Objective:To establish a rapid detection method for pathogenic microorganisms by combining loop-mediated isothermal amplification(LAMP)and clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 12a(Cas12a)(CRISPR-Cas12a)system,and to evaluate its efficacy for detecting the thermolabile hemolysin(tlh)gene of Vibrio parahaemolyticus(Vp).Methods:Using the tlh gene of Vp as the target gene,LAMP primers and CRISPR RNA(crRNA)were designed to construct and optimize the optimal concentration ratio of each component in the LAMP-CRISPR detection system.Bacillus cereus,Staphylococcus aureus,and Escherichia coli were used as control groups,and the specificity,sensitivity,reproducibility and positive conformity rate were verified to establish a rapid LAMP-CRISPR/Cas12a method for detecting the tlh gene of Vp.Results:The method specifically detected Vp,while Bacillus cereus,Staphylococcus aureus,and Escherichia coli yielded negative results.The DNA extraction concentration of Vp was 190.67 mg·L-1 with an A(260)/(A280)ratio of 1.84.Under the reaction conditions of 37℃ with 80 cycles for 40 min using quantitative PCR(qPCR)method,when the concentrations of Cas12a protein and crRNA in the LAMP-CRISPR/Cas12a system were 50 nmol·L-1,the visual brightness and relative fluorescence intensity peaks were high.The sensitivity of LAMP CRISPR/Cas12a for detecting Vp DNA concentration could reach 10-6 mg·L-1.The reproducibility test results showed that different experimenters had consistent results in different experimental environments and times.Conclusion:The established LAMP-CRISPR/Cas12a method can rapidly detect the tlh gene of Vp with high sensitivity and specificity,and can achieve short-term visual detection in the field.

OBJECTIVES: To detect prfA and hly toxin genes of Listeria monocytogenes using polymerase chain reaction (PCR) and colloidal gold technology. METHODS: L. monocytogenes DNA was extracted by boiling method. With prfA and hly of L. monocytogenes as the target genes, the 5' ends of upstream and downstream primers of prfA gene were labeled with 6-FAM and biotin, and the 5' ends of upstream and downstream primers of hly gene were labeled with digoxin and biotin, respectively, to establish the toxin gene detection method. Using cloning transformation, sequencing analysis, cloning of positive control products, the detection kid was developed and its specificity, sensitivity, reproducibility and stability were tested, followed by verification with sample testing. RESULTS: The concentration of L. monocytogenes DNA extracted by boiling method was 148.81±0.97 ng/μL, and the A₂₆₀/A₂₈₀ ratio ranged from 1.8 to 2.0. The PCR products showed a 100% homology with the gene sequences in GenBank database after cloning, transformation and sequencing. The colloidal gold strip yielded positive results only for L. monocytogenes samples without cross-reactions with Staphylococcus aureus, Escherichia coli or Bacillus cereus, and its minimum detection limit was 10^-2 ng/μL, demonstrating a 10-fold greater sensitivity of the test than agarose gel electrophoresis. The test also showed good reproducibility of the results when performed by different operators with good stability of the test strips after storage for 6 to 12 months. The test results showed that this kit could accurately and quickly detect L.monocytogenes in the test samples. CONCLUSIONS The detection kit developed in this study can simultaneously detect prfA and hly toxin genes of L. monocytogenes with good specificity, sensitivity, reproducibility and stability for use in food safety inspection.
Listeria monocytogenes/isolation & purification* ; Gold Colloid ; Bacterial Toxins/genetics* ; Polymerase Chain Reaction/methods* ; Hemolysin Proteins/genetics* ; Bacterial Proteins/genetics* ; DNA, Bacterial/genetics* ; Food Microbiology ; Heat-Shock Proteins

