1.Comparison of Efficacy and Mechanism in Warming Yang and Dispersing Cold of Aconiti Radix Lateralis Praeparata Processed by ZHANG Zhongjing's Method and Pharmacopoeia Method
Mingjie JIAO ; Qian CHEN ; Shuyu YAN ; Yiyan SONG ; Jia ZHANG ; Fei LI
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(2):207-217
ObjectiveTo investigate the therapeutic effects and mechanism of decoctions from four kinds of processed products of Aconiti Radix Lateralis Praeparata(ARLP) in deficiency-cold syndrome. MethodsA total of 36 SD rats were randomly divided into the control group, model group, Shengfupian(SFP) group, Paofuzi(PFZ) group, Heishunpian(HSP) group and Paofupian(PFP) group with 6 rats in each group. Except for the control group, rats in other groups were administered hydrocortisone sodium succinate via intramuscular injection to induce a cold deficiency syndrome model. After 14 consecutive days, each ARLP decoction pieces was administered via continuous gastric lavage at a dose of 12 g·kg-1·d-1 for 7 d, while the control and model groups received an equivalent volume of physiological saline. After the end of administration, body weight, spleen weight and thymus weight were measured for calculating the spleen and thymus indexes. Hematoxylin-eosin(HE) staining was used to observe the pathological morphology of adrenal tissue. The fully automatic biochemistry analyzer was used to measure the total cholesterol(TC), triglyceride(TG), lactic dehydrogenase(LDH) and lactate(LAC) levels in serum. Enzyme-linked immunosorbent assay(ELISA) was employed to measure the contents of 17-hydroxycorticosteroid(17-OHCS), cortisol(CORT), triiodothyronine(T3), thyroxine(T4), thyrotropin(TSH), immunoglobulin(Ig) M, IgG, cyclic adenosine monophosphate(cAMP) and cyclic guanosine monophosphate(cGMP). Western blot was used to measure the protein expression levels of protein kinase A(PKA), cAMP response element-binding protein(CREB), silent information regulator 1(Sirt1) and peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α). And high performance liquid chromatography(HPLC) was used to determine the content of major alkaloids, followed by Pearson correlation analysis with pharmacodynamic indicators. ResultsAfter modeling, compare with the control group, the model rats exhibited symptoms such as lethargy and loose stools, mild abnormalities were observed in adrenal tissue structure, and both spleen and thymus indices were significantly reduced(P<0.01). Thyroid, adrenal and immune system functions were suppressed, with decreased serum cAMP level and significantly elevated cGMP level(P<0.01). Compared with the model group, the adrenal injury by hydrocortisone sodium succinate were repaired and the spleen index were increased significantly in all four ARLP groups(P<0.05, P<0.01). The thymus index in SFP and PFZ groups were increased significantly(P<0.05). The contents of T3, TSH, 17-OHCS and CORT were increased significantly in SFP and PFZ groups(P<0.05). In addition, the content of IgG in SFP, PFZ and PFP groups were increased significantly(P<0.01), while the content of IgM in PFZ and HSP groups were also increased significantly(P<0.05). Regarding the cyclic nucleotide system, PFZ significantly elevated cAMP level while reducing cGMP level(P<0.05), exhibiting the most pronounced effect among the four decoction pieces. For energy metabolism indicators, PFZ significantly improved abnormal markers including TC, TG, LDH, and LAC(P<0.05). HSP showed marked improvement effects on TG, LDH, and LAC(P<0.05). Both PFZ and SFP significantly elevated the expression levels of PKA, CREB, Sirt1, and PGC-1α proteins(P<0.01). Additionally, the diester alkaloids in ARLP showed a strong positive correlation with TG, IgG, and CORT, a strong negative correlation with LAC, a moderate positive correlation with T4, and moderate negative correlations with cAMP and spleen index. Monomeric alkaloids showed strong positive correlations with TG and IgG, strong negative correlations with LAC, moderate positive correlations with CORT and T4, and moderate negative correlations with cAMP and spleen index. However, the content of water-soluble alkaloids showed strong positive correlations with TC, LDH, 17-OHCS, T3, TSH, and thymus index, moderate positive correlations with cAMP, CORT, T4, and spleen index, and moderate negative correlation with cGMP. ConclusionAmong different processed ARLP decoction pieces, PFZ processed according to ZHANG Zhongjing's method exhibits the most potent warming and cold-dispelling effects. Its pharmacological actions are mediated through regulating the thyroid, adrenal, immune, cyclic nucleotide systems, and material-energy metabolism pathways. Among these, water-soluble alkaloids show strong or moderate correlations with more indicators of deficiency-cold syndrome and exhibit the highest content in PFZ. Therefore, PFZ processed according to ZHANG Zhongjing's method may exert its warming and cold-dispelling effects through water-soluble alkaloids.
