Objective To establish a quality standard for the medicinal bath formula for treating exogenous fever(composed of Forsythiae Fructus,Peucedani Radix,Schizonepetae Spica,Isatidis Radix,Gypsum Fibrosum,Chrysanthemi Flos,Lophatheri Herba,etc.)based on its efficacy and indications.Methods Thin-layer chromatography(TLC)was used to establish identification method for the monarch drugs Forsythiae Fructus and Peucedani Radix.Enzyme-linked immunosorbent assay(ELISA)was employed to measure the interleukin 1β(IL-1β)regulatory activity of the main components in the formula.High-performance liquid chromatography(HPLC)was used to determine the content of active components in 10 batches of samples.Results Spots of Forsythiae Fructus and Peucedani Radix were successfully detected in the test samples.ELISA identified active components in the formula,including praeruptorin A,pulegone,rutin,praeruptorin B,forsythoside,and(R,S)-goitrin.The content determination results of 10 batches of samples showed that the content of praeruptorin A ranged from 0.493 to 0.694 mg·mL-1.Conclusion Based on its efficacy and indications,TLC identification and HPLC content determination methods were established for the medicinal bath formula for treating exogenous fever.The obtained standard can more accurately control the efficacy of the formula.

Objective:To analyze the contamination characteristics and pathogenic potential of Listeria monocytogenes (LM) in pre-packaged and refrigerated ready-to-eat cooked meat products in Chengdu City. Methods:From September 2022 to May 2023, pre-packaged and refrigerated ready-to-eat cooked meat products were collected from six districts and counties in Chengdu City. Qualitative and quantitative determination of LM was performed on these samples. Whole genome sequencing was carried out on the isolated strains. Different ST strains were selected for cell adhesion and invasion experiments. The results were expressed as adhesion rate and invasion rate. One-way ANOVA was used for comparison between groups, and the Dunnett t-test was used for pairwise comparison. Results:A total of 145 samples were collected, and LM was detected and isolated in 29 samples. The total detection rate was 20.00%, and the detection rate of braised pork was the highest (68.18%). The contamination level of LM in 9 samples (31.03%) was greater than 100.00 MPN/g. The 29 strains of Listeria monocytogenes belonged to 7 STs, including ST 3 (27.59%), ST 8 (17.24%) and ST 87 (13.79%). The strain of ST 87 carried Listeria pathogenicity islands 4 (LIPI-4), which was a highly virulent strain. The medium and high virulence strains accounted for 79.31%. Some moderately virulent and highly virulent strains of ST 8 and ST 87 were closely related to clinical patient strains. Some LM isolates of the same ST type had little SNP differences (1-6) in the same manufacturer at different stages. In vitro cell experiments showed that the highly virulent strain ST 87 had the strongest adhesion and invasion ability towards Caco-2 cells. Conclusion:The pre-packaged and refrigerated cooked meat products have a high contamination rate. Some samples have high contamination levels and carry medium and high virulence strains, posing potential health risks to human beings. LM residues continue to persist in some manufacturers′ production and processing stages.

The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P＜0.05)and IFN-λ(P＜0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.

