1.Study on the safety and efficacy of micro-perfusion device for preserving isolated porcine limbs
Pengkai LI ; Zhaodi MI ; Shen LI ; Man YUAN ; Xiwei PENG ; Jia LÜ ; Sice WANG ; Zhibo JIA ; Xiangyu SONG ; Yixuan ZHU ; Chonghui LI ; Moling XIAO ; Wenjing XU ; Jiang PENG
Organ Transplantation 2026;17(3):422-431
Objective To evaluate the safety and efficacy of a self-developed micro-normothermic machine perfusion (NMP) system (micro-perfusion device) for preserving isolated porcine limbs. Methods Five healthy Landrace pigs were selected, and their left and right forelimbs were randomly divided into the NMP group and static cold storage (SCS) group. The NMP group was perfused with the self-developed micro-perfusion device and polymerized hemoglobin perfusate for 32 hours at normothermia, while the SCS group was preserved at 4 ℃. Hemodynamic parameters such as perfusion pressure and flow were monitored. The pH value, partial pressure of oxygen (PO2), lactic acid (Lac), creatine kinase (CK) and lactate dehydrogenase (LDH) in the perfusate were measured. Hematoxylin-eosin staining was used to assess the muscle tissue structure, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling was employed to evaluate muscle cell apoptosis, and immunohistochemistry staining was applied to detect the expressions of tumor necrosis factor (TNF)-α and interleukin (IL)-6. A mixed-effects model was used to analyze the effects of time and treatment methods on tissue structure, cell apoptosis and inflammatory factors. Results The device could stably maintain a perfusion pressure of (69±15) mmHg and a flow rate of (117±42) mL/min. The pH value and electrolytes of the perfusate were generally stable, with PO2 maintained at a high level. Lac was maintained at 5.38(3.81, 6.45) mmol/L, while CK and LDH increased over time. After 32 hours of perfusion in the NMP group, both the myocyte spacing and apoptosis rate were better than those in the SCS group. Mixed-effects model analysis showed that there were statistically significant differences in the effects of NMP treatment and SCS treatment on myocyte spacing and apoptosis rate per unit time (both P < 0.05). There were no statistically significant differences in TNF-α and IL-6 between the two groups, and mixed-effects model analysis showed no statistically significant differences in the effects of NMP treatment and SCS treatment on TNF-α and IL-6 per unit time (both P > 0.05). Conclusions The micro-perfusion device used in this study may achieve 32-hour normothermic preservation in a porcine limb amputation model, maintain basic metabolism and ionic homeostasis, reduce muscle structural damage and cell apoptosis without inducing additional inflammatory responses. This technology is expected to significantly extend the time window for replantation of amputated limbs in disaster rescue and long-distance transportation, providing an important technical basis for clinical translation and subsequent replantation research.
2.Clinical efficacy of Huangkui capsules in the treatment of targeted drug-related proteinuria in patients with hepatocellular carcinoma
Miao LI ; Jia YUAN ; Chu LIU ; Maopei CHEN ; Xin XU ; Ningling GE ; Yi CHEN ; Lan ZHANG ; Rongxin CHEN ; Yan WANG
Chinese Journal of Clinical Medicine 2026;33(1):88-94
Objective To investigate the therapeutic effect of Huangkui capsules on targeted drug-related proteinuria in patients with hepatocellular carcinoma (HCC). Methods A retrospective analysis was conducted on clinical data of HCC patients with targeted drug-related proteinuria from June 2023 to December 2024 at Zhongshan Hospital, Fudan University. According to the treatment plan, patients were divided into the conventional treatment group and the Huangkui combination treatment group (Huangkui capsules combined with conventional treatment), and the clinical efficacy between the two groups was compared. The logistic regression analysis was used to identify the main factors affecting treatment efficacy. Results The Huangkui combination treatment group (n=29) showed a significantly higher overall effective rate (79.3% vs 42.3%, P=0.005), and an earlier proteinuria improvement (median time: 3 months vs 6 months, P=0.008) than the conventional treatment group (n=26) . The multivariate logistic regression analysis showed angiotensin-converting enzyme inhibitor (ACEI) or angiotensin Ⅱ receptor blocker (ARB) using (OR=0.190, 95%CI 0.045-0.808, P=0.025), targeted drug adjustment (OR=0.132, 95%CI 0.030-0.581, P=0.007), and Huangkui capsules using (OR=0.168, 95%CI 0.039-0.730, P=0.017) were protective factors for treatment efficacy of targeted drug-related proteinuria. Conclusions On the basis of conventional treatment, additive treatment with Huangkui capsules can alleviate targeted drug-related proteinuria faster and more effectively in HCC patients.
