ObjectiveTo investigate the mechanism of Xiezhuo Jiedu prescription in the treatment of ulcerative colitis （UC） by inhibiting ferroptosis and alleviating intestinal mucosal injury based on the nuclear factor E₂ related factor 2/solute carrier family 7 member/glutathione peroxidase 4 （Nrf2/SLC7A11/GPX4） signaling pathway. MethodsA total of 60 male SD rats were divided into a normal group， a model group， high- and low-dose Xiezhuo Jiedu prescription groups （26.64 and 13.32 g·kg^-1， respectively）， a ferroptosis inhibitor group （Ferrostatin-1， 0.005 g·kg^-1）， and a mesalazine group （0.27 g·kg^-1）， with 10 rats in each group. A UC rat model was established by intrarectal administration of trinitrobenzene sulfonic acid （TNBS）-ethanol. The normal group and the model group were intragastrically administered normal saline. The other groups were given intragastric administration according to the corresponding dosage for 7 d. The general condition， disease activity index （DAI） score， colon length， and mucosal injury index （CDMI） score were observed in each group. The pathological changes of colon tissue in each group were observed by hematoxylin-eosin （HE） staining. The intestinal mucosa and mitochondrial morphology in each group were observed by transmission electron microscopy. The expression levels of Occludin， Claudin-1， mucin 2 （MUC2）， and E-cadherin in intestinal tissue were detected by immunofluorescence （IF）. Enzyme-linked immunosorbent assay （ELISA） was used to detect the expression levels of serum tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-10 （IL-10） in each group， and a lactic acid assay kit or ELISA was employed to detect the expression levels of reactive oxygen species （ROS）， ferrous ions （Fe²⁺）， glutathione （GSH）， malondialdehyde （MDA）， 4-hydroxynonenal （4-HNE）， diamine oxidase （DAO）， and D-lactate （D-LA）. Real-time quantitative polymerase chain reaction （Real-time PCR） was applied to detect the mRNA expression levels of Nrf2， SLC7A11， GPX4， Occludin， Claudin-1， MUC2， and E-cadherin in each group， and Western blot was adopted to detect the protein expression levels of Nrf2， p-Nrf2， SLC7A11， and GPX4 in each group. ResultsCompared with the normal group， rats in the model group exhibited listlessness， sluggish response， and mucopurulent and bloody stools. The model group also showed significantly increased DAI score， colon length， CDMI score， and expression levels of TNF-α， IL-6， ROS， Fe²⁺， MDA， 4-HNE， DAO， and D-LA （P<0.01）. In addition， it presented significantly decreased IF values of Occludin， Claudin-1， MUC2， and E-cadherin and mRNA and protein expression levels of IL-10， GSH， Nrf2， p-Nrf2， SLC7A11， and GPX4 （P<0.01）. There were different degrees of improvement in each administration group after treatment， and the improvement was the most significant in the high-dose Xiezhuo Jiedu prescription group （P<0.01）. ConclusionXiezhuo Jiedu prescription may alleviate intestinal mucosal injury by inhibiting ferroptosis of intestinal epithelial cells via regulating the Nrf2/SLC7A11/GPX4 signaling pathway， thereby exhibiting efficacy in the treatment of UC.

