1.Expert Consensus on Clinical Application of Qidong Yixin Oral Liquid
Changkuan FU ; Xiaochang MA ; Mingjun ZHU ; Yue DENG ; Hongxu LIU ; Mingxue ZHANG ; Ying CHEN ; Yan ZHOU ; Ling ZHANG ; Jianhua FU ; Wei YANG ; Yu'er HU ; Ming CHEN ; Yanming XIE ; Yuanyuan LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(4):147-158
The prescription of Qidong Yixin oral liquid is derived from the experience of national medical master Ren Jixue in treating viral myocarditis (VMC). It has the functions of tonifying Qi, nourishing the heart,calming the mind, and relieving palpitations. It is used to treat VMC and angina pectoris of coronary heart disease caused by deficiency of both Qi and Yin. However,the understanding of its efficacy evidence, advantageous aspects, dosage and administration, and medication safety remains insufficient in clinical practice. Therefore,the development of the Expert Consensus on the Clinical Application of Qidong Yixin Oral Liquid (hereinafter referred to as consensus) was initiated. Consensus strictly followed the process and methods of the expert consensus on the clinical application of Chinese patent medicines of the China Association of Chinese Medicine,successively completing multiple tasks such as the consensus project initiation,determination of clinical problems,evidence search and evaluation,formation of recommendation opinions and consensus suggestions,solicitation of opinions,peer review, submission for review and release, and so on. Consensus formed a total of 10 recommendation opinions and 12 consensus suggestions,clarifying the clinical positioning,efficacy advantages,syndrome differentiation,dosage and administration,combination therapy,timing of medication,adverse reactions,contraindications, and precautions of Qidong Yixin oral liquid,indicating that it has good clinical advantages and safety in the treatment of VMC and angina pectoris of coronary heart disease,providing norms and references for physicians to safely and rationally apply Qidong Yixin oral liquid. Consensus was reviewed and approved for release by the Standardization Office of the China Association of Chinese Medicine on December 23, 2024. Standard number:GSCACM-376-2024.
2.Effect of Optimized New Shengmai Powder (优化新生脉散方) on Exercise Tolerance in Patients with Chronic Heart Failure of Qi Deficiency,Blood Stasis and Fluid Retention Syndrome:A Randomized,Double-Blind,Placebo-Controlled Trial
Xianliang WANG ; Jingyi ZHANG ; Zhao GE ; Tongzuo LIU ; Maozhe ZHANG ; Shuai WANG ; Zhiqiang ZHAO ; Yingfei BI ; Ruijuan ZHOU ; Ying ZHENG ; Jingyuan MAO
Journal of Traditional Chinese Medicine 2026;67(4):425-431
ObjectiveTo evaluate the effects and safety of the optimized new Shengmai Powder (优化新生脉散方) on exercise tolerance in patients with chronic heart failure (CHF) of qi deficiency, blood stasis, and fluid retention syndrome. MethodsA randomized, double-blind, placebo-controlled trial was conducted. A total of 78 CHF patients with qi deficiency, blood stasis, and fluid retention syndrome were recruited and randomly assigned to a treatment group (39 cases) and a control group (39 cases). On the basis of conventional western medical therapy, patients in the treatment group additionally received the optimized new Shengmai Powder granules, while the control group was given an oral placebo of optimized new Shengmai Powder granules. Patients in both groups took 30.6 g each time, twice a day, mixed with water for administration, with a total treatment course of 4 weeks. The primary outcomes were 6-minute walk distance (6MWD) and peak oxygen uptake (Peak VO2) measured by cardiopulmonary exercise testing. Secondary outcomes included New York Heart Association (NYHA) functional classification, B-type natriuretic peptide (BNP) levels, cardiac function indexes including left ventricular ejection fraction (LVEF), left ventri-cular end-systolic diameter (LVESD) and left ventricular end-diastolic diameter (LVEDD), Minnesota Living with Heart Failure Questionnaire (MLHFQ) scores, and scores of four diagnostic information of traditional Chinese medicine (TCM). All indicators were assessed once before and after treatment respectively. Safety indicators were evaluated, and adverse events during the trial were recorded. ResultsAll patients in both groups were included in the full ana-lysis set (FAS) and safety set (SS). Compared with baseline, the 6MWD and Peak VO₂ of cardiopulmonary exercise