Toxoplasma gondii is a single-celled parasite that infects nearly all warm-blooded animals, including humans (Montoya and Liesenfeld, 2004). It occurs worldwide and can persist for a lifetime in mammals. Humans get infected by eating undercooked meat of animals containing the tissue cysts of this parasite. In immune-competent individuals, T. gondii infection usually does not cause significant clinical symptoms, whereas in pregnant or immunocompromised individuals, T. gondii infection (toxoplasmosis) can cause more serious problems like abortion and even death (Dunn et al., 1999; Wang et al., 2017). A combination of pyrimethamine and sulfadiazine is usually used to treat toxoplasmosis, although it is generally inefficient and causes side effects (Alday and Doggett, 2017). Worse still, there is a lack of vaccines to prevent T. gondii infection in humans or animals.
Toxoplasma/enzymology* ; Animals ; Humans ; Toxoplasmosis ; Mitochondria/enzymology* ; Protozoan Proteins/metabolism*

OBJECTIVES: To investigate the therapeutic mechanism of Wendan Decoction for phlegm syndrome in rats with metabolic syndrome (MS). METHODS: Forty Wistar rats were randomly divided into normal control group (n=8) and 3 phlegm syndrome model groups (induced by high-fat, high-sugar, and high-salt feeding and a single-dose intraperitoneal STZ injection; n=24) treated with daily gavage of saline, Wendan Decoction (3.6 g/kg), or metformin (0.1 g/kg) for 4 weeks. General conditions and glucose and lipid metabolism parameters of the rats were monitored, and serum LPS, liver histopathology, hepatic expressions of FXR, CYP7A1 and FGFR4 and ileal expressions of FXR and FGF15 were examined. Gut microbiota structure was analyzed using 16S rDNA sequencing, and serum bile acids were quantified with UHPLC-MS/MS. RESULTS: The rat models of phlegm syndrome exhibited severe hepatic steatosis and necrosis, increased body weight, abdominal circumference, Lee's index, FBG, FINS, HOMA-IR, TG, TC, LDL and LPS, and decreased HDL level. The abundance of Bacteroidetes, Megamonas, and Bacteroides in gut microbiota increased while Firmicutes, Lachnospiraceae_NK4A136_group, isohyodeoxycholic acid, and glycohyodeoxycholic acid decreased significantly; hepatic FXR and FGFR4 expressions and ileal FXR and FGF15 expressions decreased while hepatic CYP7A1 expression increased significantly in the rat models. Treatment with Wendan Decoction effectively alleviated hepatic pathology, reduced body weight and abdominal circumference, improved glucose and lipid metabolic profiles and gut microbiota structure, and reversed the changes in hepatic and ileal protein expressions. Correlation analysis revealed that Firmicutes and Lachnospiraceae_NK4A136_group were positively correlated while Bacteroidetes, Megamonas and Bacteroides were negative correlated with the levels of isohyodeoxycholic acid and hyodeoxycholic acid. CONCLUSIONS Wendan Decoction can significantly improve metabolic profiles in rats with phlegm syndrome of MS possibly by regulating the intestinal flora-bile acid axis to modulate the intestinal flora structure and maintain bile acid homeostasis via the FXR signaling pathway.
Animals ; Gastrointestinal Microbiome/drug effects* ; Metabolic Syndrome/microbiology* ; Bile Acids and Salts/metabolism* ; Rats, Wistar ; Drugs, Chinese Herbal/therapeutic use* ; Rats ; Male ; Fibroblast Growth Factors/metabolism* ; Liver/metabolism* ; Cholesterol 7-alpha-Hydroxylase/metabolism* ; Receptors, Cytoplasmic and Nuclear/metabolism*

