Objective To investigate the underlying mechanism of Hongyu Decoction in the treatment of inflammatory bowel disease(IBD)based on serum amino acid metabolism.Methods A total of 50 male C57BL/6J mice were divided into control group,model group,sulfasalazine(SASP)group and Hongyu Decoction low-and high-dosage groups,with 10 mice in each group.Except for the control group,the rest groups were treated with 3%dextran sulfate sodium freely for seven consecutive days to establish the IBD model.The SASP group was given SASP solution at a dosage of 200 mg/kg by gavage,while the Hongyu Decoction low-and high-dosage groups were given Hongyu Decoction freeze-dried powder solution at dosages of 0.5 and 2.0 g/kg respectively by gavage for 11 consecutive days.The general condition was monitored and DAI score were calculated.After the mice were sacrificed,the length of colons was measured,ELISA was used to detect serum TNF-α,IL-1β,IL-17 contents,HE staining was used to observe the morphology of colon tissue,and Alizarin blue staining was used to evaluate the secretion of mucin levels by intestinal epithelial goblet cells,UPLC-MS and multivariate statistical methods were used to analyze the serum amino acid metabolism profiles of mice in the control group,model group and Hongyu Decoction high-dosage group,and differential amino acids were screened.Pathway enrichment analysis was performed on differential amino acids using MetaboAnalyst 5.0.Results Compared with the control group,the DAI score of the model group mice significantly increased(P<0.001),the colon length was significantly shortened(P<0.001),and the serum contents of TNF-α,IL-1β and IL-17 significantly increased(P<0.001);with widespread ulceration of colon tissue,disappearance of mucosal epithelium and intestinal crypt structure,infiltration of lymphocytes and neutrophils,congestion and edema of intestinal mucosa,proliferation of submucosal connective tissue,reduced number of colonic goblet cells and mucin secretion.Compared with the model group,the Hongyu Decoction high-dosage group showed a significant decrease in DAI score(P<0.001),while the SASP group and the Hongyu Decoction low-and high-dosage groups showed a significant increase in colon length(P<0.05,P<0.001),and the serum contents of TNF-α,IL-1β and IL-17 were significantly reduced(P<0.001);the colonic villi were relatively intact,the glands were clear and arranged neatly,the branches of the intestinal crypts were obvious,no obvious edema was observed,and the number of colonic goblet cells and mucin secretion increased.Seven potential biomarkers for IBD in mice were identified through serum metabolomics screening,including tryptophan,serine,glutamate,valine,histidine,methionine and phenylalanine;two potential biomarkers for the treatment of IBD with Hongyu Decoction were identified,namely valine and tyrosine.The results of pathway enrichment analysis showed that the differential amino acids in the model group mainly involved the biosynthesis of phenylalanine,tyrosine and tryptophan,histidine metabolism,cysteine and methionine metabolism,phenylalanine metabolism and arginine biosynthesis;the differential amino acids in Hongyu Decoction high-dosage group mainly involved the biosynthesis of phenylalanine,tyrosine and tryptophan,as well as the metabolism of alanine,aspartate and glutamate,and the biosynthesis of arginine.Conclusion Hongyu Decoction can improve intestinal epithelial damage and inflammatory cell infiltration in IBD mice,protect the integrity of the intestinal mucosal barrier,and may be related to regulating amino acid metabolism in the body.