3.Preoperative evaluation of lung function in patients with lung cancer using two-phase dual-energy CT perfusion imaging
Lifang LING ; Yizhen JIA ; Qinmin HAO ; Wenzheng XU ; Zhibo WANG ; Jun WANG ; Liang CHEN ; Mei YUAN
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(01):79-86
Objective To explore the application value of dual-phase dual-energy CT (DECT) perfusion imaging in preoperative lung function assessment of lung cancer patients. Methods Data were collected from patients with stageⅠA non-small cell lung cancer who underwent surgical treatment in the Department of Thoracic Surgery, the First Affiliated Hospital of Nanjing Medical University, from November 2022 to June 2024. All patients underwent DECT perfusion imaging and pulmonary function testing (PFT) before surgery. PFT observation indicators included ventilation function indicators such as forced expiratory volume in one second (FEV1), forced vital capacity (FVC), 1-second rate (FEV1/FVC), maximal voluntary ventilation (MVV), and diffusion function indicators such as diffusing capacity for carbon monoxide (DLCO) and DLCO per liter of alveolar volume (DLCO/VA). The software eXamine was used to obtain quantitative parameters of DECT perfusion imaging, including volume parameters and perfusion parameters of both lungs and each lung lobe. The correlation between the volume parameters and perfusion parameters of both lungs and the ventilation and diffusion function indicators of the patients, as well as the differences in quantitative parameters of each lung lobe, was analyzed. Results The end-inspiration lung volume and biphasic volume difference were strongly positively correlated with FEV1 and FVC (r=0.636, r=0.682, r=0.614, r=0.624, P<0.001) and moderately positively correlated with MVV and DLCO (r=0.499, r=0.514, r=0.549, r=0.447, P<0.001); the end-expiration lung volume was weakly negatively correlated with DLCO/VA (r=−0.295, P=0.026); the volume ratio was positively correlated with FEV1, FVC, MVV, and MVV% (r=0.424, r=0.399, r=0.415, r=0.310, P<0.05); the end-inspiration iodine content was weakly positively correlated with DLCO/VA% (rs=0.292, P=0.030); the end-expiration iodine content was weakly positively correlated with FEV1, FVC, MVV, DLCO%, and DLCO/VA (r=0.307, r=0.299, r=0.295, r=0.366, r=0.320, P<0.05) and moderately positively correlated with DLCO (r=0.439, P<0.001); the end-inspiration iodine concentration was negatively correlated with FEV1, FVC, MVV, and MVV% (rs=−0.407, rs=−0.426, rs=−0.352, rs=−0.277, P<0.05); the end-expiratory phase iodine concentration was moderately positively correlated with DLCO/VA (r=0.403, P=0.002); both the iodine concentration difference and the iodine concentration ratio were moderately positively correlated with FEV1, FEV1%, FVC, MVV, MVV% (P<0.05). The lung volume and iodine concentration ratio values were both highest in the left upper lung lobe and lowest in the right middle lung lobe; the differences in lung volume, lung volume ratio, intrapulmonary iodine content, and intrapulmonary iodine concentration were all highest in the lower lobes of both lungs and lowest in the middle lobe of the right lung. Conclusion Dual-phase DECT perfusion imaging can accurately assess overall lung function and quantify regional lung function.