test in the treatment group significantly increased after treatment, while the MLHFQ scores, serum BNP levels and scores of TCM four diagnostic information significantly decreased, and the NYHA cardiac function grade significantly improved (P<0.01). After treatment, the 6MWD and Peak VO₂ of cardiopulmonary exercise test, as well as their changes from baseline in the treatment group were higher than those in the control group; the MLHFQ scores, serum BNP levels and scores of TCM four diagnostic information in the treatment group were lower than those in the control group; and the improvement of NYHA cardiac function grade in the treatment group was superior to that in the control group (P<0.01). There was no statistically significant differences in all indicators after treatment in the control group (P>0.05). The incidence of adverse events was 5.1% (2/39) in the treatment group and 2.6% (1/39) in the control group, with no statistically significant difference between groups (P>0.05). ConclusionOn the basis of conventional western medicine treatment, the addition of the optimized new Shengmai Powder can further improve exercise tolerance, cardiac function and quality of life in patients with CHF of qi deficiency, blood stasis and fluid retention syndrome, and show good safety.
3.Mechanisms of Qiaobai cold compress solution in improving acne vulgaris based on transcriptomics and experiment
Zhenjiang XIE ; Weina ZHU ; Liangliang CAO ; Fuqiong ZHOU ; Shupan ZHANG ; Bingwen ZHOU ; Yinsheng CHEN ; Wen LI ; Ying ZHAO
China Pharmacy 2026;37(4):425-430
OBJECTIVE To investigate the mechanism by which Qiaobai cold compress solution (QBCS) improves acne vulgaris (AV) based on transcriptomics and animal experiments. METHODS Rats were randomly divided into a blank control group ( n =6) and a modeling group ( n =30). AV models were established in the modeling group by topical application of oleic acid to the inner surface of both ears, combined with subcutaneous injection of Cutibacterium acnes suspension into the auricle. Successfully modeled rats were further divided into the model group, positive control group (Tretinoin cream, 0.045 g/kg), and QBCS low-, medium-, high-dose groups [3.55, 7.11, 14.22 g/kg (calculated by the amount of crude drug) ] , with 6 rats in each group. Rats in each d rug group were treated with the corresponding drugs once daily for 14 consecutive days. After the final administration, changes in the appearance of the ears and histopathological changes in the ear tissues were observed, and serum levels of inflammatory factors, including tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β, were measured. Auricular tissues from the blank control group, model group and QBCS medium-dose group were collected for transcriptome sequencing. Differential expressed genes (DEGs) were screened and subjected to Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, followed by validation using real-time quantitative polymerase chain reaction and Western blot assay. RESULTS Compared with the model group, rats in all QBCS groups showed alleviated auricular acne symptoms, with reduced epidermal thickening, sebaceous gland hyperplasia, and inflammatory cell infiltration. Serum levels of TNF-α (except for the QBCS low-dose group), IL-6 (except for the QBCS low-dose group) and IL-1β were significantly decreased ( P <0.05). A total of 590 DEGs were identified (blank control group vs. model group), and 596 DEGs were identified (model group vs. QBCS medium-dose group). Above DEGs (blank control group vs. model group) were mainly enriched in Toll-like receptor (TLR) and nuclear factor-kappa B (NF-κB) signaling pathways, etc. Validation experiments showed that, compared with model group, low-, medium- and high-dose of QBCS reduced, to varying degrees, the mRNA expression of TNF-α, TLR2, interferon-γ and CXC chemokine ligand 8 in the auricular tissues of AV rats, increased the mRNA expression of peroxisome-proliferator-activated receptor gamma and tumor protein 53, and inhibited the phosphorylation of NF-κB p65 protein as well as the expressions of TLR2 and myeloid differentiation primary response protein 88(MyD88) ( P <0.05). CONCLUSIONS QBCS can alleviate auricular inflammation and skin lesions in AV rats. This effect may be related to inhibition of the TLR/MyD88/NF-κB signaling pathway, thereby suppressing the expression of downstream inflammatory factors such as TNF-α.
