Objective To establish a rapid detection method for Clostridium botulinum in food.Methods A rapid detection method based on RPA-CRISPR/Cas12a was developed by integrating recombinase polymerase amplification(RPA)with the CRISPR-Cas12a system.After the reaction conditions were optimized,the method's sensitivity,specificity,and usefulness were methodically confirmed.Results and Conclusion The optimized method achieved detection within 1 hour,with a limit of detection(LOD)of 1.91 copies/μL.No cross-reactivity was observed with non-target pathogens.The RPA-CRISPR/Cas12a-based detection method developed in this study exhibits high specificity,sensitivity,and operational simplicity and may provide a feasible solution for the rapid detection of foodborne pathogens.

Objective To observe contrast-enhanced ultrasound(CEUS)manifestations of nontumorous intrahepatic vascular shunts.Methods Data of 11 patients with intrahepatic vascular shunts were retrospectively analyzed,including 3 cases of intrahepatic arteriovenous shunts,3 cases of hepatic arterioportal shunts,1 case of intrahepatic arterioportal shunt complicated with intrahepatic arteriovenous shunt and 4 cases of intrahepatic portosystemic shunts.The conventional ultrasound and CEUS manifestations were observed.Results In 3 cases of intrahepatic arteriovenous shunts and 3 cases of hepatic arterioportal shunts,CEUS showed early enhancement of draining veins,early enhancement of local liver parenchyma without washout,while conventional ultrasound showed partial shunts in 2 cases with continuous enhancement showed on CEUS.The findings of 1 case of intrahepatic arterioportal shunt complicated with intrahepatic arteriovenous shunt were similar to the above two.Among 4 cases of intrahepatic portosystemic shunts,conventional ultrasound displayed all shunts,continuous enhancement of the draining hepatic vein and shunts were observed with CEUS,and transient low enhancement of local liver parenchyma were noticed in 2 cases.Conclusion Nontumorous intrahepatic vascular shunts could be divided into microscopic shunts that ultrasound could not directly display and shunts ultrasound could display.The former was characterized by early enhancement of the draining vein(portal vein or hepatic vein)on CEUS,some with early enhancement and no washout of local liver parenchyma.The latter was characterized by continuous enhancement of drainage vein and shunts,with or without transient low enhancement of local liver parenchyma.

The purpose of this study was to investigate the effect of medium-chain fatty acids(MC-FAs)caprylic acid(C8∶0)on lipid metabolism of calf hepatocytes.Primary calf hepatocytes were extracted and cultured,and 1.2 mmol/L nonesterified fatty acids(NEFAs)were added to the hep-atocytes to construct a model of hepatic lipid deposition in primary calf hepatocytes,Five process-ing groups have been set up:Control group(Ctrl),NEFA added group(NEFA),C8∶0 1.2 mmol/L treatment group(C8∶0 1.2),NEFA+C8∶0 0.2 mmol/L treatment group(NEFA+C8∶00.2),C8∶0 0.2 mmol/L treatment group(C8∶0 0.2).Stimulate calf liver cells for 12 hours,and the levels of triglyceride(TG),lipid oxidation(MDA),hydrogen peroxide(H2O2)and total SOD activity were detected by biochemical kit,and FAS,a protein related to lipid synthesis,was detec-ted by Western blot.The results showed that compared with the control group,the concentrations of TG,MDA and H2O2 in NEFA group increased significantly(P＜0.01),and the activity of SOD decreased significantly(P＜0.05).The protein expression levels of FAS,ACC1,DGAT2 and SREBP-1C were significantly up-regulated(P＜0.01),while the expression level of CPT1A was significantly down-regulated(P＜0.01).Compared with the NEFA group,the protein expression levels of SREBP-1C and DGAT2 in the NEFA+C8∶0(concentration 0.2 mmol/L)group de-creased significantly(P＜0.05),and the protein expression level of fatty acid β-oxidation related molecule CPT1A was slightly higher than that in the NEFA group,but there was no statistical sig-nificance(P＞0.05),and the MDA level in hepatocytes decreased significantly(P＜0.05).In a word,the results of this study show that C8∶0 has antioxidant effect,which can effectively reduce the liver injury caused by oxidative stress,regulate the expression of liver fat gene,and then pro-tect liver injury.