4.QingNangTCM: a parameter-efficient fine-tuning large language model for traditional Chinese medicine
Xuming TONG ; Liyan LIU ; Yanhong YUAN ; Xiaozheng DING ; Huiru JIA ; Xu YANG ; Sio Kei IM ; Mini Han WANG ; Zhang XIONH ; Yapeng WANG
Digital Chinese Medicine 2026;9(1):1-12
Objective:
To develop QingNangTCM, a specialized large language model (LLM) tailored for expert-level traditional Chinese medicine (TCM) question-answering and clinical reasoning, addressing the scarcity of domain-specific corpora and specialized alignment.
Methods:
We constructed QnTCM_Dataset, a corpus of 100 000 entries, by integrating data from ShenNong_TCM_Dataset and SymMap v2.0, and synthesizing additional samples via retrieval-augmented generation (RAG) and persona-driven generation. The dataset comprehensively covers diagnostic inquiries, prescriptions, and herbal knowledge. Utilizing P-Tuning v2, we fine-tuned the GLM-4-9B-Chat backbone to develop QingNangTCM. A multi-dimensional evaluation framework, assessing accuracy, coverage, consistency, safety, professionalism, and fluency, was established using metrics such as bilingual evaluation understudy (BLEU), recall-oriented understudy for gisting evaluation (ROUGE), metric for evaluation of translation with explicit ordering (METEOR), and LLM-as-a-Judge with expert review. Qualitative analysis was conducted across four simulated clinical scenarios: symptom analysis, disease treatment, herb inquiry, and failure cases. Baseline models included GLM-4-9B-Chat, DeepSeek-V2, HuatuoGPT-II (7B), and GLM-4-9B-Chat (freeze-tuning).
Results:
QingNangTCM achieved the highest scores in BLEU-1/2/3/4 (0.425/0.298/0.137/0.064), ROUGE-1/2 (0.368/0.157), and METEOR (0.218), demonstrating a balanced and superior normalized performance profile of 0.900 across the dimensions of accuracy, coverage, and consistency. Although its ROUGE-L score (0.299) was lower than that of HuatuoGPT-II (7B) (0.351), it significantly outperformed domain-specific models in expert-validated win rates for professionalism (86%) and safety (73%). Qualitative analysis confirmed that the model strictly adheres to the “symptom-syndrome-pathogenesis-treatment” reasoning chain, though occasional misclassifications and hallucinations persisted when dealing with rare medicinal materials and uncommon syndromes.
Conclusion
Combining domain-specific corpus construction with parameter-efficient prompt tuning enhances the reasoning behavior and domain adaptation of LLMs for TCM-related tasks. This work provides a technical framework for the digital organization and intelligent utilization of TCM knowledge, with potential value for supporting diagnostic reasoning and medical education.