Objective:To investigate the value of coronary CTA (CCTA)-based traditional features and radiomics of plaque in the identification of culprit lesions that caused acute myocardial infarction (AMI).Methods:This was a retrospective multicenter study. From July 2016 to November 2023, a total of 344 patients from the Affiliated Hospital of Qingdao University (training cohort, n=184), Shandong Provincial Hospital Affiliated to Shandong First Medical University (validation cohort, n=88) and Qilu Hospital of Shandong University (test cohort, n=72) who received percutaneous coronary intervention (PCI) due to AMI and underwent CCTA within 48 hours of AMI were enrolled. The culprit plaques and non-culprit plaques were identified using a combination of electrocardiogram, CCTA, and angiographic findings. The vessel, plaque location, plaque type, Coronary Artery Disease-Reporting and Data System (CAD-RADS) score, high-risk plaque characteristics, plaque length, plaque volume, and burden were analyzed, and 1 904 radiomics features were extracted for each plaque. The traditional imaging model, the radiomics model, and the combined model were established by using multivariate Logistic regression analysis. The area under the receiver operating characteristic curve (AUC) was used to evaluate the performance of each model in identifying culprit lesions. The DeLong test was used for the comparison of AUC between every two models. The net reclassification index (NRI) was used to evaluate the incremental value of the combined model to the traditional imaging model and the radiomics model. The decision curve analysis (DCA) was used to assess the clinical net benefit of these models. A correlation heatmap was used to evaluate the correlation between the radiomics score and traditional CCTA factors. The interpretable analysis of the decision process of the combined model was performed by the Shapley Additive exPlanations (SHAP). Results:In the validation cohort and the test cohort, the AUC of the traditional imaging model developed by the vessel, plaque type, positive remodeling and CAD-RADS score was 0.898 (95% CI 0.869-0.922) and 0.881 (95% CI 0.848-0.910), respectively. The radiomics model developed by six radiomics features was 0.863 (95% CI 0.831-0.891) and 0.863 (95% CI 0.827-0.864), respectively. The AUC of the combined model was 0.930 (95% CI 0.905-0.950)and 0.919 (95% CI 0.889-0.942), respectively. In the validation cohort and the test cohort, the AUC of the combined model was higher than that of the traditional imaging model ( Z=4.013, 4.272, P<0.001) and that of the radiomics model ( Z=4.819, 3.784, P<0.001), respectively. In the validation cohort, the combined model yielded an NRI of 20.43% (95% CI 10.43%-30.44%, P<0.001) and 20.21% (95% CI 9.62%-30.80%, P<0.001) for identifying culprit lesions compared with the traditional imaging model and the radiomics model, respectively. In the test cohort, the combined model yielded an NRI of 28.05% (95% CI 16.72%-39.38%, P<0.001) and 23.57% (95% CI 13.58%-33.56%, P<0.001) for identifying culprit lesions compared with the traditional imaging model and the radiomics model, respectively. DCA showed the combined model had the highest clinical net benefit. The correlation heatmap showed the radiomics score was not correlated or only weakly correlated with traditional CCTA factors. SHAP indicated the radiomics and CAD-RADS score contributed significantly to the model. Conclusion:The CCTA-based traditional features and radiomics of plaque have favorable performance for the identification of culprit plaques in patients with AMI.

Objective To investigate the therapeutic mechanism of Wendan Decoction for phlegm syndrome in rats with metabolic syndrome(MS).Methods Forty Wistar rats were randomly divided into normal control group(n=8)and 3 phlegm syndrome model groups(induced by high-fat,high-sugar,and high-salt feeding and a single-dose intraperitoneal STZ injection;n=24)treated with daily gavage of saline,Wendan Decoction(3.6 g/kg),or metformin(0.1 g/kg)for 4 weeks.General conditions and glucose and lipid metabolism parameters of the rats were monitored,and serum LPS,liver histopathology,hepatic expressions of FXR,CYP7A1 and FGFR4 and ileal expressions of FXR and FGF15 were examined.Gut microbiota structure was analyzed using 16S rDNA sequencing,and serum bile acids were quantified with UHPLC-MS/MS.Results The rat models of phlegm syndrome exhibited severe hepatic steatosis and necrosis,increased body weight,abdominal circumference,Lee's index,FBG,FINS,HOMA-IR,TG,TC,LDL and LPS,and decreased HDL level.The abundance of Bacteroidetes,Megamonas,and Bacteroides in gut microbiota increased while Firmicutes,Lachnospiraceae_NK4A136_group,isohyodeoxycholic acid,and glycohyodeoxycholic acid decreased significantly;hepatic FXR and FGFR4 expressions and ileal FXR and FGF15 expressions decreased while hepatic CYP7A1 expression increased significantly in the rat models.Treatment with Wendan Decoction effectively alleviated hepatic pathology,reduced body weight and abdominal circumference,improved glucose and lipid metabolic profiles and gut microbiota structure,and reversed the changes in hepatic and ileal protein expressions.Correlation analysis revealed that Firmicutes and Lachnospiraceae_NK4A136_group were positively correlated while Bacteroidetes,Megamonas and Bacteroides were negative correlated with the levels of isohyodeoxycholic acid and hyodeoxycholic acid.Conclusion Wendan Decoction can significantly improve metabolic profiles in rats with phlegm syndrome of MS possibly by regulating the intestinal flora-bile acid axis to modulate the intestinal flora structure and maintain bile acid homeostasis via the FXR signaling pathway.