Objective To investigate the therapeutic effect of Sheng Mai San mixed Jiawei Sheng Xian Tang on patients with heart failure with reduced ejection fraction(HFrEF)of Qi deficiency blood stasis type,and its impact on the immune inflammatory response of patients.Methods Ninety patients with HFrEF of Qi deficiency blood stasis type were selected and divided into two groups according to the treatment plan.Patients of the conventional group(42 cases)took sacubitril/valsartan sodium tablets orally.Patients of the combined group(48 cases)were treated with Sheng Mai San combined with Jiawei Sheng Xian Tang on the basis of the conventional group,one dose per day,and divided it in two servings used in the morning and evening.The symptoms of the patients were evaluated by using traditional Chinese medicine(TCM)syndrome score before and after the treatment respectively.Echocardiography technology was used to measure the left ventricular end-diastolic diameter(LVEDD)and left ventricular ejection fraction(LVEF).Enzyme-linked immunosorbent assay was used to detect pentaggrin 3(PTX3),tumor necrosis factor-α(TNF-α),galectin-3(Gal-3),interleukin(IL)-8,IL-6,IL-33 and angiotensin Ⅱ(Ang Ⅱ).N-terminal pro-brain natriuretic peptide(NT-proBNP)was detected by immunofluorescence analysis.The therapeutic effect was evaluated based on the symptoms and the New York Heart Association(NYHA)cardiac function classification in the United States.Results After treatment,the levels of TCM syndrome scores,LVEDD,TNF-α,PTX3,IL-8,IL-6,IL-33,NT-proBNP,Gal-3 and AngⅡ decreased in both groups,and those in the combined group were lower than those in the conventional group(P＜0.05).The LVEF increased,and which were higher in the combined group than those of the conventional group(P＜0.05).After the treatment,the total effective rate of the combined group was higher than that of the conventional group(P＜0.05).Conclusion The Sheng Mai San mixed Jiawei Sheng Xian Tang as adjuvant therapy can effectively reduce the levels of immune inflammatory cytokines in HFrEF patients of Qi deficiency blood stasis type,improve their clinical symptoms and enhance therapeutic effect.

The specific mechanisms of interferon regulatory factor 8(IRF8)in porcine intestinal in-nate immunity and resistance to enteric virus infection remain to be elucidated.To investigate the immunoregulatory role of IRF8,establishing an IRF8 gene knockout porcine intestinal epithelial cell(IPEC-J2)monoclonal cell line is of significant importance.This study initially aimed to obtain recombinant adeno-associated virus rAAV-sgIRF8-eGFP capable of knocking out the IRF8 gene through co-transfection of HEK-293T cells with three plasmids.Subsequently,IPEC-J2 cells were infected with the virus,and those expressing eGFP were selected by flow cytometry and cultured to form monoclonal cell lines.These cell lines were then identified by Sanger sequencing and West-ern blot techniques.Lastly,qPCR analysis was used to measure the expression levels of interferon factors IFN-α,IFN-β,IFN-γ and IFN-λ,providing preliminary insights into the impact of IRF8 gene knockout on IPEC-J2 cell immunity.The results demonstrated successful generation of rAAV-sgIRF8-eGFP,which successfully infected IPEC-J2 cells leading to eGFP fluorescence.Flow cytometry followed by cell culture led to the establishment of two monoclonal cell lines,IRF8-KO1 and IRF8-KO3.Sanger sequencing revealed a five-base deletion in IRF8-KO1 and a seven-base dele-tion in IRF8-KO3.Western blot confirmed the absence of IRF8 protein expression in IRF8-KO1,making it an ideal candidate monoclonal cell line.qPCR analysis of interferon factors indicated sig-nificant decrease in IFN-γ(P＜0.05)and IFN-λ(P＜0.01)transcription level in IRF8-knockout cells,while the transcription levels of IFN-α and IFN-β remained relatively unchanged.This study successfully established an IRF8 gene knockout IPEC-J2 monoclonal cell line,providing a founda-tion for further research on IRF8-related porcine intestinal immune regulation and mechanisms of intestinal virus infection.