Background and Aims:Colorectal cancer(CRC)has a complex pathogenesis,and current treatments remain limited in efficacy for advanced metastatic disease.Dasatinib is a multi-target tyrosine kinase inhibitor that has shown potential antitumor activity in various solid tumors.This study aimed to evaluate the causal relationships between dasatinib-related target genes and CRC based on genetic variation,and to explore the mediating role of immune cells,thereby providing genetic epidemiological evidence for the prevention and targeted therapy of CRC.Methods:Dasatinib-related target genes were identified through DrugBank,and the corresponding eQTLs,GWAS data for CRC(ebi-a-GCST90018808),and 731 immune-cell traits were obtained from the IEU OpenGWAS database.A two-sample Mendelian randomization(MR)framework with a two-step mediation approach was applied:first,to assess the causal relationship between dasatinib target genes(as exposures)and CRC;second,to evaluate the causal effects between target genes and immune cells,as well as between immune cells and CRC;and finally,to calculate the proportion of mediated effects.Wald ratio,inverse-variance weighted(IVW),MR-Egger,MR-PRESSO,Cochran's Q,I2,and leave-one-out analyses were used to examine heterogeneity,horizontal pleiotropy,and robustness.Results:MR results showed that dasatinib-associated inhibition of ABL1 was significantly associated with a reduced risk of CRC(OR=0.511 0,95%CI=0.323 1-0.808 0,P=0.004 1).Inhibition of YES1 was also associated with decreased CRC risk(IVW OR=0.889 9,95%CI=0.811 6-0.975 8,P=0.013 1),with no evident heterogeneity or horizontal pleiotropy among the corresponding SNPs.Further analysis revealed that dasatinib-related inhibition of YES1 significantly reduced the levels of IgD-CD24-AC level(OR=0.818 0,95%CI=0.678 2-0.986 7,P=0.035 7),and this immune cell subset itself was identified as a risk factor for CRC(OR=1.105 7,95%CI=1.029 6-1.187 5,P=0.005 7).Mediation analysis indicated that IgD-CD24-AC accounted for-9.89%and 17.31%of the mediation effects in the ABL1→CRC and YES1→CRC pathways,respectively.Conclusion:Genetic evidence from MR suggests dasatinib-target genes ABL1 and YES1 are causally linked to reduced CRC risk,with IgD-CD24-AC partially mediating the YES1-related protective effect.These findings point to immune-mediated mechanisms underlying dasatinib's potential influence on CRC risk;further experimental validation and replication across populations are warranted.

Objective:To investigate the prevalence,related factors,and career implications of occupational burnout among resident physicians during standardized training in Inner Mongolia.Methods:This cross-sectional study used convenience sampling to enroll 2 891 resident physicians,assessing them with a self-developed general information questionnaire,socio-psychological scales,and a universal burnout inventory.Latent profile analysis clas-sified participants based on burnout dimensions,while logistic regression models examined the factors influencing burnout and their relationship with career choices.Results:Resident physicians were divided into low(59.0％),moderate(28.9％),and high(12.1％)burnout subgroups.Logistic regression showed that,compared to the low burnout group,significant risk factors included professional master's students(OR=0.44),working＞70 hours weekly(OR=0.63),＞4 night shifts monthly(OR=0.66),anxiety(mild OR=0.49;moderate OR=0.26;se-vere OR=0.14),depression(mild OR=0.38;moderate OR=0.24;severe OR=0.11),insomnia(mild OR=0.51;moderate OR=0.44;severe OR=0.38),low social support(OR=0.23),and low happiness index(OR=0.42).Male residents(OR=1.25)were less likely to experience burnout.High burnout residents were 5.11 times more likely to quit the medical profession.Conclusion:Training intensity and socio-psychological factors signifi-cantly contribute resident physicians' burnout levels,with severe burnout increase career attrition.