5.Mechanism of Xixintang in Protecting Blood-brain Barrier in Alzheimer's Disease Model Rats Based on AQP4 Polarization
Siyuan JIA ; Yongchang DIWU ; Yuan TIAN ; Jie GAO ; Meirong WU ; Dengkun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(9):1-10
ObjectiveThis paper aims to investigate the effects of Xixintang on aquaporin-4 (AQP4) polarity distribution, blood-brain barrier (BBB) function, and neuroinflammationin rats with Alzheimer's disease (AD), thereby revealing the potential mechanism through which this formula protects the BBB by regulating AQP4 polarization. The aim is to provide a scientific basis for clinical treatment. MethodsSixty Sprague-Dawley (SD) rats were randomly divided into a normal group, a model group, a probiotic group, a donepezil group, and an Xixintang group. The model was established by intraperitoneal injection of D-galactose (D-Gal) combined with bilateral intracerebroventricular injection of amyloid-β25-35 (Aβ25-35). The probiotic group (30.85 mg·kg-1), donepezil group (0.88 mg·kg-1), and Xixintang group (1.174 g·kg-1) received daily gavage administration, while the normal and model groups received intragastric administration with an equal volume of normal saline for one month. Cognitive ability was assessed by using the Morris water maze. BBB permeability was detected via Evans blue extravasation. The contents of interleukin-6 (IL-6), amyloid-β1-42 (Aβ1-42), and tumor necrosis factor-α (TNF-α) in the hippocampal tissues were measured by enzyme-linked immunosorbent assay (ELISA). The protein expressions of zonula occludens-1 (ZO-1), occludin, tissue inhibitor of metalloproteinase-1 (TIMP-1), matrix metalloproteinase-9 (MMP-9), and AQP4 in the hippocampal tissues were detected by western blot. The expression and co-localization levels of Aβ1-42, ionized calcium-binding adapter molecule 1 (IBA1), and AQP4/platelet endothelial cell adhesion molecule 31 (CD31) in the hippocampal region were examined by immunofluorescence. ResultsCompared with the normal group, the model group exhibited a significant decline in cognitive ability (P<0.01) and a marked increase in Evans blue extravasation in the brain (P<0.01). The expressions of ZO-1, occludin, and TIMP-1 were significantly decreased (P<0.01), while the expressions of AQP4 and MMP-9 were significantly increased (P<0.01). The co-localization level of AQP4/CD31 was significantly reduced (P<0.01), and the expressions of Aβ1-42, IL-6, TNF-α, and IBA1 were significantly elevated (P<0.01). Compared with the model group, the Xixintang group showed significant improvement in cognitive ability (P<0.01) and a significant reduction in Evans blue extravasation in the brain (P<0.01). The expressions of occludin, TIMP-1, and ZO-1 were significantly increased (P<0.05, P<0.01), while the expressions of AQP4 and MMP-9 were significantly decreased (P<0.05). The co-localization level of AQP4/CD31 was significantly enhanced (P<0.01), and the expressions of Aβ1-42, IL-6, TNF-α, and IBA1 were significantly reduced (P<0.05, P<0.01). ConclusionXixintang may improve cognitive function and alleviate AD pathology in AD model rats by regulating AQP4 polarity distribution, thereby breaking the vicious cycle of "Aβ deposition-neuroinflammation-BBB damage" and restoring the homeostasis of the microenvironment in the brain.
6.Effect of Xixintang on Colonic Mucosal Barrier and TLR4 /NF-κB p65 Signaling Pathway in AD Model Rats Induced by D-galactose Combined with Aβ25-35
Yuan TIAN ; Yongchang DIWU ; Siyuan JIA ; Jie GAO ; Meirong WU ; Dengkun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):1-11
ObjectiveThis study aims to investigate whether Xixintang could ameliorate cognitive dysfunction in an Alzheimer's disease (AD) rat model induced by D-galactose and β-amyloid (Aβ25-35), by means of repairing the colonic mucosal barrier, regulating the Toll-like receptor 4 (TLR4)/nuclear factor-κB p65 (NF-κB p65) signaling pathway, and intervening in the pathological process mediated by the gut-brain axis. MethodsSixty specific pathogen-free (SPF) male Sprague-Dawley (SD) rats were randomly divided to five groups (n=12): A control group, a model group, a donepezil group, an Xixintang group, and a probiotic group. Except for those in the control group, rats in all other groups received daily intraperitoneal injections of D-galactose for six consecutive weeks. Subsequently, aggregated Aβ25-35 was injected stereotactically into the bilateral ventricles to