Objective To investigate the therapeutic effect of Sheng Mai San mixed Jiawei Sheng Xian Tang on patients with heart failure with reduced ejection fraction(HFrEF)of Qi deficiency blood stasis type,and its impact on the immune inflammatory response of patients.Methods Ninety patients with HFrEF of Qi deficiency blood stasis type were selected and divided into two groups according to the treatment plan.Patients of the conventional group(42 cases)took sacubitril/valsartan sodium tablets orally.Patients of the combined group(48 cases)were treated with Sheng Mai San combined with Jiawei Sheng Xian Tang on the basis of the conventional group,one dose per day,and divided it in two servings used in the morning and evening.The symptoms of the patients were evaluated by using traditional Chinese medicine(TCM)syndrome score before and after the treatment respectively.Echocardiography technology was used to measure the left ventricular end-diastolic diameter(LVEDD)and left ventricular ejection fraction(LVEF).Enzyme-linked immunosorbent assay was used to detect pentaggrin 3(PTX3),tumor necrosis factor-α(TNF-α),galectin-3(Gal-3),interleukin(IL)-8,IL-6,IL-33 and angiotensin Ⅱ(Ang Ⅱ).N-terminal pro-brain natriuretic peptide(NT-proBNP)was detected by immunofluorescence analysis.The therapeutic effect was evaluated based on the symptoms and the New York Heart Association(NYHA)cardiac function classification in the United States.Results After treatment,the levels of TCM syndrome scores,LVEDD,TNF-α,PTX3,IL-8,IL-6,IL-33,NT-proBNP,Gal-3 and AngⅡ decreased in both groups,and those in the combined group were lower than those in the conventional group(P＜0.05).The LVEF increased,and which were higher in the combined group than those of the conventional group(P＜0.05).After the treatment,the total effective rate of the combined group was higher than that of the conventional group(P＜0.05).Conclusion The Sheng Mai San mixed Jiawei Sheng Xian Tang as adjuvant therapy can effectively reduce the levels of immune inflammatory cytokines in HFrEF patients of Qi deficiency blood stasis type,improve their clinical symptoms and enhance therapeutic effect.

Objective:To develop a Recurrence Fear Scale for Patients with Atrial Fibrillation after Catheter Ablation and conduct psychometric testing for its reliability and validity.Methods:Based on the common-sense model of self-regulation, the scale was developed through a literature review and semi-structured interviews to construct an item pool. The scale draft was formed through expert consultation and a pre-survey. Using convenience sampling, 361 patients with atrial fibrillation who had undergone catheter ablation and were either hospitalized or attending outpatient follow-up at the Department of Cardiology, Affiliated Hospital of Qingdao University, between January and March 2024, were selected for questionnaire survey. Item analysis and reliability and validity tests were performed on the scale.Results:Exploratory factor analysis identified five common factors, with a cumulative variance contribution rate of 74.375%. Confirmatory factor analysis showed good model fit. The final scale included five dimensions and 24 items: trigger factors, recurrence risk perception, disease uncertainty, psychological burden, and coping strategies. The content validity index at the scale level was 0.864, and the item-level content validity index ranged from 0.684 to 0.947. The Cronbach's α coefficient of the scale was 0.919, the split-half reliability coefficient was 0.834, and the test-retest reliability coefficient was 0.872.Conclusions:The development of the Recurrence Fear Scale for Patients with Atrial Fibrillation after Catheter Ablation is scientific and standardized, and the scale has good reliability and validity. It can effectively assess the level of recurrence fear in these patients.

Objective:To analyze the contamination characteristics and pathogenic potential of Listeria monocytogenes (LM) in pre-packaged and refrigerated ready-to-eat cooked meat products in Chengdu City. Methods:From September 2022 to May 2023, pre-packaged and refrigerated ready-to-eat cooked meat products were collected from six districts and counties in Chengdu City. Qualitative and quantitative determination of LM was performed on these samples. Whole genome sequencing was carried out on the isolated strains. Different ST strains were selected for cell adhesion and invasion experiments. The results were expressed as adhesion rate and invasion rate. One-way ANOVA was used for comparison between groups, and the Dunnett t-test was used for pairwise comparison. Results:A total of 145 samples were collected, and LM was detected and isolated in 29 samples. The total detection rate was 20.00%, and the detection rate of braised pork was the highest (68.18%). The contamination level of LM in 9 samples (31.03%) was greater than 100.00 MPN/g. The 29 strains of Listeria monocytogenes belonged to 7 STs, including ST 3 (27.59%), ST 8 (17.24%) and ST 87 (13.79%). The strain of ST 87 carried Listeria pathogenicity islands 4 (LIPI-4), which was a highly virulent strain. The medium and high virulence strains accounted for 79.31%. Some moderately virulent and highly virulent strains of ST 8 and ST 87 were closely related to clinical patient strains. Some LM isolates of the same ST type had little SNP differences (1-6) in the same manufacturer at different stages. In vitro cell experiments showed that the highly virulent strain ST 87 had the strongest adhesion and invasion ability towards Caco-2 cells. Conclusion:The pre-packaged and refrigerated cooked meat products have a high contamination rate. Some samples have high contamination levels and carry medium and high virulence strains, posing potential health risks to human beings. LM residues continue to persist in some manufacturers′ production and processing stages.