Objective:To investigate the prevalence,related factors,and career implications of occupational burnout among resident physicians during standardized training in Inner Mongolia.Methods:This cross-sectional study used convenience sampling to enroll 2 891 resident physicians,assessing them with a self-developed general information questionnaire,socio-psychological scales,and a universal burnout inventory.Latent profile analysis clas-sified participants based on burnout dimensions,while logistic regression models examined the factors influencing burnout and their relationship with career choices.Results:Resident physicians were divided into low(59.0％),moderate(28.9％),and high(12.1％)burnout subgroups.Logistic regression showed that,compared to the low burnout group,significant risk factors included professional master's students(OR=0.44),working＞70 hours weekly(OR=0.63),＞4 night shifts monthly(OR=0.66),anxiety(mild OR=0.49;moderate OR=0.26;se-vere OR=0.14),depression(mild OR=0.38;moderate OR=0.24;severe OR=0.11),insomnia(mild OR=0.51;moderate OR=0.44;severe OR=0.38),low social support(OR=0.23),and low happiness index(OR=0.42).Male residents(OR=1.25)were less likely to experience burnout.High burnout residents were 5.11 times more likely to quit the medical profession.Conclusion:Training intensity and socio-psychological factors signifi-cantly contribute resident physicians' burnout levels,with severe burnout increase career attrition.

Background and Aims:Colorectal cancer(CRC)has a complex pathogenesis,and current treatments remain limited in efficacy for advanced metastatic disease.Dasatinib is a multi-target tyrosine kinase inhibitor that has shown potential antitumor activity in various solid tumors.This study aimed to evaluate the causal relationships between dasatinib-related target genes and CRC based on genetic variation,and to explore the mediating role of immune cells,thereby providing genetic epidemiological evidence for the prevention and targeted therapy of CRC.Methods:Dasatinib-related target genes were identified through DrugBank,and the corresponding eQTLs,GWAS data for CRC(ebi-a-GCST90018808),and 731 immune-cell traits were obtained from the IEU OpenGWAS database.A two-sample Mendelian randomization(MR)framework with a two-step mediation approach was applied:first,to assess the causal relationship between dasatinib target genes(as exposures)and CRC;second,to evaluate the causal effects between target genes and immune cells,as well as between immune cells and CRC;and finally,to calculate the proportion of mediated effects.Wald ratio,inverse-variance weighted(IVW),MR-Egger,MR-PRESSO,Cochran's Q,I2,and leave-one-out analyses were used to examine heterogeneity,horizontal pleiotropy,and robustness.Results:MR results showed that dasatinib-associated inhibition of ABL1 was significantly associated with a reduced risk of CRC(OR=0.511 0,95%CI=0.323 1-0.808 0,P=0.004 1).Inhibition of YES1 was also associated with decreased CRC risk(IVW OR=0.889 9,95%CI=0.811 6-0.975 8,P=0.013 1),with no evident heterogeneity or horizontal pleiotropy among the corresponding SNPs.Further analysis revealed that dasatinib-related inhibition of YES1 significantly reduced the levels of IgD-CD24-AC level(OR=0.818 0,95%CI=0.678 2-0.986 7,P=0.035 7),and this immune cell subset itself was identified as a risk factor for CRC(OR=1.105 7,95%CI=1.029 6-1.187 5,P=0.005 7).Mediation analysis indicated that IgD-CD24-AC accounted for-9.89%and 17.31%of the mediation effects in the ABL1→CRC and YES1→CRC pathways,respectively.Conclusion:Genetic evidence from MR suggests dasatinib-target genes ABL1 and YES1 are causally linked to reduced CRC risk,with IgD-CD24-AC partially mediating the YES1-related protective effect.These findings point to immune-mediated mechanisms underlying dasatinib's potential influence on CRC risk;further experimental validation and replication across populations are warranted.