4.Mechanism of Diaphragmatic Dysfunction in Chronic Obstructive Pulmonary Disease and Treatment with Traditional Chinese Medicine: A Review
Yuanyuan YING ; Xiaoqing ZHOU ; Kaiwen NI ; Zhen WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):285-296
Chronic obstructive pulmonary disease(COPD) is a common chronic respiratory disorder frequently accompanied by diaphragmatic dysfunction during its course, which significantly increases respiratory burden and impairs quality of life. As the primary inspiratory muscle, the diaphragm is prone to fatigue, atrophy, inflammation, and fibrosis during the long-term progression of COPD. Its pathological mechanisms involve multiple pathways such as inflammatory responses, oxidative stress, mitochondrial dysfunction, apoptosis, ion channel abnormalities, epigenetic regulation, autophagy disorder, and protein metabolism imbalance. In recent years, traditional Chinese medicine(TCM) has demonstrated multi-targeted and systemic regulatory advantages in improving diaphragmatic function in COPD. However, related studies remain fragmented, and integrated mechanistic understanding is lacking. This paper focuses on the mechanism-target-TCM intervention framework, systematically summarizing the molecular mechanisms of diaphragmatic dysfunction, while incorporating the TCM theory of Zongqi(ancestral Qi). It highlights the therapeutic effects of Chinese herbal formulas, single herbs, and active components in modulating inflammation, oxidative stress, mitochondrial function, ion channels, epigenetic processes, autophagy, and protein homeostasis. Additionally, the review outlines existing challenges, including insufficient study volume, unbalanced selection of herbal prescriptions, limited mechanistic depth, inconsistent disease models and experimental designs, lack of standardized diaphragmatic function assessment, and weak clinical validation. Future research should strengthen the integration of TCM and modern medicine, identify additional therapeutic targets, deepen mechanistic research, and establish unified and standardized experimental systems to advance the theoretical foundation and clinical application of TCM in the prevention and treatment of COPD-related diaphragmatic dysfunction.
5.Mechanism of Diaphragmatic Dysfunction in Chronic Obstructive Pulmonary Disease and Treatment with Traditional Chinese Medicine: A Review
Yuanyuan YING ; Xiaoqing ZHOU ; Kaiwen NI ; Zhen WANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(8):285-296
Chronic obstructive pulmonary disease(COPD) is a common chronic respiratory disorder frequently accompanied by diaphragmatic dysfunction during its course, which significantly increases respiratory burden and impairs quality of life. As the primary inspiratory muscle, the diaphragm is prone to fatigue, atrophy, inflammation, and fibrosis during the long-term progression of COPD. Its pathological mechanisms involve multiple pathways such as inflammatory responses, oxidative stress, mitochondrial dysfunction, apoptosis, ion channel abnormalities, epigenetic regulation, autophagy disorder, and protein metabolism imbalance. In recent years, traditional Chinese medicine(TCM) has demonstrated multi-targeted and systemic regulatory advantages in improving diaphragmatic function in COPD. However, related studies remain fragmented, and integrated mechanistic understanding is lacking. This paper focuses on the mechanism-target-TCM intervention framework, systematically summarizing the molecular mechanisms of diaphragmatic dysfunction, while incorporating the TCM theory of Zongqi(ancestral Qi). It highlights the therapeutic effects of Chinese herbal formulas, single herbs, and active components in modulating inflammation, oxidative stress, mitochondrial function, ion channels, epigenetic processes, autophagy, and protein homeostasis. Additionally, the review outlines existing challenges, including insufficient study volume, unbalanced selection of herbal prescriptions, limited mechanistic depth, inconsistent disease models and experimental designs, lack of standardized diaphragmatic function assessment, and weak clinical validation. Future research should strengthen the integration of TCM and modern medicine, identify additional therapeutic targets, deepen mechanistic research, and establish unified and standardized experimental systems to advance the theoretical foundation and clinical application of TCM in the prevention and treatment of COPD-related diaphragmatic dysfunction.