Objective:To develop a Recurrence Fear Scale for Patients with Atrial Fibrillation after Catheter Ablation and conduct psychometric testing for its reliability and validity.Methods:Based on the common-sense model of self-regulation, the scale was developed through a literature review and semi-structured interviews to construct an item pool. The scale draft was formed through expert consultation and a pre-survey. Using convenience sampling, 361 patients with atrial fibrillation who had undergone catheter ablation and were either hospitalized or attending outpatient follow-up at the Department of Cardiology, Affiliated Hospital of Qingdao University, between January and March 2024, were selected for questionnaire survey. Item analysis and reliability and validity tests were performed on the scale.Results:Exploratory factor analysis identified five common factors, with a cumulative variance contribution rate of 74.375%. Confirmatory factor analysis showed good model fit. The final scale included five dimensions and 24 items: trigger factors, recurrence risk perception, disease uncertainty, psychological burden, and coping strategies. The content validity index at the scale level was 0.864, and the item-level content validity index ranged from 0.684 to 0.947. The Cronbach's α coefficient of the scale was 0.919, the split-half reliability coefficient was 0.834, and the test-retest reliability coefficient was 0.872.Conclusions:The development of the Recurrence Fear Scale for Patients with Atrial Fibrillation after Catheter Ablation is scientific and standardized, and the scale has good reliability and validity. It can effectively assess the level of recurrence fear in these patients.

BACKGROUND:Lumbar fixed-point rotation operation needs collaborative operation of the doctor's hands,and outputs rotation and thumb thrust.Lumbar disc herniation can be treated through disc displacement and adjusting stress distribution.However,the mechanical effects of thumb thrust and the biomechanical effects of loading direction on manipulative effects remain unclear. OBJECTIVE:To compare the biomechanical difference of lumbar fixed-point rotation manipulation for treating lumbar disc herniation under different thrust directions. METHODS:The L3-5 normal three-dimensional finite element model was constructed and validity was verified.According to the intervertebral disc degeneration Pfirrmann grade,intervertebral disc degeneration was simulated by modifying the L4/5 intervertebral space height,the volume of the nucleus pulposus,as well as the material parameters of the annulus fibrosus,nucleus pulposus,and ligament.Finally,the pathological model of L4/5 moderate disc degeneration with left para-central herniation was constructed,and then the pathological models were used as research objects.Simulation technique:spinning to the right;taking the condition on changing the direction of the thumb thrust to establish three modes of operation(M1:thumb push to the left;M2:thumb push to the right;M3:no thrust push).The protrusion displacement and the disc stress,and the stress and strain of the facet joint cartilage were compared in the three operating modes. RESULTS AND CONCLUSION:(1)Maximum displacement value of L4/5 disc herniation:displacement was 2.672 3 mm for M1,1.156 1 mm for M2,1.826 4 mm for M3,M1＞M3＞M2.(2)The maximum Von Mises stress of L4/5 discs was 1.846 7 MPa for M1,0.419 0 MPa for M2,and 1.257 9 MPa for M3,M1＞M3＞M2.(3)L4/5 bilateral small cartilage produced different degrees of contact stress changes:It was 0.485 5 MPa for M1,0.026 7 MPa for M2,and 0.441 4 MPa for M3,M1＞M3＞M2.Right cartilage contact force was 0.000 5 MPa for M1,0.025 9 MPa for M2,and 0.001 3 MPa for M3,M2＞M3＞M1;the left greater than the right,M1 had the highest value;cartilage strain was consistent with contact stress changes.(4)Different operation modes will have some biomechanical influences on the diseased intervertebral disc and accessory structure.The M1 operation mode can maximize the displacement of protrusion,disc stress and left joint cartilage contact,which can better promote disc displacement,balance stress distribution and reduce facet joint disorder,so the operation is better.