BACKGROUND: There are few multi-city studies on the association between temperature and mortality in basin climates. This study was based on the Sichuan Basin in southwest China to assess the association of basin temperature with non-accidental mortality in the population and with the temperature-related mortality burden. METHODS: Daily mortality data, meteorological and air pollution data were collected for four cities in the Sichuan Basin of southwest China. We used a two-stage time-series analysis to quantify the association between temperature and non-accidental mortality in each city, and a multivariate meta-analysis was performed to obtain the overall cumulative risk. The attributable fractions (AFs) were calculated to access the mortality burden attributable to non-optimal temperature. Additionally, we performed a stratified analyses by gender, age group, education level, and marital status. RESULTS: A total of 751,930 non-accidental deaths were collected in our study. Overall, 10.16% of non-accidental deaths could be attributed to non-optimal temperatures. A majority of temperature-related non-accidental deaths were caused by low temperature, accounting for 9.10% (95% eCI: 5.50%, 12.19%), and heat effects accounted for only 1.06% (95% eCI: 0.76%, 1.33%). The mortality burden attributable to non-optimal temperatures was higher among those under 65 years old, females, those with a low education level, and those with an alternative marriage status. CONCLUSIONS Our study suggested that a significant association between non-optimal temperature and non-accidental mortality. Those under 65 years old, females, and those with a low educational level or alternative marriage status had the highest attributable burden.
Female ; Humans ; China/epidemiology* ; Cities ; Cold Temperature ; Hot Temperature ; Mortality ; Temperature ; Time Factors ; Middle Aged ; Male

BACKGROUND:Plastic as a durable,inexpensive,easy to manufacture organic synthetic polymer materials are widely used.At the same time,plastic resistance to high temperatures,acid and alkali resistance,corrosion-resistant properties make it difficult to degrade in nature,and ultimately forming a huge number of microplastic pollution threatening human health. OBJECTIVE:To investigate the effects of microplastic exposure on growth and development and hepatic lipid metabolism in mice. METHODS:Twenty C57BL/6J male mice were adaptively fed for one week,and then randomly divided into normal and microplastic groups(n=10 per group).Mice in the normal group were given a normal diet and water,for 4 weeks.Mice in the microplastic group were given a normal diet and free drinking of microplastic(polystyrene)water with a concentration of 1 000 μg/L,for 4 weeks.At 2 and 4 weeks of drinking,body mass and grip strength,blood lipids and liver and kidney function,ultrasonic morphology and pathological morphology of liver and lipid deposition were detected. RESULTS AND CONCLUSION:(1)With the extension of time,the body mass of mice in the two groups gradually increased,and the body mass of mice in the microplastic group was greater than that in the normal group after 2,4 weeks of drinking water(P<0.05).With the extension of time,the grip strength of mice in the normal group gradually increased,and the grip strength of mice in the microplastic group first decreased and then increased,and the grip strength of mice in the microplastic group was lower than that in the normal group after drinking water for 4 weeks(P<0.05).(2)Liver ultrasound examination showed that compared with the normal group,the ultrasonic echo signal of the liver in the microplastic group was enhanced after 2 and 4 weeks of drinking water.(3)Hematoxylin-eosin staining showed that the morphology of liver cells in the microplastic group did not change significantly after 2 and 4 weeks of drinking water,but inflammatory cell infiltration could be seen.Oil red O staining showed that obvious lipid deposition was observed in the liver of microplastic group after 2 and 4 weeks of drinking water.(4)Compared with the normal group,the levels of serum high density lipoprotein cholesterol,triacylglycerol,and aspartate aminotransferase in the microplastic group were decreased after 2 weeks of drinking water(P<0.05),and the serum triacylglycerol concentration was decreased after 4 weeks of drinking water(P<0.05).(5)These findings confirm that microplastics may cause weight gain,loss of physical strength,and abnormal hepatic lipid metabolism in mice.