6.Application Analysis of Animal Models of Diarrhea-predominant Irritable Bowel Syndrome Based on Data Mining
Fangli LUO ; Luqiang SUN ; Yujun HOU ; Siqi WANG ; Ying LI ; Siyuan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(2):219-226
ObjectiveBased on literature data mining, this study explores the modeling elements of diarrhea-predominant irritable bowel syndrome (IBS-D) animal models in China and abroad, providing references and suggestions for improving modeling methods and evaluation indicators. MethodsRelevant literature on IBS-D animal experiments from 2014 to 2024 was retrieved through computer searches in databases such as China National Knowledge Infrastructure (CNKI), Wanfang Data, VIP, Chinese Medical Journals Full-text Database, and PubMed. Information on experimental animal species, gender, body weight, modeling methods, modeling periods, intervention controls, modeling standards, and detection indicators was organized. Microsoft Excel 2021 software was used to establish a database and perform statistical analysis to examine the characteristics of IBS-D animal models. ResultsA total of 398 articles that met the inclusion criteria were reviewed. The IBS-D animal models were predominantly established using SD rats, Wistar rats, and C57BL/6 mice. Male animals were more commonly used, with rats typically aged 6-8 weeks and mice aged 4-6 weeks. In terms of interventions, piverium bromide was the main Western medicine, Tongxieyaofang was the primary Chinese medicine, and electroacupuncture was the primary acupuncture method. Among the modeling methods, the multi-factor combined composite modeling approach was the most common. Modeling periods were mainly concentrated between 1-14 days and 15-30 days. The success criteria for modeling were mainly evaluated based on the animal's general condition, fecal appearance, visceral sensitivity, gastrointestinal motility, behavior, and pathology. Detection indicators included apparent indexes, pathological markers, biochemical indicators, oxidative stress, brain-gut peptides, neurotransmitters, inflammatory factors, immune function, intestinal permeability, autophagy, apoptosis, proteins related to relevant signaling pathways, intestinal microbiota and its metabolites, etc. ConclusionThere are various methods for establishing IBS-D animal models, but no unified and universally accepted method has been established. The operation of the same modeling methods and the evaluation standards of the models vary across studies. Based on the results of data mining, the authors suggest that the multi-factor combined composite modeling approach most closely reflects the pathophysiological processes of IBS-D, better simulating the complex clinical symptoms of IBS-D patients, such as abdominal pain and diarrhea, and has a high degree of clinical relevance. This method is relatively recommended. While animal models in general align with Western medicine standards, models incorporating traditional Chinese medicine (TCM) syndromes are relatively few. Therefore, one of the future directions for research is to establish IBS-D animal models that meet the combined clinical disease and syndrome requirements of both Western and Chinese medicine.