establish the AD model. During the intervention periods, the rats in all groups were administered their respective drugs and normal saline by gavage. The Morris water maze test was used to assess the capacity for spatial learning and memory. Hematoxylin-eosin (HE) staining was employed to observe the histopathological changes in the colon tissues. Immunofluorescence was used to detect Aβ1-41 deposition in the hippocampal region and Mucin 2 (MUC2) expression in the colonic mucosa. Western blot was performed to measure the protein expression levels of FFAR2,TLR4, NF-κB p65, occludin (OCLN), zonula occludens-1 (ZO-1), and MUC2 in the colonic tissues. Enzyme-linked immunosorbent assay (ELISA) was used to determine the contents of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), serum amyloid A (SAA), and Aβ1-42 in the hippocampal region from the colonic tissues. The lipopolysaccharide (LPS) concentrations in colon tissues of rats were measured by using a dynamic chromogenic limulus assay. ResultsCompared with those in the control group, the rats in the model group exhibited a significantly prolonged escape latency and a markedly shorter duration in the target quadrant (P<0.01). The integrity of the colonic mucosal structure was compromised, with disordered gland arrangement and a reduced number of goblet cells. The Aβ1-42 deposition in the hippocampal region was significantly increased (P<0.01). The protein expression levels of TLR4 and NF-κB p65 in colonic tissues were significantly upregulated (P<0.01), while those of occludin and ZO-1 were downregulated (P<0.01). The contents of inflammatory factors such as IL-6, TNF-α, and SAA were significantly elevated (P<0.01), and the LPS level in the serum was markedly increased (P<0.01). In comparison to those in the model group, the rats in the Xixintang group showed a significantly shortened escape latency and a prolonged duration in the target quadrant (P<0.01). The colonic mucosal structure was ameliorated, with neat gland arrangement and an increased number of goblet cells. The Aβ1-42 deposition in the hippocampal region was reduced (P<0.01). The protein expressions of TLR4 and NF-κB p65 in the colon tissues were decreased (P<0.05,P<0.01), while the protein levels of occludin and ZO-1 were increased (P<0.01). The contents of IL-6, TNF-α, and serum amyloid A (SAA) were decreased (P<0.01), and the LPS level was reduced (P<0.01). ConclusionXixintang can significantly ameliorate cognitive dysfunction of AD model rats, by means of restoring the colonic mucosal barrier structure, reducing cerebral Aβ deposition, and suppressing peripheral and central inflammatory response. Its mechanism of action may be closely associated with the suppression of the TLR4/NF-κB signaling pathway activation, reduction of endotoxin levels, and regulation of the gut-brain axis.
7.Effect of Xixintang on Colonic Mucosal Barrier and TLR4 /NF-κB p65 Signaling Pathway in AD Model Rats Induced by D-galactose Combined with Aβ25-35
Yuan TIAN ; Yongchang DIWU ; Siyuan JIA ; Jie GAO ; Meirong WU ; Dengkun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):1-11
ObjectiveThis study aims to investigate whether Xixintang could ameliorate cognitive dysfunction in an Alzheimer's disease (AD) rat model induced by D-galactose and β-amyloid (Aβ25-35), by means of repairing the colonic mucosal barrier, regulating the Toll-like receptor 4 (TLR4)/nuclear factor-κB p65 (NF-κB p65) signaling pathway, and intervening in the pathological process mediated by the gut-brain axis. MethodsSixty specific pathogen-free (SPF) male Sprague-Dawley (SD) rats were randomly divided to five groups (n=12): A control group, a model group, a donepezil group, an Xixintang group, and a probiotic group. Except for those in the control group, rats in all other groups received daily intraperitoneal injections of D-galactose for six consecutive weeks. Subsequently, aggregated Aβ25-35 was injected stereotactically into the bilateral ventricles to establish the AD model. During the intervention periods, the rats in all groups were administered their respective drugs and normal saline by gavage. The Morris water maze test was used to assess the capacity for spatial learning and memory. Hematoxylin-eosin (HE) staining was employed to observe the histopathological changes in the colon tissues. Immunofluorescence was used to detect Aβ1-41 deposition in the hippocampal region and Mucin 2 (MUC2) expression in the colonic mucosa. Western