ObjectiveThis study aims to explore the potential mechanism of the Xiezhuo Jiedu formula in regulating pyroptosis for the treatment of ulcerative colitis （UC） using bioinformatics and in vivo animal experiments. MethodsDifferentially expressed genes （DEGs） in colon tissues of UC patients were retrieved from the Gene Expression Omnibus （GEO） database. Pyroptosis-related genes were obtained from the GEO and GeneCards databases. The intersection of these datasets yielded pyroptosis-related DEGs （Pyro-DEGs）. Pyro-DEGs were subjected to Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis using the Metascape database. A protein-protein interaction （PPI） network was constructed using the STRING database. Least absolute shrinkage and selection operator （LASSO） prediction model and receiver operating characteristic （ROC） analysis were conducted to identify core Pyro-DEGs with diagnostic and therapeutic potential. Immune infiltration analysis of the UC datasets was performed using the deconvolution method （CIBERSORT）， along with correlation analysis with core Pyro-DEGs. Sixty male Sprague-Dawley （SD） rats were randomly divided into a control group， a model group， high-， medium-， and low-dose groups of Xiezhuo Jiedu formula （26.64， 13.32， 6.66 g·kg^-1）， and a mesalazine group （0.27 g·kg^-1）， with 10 rats in each group. UC was established by intrarectal administration of 3，5-trinitrobenzenesulfonic acid （TNBS） dissolved in ethanol. The control and model groups were given distilled water by gavage， while the treatment groups were administered the corresponding drugs for 7 consecutive days. Hematoxylin-eosin （HE） staining was used to observe the colon histopathology. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of inflammatory factors such as interleukin-1β （IL-1β）， IL-10， IL-18， and transforming growth factor-β （TGF-β）. Immunohistochemistry （IHC） and Western blot were applied to detect the expression of Caspase-1， gap junction alpha-1 protein （GJA1）， peroxisome proliferator-activated receptor gamma （PPARG）， and S100 calcium-binding protein A8 （S100A8）. Real-time quantitative polymerase chain reaction （Real-time PCR） was utilized to measure mRNA expression of Caspase-1， GJA1， PPARG， and S100A8. Western blot was performed to assess protein expression levels of Caspase-1， GJA1， PPARG， and S100A8. ResultsGEO datasets GSE87466 and GSE87473 yielded 64 Pyro-DEGs. KEGG analysis indicated that these genes were enriched in the NOD-like receptor signaling pathway， tumor necrosis factor （TNF） signaling pathway， and hypoxia-inducible factor 1 （HIF-1） signaling pathway. Four core Pyro-DEGs （Caspase-1， GJA1， PPARG， and S100A8） were identified. Immune infiltration analysis showed that expression of these genes was positively correlated with mast cells， neutrophils， M0 macrophages， M1 macrophages， and dendritic cells. Animal experimental results indicated that compared with the control group， the model group had significantly increased levels of IL-1β and IL-18， significantly decreased levels of IL-10 and TGF-β. The model group showed enhanced Caspase-1， GJA1， and S100A8 staining， and significantly increased mRNA and protein expression of Caspase-1， GJA1， and S100A8 （P<0.01）. In contrast， the expression of PPARG was reduced in the model group （P<0.01）. After treatment， all dosage groups showed varying degrees of improvement （P<0.05， P<0.01）， with the high-dose group showing the most significant improvement （P<0.01）. ConclusionCaspase-1， GJA1， PPARG， and S100A8 are core Pyro-DEGs closely associated with the pathogenesis of UC. These genes may collaborate with immune cells such as mast cells， neutrophils， and M0 macrophages to mediate disease development. The Xiezhuo Jiedu formula may regulate the expression of core Pyro-DEGs through the NOD-like receptor， TNF， and HIF-1 core signaling pathways， thereby modulating immune homeostasis in UC rats and effectively alleviating UC.

Objective This study aims to explore the layout and optimization of management disciplines in"Double First-Class"Traditional Chinese Medicine(TCM)universities,providing talent support and management guidance for the devel-opment of TCM.Methods Based on an overview of the development of management disciplines in TCM universities,six"Doub-le First-Class"TCM universities were selected as research subjects.Data were collected and compared using web retrieval and lit-erature research methods.Results Management disciplines in TCM universities still face challenges,such as weak foundational infrastructure and difficulties in interdisciplinary integration,insufficient faculty and suboptimal curriculum systems,and a single teaching model with a disconnect between theory and practice.Conclusion This paper proposes targeted suggestions and strate-gies for optimizing the layout of management disciplines in TCM universities,including strengthening support for management dis-ciplines and deepening TCM management characteristics,optimizing curriculum design and enhancing faculty resources,and in-novating teaching models to promote the deep integration of theory and practice.

Objective This study aims to explore the layout and optimization of management disciplines in"Double First-Class"Traditional Chinese Medicine(TCM)universities,providing talent support and management guidance for the devel-opment of TCM.Methods Based on an overview of the development of management disciplines in TCM universities,six"Doub-le First-Class"TCM universities were selected as research subjects.Data were collected and compared using web retrieval and lit-erature research methods.Results Management disciplines in TCM universities still face challenges,such as weak foundational infrastructure and difficulties in interdisciplinary integration,insufficient faculty and suboptimal curriculum systems,and a single teaching model with a disconnect between theory and practice.Conclusion This paper proposes targeted suggestions and strate-gies for optimizing the layout of management disciplines in TCM universities,including strengthening support for management dis-ciplines and deepening TCM management characteristics,optimizing curriculum design and enhancing faculty resources,and in-novating teaching models to promote the deep integration of theory and practice.