7.Plasma miRNA testing in the differential diagnosis of very early-stage hepatocellular carcinoma: a multicenter real-world study
Jie HU ; Ying XU ; Ao HUANG ; Lei YU ; Zheng WANG ; Xiaoying WANG ; Xinrong YANG ; Zhenbin DING ; Qinghai YE ; Yinghong SHI ; Shuangjian QIU ; Huichuan SUN ; Qiang GAO ; Jia FAN ; Jian ZHOU
Chinese Journal of Clinical Medicine 2025;32(3):350-354
Objective To explore the application of plasma 7 microRNA (miR7) testing in the differential diagnosis of very early-stage hepatocellular carcinoma (HCC). Methods This study is a multicenter real-world study. Patients with single hepatic lesion (maximum diameter≤2 cm) who underwent plasma miR7 testing at Zhongshan Hospital, Fudan University, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Anhui Provincial Hospital, and Peking University People’s Hospital between January 2019 and December 2024 were retrospectively enrolled. Patients were divided into very early-stage HCC group and non-HCC group, and the clinical pathological characteristics of the two groups were compared. The value of plasma miR7 levels, alpha-fetoprotein (AFP), and des-gamma-carboxy prothrombin (DCP) in the differential diagnosis of very early-stage HCC was evaluated using receiver operating characteristic (ROC) curves and area under the curve (AUC). In patients with both negative AFP and DCP (AFP<20 ng/mL, DCP<40 mAU/mL), the diagnostic value of plasma miR7 for very early-stage HCC was analyzed. Results A total of 64 528 patients from 4 hospitals underwent miR7 testing, and 1 682 were finally included, of which 1 073 were diagnosed with very early-stage HCC and 609 were diagnosed with non-HCC. The positive rate of miR7 in HCC patients was significantly higher than that in non-HCC patients (67.9% vs 24.3%, P<0.001). ROC curves showed that the AUCs for miR7, AFP, and DCP in distinguishing HCC patients from the non-HCC individuals were 0.718, 0.682, and 0.642, respectively. The sensitivities were 67.85%, 43.71%, and 44.45%, and the specificities were 75.70%, 92.78%, and 83.91%, respectively. The pairwise comparison of AUCs showed that the diagnostic efficacy of plasma miR7 detection was significantly better than that of AFP or DCP (P<0.05). Although its specificity was slightly lower than AFP and DCP, the sensitivity was significantly higher. Among patients negative for both AFP and DCP, miR7 maintained an AUC of 0.728 for diagnosing very early-stage HCC, with 67.82% sensitivity and 77.73% specificity. Conclusions Plasma miR7 testing is a potential molecular marker with high sensitivity and specificity for the differential diagnosis of small hepatic nodules. In patients with very early-stage HCC lacking effective molecular markers (negative for both AFP and DCP), miR7 can serve as a novel and effective molecular marker to assist diagnosis.
8.Downregulation of LINC00638 contributes to the pathogenesis of rheumatoid arthritis-associated interstitial lung disease via inhibiting the Nrf2/ARE signaling pathway
Zhuojun LIAO ; Naiwang TANG ; Jiahui CHEN ; Xueying SUN ; Jiamin LU ; Qin WU ; Ronghuan YU ; Ying ZHOU
Chinese Journal of Clinical Medicine 2025;32(3):421-431
Objective To identify long non-coding RNA (lncRNA) associated with rheumatoid arthritis-associated interstitial lung disease (RA-ILD) and investigate their mechanisms. Methods Peripheral blood samples were collected from RA-ILD patients (n=3), RA patients without lung involvement (n=3), and healthy controls (n=3). Next-generation sequencing was performed to screen differentially expressed lncRNA. A human fibrotic lung cell model was established by inducing the MRC-5 cell line with transforming growth factor-β (TGF-β). Following siRNA-mediated knockdown of target genes, changes in inflammatory and oxidative stress-related genes were analyzed via real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Western blotting and dual-luciferase reporter (DLR) assays were used to validate protein expression, ubiquitination levels, and nuclear translocation of oxidative stress regulators, and antioxidant response element (ARE) transcriptional activity. Rescue experiments were conducted to confirm the role of target lncRNA in oxidative stress and inflammation in fibrotic lung cells. Results High-throughput sequencing revealed significant downregulation of LINC00638 in RA-ILD patients. Knockdown of LINC00638 markedly reduced transcriptional levels of interleukin (IL)-4, nuclear factor erythroid-2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and superoxide dismutase 2 (SOD2), while increasing IL-6, IL-1β, interferon-γ (IFN-γ), and reactive oxygen species (ROS) levels. Furthermore, LINC00638 knockdown decreased Nrf2 protein expression, increased its ubiquitination, reduced nuclear translocation, and suppressed ARE transcriptional activity. In MRC-5 cells, LINC00638 knockdown combined with N-acetylcysteine treatment restored Nrf2 and HO-1 levels while reducing IL-6 expression. Conclusions LINC00638 suppresses inflammatory responses in RA-ILD by activating the Nrf2/ARE antioxidant signaling pathway, suggesting its potential as a therapeutic target for diagnosis and treatment.