blot was performed to measure the protein expression levels of FFAR2,TLR4, NF-κB p65, occludin (OCLN), zonula occludens-1 (ZO-1), and MUC2 in the colonic tissues. Enzyme-linked immunosorbent assay (ELISA) was used to determine the contents of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), serum amyloid A (SAA), and Aβ1-42 in the hippocampal region from the colonic tissues. The lipopolysaccharide (LPS) concentrations in colon tissues of rats were measured by using a dynamic chromogenic limulus assay. ResultsCompared with those in the control group, the rats in the model group exhibited a significantly prolonged escape latency and a markedly shorter duration in the target quadrant (P<0.01). The integrity of the colonic mucosal structure was compromised, with disordered gland arrangement and a reduced number of goblet cells. The Aβ1-42 deposition in the hippocampal region was significantly increased (P<0.01). The protein expression levels of TLR4 and NF-κB p65 in colonic tissues were significantly upregulated (P<0.01), while those of occludin and ZO-1 were downregulated (P<0.01). The contents of inflammatory factors such as IL-6, TNF-α, and SAA were significantly elevated (P<0.01), and the LPS level in the serum was markedly increased (P<0.01). In comparison to those in the model group, the rats in the Xixintang group showed a significantly shortened escape latency and a prolonged duration in the target quadrant (P<0.01). The colonic mucosal structure was ameliorated, with neat gland arrangement and an increased number of goblet cells. The Aβ1-42 deposition in the hippocampal region was reduced (P<0.01). The protein expressions of TLR4 and NF-κB p65 in the colon tissues were decreased (P<0.05,P<0.01), while the protein levels of occludin and ZO-1 were increased (P<0.01). The contents of IL-6, TNF-α, and serum amyloid A (SAA) were decreased (P<0.01), and the LPS level was reduced (P<0.01). ConclusionXixintang can significantly ameliorate cognitive dysfunction of AD model rats, by means of restoring the colonic mucosal barrier structure, reducing cerebral Aβ deposition, and suppressing peripheral and central inflammatory response. Its mechanism of action may be closely associated with the suppression of the TLR4/NF-κB signaling pathway activation, reduction of endotoxin levels, and regulation of the gut-brain axis.
8.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.
9.Mechanistic Interpretation of Zheng’s San Qi San Powder in Treating Skeletal Muscle Injury via Bioinformatics Prediction, Chemical Analysis and Experimental Verification
Ding-Rui WANG ; Yun-Xin LIU ; Jun-Jie XU ; Liu YANG ; Jia-Hao LÜ ; Cheng-Yuan XING ; Lei LÜ ; Bei-Bei QIE
Progress in Biochemistry and Biophysics 2026;53(4):1028-1047
ObjectiveZheng’s San Qi San (ZSQS) power, a classic traditional Chinese medicine (TCM) formula, is used for treating soft tissue injuries involving muscles, tendons, and ligaments. However, its underlying therapeutic mechanisms remain unclear. This study aimed to screen and identify pharmaceutically active ingredients and their candidate biomolecule targets, and further elucidate the molecular mechanism of ZSQS in the treatment of skeletal muscle injury. MethodsNetwork pharmacology was employed to construct “ZSQS-component-target”, “protein-protein interaction (PPI)” and “active ingredient-core protein-pathway” networks to predict the key active ingredients and potential core targets of ZSQS for skeletal muscle injury. The predicted results were then validated via microarray data from the GEO database. Molecular docking was then performed to assess the binding ability between the screened active ingredients of ZSQS and the candidate core targets. Moreover, liquid chromatography-mass spectrometry (LC-MS) was used for qualitative and quantitative analysis to verify the active components of the drug and ZSQS serum. Finally, an animal model of eccentric exercise-induced skeletal muscle injury and a myotube cell model of oxidative stress-induced injury were established to validate the effects of ZSQS and its interventional effects on the biological functions of critical targets, thereby demonstrating the potential therapeutic mechanism of ZSQS. ResultsAmong the 111 active components identified in ZSQS and their corresponding 204 targets related to the skeletal muscle injury repair process, 14 core targets (including AKT1) and 4 core active components (quercetin, luteolin, kaempferol, and