To explore the impacts of TGR5 on liver lipid metabolism and bile acid synthesis in dairy cows with fatty liver.Liver tissues of healthy cows and cows with fatty liver were collected through puncture technique.The protein and mRNA expressions of lipid synthesis-related factors ACC1,FAS,SREBF1,lipid oxidation factor CPT1A,and bile acid synthesis-related factors CYP8B1,CYP7B1,CYP27A1 were detected by Western blot and fluorescent quantitative PCR.Moreover,the mRNA levels of CYP7B1 were determined.Primary hepatocytes of 1-day-old calves were extracted and cultured in vitro,and four treatment groups were established,namely Control,NEFA,INT-777,and the INT-777+NEFA group.The concentration of NEFA group was 1.2 mmol/L,the con-centration of INT-777 group was 1 μmol/L,and the concentration of INT-777+NEFA group was 1.2 mmol/L NEFA and 1 μmol/L INT-777 simultaneously.After 12 h of stimulation,cells were collected,and the protein and mRNA levels of ACC1,FAS,SREBF1,CPT1A,CYP8B1,CYP7A1,CYP27A1,and the mRNA levels of CYP7B1 were detected by Western blot and fluorescent quanti-tative PCR.The content of lipid droplets and TG in the cells were detected by flow cytometry and kit.The results demonstrated that compared with healthy cows,the protein and mRNA expressions of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the liver tissues of fatty liver cows were upreg-ulated,while the protein and mRNA levels of CPT1 A,CYP27A1,TGR5,and the mRNA levels of CYP7B1 were downregulated.In vitro experiments revealed that compared with the Control group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the NEFA group were upregulated,and the protein and mRNA levels of CPT1A,CYP27A1,and TGR5,as well as the mRNA level of CYP7B1,were downregulated.Compared with the NEFA group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,CYP7A1 were downregulated in the INT-777+NEFA group,while the protein and mRNA levels of CPT1A CYP27A1,and TGR5 as well as the mRNA level of CYP7B1,were upregulated.The results of flow cytometry and the kit indicated that the lipid droplets and TG content in the NEFA group were upregulated compared with the Control group,while the lipid droplets and TG content in the INT-777+NEFA group were downregulated compared with the NEFA group.The above results suggested that the addition of TGR5 agonist promoted the expression of TGR5 and ameliorated the abnormal lipid metabolism and bile acid synthesis in the liver of dairy cows with fatty liver.