In recent years, the relationship between intestinal flora dysbiosis and abnormal bile acid metabolism and anxiety has received widespread attention. This article discussed the biological mechanism of the pathogenesis of anxiety from the perspective of intestinal flora and bile acid metabolism, in order to provide new ideas and theoretical basis for the TCM prevention and treatment of anxiety. According to TCM, spleen and stomach belong to earth, liver and gallbladder belong to wood, when spleen qi is healthy, the normal distribution of water and grain essence can be achieved, so that the liver can be nourished, qi and blood is sufficient, and its excretory function is normal, and the bile is sufficient. Once the spleen is not healthy, the distribution of water and grain essence is good, affecting the metabolism of fluids, resulting in phlegm and dampness, the qi is not smooth, can affect the liver's excretory function; liver and wood depression for a long time, transgressing the spleen and earth, qi and blood lack of biochemical sources, the formation of soil congestion and wood depression of the pathological phenomenon will appear. From the viewpoint of modern medicine and molecular biology, changes in the structure of intestinal flora affect the organism through neurological, endocrine, immune and metabolic pathways, which is consistent with the pathogenesis of "congestion of the earth"; disorders in the metabolism of bile acids can lead to changes in neurotransmitters and synaptic structure in the brain, causing anxiety symptoms, which is consistent with the characteristics of "wood depression". The bile acid metabolism disorder can cause neurotransmitter and synaptic structure changes in the brain, causing anxiety symptoms, which is consistent with the characteristics of "wood depression". It is important to regulate the intestinal flora and bile acid metabolism pathway to ease the liver and strengthen the spleen to improve anxiety.

ObjectiveTo investigate a suspected outbreak of healthcare-associated infection with carbapenem-resistant Klebsiella pneumoniae (CRKP) in a geriatric emergency ward, and to provide references for the prevention and control of multidrug-resistant bacteria in a hospital in Shanghai. MethodsOn-site epidemiological investigation, combined with environmental hygiene monitoring and pulsed field gel electrophoresis (PFGE) molecular typing method, were adopted to investigate a suspected outbreak of CRKP infection in the geriatric emergency ward of a hospital from October to November 2022, aiming at finding out factors caused the outbreak before taking corresponding control measures. ResultsA total of 3 cases of healthcare-associated CRKP infection were identified, of which 2 cases were homologous to a previous case of community-associated CRKP infection. What’s more, the 2 cases lived in the same ward with the latter and with adjacent beds, but the third case was non-homologous to the community-associated infection case. A total of 46 samples were collected from the environmental surfaces and the hands of healthcare workers, of which 7 samples tested positive for CRKP and were identical to the strains from the 2 healthcare-associated infection cases and the 1 community-associated infection case, originating from the bedrails, bedside tables, surface of non-invasive ventilator, bed curtains and panels of monitoring equipment, with a detection rate of 15.22%. But none of the 11 samples from the hands of healthcare workers tested positive for CRKP. The outbreak was effectively controlled after taking specific prevention and control measures such as strengthening personnel management, intensifying environmental cleaning and disinfection and strictly enforcing hand hygiene among healthcare workers. Subsequently, no similar new cases were reported during the 14-day follow-up period. ConclusionIncomplete environmental cleaning and disinfection, as well as inadequate enforcement of hand hygiene among heatheare workers may have contributed to the suspected outbreak of CRKP in the geriatric emergency ward. Early warning and timely investigation of suspected outbreaks of multidrug-resistant bacteria are crucial for preventing and controlling such outbreaks in hospitals.