Objective The CRISPR/Cas9 system was utilized to generate an Apoe knockout mice model to support further investigations of the role of Apoe in lipid metabolism and atherosclerosis.Methods Two single guide RNAs designed for Apoe in C57BL/6J mice were co-injected with Cas9 mRNA into fertilized eggs,followed by transplantation into ICR recipient mice to obtain F0 generation mice.KO mice were identified by polymerase chain reaction(PCR)screening of tail DNA.Apoe mRNA expression in various tissues was assessed by quantitative real-time PCR and lipid indexes were measured in serum samples.Lipid accumulation in the inner lining of aortic vessels was detected by oil red O staining.Results PCR and sequencing confirmed the successful construction of Apoe KO mice(C57BL/6-Apoeem1/Nifdc).Apoe mRNA levels were significantly reduced in the liver,brain,spleen,kidney,and lung tissues of Apoe KO homozygous mice(Apoe-/-),as shown by reverse transcription quantitative real-time PCR.Serum total cholesterol and low-density lipoprotein cholesterol levels were increased in Apoe-/-mice,and high-density lipoprotein cholesterol levels were decreased in male Apoe-/-mice.Extensive lipid plaques were observed in the inner lining of the arteries in Apoe-/-mice compared with WT mice,under normal chow consumption conditions.Conclusions This study successfully established an Apoe KO mice model exhibiting a typical abnormal lipid metabolism phenotype with arterial lipid accumulation,even without a high-fat diet intervention.This work provides background data for the Apoe KO mice resource and a new model for the study of abnormal lipid metabolism.

Objective We generated ob mice(C57BL/6N-Lepem1/Nifdc)with Lep gene knockout(ob/ob)using the CRISPR/Cas9 system,to establish a suitable animal model for preclinical drug evaluation for clinical diseases such as diabetes.Methods According to the principle of CRISPR/Cas9 target design,single guide RNA targeting the mouse Lep gene was designed for transcription in vitro,and microinjected with Cas9 mRNA into mouse zygotes.Mouse tail DNA was extracted and detected by polymerase chain reaction and sequencing,followed by mating of positive and wild-type mice.Blood biochemistry and liver pathology were assessed in homozygous ob mice.Results Eight positive mice were identified and a stable mouse strain was selected for further breeding.Serum triglycerides,total cholesterol,and alanine aminotransferase levels were significantly higher in homozygous ob mice than in wild-type mice,and liver pathology showed inflammatory infiltration and lipid vacuolar transformations.Conclusions We successfully established a Lep gene knockout mouse model,which will provide an important addition to the national rodent experimental animal database and an animal model for preclinical drug evaluation.

Objective The CRISPR/Cas9 system was utilized to generate an Apoe knockout mice model to support further investigations of the role of Apoe in lipid metabolism and atherosclerosis.Methods Two single guide RNAs designed for Apoe in C57BL/6J mice were co-injected with Cas9 mRNA into fertilized eggs,followed by transplantation into ICR recipient mice to obtain F0 generation mice.KO mice were identified by polymerase chain reaction(PCR)screening of tail DNA.Apoe mRNA expression in various tissues was assessed by quantitative real-time PCR and lipid indexes were measured in serum samples.Lipid accumulation in the inner lining of aortic vessels was detected by oil red O staining.Results PCR and sequencing confirmed the successful construction of Apoe KO mice(C57BL/6-Apoeem1/Nifdc).Apoe mRNA levels were significantly reduced in the liver,brain,spleen,kidney,and lung tissues of Apoe KO homozygous mice(Apoe-/-),as shown by reverse transcription quantitative real-time PCR.Serum total cholesterol and low-density lipoprotein cholesterol levels were increased in Apoe-/-mice,and high-density lipoprotein cholesterol levels were decreased in male Apoe-/-mice.Extensive lipid plaques were observed in the inner lining of the arteries in Apoe-/-mice compared with WT mice,under normal chow consumption conditions.Conclusions This study successfully established an Apoe KO mice model exhibiting a typical abnormal lipid metabolism phenotype with arterial lipid accumulation,even without a high-fat diet intervention.This work provides background data for the Apoe KO mice resource and a new model for the study of abnormal lipid metabolism.

Objective We generated ob mice(C57BL/6N-Lepem1/Nifdc)with Lep gene knockout(ob/ob)using the CRISPR/Cas9 system,to establish a suitable animal model for preclinical drug evaluation for clinical diseases such as diabetes.Methods According to the principle of CRISPR/Cas9 target design,single guide RNA targeting the mouse Lep gene was designed for transcription in vitro,and microinjected with Cas9 mRNA into mouse zygotes.Mouse tail DNA was extracted and detected by polymerase chain reaction and sequencing,followed by mating of positive and wild-type mice.Blood biochemistry and liver pathology were assessed in homozygous ob mice.Results Eight positive mice were identified and a stable mouse strain was selected for further breeding.Serum triglycerides,total cholesterol,and alanine aminotransferase levels were significantly higher in homozygous ob mice than in wild-type mice,and liver pathology showed inflammatory infiltration and lipid vacuolar transformations.Conclusions We successfully established a Lep gene knockout mouse model,which will provide an important addition to the national rodent experimental animal database and an animal model for preclinical drug evaluation.