9.Optimization of Processing Technology of Honey Bran-fried Rosae Laevigatae Fructus and Analysis of Its Mechanism in Treatment of Ulcerative Colitis
Bin LIU ; Lingyun ZHONG ; Hongbing LUO ; Qi DENG ; Fuyu XU ; Simin ZHONG ; Ying ZHOU ; Xide YE ; Feipeng GONG ; Yuncheng GU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):216-224
ObjectiveTo optimize the processing technology of honey bran-fried Rosae Laevigatae Fructus(h-RLF), formulate relevant quality standards, and explore its improving effect and mechanism on mice with ulcerative colitis(UC) induced by dextran sodium sulfate(DSS). MethodsTaking the content of polysaccharides and water-soluble extract as the indexes, L9(34) orthogonal test was used to optimize parameters of the amount of honey bran, frying time and frying temperature. The quality of 15 batches of h-RLF decoction pieces was evaluated according to the optimized process, and the inspection limit standard was preliminarily drawn up. Eighty SPF male Kunming mice were randomly divided into 8 groups, including the blank group, model group, mesalazine group(0.13 g·kg-1), RLF group(3.77 g·kg-1), bran-fried RLF group(3.77 g·kg-1), h-RLF low, medium and high dose groups(1.89, 3.77, 7.54 g·kg-1), with 10 mice in each group. The mice in the blank group were free to drink pure water, and the other groups were free to drink 3% DSS solution for 7 days to prepare UC mouse model. Each treatment group was given corresponding drugs by intragastric administration, and the blank and model groups were given equal volume of normal saline. The body weight of mice was recorded daily and the disease activity index(DAI) was calculated. After the administration, the colon tissues of mice were collected to observe the pathological changes by hematoxylin-eosin(HE) staining. The levels of tumor necrosis factor(TNF)-α, interleukin(IL)-1β, IL-6 and IL-10 in the colon of mice were detected by enzyme-linked immunosorbent assay(ELISA). Western blot was used to detect the expression levels of phosphorylation nuclear transcription factor-κB p65(p-NF-κB p65), Toll-like receptor 4(TLR4), p-p38 mitogen-activated protein kinase(p-p38 MAPK), p-extracellular signal-regulated kinase(p-ERK) and p-c-Jun N-terminal kinase(p-JNK) proteins in colon tissues. ResultsThe optimum processing technology of h-RLF was 20 g honey bran per 100 g RLF, and stir-frying at 200 ℃ for 8 min. The limit standard under the examination of h-RLF was preliminarily formulated as follows:the polysaccharide content should not be less than 25% based on anhydrous glucose(C6H12O6), the content of water-soluble extract should not be less than 38%, the moisture content should not be more than 12.0%, the total ash content should not be more than 5.0%, and the acid-insoluble ash content should not be more than 1.0%. The cluster heat map analysis showed that the quality of RLF from Huanggang, Hubei province was better. Animal experiments showed that compared with the blank group, the DAI score of the model group was significantly increased, the levels of TNF-α, IL-1β and IL-6 in the colon tissue were significantly increased, the IL-10 level was significantly decreased, the colonic mucosa was seriously damaged, accompanied by a large number of inflammatory cell infiltration, tissue congestion and a significant reduction in glands, and the expression levels of p-NF-κB p65, TLR4, p-p38 MAPK, p-ERK and p-JNK proteins were significantly increased(P<0.01). Compared with the model group, each administration group could alleviate the symptoms of colonic ulcer, the structure of colonic crypt was basically intact, and the glands were arranged in an orderly manner. Among them, the high-dose group of h-RLF had a better effect, which could significantly reduce the DAI score and the levels of TNF-α, IL-1β and IL-6 in colon tissue(P<0.01), and significantly increase the level of IL-10(P<0.01), alleviate the colonic mucosal injury, and effectively inhibit the expression levels of p-NF-κB p65, TLR4, p-p38 MAPK, p-ERK and p-JNK proteins(P<0.01). ConclusionThe key parameters of the processing technology of h-RLF are determined, and the optimized technology is stable and feasible. The established quality standard is simple and reliable, and can be used for the quality control. h-RLF can effectively alleviate DSS-induced UC, and its mechanism may be related to inhibiting the activation of NF-κB/TLR4/MAPK pathway.
