Splenic proportions of M1 macrophages, M2 macrophages, T helper 17 (Th17) cells, and regulatory T (Treg) cells were analyzed by flow cytometry. Experiment 2: RAW 264.7 cells were stimulated with lipopolysaccharide (LPS) to establish an M1 macrophage polarization model, and then treated with SYG-containing serum for 24 h. The cells were divided into four groups, including blank serum group, high-dose SYG-containing serum group, LPS group, and LPS plus high-dose SYG-containing serum group. Combined with asignal transducer and activator of transcription 3 (Stat3) inhibitor for 24 h, the cells were divided into a lipopolysaccharide group, a high-dose SYG-containing serum group, an inhibitor group, and high-dose SYG-containing serum plus inhibitor group. The expression levels of inducible nitric oxide synthase (iNOS) and interleukin-1β (IL-1β) in each group were detected by qRT-PCR. The protein levels of iNOS, IL-1β, Stat3, and phosphorylated Stat3 (p-Stat3) were detected by Western Blotting. ResultsExperiment 1: Compared to the control group, the model group showed significantly decreased E/A ratio, and increased cardiac BNP expression, plasma NT-proBNP,HW/TL ratio, body weight, TC, LDL-C, SBP, and DBP, as well as shortened exhaustion distance and exhaustion time (P<0.05 or P<0.01). Compared to the model group, the E/A ratio increased in the low-, medium-, and high-dose SYG groups; cardiac BNP expression decreased in the low- and high-dose groups, and plasma NT-proBNP levels decreased in the high-dose group; the HW/TL ratio and body weight decreased, while exhaustion distance and exhaustion time increased in the medium- and high-dose groups; TG levels decreased in the low- and medium-dose groups, and LDL-C levels decreased in the medium- and high-dose groups; SBP and DBP decreased in the low-, medium-, and high-dose groups (P<0.05 or P<0.01). Compared to the low-dose SYG group, body weight decreased, and both exhaustion distance and exhaustion time increased in the medium- and high-dose groups (P<0.05 or P<0.01). Compared to the medium-dose SYG group, the exhaustion time increased in the high-dose SYG group (P<0.01). Experiment 2: In the LPS‑induced M1 macrophage polarization model, compared to the blank serum group, the LPS group showed significantly increased mRNA and protein expression of iNOS and IL‑1β, as well as the p‑Stat3/Stat3 ratio (P<0.01). Compared to the LPS group, the high‑dose SYG-containing serum plus LPS group showed significantly decreased mRNA and protein expression of iNOS and IL‑1β, as well as the p‑Stat3/Stat3 ratio (P<0.05 or P<0.01). Compared to the LPS group, the inhibitor group also showed significantly reduced these parameters (P<0.05 or P<0.01). ConclusionSYG can improve cardiac function, ventricular remodeling, exercise capacity, and lipid metabolism in HFpEF mice, while inhibiting inflammatory responses. Its mechanism may be related to the regulation of the Stat3 signaling pathway and inhibition of M1 macrophage polarization.
10.Liuwei Dihuangwan Reduces Valproic Acid-induced ASD-like Behaviors in Rats by Modulating mTOR-mediated Synaptic Plasticity in Prefrontal Cortex-hippocampus Circuit
Ji WU ; Niya YIN ; Lei OUYANG ; Siyu HE ; Fangyu PEI ; Ying XIE ; Fan LI ; Qian LEI ; Sha ZHAO ; Di ZHANG ; Qinquan ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(22):1-11
ObjectiveTo investigate the effects of Liuwei Dihuangwan on mammalian target of rapamycin (mTOR)-mediated synaptic plasticity in the prefrontal cortex-hippocampus circuit in the rat model of valproic acid (VPA)-induced autism spectrum disorder (ASD) and to explore its potential mechanism for reducing behavioral abnormalities in ASD rats. MethodsThirteen SD pregnant rats were randomized into the VPA group (10 rats) and the control group (3 rats). On gestational day 13, the VPA group was intraperitoneally injected with VPA solution (600 mg·kg-1), while the control group was injected with an equal volume of normal saline. Male pups of 21 days old from both groups underwent behavioral tests to screen the ASD model rats (32 rats). These rats were then randomly assigned to the model (distilled water), vitamin D (1 480 U·kg-1), low-dose (0.75 g·kg-1) Liuwei Dihuangwan, and high-dose (1.5 g·kg-1) Liuwei Dihuangwan groups, and normal male pups were selected as the blank group (distilled water), with 8 rats in each group. Each group was treated with the corresponding drug or an equal volume of distilled water via gavage daily. After drug administration, behavioral tests were conducted again, followed by anesthesia and tissue collection. Nissl staining was carried out to observe the morphological changes of neurons in the prefrontal cortex and hippocampus. Transmission electron microscopy was used to observe changes in hippocampal synaptic structure. Immunohistochemistry and Western blot were employed to determine the expression of mTOR, phosphatidylinositol 3-kinase (PI3K), and protein kinase B (Akt) in the prefrontal cortex, as well as synaptophysin-1 (SYP-1), postsynaptic density 95 (PSD95), and N-methyl-D-aspartate receptors (NMDARs) in the hippocampus. ResultsCompared with the control group, the model group exhibited severe growth retardation and developmental deformities, with abnormal behavioral performance and pathological damage to neurons and synapses in the prefrontal cortex and hippocampus. Compared with the model group, all treatment groups showed reduced behavioral abnormalities and pathological damage to neurons and synapses in the prefrontal cortex and hippocampus, with the most significant effects in the high-dose Liuwei Dihuangwan group. Immunohistochemistry and Western blotting results showed that compared with the control group, the model group exhibited significantly increased protein levels and phosphorylation ratios of p-mTOR, p-Akt, and p-PI3K in the prefrontal cortex (P<0.05, P<0.01), and decreased protein levels of SYP-1 and PSD95 and increased protein level of NMDARs in the hippocampus (P<0.01). Compared with those in the model group, the protein levels of p-mTOR, p-Akt, and p-PI3K in the prefrontal cortex decreased in the vitamin D and high-dose Liuwei Dihuangwan groups (P<0.05), and the p-mTOR/mTOR, p-Akt/Akt, and p-PI3K/PI3K ratios in the prefrontal cortex declined in the vitamin D group (P<0.05). The high-dose Liuwei Dihuangwan group showed a reduction in p-PI3K/PI3K ratio (P<0.05). The protein level of SYP-1 and PSD95 in the hippocampus increased in the vitamin D, low-dose Liuwei Dihuangwan, and high-dose Liuwei Dihuangwan groups (P<0.05, P<0.01), while the protein level of NMDARs decreased in the vitamin D and high-dose Liuwei Dihuangwan groups (P<0.01). ConclusionLiuwei Dihuangwan significantly reduces VPA-induced ASD-like behaviors and enhances the regeneration and repair of neurons in the prefrontal cortex and synaptic structures in the hippocampus by downregulating of PI3K/Akt/mTOR expression in the prefrontal cortex, upregulating SYP-1 and PSD95 expression in the hippocampus, and inhibiting NMDARs expression.


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