ObjectiveThrough improving the potential of vascular endothelial growth factor receptor (VEGFR)-humanized mouse model (hKDR^+/+) with C57BL/6N background to allow the growth of different mouse tumor cell lines, to establish novel tumor-bearing mouse models which can support in vivo tumorigenesis of different mouse tumor cell lines and be used to evaluate drugs targeting VEGFR.MethodsFirstly, a method to evaluate the in vivo activity of antibody targeting VEGFR based on the hKDR^+/+ humanized mouse model was established. Recombinant activating gene 1 (Rag1) knockout mice (Rag1^-/-) were established using CRISPR/Cas9 technology. Then these Rag1^-/- mice were crossed with hKDR^+/+ mice to get a double gene modified homozygous hKDR^+/+/Rag1^-/- mouse model by screening. Finally, tumor cell lines derived from different mouse strains were tested on the double gene-modified mouse model to compare the differences in tumor growth. ResultsAntibodies designed for VEGFR showed significant anti-tumor activity in hKDR^+/+ mice, which significantly reduced tumor volume and weight compared with the PBS group (P<0.01, P<0.05). The number of B cells and T cells in the peripheral blood of Rag1^-/- mice and hKDR^+/+/Rag1^-/- mice decreased (P<0.05, P<0.001). Tumors were observed in hKDR^+/+/Rag1^-/-, Rag1^-/-, wild-type, and hKDR^+/+ mice after 7 d of inoculation of MC38 cells derived from C57BL/6 mice. Tumors were only observed in groups of hKDR^+/+/Rag1^-/- and Rag1^-/- mice, but not in the wild-type and hKDR^+/+ mice after 10 d of inoculation with CT26 cells derived from BALB/c mice. After 3 weeks of inoculation, the tumor volume of hKDR^+/+/Rag1^-/- mice was significantly larger than that of Rag1^-/- mice (P<0.01). ConclusionRag1 knockout mice were obtained and a novel hKDR^+/+/Rag1^-/- double genes modified mouse model was further screened. The tumor cell lines from different mouse strain origins were more prone to growth in mice with Rag1 gene deficiency. The results suggest that the reduced immune response of hKDR^+/+ humanized mice will improve the capacity of supporting the growth of mouse tumor lines in the model. As a result, more tumor-bearing mouse models may be constructed for the evaluation of drugs targeting VEGFR in this way.

Objective To investigate the effect of minitype titanium plate fixation through transoral approach in the treatment of unstable atlas fractures.Methods A retrospective case series study was made on 21 patients with unstable atlas fractures treated by minitype titanium plate fixation through transoral approach from June 2008 to June 2014.There were 15 males and 6 females,at age of (40.9 ± 10.6)years (range,21 to 57 years).Anterior 1/2 Jefferson fractures were seen in 12 patients and 1/2 ring Jefferson fractures in 9 patients.Preoperative visual analogue score (VAS) was 4-9 points [(7.6 ± 1.3) points].Before operation,degree of mobility of the cervical vertebra was (15.4 ± 3.9) °in bending,(10.8 ± 2.5) °in extending,(18.3 ± 3.1) ° in left-bending,(18.9 ± 2.7) ° in right-bending,(21.8 ± 5.8) °in left-rotation and (22.4 ± 4.6) ° in right-rotation.Operation time,intraoperative blood loss,VAS,cervical mobility and bone healing were detected after operation.Results Operation time was (86.3 ±25.3)m in,and intraoperative blood loss was (120.5 ± 33.3)ml.VAS was improved to 0-2 points [(1.6 ± 0.4) points] at postoperative 3 days (P ＜ 0.05).All patients were followed up for 12 to 48 months[(23.7 ±5.9) months].VAS was improved to 0-2 points[(0.6 ± 0.1) points] at postoperative 3 months (P ＜ 0.05).Degree of mobility of the cervical vertebra was improved significantly at postoperative 3 months,with the bending of(38.6 ± 4.5) °,extending of (39.3 ± 4.0) °,left-bending of (39.2 ± 4.0) °,right-bending of (39.2 ± 2.9) °,left-rotation of (66.8 ± 8.8) ° and right-rotation of (66.3 ± 9.2) ° (P ＜ 0.05).Postoperatively,there were no surgical wound incision infections and vertebral artery or spinal injuries,Bone union was found in all patients,without the occurrence of implant loosening or breakage and the dysfunction of the cervical vertebra.Conclusion Minitype titanium plate fixation through transoral approach is associated with less trauma,high healing rate and preservation of the activity of cervical vertebra in the treatment of unstable atlas fractures.