Eimeria labbeana is a major pathogen of pigeon coccidiosis,causing damage to the intes-tinal epithelial cells of pigeons,which leads to gut injury,diarrhea,decreased production perform-ance,and even death.There have been no research on the pathogen characteristics of isolated strains in China.In this study,1 008 fecal samples were collected from nine cities in Zhejiang Prov-ince,including Hangzhou,Ningbo,Wenzhou,Shaoxing,Huzhou,Jinhua,Quzhou,Taizhou and Lishui.The samples were examined using the McMaster counting method to quantify oocysts per gram of feces,and coccidia species in positive samples were identified through microscopy.Patho-gens were isolated and purified with methods of single oocyst pickout under the microscopy and passage in coccidia-free pigeon,followed by a detailed study of their characteristics.Our findings demonstrated an overall infection rate of 55.8％(562/1 008)for pigeon coccidia in the surveyed ar-eas,with E.labbeana present across all farms.A strain of E.labbeana isolated from Zhejiang was successfully obtained and designated E.labbeana-ZJ.PCR identification and sequence alignment showed that this Zhejiang isolate shared a 99.67％similarity in the 18S rRNA gene sequence with the Australian strain(KT305927.1)and clustered into a small subgroup.Pathogenicity and oocyst shedding patterns were assessed through animal infection experiments,revealing a 4 days latent period,with peak infection occurring on the 8th day.Following infection,notable clinical symptoms emerged,with significant intestinal damage,and changes in body weight,indicating moderate path-ogenicity.The results enriched the epidemiological survey data of pigeon coccidiosis in China,and provide a new basis for further research and effective control measures against pigeon coccidiosis.

Eimeria labbeana is a major pathogen of pigeon coccidiosis,causing damage to the intes-tinal epithelial cells of pigeons,which leads to gut injury,diarrhea,decreased production perform-ance,and even death.There have been no research on the pathogen characteristics of isolated strains in China.In this study,1 008 fecal samples were collected from nine cities in Zhejiang Prov-ince,including Hangzhou,Ningbo,Wenzhou,Shaoxing,Huzhou,Jinhua,Quzhou,Taizhou and Lishui.The samples were examined using the McMaster counting method to quantify oocysts per gram of feces,and coccidia species in positive samples were identified through microscopy.Patho-gens were isolated and purified with methods of single oocyst pickout under the microscopy and passage in coccidia-free pigeon,followed by a detailed study of their characteristics.Our findings demonstrated an overall infection rate of 55.8％(562/1 008)for pigeon coccidia in the surveyed ar-eas,with E.labbeana present across all farms.A strain of E.labbeana isolated from Zhejiang was successfully obtained and designated E.labbeana-ZJ.PCR identification and sequence alignment showed that this Zhejiang isolate shared a 99.67％similarity in the 18S rRNA gene sequence with the Australian strain(KT305927.1)and clustered into a small subgroup.Pathogenicity and oocyst shedding patterns were assessed through animal infection experiments,revealing a 4 days latent period,with peak infection occurring on the 8th day.Following infection,notable clinical symptoms emerged,with significant intestinal damage,and changes in body weight,indicating moderate path-ogenicity.The results enriched the epidemiological survey data of pigeon coccidiosis in China,and provide a new basis for further research and effective control measures against pigeon coccidiosis.