Objective:To analyze the association between unhealthy lifestyles and depressive symptoms, anxiety symptoms and stress among secondary school students in Jiangsu Province.Methods:From September to November 2023, a multistage stratified cluster random sampling method was used to select secondary school students from 13 districts and cities in Jiangsu Province. A questionnaire survey was conducted on their unhealthy lifestyles (low physical activity, smoking, drinking, internet addiction, poor sleep quality, and unhealthy diet), as well as their depressive symptoms, anxiety symptoms and stress. The multivariate logistic regression model and mixed graph model were used to construct a network and analyze the association between unhealthy lifestyles and depressive symptoms, anxiety symptoms and stress.Results:A total of 81 414 secondary school students were finally included in this study, including 39 725 (48.79%) female students and 41 689 (51.21%) male students. The prevalence of depressive symptoms, anxiety symptoms and stress were 18.55%, 32.09% and 12.91%, respectively. The multivariate logistic regression model showed that after adjusting for age, gender, urban-rural status, residential status, and family type, compared with students without unhealthy lifestyles, students with low physical activity, smoking, alcohol consumption, internet addiction, poor sleep quality, and unhealthy diet had a significantly increased risk of depressive symptoms ( OR=1.12, 95% CI:1.07-1.17; OR=1.60, 95% CI: 1.49-1.72; OR=1.79, 95% CI: 1.71-1.88; OR=3.05, 95% CI: 2.77-3.36; OR=6.66, 95% CI: 6.40-6.93; OR=1.29, 95% CI: 1.24-1.34) and a significantly increased risk of anxiety symptoms ( OR=1.09, 95% CI: 1.05-1.13; OR=1.42, 95% CI: 1.33-1.52; OR=1.76, 95% CI: 1.69-1.83; OR=2.40, 95% CI: 2.17-2.65; OR=5.79, 95% CI: 5.59-6.00; OR=1.16, 95% CI: 1.12-1.21). Students who smoked, drank alcohol, had internet addiction, and had poor sleep quality had a significantly increased risk of stress ( OR=1.49, 95% CI: 1.38-1.61; OR=1.79, 95% CI: 1.70-1.89; OR=2.25, 95% CI: 2.04-2.48; OR=6.74, 95% CI: 6.43-7.06). The node with poor sleep quality (bridge strength=0.48) in the network constructed by the mixed graph model had the strongest centrality of the bridge connecting unhealthy lifestyles with depressive symptoms, anxiety symptoms and stress. Conclusion:Low physical activity, smoking, alcohol consumption, internet addiction, poor sleep quality, and unhealthy diet increase the risk of depressive symptoms and anxiety symptoms among Jiangsu Province secondary school students. Smoking, alcohol consumption, internet addiction, and poor sleep quality increase the risk of stress among Jiangsu Province secondary school students. Sleep quality is an important intervention target for Jiangsu Province secondary school students to alleviate their negative emotions.

Objective:To explore the value of ultrasound imaging characteristics combined with clinical indicators in assessing the prognosis of patients with pancreatic ductal adenocarcinoma (PDAC).Methods:A retrospective analysis was conducted for patients who underwent pancreatic contrast-enhanced ultrasound (CEUS) from September 2017 to October 2023 at Peking Union Medical College Hospital and were diagnosed with PDAC based on pathological findings. Various parameters were recorded, including CA19-9 levels, tumor size, location, morphologic features, echogenicity, presence of internal cystic components, dilatation of the main pancreatic duct, peripheral vascular invasion, CEUS characteristics, presence or absence of liver metastasis, and treatment methods. In April 2024, patient survival information was obtained through telephone follow-up or review of medical records. Based on the results of the cox regression model analysis, a nomogram model of the risk of death was developed. The receiver operating characteristic (ROC) curves were applied to evaluate the predictive efficacy of the model. The calibration curves were plotted to evaluate the accuracy of the model, and clinical decision curves were used to evaluate the clinical benefit of the model.Results:This study included a total of 207 patients with PDAC. As of April 2024, 71 patients were alive and 136 died, with a median survival time of 14 months (95% CI: 12 -17). Multivariate analysis confirmed that the elevated CA19-9 ( HR=1.689, 95% CI: 1.102-2.588), tumor size >4 cm ( HR=1.641, 95% CI: 1.159-2.322), taller-than-wide shapes ( HR=1.450, 95% CI: 1.019-2.065), incomplete hypo-enhancement ( HR=1.618, 95% CI: 1.100-2.380), and liver metastasis ( HR=1.687, 95% CI: 1.175-2.423) were independent risk factors for survival in patients with PDAC. A nomogram model was further constructed for 6-month, 12-month and 3-year survival of patients with PDAC. The areas under the ROC curve were 0.679, 0.705 and 0.815, respectively. The calibration curves suggested that the model was more accurate, and the clinical decision curves showed that the model had a better clinical benefit. Conclusion:The combined use of ultrasound imaging characteristics and clinical indicators could effectively predict the prognosis of PDAC patients. Specifically, tumor size >4 cm, taller-than-wide shapes, incomplete hypo-enhancement, elevated CA19-9, and the presence of liver metastasis are correlated with poorer survival outcomes. The nomogram model constructed on the basis of these factors can be used to assess the survival of patients with PDAC.