2.Influence of pterygium thickness and area on corneal refractive status
Xiaodong CHENG ; Jie WANG ; Song GAO ; Yanhong LU ; Yanbo MA ; Xinming CUI ; Xihui CHEN
International Eye Science 2026;26(1):152-156
AIM: To investigate the influence of pterygium thickness and area on corneal refractive status.METHODS: Prospective longitudinal study. A total of 60 cases(60 eyes)of pterygium patients admitted to our hospital from January 2024 to September 2024 were randomly selected. All patients underwent pterygium excision combined with pedicle conjunctival flap transplantation for treatment. Optical coherence tomography(OCT)was used to measure the preoperative thickness of patient's pterygium, and a digital slit lamp microscope was used to measure the area of pterygium. The corneal refractive status(degree of corneal astigmatism and average curvature)and changes in uncorrected visual acuity of patients before surgery, 1 d, 1, and 3 mo after surgery were compared. The relationship between preoperative thickness and area of pterygium in patients and corneal refractive status indicators at different postoperative time points were analyzed, and Logistic regression was used to analyze the impact of pterygium thickness and area on postoperative visual improvement in patients.RESULTS: All patients completed follow-up after surgery for 3 mo. At 3 mo after surgery, visual acuity improved in 21 eyes(35%). The results of bivariate Pearson correlation analysis showed that the thickness and area of pterygium positively correlated with the degree of corneal astigmatism and uncorrected visual acuity before surgery and 1 d, 1, and 3 mo after surgery(all P<0.05), and negatively correlated with the average corneal curvature before surgery and 1 d, 1, and 3 mo after surgery(all P<0.05). Logistic regression analysis showed that the thickness and area of pterygium before surgery, high degree of corneal astigmatism, and low uncorrected visual acuity(large LogMAR value)were all risk factors for poor postoperative visual improvement in patients(OR>1, P<0.05). The large average corneal curvature before surgery was a protective factor for poor postoperative visual improvement in patients(OR<1, P<0.05).CONCLUSION: The increase in thickness and area of pterygium can, to some extent, improve corneal astigmatism, reduce the average curvature of the cornea, and affect postoperative visual recovery.
3.Expert consensus on neoadjuvant PD-1 inhibitors for locally advanced oral squamous cell carcinoma (2026)
LI Jinsong ; LIAO Guiqing ; LI Longjiang ; ZHANG Chenping ; SHANG Chenping ; ZHANG Jie ; ZHONG Laiping ; LIU Bing ; CHEN Gang ; WEI Jianhua ; JI Tong ; LI Chunjie ; LIN Lisong ; REN Guoxin ; LI Yi ; SHANG Wei ; HAN Bing ; JIANG Canhua ; ZHANG Sheng ; SONG Ming ; LIU Xuekui ; WANG Anxun ; LIU Shuguang ; CHEN Zhanhong ; WANG Youyuan ; LIN Zhaoyu ; LI Haigang ; DUAN Xiaohui ; YE Ling ; ZHENG Jun ; WANG Jun ; LV Xiaozhi ; ZHU Lijun ; CAO Haotian
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(2):105-118
Oral squamous cell carcinoma (OSCC) is a common head and neck malignancy. Approximately 50% to 60% of patients with OSCC are diagnosed at a locally advanced stage (clinical staging III-IVa). Even with comprehensive and sequential treatment primarily based on surgery, the 5-year overall survival rate remains below 50%, and patients often suffer from postoperative functional impairments such as difficulties with speaking and swallowing. Programmed death receptor-1 (PD-1) inhibitors are increasingly used in the neoadjuvant treatment of locally advanced OSCC and have shown encouraging efficacy. However, clinical practice still faces key challenges, including the definition of indications, optimization of combination regimens, and standards for efficacy evaluation. Based on the latest research advances worldwide and the clinical experience of the expert group, this expert consensus systematically