Objective To investigate the underlying mechanism of Hongyu Decoction in the treatment of inflammatory bowel disease(IBD)based on serum amino acid metabolism.Methods A total of 50 male C57BL/6J mice were divided into control group,model group,sulfasalazine(SASP)group and Hongyu Decoction low-and high-dosage groups,with 10 mice in each group.Except for the control group,the rest groups were treated with 3%dextran sulfate sodium freely for seven consecutive days to establish the IBD model.The SASP group was given SASP solution at a dosage of 200 mg/kg by gavage,while the Hongyu Decoction low-and high-dosage groups were given Hongyu Decoction freeze-dried powder solution at dosages of 0.5 and 2.0 g/kg respectively by gavage for 11 consecutive days.The general condition was monitored and DAI score were calculated.After the mice were sacrificed,the length of colons was measured,ELISA was used to detect serum TNF-α,IL-1β,IL-17 contents,HE staining was used to observe the morphology of colon tissue,and Alizarin blue staining was used to evaluate the secretion of mucin levels by intestinal epithelial goblet cells,UPLC-MS and multivariate statistical methods were used to analyze the serum amino acid metabolism profiles of mice in the control group,model group and Hongyu Decoction high-dosage group,and differential amino acids were screened.Pathway enrichment analysis was performed on differential amino acids using MetaboAnalyst 5.0.Results Compared with the control group,the DAI score of the model group mice significantly increased(P<0.001),the colon length was significantly shortened(P<0.001),and the serum contents of TNF-α,IL-1β and IL-17 significantly increased(P<0.001);with widespread ulceration of colon tissue,disappearance of mucosal epithelium and intestinal crypt structure,infiltration of lymphocytes and neutrophils,congestion and edema of intestinal mucosa,proliferation of submucosal connective tissue,reduced number of colonic goblet cells and mucin secretion.Compared with the model group,the Hongyu Decoction high-dosage group showed a significant decrease in DAI score(P<0.001),while the SASP group and the Hongyu Decoction low-and high-dosage groups showed a significant increase in colon length(P<0.05,P<0.001),and the serum contents of TNF-α,IL-1β and IL-17 were significantly reduced(P<0.001);the colonic villi were relatively intact,the glands were clear and arranged neatly,the branches of the intestinal crypts were obvious,no obvious edema was observed,and the number of colonic goblet cells and mucin secretion increased.Seven potential biomarkers for IBD in mice were identified through serum metabolomics screening,including tryptophan,serine,glutamate,valine,histidine,methionine and phenylalanine;two potential biomarkers for the treatment of IBD with Hongyu Decoction were identified,namely valine and tyrosine.The results of pathway enrichment analysis showed that the differential amino acids in the model group mainly involved the biosynthesis of phenylalanine,tyrosine and tryptophan,histidine metabolism,cysteine and methionine metabolism,phenylalanine metabolism and arginine biosynthesis;the differential amino acids in Hongyu Decoction high-dosage group mainly involved the biosynthesis of phenylalanine,tyrosine and tryptophan,as well as the metabolism of alanine,aspartate and glutamate,and the biosynthesis of arginine.Conclusion Hongyu Decoction can improve intestinal epithelial damage and inflammatory cell infiltration in IBD mice,protect the integrity of the intestinal mucosal barrier,and may be related to regulating amino acid metabolism in the body.

Objective To explore the possibility and genetic identification method of constructing a hepatocyte-specific NLRP3 gene knockout mouse model by using Cre-LoxP system gene knockout technology.Methods Phase one:mice specifically expressing the albumin promoter-Cre(AlbCre)recombinase in hepatocytes were mated with NLRP3flox/flox mice,and the hepatocyte-specific NLRP3 gene knockout mice with the genotype of NLRP3flox/flox/AlbCre+/-(hepatocyte NLRP3 knockout group)and the control mice in the same litter with the genotype of NLRP3flox/flox/AlbCre-/-(control group in the same litter)were obtained after two generations of selection and mating.The second stage was the mass reproduction stage.Mating NLRP3flox/flox/AlbCre+/-target mice with NLRP3flox/flox mice could quickly obtain a large number of experimental target mice and control mice in the same litter.The DNA was extracted from the tails of mice after numbering,and the offspring genotype was identified by PCR.qPCR and Western blot were used to detect the mRNA and protein expression levels of NLRP3 gene in the liver tissue.HE staining was used to observe the morphological changes in liver tissues,and serum liver transaminases and inflammatory factors were detected.The changes in body weight,liver-to-body ratio and special circumstances during reproduction and development of mice in the two groups were observed.Results The offspring genotype of the target mice in the F2 generation was consistent with theoretical result of NLRP3flox/flox/AlbCre+/-.The mRNA and protein levels of NLRP3 in liver tissues of mice in the hepatocyte NLRP3 knockout group were significantly lower than those in the control group in the same litter(P<0.05).The mice in the hepatocyte NLRP3 knockout group was not affected in terms of growth,development and reproduction after the NLRP3 gene knockout.There were no statistically significant differences in the body weight,liver-to-body ratio,liver tissue morphology,serum liver transaminase or inflammatory factors between the hepatocyte NLRP3 knockout group and the control group in the same litter(P>0.05).Conclusion The Cre-LoxP gene knockout technology can be used to successfully construct a hepatocyte-specific NLRP3 gene knockout mouse model,providing an important technical support for the next step of studying the function of the NLRP3 gene in the liver at the animal level.