Objective:To investigate the effect of inhibitory activity of high mobility group protein B1 (HMGB1), signal transduction and activator of transcription 3 (STAT3) on myocardial ischemia-reperfusion injury in rats.Methods:In vivo and in vitro models of MIRI were established. SD rats were randomly divided into a sham group, a model group, a glycyrrhizic acid group, and a NSC74859 group, with 6 rats in each group. Rats in the sham group were not ligation, and rats in the sham group and model group were not given medication. The rats in the glycyrrhizic acid group and the NSC74859 group were injected with HMGB1 antagonist glycyrrhizic acid or STAT3 inhibitor NSC74859 5 mg/kg in the tail vein at 12 h 30 min before ischemia/reperfusion and 30 min after ischemia, respectively. Left ventricular shortening fraction (FS) and left ventricular ejection fraction (EF) were evaluated by echocardiography, and apoptosis of cardiomyocytes was evaluated by hematoxylin-eosin (HE) and TUNEL staining. The expression levels of HMGB1, STAT3, and phosphorylated STAT3 (p-STAT3) were detected by real-time fluorescence quantitative PCR and Western Blot. The viability of H9C2 cells was determined by the MTS assay, intracellular ATP content was determined, and the mitochondrial membrane potential of H9C2 cells was measured by flow cytometry to evaluate the survival of cardiomyocytes. The action mode of HMGB1/STAT3 was studied by the immunoprecipitation method. The expression and migration of HMGB1/STAT3 in the nucleus and cytoplasm were detected by immunostaining. Results:After inhibiting the expression of HMGB1 or STAT3, EF and FS were increased, and immune infiltration and apoptosis of cardiomyocytes were decreased. Inhibition of HMGB1 expression could decrease the expression of STAT3, but inhibition of STAT3 expression didn’t affect the expression of HMGB1. Hypoxia could lead to increased expression of HMGB1 and p-STAT3, and decreased expression of STAT3. After 8 hours of hypoxia, the expression level of STAT3 suddenly increased. After reoxygenation, the expression of HMGB1 and STAT3 decreased, and the expression of p-STAT3 increased, but p-STAT3 (Ser 727) didn’t participate in this process. After ischemia-reperfusion injury, HMGB1 and STAT3 binded firmly in cardiomyocytes, but inhibition of STAT3 or HMGB1 weakened this binding. Inhibition of HMGB1 or STAT3 expression could reduce myocardial ischemia-reperfusion injury. The expression of HMGB1 in reoxygenated cardiomyocytes increased after hypoxia, and HMGB1 migrated from the nucleus to the cytoplasm.Conclusions:Inhibiting the activity of the HMGB1/STAT3 axis effectively reduces MIRI in rats.

Objective To evaluate the application value of perioperative interventional strategy guided by enhanced recovery after surgery (ERAS) in elderly recipients undergoing liver transplantation. Methods Clinical data of 405 liver transplant recipients were retrospectively analyzed. According to age, all recipients were divided into the elderly (≥60 years, n=122) and non-elderly groups (< 60 years, n=283). All patients received perioperative interventions under the guidance of ERAS. Intraoperative and postoperative indexes, incidence of postoperative complications and discharge were analyzed between two groups. Results There were no significant differences in the duration of anesthesia, operation time, anhepatic phase, hemorrhage volume, blood transfusion volume, lactic acid level before abdominal closure, ventilator-assisted time, the length of intensive care unit (ICU) stay, Caprini score, CHIPPS score, time of gastric tube, urinary tube and drainage tube removal, time to first drinking, time to first physical activity and time to first flatus between two groups (all P > 0.05). In the elderly group, the time to first feeding was later than that in the non-elderly group (P < 0.05). There were no significant differences in the incidence of fever, ascites, pulmonary infection, delayed gastric emptying, hemorrhage and inactive venous thrombosis between two groups (all P > 0.05). No significant differences were observed in the levels of aspartate aminotransferase, total bilirubin, direct bilirubin, serum creatinine before discharge and total length of hospital stay between two groups (all P > 0.05). The alanine aminotransferase level in elderly recipients was lower than that in non-elderly counterparts, and the difference was statistically significant (P < 0.05). No unplanned reoperation was performed within postoperative 30 d in two groups. There was no significant difference in the re-hospitalization rate within 30 d after discharge (P > 0.05). Conclusions ERAS-guided interventional strategy contributes to perioperative recovery of elderly recipients undergoing liver transplantation, and yields equivalent postoperative recovery between elderly and non-elderly recipients.