evaluates the application of PD-1 inhibitors in the neoadjuvant treatment of locally advanced OSCC, covering combination strategies, treatment cycles and surgical timing, efficacy assessment, use of biomarkers, management of special populations and immune related adverse events, principles for immunotherapy rechallenge, and function preservation strategies. After multiple rounds of panel discussion and through anonymous voting using the Delphi method, the following consensus statements have been formulated: 1) Neoadjuvant therapy with PD-1 inhibitors can be used preoperatively in patients with locally advanced OSCC. The preferred regimen is a PD-1 inhibitor combined with platinum based chemotherapy, administered for 2-3 cycles. 2) During the efficacy evaluation of neoadjuvant therapy, radiographic assessment should follow the dual criteria of Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1 and immune RECIST (iRECIST). After surgery, systematic pathological evaluation of both the primary lesion and regional lymph nodes is required. For combination chemotherapy regimens, PD-L1 expression and combined positive score need not be used as mandatory inclusion or exclusion criteria. 3) For special populations such as the elderly (≥ 70 years), individuals with stable HIV viral load, and carriers of chronic HBV/HCV, PD-1 inhibitors may be used cautiously under the guidance of a multidisciplinary team (MDT), with close monitoring for adverse events. 4) For patients with a poor response to neoadjuvant therapy, continuation of the original treatment regimen is not recommended; the subsequent treatment plan should be adjusted promptly after MDT assessment. Organ transplant recipients and patients with active autoimmune diseases are not recommended to receive neoadjuvant PD-1 inhibitor therapy due to the high risk of immune related activation. Rechallenge is generally not advised for patients who have experienced high risk immune related adverse events such as immune mediated myocarditis, neurotoxicity, or pneumonitis. 5) For patients with a good pathological response, individualized de escalation surgery and function preservation strategies can be explored. This consensus aims to promote the standardized, safe, and precise application of neoadjuvant PD-1 inhibitor strategies in the management of locally advanced OSCC patients.
4.Correlation study between ankyloglossia and mandibular development and lower anterior crowding
CHEN Hongji ; TU Chunhua ; MO Zhenfei ; SONG Juan
Journal of Prevention and Treatment for Stomatological Diseases 2026;34(2):148-155
Objective:
To study the association between ankyloglossia and sagittal mandibular development impairment as well as lower anterior dental crowding, providing a reference for clinical practice.
Methods:
This study was approved by the hospital's Medical Ethics Committee. A total of 100 patients aged 7-13 years were enrolled from January 2024 to January 2025, comprising 50 patients with ankyloglossia (case group) and 50 individuals with a healthy lingual frenulum (normal group). Clinical examination was performed to assess lingual frenulum length, Kotlow classification, and the simplified Hazelbaker assessment tool for lingual frenulum function (HATLFF) score. Cephalometric radiographs were used to measure the A-point-nasion-B-point (ANB) angle, sella-nasion-B-point (SNB) angle, and mandibular total length (condylion-gnathion [Co-Gn]). Dental cast analysis was conducted to evaluate lower anterior teeth crowding. Data were compared between the two groups. Pearson correlation analysis was used to examine the relationships between the lingual frenulum length, simplified HATLFF score, and cephalometric/dental cast parameters (ANB, SNB, Co-Gn, lower anterior crowding). The diagnostic value of ankyloglossia for mandibular development and lower anterior crowding was analyzed using receiver operating characteristic (ROC) curves.