Objective To explore the possibility and genetic identification method of constructing a hepatocyte-specific NLRP3 gene knockout mouse model by using Cre-LoxP system gene knockout technology.Methods Phase one:mice specifically expressing the albumin promoter-Cre(AlbCre)recombinase in hepatocytes were mated with NLRP3flox/flox mice,and the hepatocyte-specific NLRP3 gene knockout mice with the genotype of NLRP3flox/flox/AlbCre+/-(hepatocyte NLRP3 knockout group)and the control mice in the same litter with the genotype of NLRP3flox/flox/AlbCre-/-(control group in the same litter)were obtained after two generations of selection and mating.The second stage was the mass reproduction stage.Mating NLRP3flox/flox/AlbCre+/-target mice with NLRP3flox/flox mice could quickly obtain a large number of experimental target mice and control mice in the same litter.The DNA was extracted from the tails of mice after numbering,and the offspring genotype was identified by PCR.qPCR and Western blot were used to detect the mRNA and protein expression levels of NLRP3 gene in the liver tissue.HE staining was used to observe the morphological changes in liver tissues,and serum liver transaminases and inflammatory factors were detected.The changes in body weight,liver-to-body ratio and special circumstances during reproduction and development of mice in the two groups were observed.Results The offspring genotype of the target mice in the F2 generation was consistent with theoretical result of NLRP3flox/flox/AlbCre+/-.The mRNA and protein levels of NLRP3 in liver tissues of mice in the hepatocyte NLRP3 knockout group were significantly lower than those in the control group in the same litter(P<0.05).The mice in the hepatocyte NLRP3 knockout group was not affected in terms of growth,development and reproduction after the NLRP3 gene knockout.There were no statistically significant differences in the body weight,liver-to-body ratio,liver tissue morphology,serum liver transaminase or inflammatory factors between the hepatocyte NLRP3 knockout group and the control group in the same litter(P>0.05).Conclusion The Cre-LoxP gene knockout technology can be used to successfully construct a hepatocyte-specific NLRP3 gene knockout mouse model,providing an important technical support for the next step of studying the function of the NLRP3 gene in the liver at the animal level.

Objective:To evaluate the protective effectiveness (VE) of the acellular pertussis vaccine (aPV) against pertussis in children aged 2 months to 6 years.Methods:A test-negative case-control study was conducted among children aged 2 months to 6 years who sought medical care for cough and underwent pertussis nucleic acid testing at sentinel surveillance hospitals in Zhejiang Province in 2024. Cases were defined as those with positive pertussis nucleic acid test results, while controls were test-negative individuals matched 1∶1 based on propensity scores using the caliper matching method. Conditional logistic regression models were used to calculate odds ratios ( ORs) and VEs. Results:Among the 658 participants, 31.76% (209 cases) tested positive for pertussis. After propensity score matching, 203 cases and 203 controls were included in the analysis. The VE of 1-2, 3, and 4 doses of aPV against pertussis was 52.46% (95% CI:-39.82%-83.84%), 65.22% (95% CI: 6.86%-87.02%), and 72.21% (95% CI: 34.33%-88.24%), respectively. For pertussis-related hospitalization, the VE of 1-3 and 4 doses was 80.95% (95% CI:31.38%-94.71%) and 86.79% (95% CI: 51.89%-96.37%). The VE for those who completed 4 doses of vaccination and had intervals of less than 2 years, 2 years, 3 years, and 4 years or more after vaccination were 91.15% (95% CI: 67.61%-97.58%), 84.70% (95% CI: 43.71%-95.84%),56.23% (95% CI:-47.58%-87.02%), and 49.92% (95% CI:-83.74%-86.35%), respectively. Conclusion:The VE of aPV against pertussis in children aged 2 months to 6 years increases with the number of doses administered, and it is more effective in preventing hospitalization due to pertussis. The VE declines rapidly over time after the last dose. It is recommended to follow the new pertussis immunization program for timely and full vaccination.

Stress is a critical inducer in the onset and progression of many chronic diseases. Prolonged or intense stress can disrupt the overall balance between the nervous, immune, and endocrine systems. The resulting biological signals may act on corresponding receptors in the cervical spine region, leading to adverse pathological changes. The vertebral artery and the surrounding muscular and connective tissues are influenced by biomechanical abnormalities and inflammatory cascades associated with cervical spondylosis of vertebral artery type (CSA), which promotes the release of various hormones. These hormones, through the neuroendocrine-immune system, affect the central nervous system, inducing or exacerbating negative emotional feedback and thereby establishing a "central-local-central" vicious cycle. This article explores the mechanisms underlying the impact of stress on the key CSA symptoms through the neuroendocrine-immune network (NEI) theory, providing a more comprehensive framework for targeted therapeutic interventions in CSA.
Humans ; Neurosecretory Systems/immunology* ; Spondylosis/etiology* ; Vertebral Artery/immunology* ; Stress, Psychological/complications* ; Chronic Disease