To explore the impacts of TGR5 on liver lipid metabolism and bile acid synthesis in dairy cows with fatty liver.Liver tissues of healthy cows and cows with fatty liver were collected through puncture technique.The protein and mRNA expressions of lipid synthesis-related factors ACC1,FAS,SREBF1,lipid oxidation factor CPT1A,and bile acid synthesis-related factors CYP8B1,CYP7B1,CYP27A1 were detected by Western blot and fluorescent quantitative PCR.Moreover,the mRNA levels of CYP7B1 were determined.Primary hepatocytes of 1-day-old calves were extracted and cultured in vitro,and four treatment groups were established,namely Control,NEFA,INT-777,and the INT-777+NEFA group.The concentration of NEFA group was 1.2 mmol/L,the con-centration of INT-777 group was 1 μmol/L,and the concentration of INT-777+NEFA group was 1.2 mmol/L NEFA and 1 μmol/L INT-777 simultaneously.After 12 h of stimulation,cells were collected,and the protein and mRNA levels of ACC1,FAS,SREBF1,CPT1A,CYP8B1,CYP7A1,CYP27A1,and the mRNA levels of CYP7B1 were detected by Western blot and fluorescent quanti-tative PCR.The content of lipid droplets and TG in the cells were detected by flow cytometry and kit.The results demonstrated that compared with healthy cows,the protein and mRNA expressions of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the liver tissues of fatty liver cows were upreg-ulated,while the protein and mRNA levels of CPT1 A,CYP27A1,TGR5,and the mRNA levels of CYP7B1 were downregulated.In vitro experiments revealed that compared with the Control group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the NEFA group were upregulated,and the protein and mRNA levels of CPT1A,CYP27A1,and TGR5,as well as the mRNA level of CYP7B1,were downregulated.Compared with the NEFA group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,CYP7A1 were downregulated in the INT-777+NEFA group,while the protein and mRNA levels of CPT1A CYP27A1,and TGR5 as well as the mRNA level of CYP7B1,were upregulated.The results of flow cytometry and the kit indicated that the lipid droplets and TG content in the NEFA group were upregulated compared with the Control group,while the lipid droplets and TG content in the INT-777+NEFA group were downregulated compared with the NEFA group.The above results suggested that the addition of TGR5 agonist promoted the expression of TGR5 and ameliorated the abnormal lipid metabolism and bile acid synthesis in the liver of dairy cows with fatty liver.

Objective To observe contrast-enhanced ultrasound(CEUS)manifestations of nontumorous intrahepatic vascular shunts.Methods Data of 11 patients with intrahepatic vascular shunts were retrospectively analyzed,including 3 cases of intrahepatic arteriovenous shunts,3 cases of hepatic arterioportal shunts,1 case of intrahepatic arterioportal shunt complicated with intrahepatic arteriovenous shunt and 4 cases of intrahepatic portosystemic shunts.The conventional ultrasound and CEUS manifestations were observed.Results In 3 cases of intrahepatic arteriovenous shunts and 3 cases of hepatic arterioportal shunts,CEUS showed early enhancement of draining veins,early enhancement of local liver parenchyma without washout,while conventional ultrasound showed partial shunts in 2 cases with continuous enhancement showed on CEUS.The findings of 1 case of intrahepatic arterioportal shunt complicated with intrahepatic arteriovenous shunt were similar to the above two.Among 4 cases of intrahepatic portosystemic shunts,conventional ultrasound displayed all shunts,continuous enhancement of the draining hepatic vein and shunts were observed with CEUS,and transient low enhancement of local liver parenchyma were noticed in 2 cases.Conclusion Nontumorous intrahepatic vascular shunts could be divided into microscopic shunts that ultrasound could not directly display and shunts ultrasound could display.The former was characterized by early enhancement of the draining vein(portal vein or hepatic vein)on CEUS,some with early enhancement and no washout of local liver parenchyma.The latter was characterized by continuous enhancement of drainage vein and shunts,with or without transient low enhancement of local liver parenchyma.