Results:
Ankyloglossia was significantly associated with mandibular development and lower anterior crowding (P<0.05). The case group showed significantly lower values for the lingual frenulum length, simplified HATLFF score, SNB angle, and Co-Gn, while the ANB angle and lower anterior crowding index were significantly higher compared to the normal group (P<0.05). The lingual frenulum length and simplified HATLFF score were negatively correlated with the ANB angle and lower anterior crowding index, and positively correlated with the SNB angle and Co-Gn (P<0.05). ROC curve analysis indicated that the area under the curve (AUC) for the simplified HATLFF score, and ankyloglossia in predicting mandibular development deficiency and lower anterior crowding was greater than 0.700, demonstrating good diagnostic value.
Conclusion
A significant correlation exists between ankyloglossia and both mandibular development deficiency and lower anterior crowding.
5.Association between psychological help seeking intentions and non suicidal self injury among adolescents in Wuhan
SONG Yu, ZHANG Jiaxiu, ZHOU Yang, CHEN Mo
Chinese Journal of School Health 2026;47(1):94-99
Objective:
To explore the latent profiles of psychological help seeking intentions among adolescents in Wuhan, and to investigate their association with non suicidal self injury (NSSI), in order to provide a theoretical basis for constructing an early intervention system for adolescents NSSI.
Methods:
From October to December 2022, a convenient sampling method was used to select 3 975 students from grades 7 to 12 in Wuhan. A self administered questionnaire assessed psychological help seeking intentions, followed by a post survey interview using the Ottawa Self injury Inventory to evaluate NSSI behaviors. Latent profile analysis(LPA) was used to explore the potential categories of help seeking intentions in adolescents. Multinomial Logistic regression was used to analyze factors associated with the latent profiles of help seeking intentions and multiple Logistic regression was used to analyze the association between latent profiles of help seeking intentions and NSSI.
Results:
Based on latent profile analysis, four latent profiles of help seeking intentions in adolescents were identified, namely overall low, moderate, and high help seeking group, as well as family/friendfocused help seeking group, accouting for 14.1%, 20.9%, 43.1% and 21.8%, respectively. Multinomial Logistic regression analysis showed that compared to senior high school students, junior high school students were more willing to seek help from family and friends ( OR =1.56); compared to males, females were more likely to exhibit moderate help seeking intentions( OR =1.37); students with extroverted or balanced personalities were more likely to exhibit high level help seeking intentions( OR =1.50, 1.49); students with average or good family economic status, better parental relationships, and adequate mental health knowledge were more likely to exhibit moderate and high level help seeking intentions( OR =1.59, 2.02; 1.80, 2.64; 1.44, 1.55; 1.34, 1.58) (all P <0.05). Students with moderate or severe family dysfunction were less likely to seek help from family and friends or to exhibit moderate and high level help seeking intentions( OR =0.51, 0.60, 0.25; 0.22, 0.27, 0.06, all P <0.01). Students whose parents exhibited stigma towards mental illness were less likely to show high level help seeking intentions( OR= 0.78 , P <0.05). Multivariable Logistic regression analysis results indicated that compared to the overall low help seeking intentions group, reduced risk of NSSI was observed among students in the overall moderate and overall high groups, as well as in the family/friend focused help seeking group( OR =0.73, 0.60, 0.70, all P <0.05).
Conclusions
Active psychological help seeking intentions can reduce NSSI behaviors in adolescents. Interventions should focus on improving family support environment to enhance adolescents intentions to seek psychological help.