Objective:To investigate the value of coronary CTA (CCTA)-based traditional features and radiomics of plaque in the identification of culprit lesions that caused acute myocardial infarction (AMI).Methods:This was a retrospective multicenter study. From July 2016 to November 2023, a total of 344 patients from the Affiliated Hospital of Qingdao University (training cohort, n=184), Shandong Provincial Hospital Affiliated to Shandong First Medical University (validation cohort, n=88) and Qilu Hospital of Shandong University (test cohort, n=72) who received percutaneous coronary intervention (PCI) due to AMI and underwent CCTA within 48 hours of AMI were enrolled. The culprit plaques and non-culprit plaques were identified using a combination of electrocardiogram, CCTA, and angiographic findings. The vessel, plaque location, plaque type, Coronary Artery Disease-Reporting and Data System (CAD-RADS) score, high-risk plaque characteristics, plaque length, plaque volume, and burden were analyzed, and 1 904 radiomics features were extracted for each plaque. The traditional imaging model, the radiomics model, and the combined model were established by using multivariate Logistic regression analysis. The area under the receiver operating characteristic curve (AUC) was used to evaluate the performance of each model in identifying culprit lesions. The DeLong test was used for the comparison of AUC between every two models. The net reclassification index (NRI) was used to evaluate the incremental value of the combined model to the traditional imaging model and the radiomics model. The decision curve analysis (DCA) was used to assess the clinical net benefit of these models. A correlation heatmap was used to evaluate the correlation between the radiomics score and traditional CCTA factors. The interpretable analysis of the decision process of the combined model was performed by the Shapley Additive exPlanations (SHAP). Results:In the validation cohort and the test cohort, the AUC of the traditional imaging model developed by the vessel, plaque type, positive remodeling and CAD-RADS score was 0.898 (95% CI 0.869-0.922) and 0.881 (95% CI 0.848-0.910), respectively. The radiomics model developed by six radiomics features was 0.863 (95% CI 0.831-0.891) and 0.863 (95% CI 0.827-0.864), respectively. The AUC of the combined model was 0.930 (95% CI 0.905-0.950)and 0.919 (95% CI 0.889-0.942), respectively. In the validation cohort and the test cohort, the AUC of the combined model was higher than that of the traditional imaging model ( Z=4.013, 4.272, P<0.001) and that of the radiomics model ( Z=4.819, 3.784, P<0.001), respectively. In the validation cohort, the combined model yielded an NRI of 20.43% (95% CI 10.43%-30.44%, P<0.001) and 20.21% (95% CI 9.62%-30.80%, P<0.001) for identifying culprit lesions compared with the traditional imaging model and the radiomics model, respectively. In the test cohort, the combined model yielded an NRI of 28.05% (95% CI 16.72%-39.38%, P<0.001) and 23.57% (95% CI 13.58%-33.56%, P<0.001) for identifying culprit lesions compared with the traditional imaging model and the radiomics model, respectively. DCA showed the combined model had the highest clinical net benefit. The correlation heatmap showed the radiomics score was not correlated or only weakly correlated with traditional CCTA factors. SHAP indicated the radiomics and CAD-RADS score contributed significantly to the model. Conclusion:The CCTA-based traditional features and radiomics of plaque have favorable performance for the identification of culprit plaques in patients with AMI.

Breast cancer remains a leading cause of mortality in women worldwide.Triple-negative breast cancer(TNBC)is a particularly aggressive subtype characterized by rapid progression,poor prognosis,and lack of clear therapeutic targets.In the clinic,delineation of tumor heterogeneity and development of effective drugs continue to pose considerable challenges.Within the scope of our study,high hetero-geneity inherent to breast cancer was uncovered based on the landscape constructed from both tumor and healthy breast tissue samples.Notably,TNBC exhibited significant specificity regarding cell prolif-eration,differentiation,and disease progression.Significant associations between tumor grade,prog-nosis,and TNBC oncogenes were established via pseudotime trajectory analysis.Consequently,we further performed comprehensive characterization of the TNBC microenvironment.A crucial epithelial subcluster,E8,was identified as highly malignant and strongly associated with tumor cell proliferation in TNBC.Additionally,epithelial-mesenchymal transition(EMT)-associated fibroblast and M2 macrophage subclusters exerted an influence on E8 through cellular interactions,contributing to tumor growth.Characteristic genes in these three cluster cells could therefore serve as potential therapeutic targets for TNBC.The collective findings provided valuable insights that assisted in the screening of a series of therapeutic drugs,such as pelitinib.We further confirmed the anti-cancer effect of pelitinib in an orthotopic 4T1 tumor-bearing mouse model.Overall,our study sheds light on the unique characteristics of TNBC at single-cell resolution and the crucial cell types associated with tumor cell proliferation that may serve as potent tools in the development of effective anti-cancer drugs.