Objective:To investigate the application value of scenario simulation combined with standardized patient teaching in the training of doctor-patient communication skills among interns.Methods:A total of 110 clinical medicine interns were selected from Qilu Hospital of Shandong University and were divided into experimental group and control group using a random number table, with 55 interns in each group. The interns in the control group received traditional lecturing, and those in the experimental group received scenario simulation combined with standardized patient case-based teaching. The SEGUE Communication Skill Evaluation Scale and Patient Satisfaction Questionnaire were used to assess the improvement in communication skills in both groups, and Physician Self-Evaluation Scale and Course Satisfaction Questionnaire were used to assess the effectiveness of the course and the degree of satisfaction with the course. SPSS 26.0 was used for the t-test, the Mann-Whitney U test, and the rank sum test. Results:After training, both groups showed significant improvements in communication skills and patient satisfaction [experimental group in terms of communication skills: (55.38±13.11) vs. (74.82±6.75), P<0.001; experimental group in terms of patient satisfaction: 39.00 (39.00, 42.00) vs. 81.00 (79.00, 83.00), P<0.01; control group in terms of communication skills: (56.53±12.34) vs. (65.45±10.18), P<0.001; control group in terms of patient satisfaction: 39.00 (39.00, 42.00) vs. 73.00 (68.00, 77.00), P<0.001], and the experimental group had significantly higher scores than the control group [communication skills: (74.82±6.75) vs. (65.45±10.18), P<0.001; patient satisfaction: 81.00 (79.00, 83.00) vs. 73.00 (68.00, 77.00), P<0.001]. The Physician Self-Evaluation Scale showed that the experimental group had a significant improvement compared with the control group ( P<0.05). The results of the Course Satisfaction Questionnaire showed that the degree of overall satisfaction of the course was only 87.28% in the control group, while all the interns in the experimental group were satisfied with the course ( P<0.001); the new teaching method showed great advantages ( P<0.001). Conclusions:In the training of doctor-patient communication skills among interns, the application of scenario simulation combined with standardized patient case-based teaching can significantly improve their abilities of the application of knowledge application, humanistic concern, and communication skills, and therefore, it is an effective and promising method for the training of doctor-patient communication skills.

The purpose of this study was to investigate the effect of medium-chain fatty acids(MC-FAs)caprylic acid(C8∶0)on lipid metabolism of calf hepatocytes.Primary calf hepatocytes were extracted and cultured,and 1.2 mmol/L nonesterified fatty acids(NEFAs)were added to the hep-atocytes to construct a model of hepatic lipid deposition in primary calf hepatocytes,Five process-ing groups have been set up:Control group(Ctrl),NEFA added group(NEFA),C8∶0 1.2 mmol/L treatment group(C8∶0 1.2),NEFA+C8∶0 0.2 mmol/L treatment group(NEFA+C8∶00.2),C8∶0 0.2 mmol/L treatment group(C8∶0 0.2).Stimulate calf liver cells for 12 hours,and the levels of triglyceride(TG),lipid oxidation(MDA),hydrogen peroxide(H2O2)and total SOD activity were detected by biochemical kit,and FAS,a protein related to lipid synthesis,was detec-ted by Western blot.The results showed that compared with the control group,the concentrations of TG,MDA and H2O2 in NEFA group increased significantly(P＜0.01),and the activity of SOD decreased significantly(P＜0.05).The protein expression levels of FAS,ACC1,DGAT2 and SREBP-1C were significantly up-regulated(P＜0.01),while the expression level of CPT1A was significantly down-regulated(P＜0.01).Compared with the NEFA group,the protein expression levels of SREBP-1C and DGAT2 in the NEFA+C8∶0(concentration 0.2 mmol/L)group de-creased significantly(P＜0.05),and the protein expression level of fatty acid β-oxidation related molecule CPT1A was slightly higher than that in the NEFA group,but there was no statistical sig-nificance(P＞0.05),and the MDA level in hepatocytes decreased significantly(P＜0.05).In a word,the results of this study show that C8∶0 has antioxidant effect,which can effectively reduce the liver injury caused by oxidative stress,regulate the expression of liver fat gene,and then pro-tect liver injury.