6.Mechanisms of Antidepressant Effect of Zhizi Houpotang and Its Herbal Pairs Based on NLRP3/GSDMD Signaling Pathway
Chang CHEN ; Ziwen GUO ; Tingyu SONG ; Yan WANG ; Baomei XIA ; Weiwei TAO
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):72-80
ObjectiveTaking classical herbal pair compatibility research as the entry point, this study aimed to deeply investigate the material basis and compatibility rules underlying the antidepressant effects of the traditional Chinese medicine (TCM) formula Zhizi Houpotang, and to elucidate its antidepressant mechanism, with a particular focus on its regulation of neuroinflammatory responses mediated by the NOD-like receptor protein 3 (NLRP3)/gasdermin D (GSDMD) signaling pathway and the consequent improvement of neuronal synaptic plasticity. MethodsC57BL/6J mice were randomly divided into a blank control group, a chronic unpredictable mild stress (CUMS) depression model group, a Zhizi Houpotang full-formula group (6 g·kg-1·d-1), a Magnoliae Officinalis Cortex (MOC)-Aurantii Fructus Immaturus (AFI) herbal pair group (4.2 g·kg-1·d-1), a Gardeniae Fructus (GF)-MOC herbal pair group (4.2 g·kg-1·d-1), a GF-AFI herbal pair group (3.6 g·kg-1·d-1), and a positive drug group (fluoxetine, 12 mg·kg-1·d-1). Depressive-like behaviors in mice were evaluated using behavioral tests. Immunofluorescence staining was used to label and quantify the expression of the microglial marker ionized calcium-binding adaptor molecule 1 (Ibal) and the purinergic receptor P2X ligand-gated ion channel 7 (P2RX7) in the prefrontal cortex (PFC). Enzyme-linked immunosorbent assay (ELISA) was applied to detect the levels of inflammatory cytokines interleukin-1β (IL-1β) and interleukin-18 (IL-18) in serum and PFC tissues. Western blot was employed to determine the expression of pannexin 1 (Panx1), P2RX7, NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), Caspase-1, GSDMD, postsynaptic density protein 95 (PSD95), and the presynaptic protein Synapsin 1 in PFC tissues. Golgi staining was used to assess dendritic spine density of neurons in the PFC. ResultsCompared with the blank control group, the depression model group exhibited significant depressive-like behaviors. In addition, the immunofluorescence areas of Ibal and P2RX7 in the PFC were significantly increased (P<0.01), the levels of IL-1β and IL-18 in serum and the PFC were significantly elevated (P<0.01), and the protein expression levels of Panx1, P2RX7, NLRP3, ASC, Caspase-1, and GSDMD in the PFC were significantly upregulated (P<0.01). In contrast, the protein expression levels of PSD95 and Synapsin 1 were significantly downregulated (P<0.01), and neuronal dendritic spine density was significantly reduced (P<0.01). Compared with the model group, the Zhizi Houpotang full-formula group and the GF-MOC herbal pair group showed significant improvement in all the above indicators (P<0.01). The GF-AFI herbal pair group improved all the above indicators except P2RX7, Caspase-1, GSDMD, and PSD95 (P<0.05, P<0.01). In contrast, the MOC-AFI herbal pair group showed no statistically significant improvement in any of the above indicators compared with the model group. ConclusionZhizi Houpotang and its key herbal pair, GF-MOC, can effectively ameliorate CUMS-induced depressive-like behaviors in mice. Its core antidepressant mechanism may involve inhibition of P2RX7/Panx1 signaling, thereby blocking the NLRP3/GSDMD-mediated pyroptosis pathway and significantly reducing the release of inflammatory cytokines IL-1β and IL-18. Simultaneously, it upregulates the expression of synapse-related proteins PSD95 and Synapsin 1 and increases dendritic spine density, promoting the recovery of synaptic plasticity. These results suggest that GF plays a key role in the antidepressant effects of this formula, and that the compatibility of GF with MOC may represent the principal herbal pair combination responsible for its core therapeutic action.
7.Research progress on the tumor microenvironment of liver cancer
Tianhao SONG ; Weiheng XU ; Yan WANG ; Li CHEN
Journal of Pharmaceutical Practice and Service 2026;44(2):65-70
Liver cancer, one of the most common primary malignancies in humans, is a malignant tumor characterized by multifactorial induction, polygenic involvement, and intricate molecular mechanisms. This disease is characterized by its treatment challenges and poor prognosis, which are closely related to its unique tumor microenvironment composition. The tumor microenvironment of liver cancer is a dynamic ecosystem composed of heterogeneous cellular populations, soluble cytokines, and remodeled extracellular matrix. In recent years, significant progress has been made in the study of the tumor microenvironment of liver cancer, revealed an important role in the occurrence, development, and treatment of liver cancer. The key regulatory elements of the tumor microenvironment in liver cancer were systematically summarized, such as activation of hepatic stellate cells, dysfunction of immune cells, abnormalities of platelet, and remodeling of the extracellular matrix, which provided theoretical foundations for prevention and treatment strategies against liver cancer.