β‑sitosterol) were screened out, while the corresponding metabolites of quercetin, luteolin and kaempferol were detected in the ZSQS serum. Among these targets, 5 candidate genes (IL-6, CASP3, HIF1A, STAT3, and JUN) overlapped with the differential expression screening results with GEO data, and IL-6 was confirmed to be enriched in the PI3K/AKT pathway. Combined with the prediction results of the AKT expression levels, these findings suggest that the phosphorylation level of AKT1 plays a core role in the therapeutic mechanism of ZSQS. Molecular docking analysis further revealed that the PH domain of AKT1 had high binding energy with all 4 core active components, as verified by LC-MS. Finally, animal model studies have shown the promoting effect of ZSQS administration on skeletal muscle injury repair and its possible antioxidant damage mechanism. Cell model studies further demonstrated that ZSQS-containing serum, core active ingredient combination therapy, and quercetin monomer could increase the phosphorylation level of AKT, promote the nuclear translocation of Nrf2, upregulate the expression of downstream antioxidant enzymes (SOD, GPx, and GR), and inhibit the expression of inflammatory factors (IL-6 and TNF-α), thereby alleviating oxidative stress and the inflammatory response. ConclusionZSQS alleviates skeletal muscle injury mainly by activating the AKT/Nrf2 signaling pathway, enhancing cellular antioxidant and anti-inflammatory capabilities. The results of this study provide a scientific basis for the clinical application and modernized development of ZSQS.
10.Identification of Lonicera japonica TPS gene family and expression analysis under aphid damage.
Gang WANG ; Yuan CUI ; Qi-Dong LI ; Lu-Yao HUANG ; Zhen-Hua LIU ; Jia LI
China Journal of Chinese Materia Medica 2025;50(8):2116-2129
This study explores the basic characteristics and potential functions of the terpene synthase(TPS) gene family members in Lonicera japonica. The L. japonica TPS(LjTPS) gene family was identified and functionally analyzed using bioinformatics methods. The results showed that a total of 70 members of the LjTPS gene family were identified in L. japonica, with protein lengths ranging from 130 to 1 437 amino acids. Most of these proteins were hydrophilic, and they were unevenly distributed across nine chromosomes. Phylogenetic analysis showed that the LjTPS gene family members were divided into six subfamilies, mainly consisting of members from the TPS-a, TPS-b, and TPS-e subfamilies. Promoter cis-acting element analysis showed that LjTPS members contained a large number of stress-responsive cis-acting elements. Aphid inoculation experiments showed that key enzyme genes in the MVA pathway for terpenoid backbone synthesis in L. japonica, such as HMGS, HMGR, MK, MPD, and the key enzyme gene in the DXP pathway, DXS, exhibited an initial increase followed by a decrease under aphid stress. The qRT-PCR analysis showed that the expression levels of the α-farnesene synthase genes LjTPS34 and LjTPS39 were down-regulated, while the expression levels of(E)-β-caryophyllene synthase genes LjTPS15 and LjTPS17 were up-regulated 12 h before aphid feeding, then began to decline. Farnesyl pyrophosphate synthase(FPS), which interacted with these genes, also displayed a pattern of increasing followed by decreasing expression. The expression of linalool synthase genes LjTPS12 and LjTPS33 was significantly up-regulated after 72 h of aphid feeding(P<0.000 1), reaching 24.39 and 22.64 times the initial expression, respectively. This pattern was in close alignment with the trend of linalool content in L. japonica. This study provides a theoretical foundation for future research on the interaction between L. japonica and pests, as well as on the functional roles of the LjTPS gene family.
Animals
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Aphids/physiology*
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Alkyl and Aryl Transferases/chemistry*
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Lonicera/parasitology*
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Phylogeny
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Plant Proteins/chemistry*
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Gene Expression Regulation, Plant
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Multigene Family
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Terpenes/metabolism*

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