10.Protective effect of Sini Decoction in attenuating cryopreservation-induced injury of rats' sciatic nerves based on apoptosis and oxidative stress.
Kang YANG ; Jun LIU ; Lin-Lan ZHOU ; Yun-Xiao LIU ; Chun-Lin DU ; Xiao-Zhi MEI ; Ying-Ru HUANG
China Journal of Chinese Materia Medica 2025;50(5):1351-1362
Cryopreservation is the primary technique for in vitro preservation of allogeneic tissue. However, its success is often hindered by factors such as low temperature, ischemia, and hypoxia. This study investigated the potential of Sini Decoction, known for its antioxidant and anti-apoptotic properties, to reduce cryopreservation-induced injury in rats' sciatic nerves. Sini Decoction was prepared according to the Chinese Pharmacopoeia, and its cytotoxicity on Rsc96 cells was assessed by using the CCK-8 method. Sini Decoction at concentrations of 4, 8, and 16 mg·mL~(-1), termed as low-(SL), medium-(SM), and high-(SH) doses group, was used for cryopreservation of rats' sciatic nerves. A normal control(NC) group and a fresh nerve control(fresh) group were set. Flow cytometry and TUNEL staining were used to detect the apoptosis of neural tissue cells after cryopreservation. Western blot was used to detect the expression of apoptosis-related proteins(Bcl-2, Bax, caspase-3, and caspase-8) and nerve regeneration proteins(NGF and BDNF) in vitro after cryopreservation. Oxidative damage of neural tissue after cryopreservation was evaluated by measuring levels of GSH, SOD, MDA, ROS, and ATP. Cryopreserved nerves were then used for allogeneic transplantation. One week after transplantation, CD4~+ and CD8~+ fluorescent double staining assessed inflammatory cell invasion in the transplanted nerve segment, and ELISA evaluated the expression of serum inflammatory factors(IL-1, IFN-γ, and TNF-α) in recipients. Twenty weeks after transplantation, electrophysiology and NF200 neurofilament staining were used to evaluate nerve regeneration. RESULTS:: showed that Sini Decoction at concentrations of below 32 mg·mL~(-1) exhibited no cytotoxicity to Rsc96 cells. During in vitro nerve cryopreservation, Sini Decoction significantly reduced cell apoptosis, ROS, and MDA production compared to the NC group. In the SH group, the protein expression of NGF and BDNF in vitro, as well as ATP, SOD, and GSH production, were significantly increased. In the rejection reaction one week after transplantation, compared to the fresh nerve transplantation group, the SL and SM groups showed reduced CD4~+ and CD8~+ T cell invasion in the transplanted nerve segment and down-regulated IL-1, IFN-γ, and TNF-α expression in recipient serum. Twenty weeks after transplantation, the electrophysiological test results of CMAP, NCV, and NF200 neurofilament protein fluorescent staining in the SM and SH groups were superior to those in the NC and fresh groups. These findings indicate that Sini Decoction offers protective benefits in the cryopreservation of rats' sciatic nerves and holds significant potential for the in vitro preservation of tissue and organs.
Animals
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Apoptosis/drug effects*
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Rats
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Oxidative Stress/drug effects*
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Sciatic Nerve/cytology*
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Cryopreservation
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Drugs, Chinese Herbal/administration & dosage*
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Male
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Rats, Sprague-Dawley
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Protective Agents/pharmacology*

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