8.Analysis of the changes in intestinal microbiota of patients with moderate to severe acne based on 16S rRNA high-throughput sequencing technology
Shichao JIANG ; Xiaomeng WANG ; Zheng CHEN ; Song QIAO ; Fan YANG ; Birong GUO
Acta Universitatis Medicinalis Anhui 2026;61(1):98-103
ObjectiveTo explore the relationship between acne vulgaris and gut microbiota. MethodsA total of 29 clinical cases diagnosed with moderate-to-severe acne vulgaris and 26 healthy individuals as control subjects were recruited. Fecal specimens were collected from all participants, and further analysis of gut microbial communities was performed by leveraging high-throughput sequencing techniques that target the hypervariable regions of 16S rRNA genes. ResultsAssociations between acne vulgaris and alterations in gut microbiota were identified. At the phylum level, the relative abundance of Bacteroidota exhibited a statistically significant elevation in the acne vulgaris cohort when compared with the healthy control group (P<0.01), while Cyanobacteria was significantly lower in the acne group (P<0.01). At the genus level, the top five different bacterial taxa in both groups were Bacteroides, Escherichia⁃Shigella, Klebsiella, Roseburia, and Parabacteroides. Among them, Bacteroides, Roseburia, and Parabacteroides were more abundant in acne patients. Linear discriminant analysis identified five biomarkers all belonging to the Bacteroidota phylum in the acne and control groups. These biomarkers belong to the phylum Bacteroidetes. ConclusionThere are significant differences in the composition of intestinal microbiota between acne patients and healthy people. Changes in the richness of specific bacterial genera may become new targets for the diagnosis and treatment of acne.
9.Pharmacodynamic Substances and Mechanisms of Xinglou Chengqi Tang in Treating Post-stroke Complications: A Review
Yujin ZHANG ; Xiangzhuo LIU ; Zhouyang CHEN ; Zihao SONG ; Xinyi LIU ; Yizhi YAN ; Chaoya LI ; Yingyan FANG ; Shasha YANG ; Xueqin CHENG ; Zhou XIE ; Sijie TAN ; Peng ZENG ; Yue ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(1):327-337
Stroke is the leading cause of death and disability among adults in China, and its common complications include digestive system abnormalities, cognitive impairment, depression, stroke-associated pneumonia, and hemiplegia. The combination of traditional Chinese and Western medicine has great potential in treating post-stroke complications. Xinglou Chengqitang (XLCQT) is a representative prescription of alleviating the disease in the upper part by treating the lower part. It has definite therapeutic effect and high safety. Clinically, XLCQT is often used to treat stroke and its complications. However, the quantity and quality of clinical trials of XLCQT in treating post-stroke complications need to be improved. Additionally, since the basic research is weak, the material basis and multi-target mechanism for the efficacy of this prescription are unknown. This article reviews XLCQT in terms of the pharmacodynamic basis, medicinal properties, safety evaluation, and progress in clinical research and mechanisms in treating post-stroke complications. This article summarizes 22 key active ingredients of XLCQT in treating acute stroke complicated with syndrome of phlegm heat and fu-organ excess. Among these key active ingredients, resveratrol, kaempferol, luteolin, chrysoeriol, apigenin, (+)-catechin, and adenosine have good pharmacokinetic properties and high bioavailability. The mechanisms of XLCQT in treating post-stroke complications are complex, including inflammatory response, brain-gut axis, hypothalamic-pituitary-adrenal (HPA) axis, intestinal flora, neurotrophic factors, autophagy, oxidative stress, and free radical damage. This review helps to deeply understand the pharmacodynamic basis and mechanisms of XLCQT in treating post-stroke complications and provides a theoretical basis for the clinical application of XLCQT against post-stroke complications and the development of drugs.
10.Effect of Shixiaosan on Neurological Function and Ferroptosis in Rats with Cerebral Ischemia-reperfusion Injury Based on Nrf2/SLC7A11/GPX4 Signaling Pathway
Ying WEI ; Lixia WANG ; Junjun YIN ; Xiaohong CHEN ; Lijuan SONG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(7):22-31
ObjectiveTo investigate whether Shixiaosan can improve neurological function and inhibit ferroptosis in rats with cerebral ischemia-reperfusion injury (CIRI) by regulating the nuclear factor E2-related factor 2 (Nrf2)/solute carrier family 7 member 11 (SLC7A11)/glutathione peroxidase 4 (GPX4) pathway. MethodsA rat model of CIRI was established using the intraluminal filament method. Briefly, cervical blood vessels were separated, branches of the external carotid artery were ligated, and the common carotid artery and internal carotid artery were clamped. A nylon filament was inserted through the opening of the external carotid artery to the origin of the middle cerebral artery to block blood flow and induce cerebral ischemia. After 60-120 min of ischemia, the filament was withdrawn to restore blood flow, and the external carotid artery incision was ligated. The rats were divided into a CIRI group, a Shixiaosan low-dose (-L) group (intragastric administration of 1.26 g·kg-1 Shixiaosan), a Shixiaosan high-dose (-H) group (intragastric administration of 2.52 g·kg-1 Shixiaosan), a donepezil hydrochloride tablet (DON) group (intragastric administration of 0.45 mg·kg-1 DON), and a Shixiaosan -H + Nrf2 inhibitor (ML385) group (intragastric administration of 2.52 g·kg-1 Shixiaosan combined with intraperitoneal injection of 30 mg·kg-1 ML385). An additional 12 rats underwent cervical artery separation followed by incision suturing and served as the control group. Equal volumes of double-distilled water were administered to the CIRI and control groups. Neurological function impairment was assessed using the modified Garcia JH score. Magnetic resonance imaging was used to determine the cerebral infarct volume ratio. Hematoxylin-eosin (HE) staining and Prussian blue staining were performed to observe neuronal injury and iron accumulation in the ischemic penumbra, respectively. Transmission electron microscopy was used to examine the ultrastructure of neuronal mitochondria in the ischemic penumbra. Commercial kits were used to measure ferrous iron (Fe2+), malondialdehyde (MDA), reduced glutathione (GSH) content, and reactive oxygen species (ROS) activity in the ischemic penumbra. The BODIPY (581/591) C11 fluorescent probe was used to detect intracellular lipid peroxidation levels. Western blot was performed to detect protein expression levels of Nrf2, SLC7A11, GPX4, transferrin receptor 1 (TFRC), ferritin heavy chain (FHC), and ferritin light chain (FLC) in the ischemic penumbra. ResultsCompared with the control group, the CIRI group exhibited neuronal injury in the ischemic penumbra, characterized by reduced neuron numbers, nucleolar shrinkage, and interstitial edema. Marked iron accumulation was observed in the tissue. Neuronal mitochondria showed atrophy and rupture, with reduced mitochondrial cristae and increased membrane density. The cerebral infarct volume ratio, Fe2+ content, MDA content, ROS activity, and lipid peroxidation levels were increased, whereas the modified Garcia JH score, GSH content, and protein expression levels of Nrf2, SLC7A11, GPX4, FHC, and FLC were decreased, and TFRC protein expression was increased (P<0.05). Compared with the CIRI group, the Shixiaosan -L group, Shixiaosan -H group, and DON group showed attenuated neuronal injury in the ischemic penumbra, reduced iron accumulation, alleviated mitochondrial damage, decreased cerebral infarct volume ratio, Fe2+ and MDA contents, ROS activity, and lipid peroxidation levels, as well as increased modified Garcia JH scores, GSH content, and protein expression levels of Nrf2, SLC7A11, GPX4, FHC, and FLC, while TFRC protein expression was decreased (P<0.05). The magnitude of changes in all indicators was greater in the Shixiaosan -H group than in the Shixiaosan -L group (P<0.05). Compared with the Shixiaosan -H group, all measured indicators in the Shixiaosan -H + ML385 group showed opposite trends (P<0.05). ConclusionShixiaosan may inhibit ferroptosis and restore neurological function in rats with CIRI by activating the Nrf2/SLC7A11/GPX4 pathway.


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