Objective To prepare and characterize the mouse polyclonal antibody against the dense granule protein 24 (GRA24) of Toxoplasma gondii, and explore its preliminary applications. Methods The GRA24 coding sequences of different T. gondii strains were aligned using the MEGA-X software, and the dominant peptide of the GRA24 protein was analyzed with the Protean software. The base sequence encoding this peptide was amplified using PCR assay and ligated into the pET-28a vector, and the generated GRA24 truncated protein was transformed into Escherichia coli BL21. After induction by isopropyl-beta-D-thiogalactopyranoside (IPTG), the expression and purification of the recombinant GRA24 protein was analyzed using sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE). BALB/c mice were immunized by subcutaneous injection with the purified recombinant GRA24 truncated protein to generate the polyclonal antibody, and the titer of the polyclonal antibody was measured using enzyme linked immunosorbent assay (ELISA). The specificity of the polyclonal antibody was tested using Western blotting, and the intracellular localization of the polyclonal antibody was investigated using immunofluorescence assay (IFA). Results SDS-PAGE showed successful construction of the recombinant expression plasmid, and Coomassie brilliant blue staining showed the generation of the high-purity recombinant GRA24 truncated protein. ELISA measured that the titer of the polyclonal antibody against the GRA24 truncated protein was higher than 1:208 400, and Western blotting showed that the polyclonal antibody was effective to recognize the endogenous GRA24 proteins of different T. gondii strains and specifically recognize the recombinant GRA24 truncated protein. Indirect IFA showed that the GRA24 protein secreted 16 hour following T. gondii invasion in host cells. Conclusions The polyclonal antibody against the T. gondii GRA24 protein has been successfully prepared, which has a widespread applicability, high titers and a high specificity. This polyclonal antibody is available for Western blotting and IFA, which provides the basis for investigating the function of the GRA24 protein.

An innovative on-site real-time avian influenza virus(AIV)detection method was estab-lished by integratingrecombinase-aided amplification(RAA)with the clustered regularly inter-spaced short palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)system.After analy-zing 120 sequences of the M gene of avian influenza viruses of different subtypes publicly available on NCBI,the RAA primers and crRNA were designed based on the identified highly conserved segment and used for RAA nucleic acid amplification.After the amplified products were transferred to a CRISPR/Cas13a detection system,the fluorescence values were monitored throughout the re-action process to indicate the results.The sensitivity and specificity of the RAA-CRISPR/Cas13a method were validated using gradient dilutions(106-100 copies/μL)of positive plasmids and sev-en other avian viruses.Fifty clinical samples were tested using this method and compared with the national standard fluorescence RT-PCR method.The results indicated that the detection limit for RAA-CRISPR/Cas13a method was 102 copies/μL,a two-fold improvement over the standard RAA.Specificity assay showed the established method only detected AIV with no cross-reactivity with other seven avian viruses.Compared to the national standard fluorescence RT-PCR method,this method exhibited 100％specificity,95.24％accuracy,and 98.00％consistency in detection of clinical samples.In conclusion,a universal and rapid RAA-CRISPR/Cas13a for detection of AIV was established with the capacity of achieving detection within 60 minutes at 37 ℃,which provides a rapid,sensitive,and specific on-site detection method for AIV.

Objective:Cardiac arrest (CA) represents a significant public health challenge, posing a substantial threat to individual health and survival. To enhance the survival rates of patients experiencing out-of-hospital cardiac arrest (OHCA), Baoan District in Shenzhen City has undertaken exploratory initiatives and practical interventions, yielding promising preliminary outcomes.Methods:1.Innovate emergency medical services by developing a "four-circle integration" system that connects to the hospital. This system encompasses the social emergency medical system, the out-of-hospital emergency medical system, the in-hospital emergency medical service system, and the intensive care treatment system. 2.Develop a comprehensive model for the construction of a social emergency medical training system, characterized by party leadership, government oversight, departmental coordination, professional guidance, technological support, and community involvement, termed the "Baonan Model." Additionally, establish evaluation criteria to assess the effectiveness of the social emergency medical training system in Baonan District; 3. Develop a cardiac arrest registration system and a social emergency medical training management system for Baonan District; 4. Enhance the proficiency in treatment techniques and the quality of cardiopulmonary resuscitation among emergency medical professionals; 5. Strengthen and advance the development of a "five-minute social rescue network" to address the critical "emergency window period." .Result:In Baonan District, 9.18% of the public is trained in emergency medical skills. The bystander CPR rate for OHCA is 26.11%, AED use is at 4.78%, the 30-day survival rate is 6.31%, and the discharge survival rate is 4.44%.Conclusion:The implementation of the aforementioned measures can substantially enhance the survival rate of patients experiencing OHCA at the time of discharge.

Salmonella is an important foodborne pathogen and one of the main causes of diarrhea. Every year, about 550 million people suffer from diarrhea due to Salmonella infection, of which about 230 000 die. It has become a major global public safety issue. The application fields of Salmonella detection involve food safety, water quality monitoring, animal husbandry, public health monitoring, and medical diagnosis. The detection requirements mainly come from three aspects: pathogen identification, serotype identification, drug resistance and virulence identification. In recent years, the detection technology for Salmonella has made rapid progress, especially the emergence and development of emerging molecular detection technologies, providing new perspectives for Salmonella detection in different scenarios. However, due to the diversity of Salmonella serotypes and the complexity of detection scenarios, existing detection technologies still have some pain points (such as long detection time, cumbersome operation steps, low scene adaptability, etc.). This article will elaborate on the application of several emerging molecular detection technologies with distinct characteristics, such as CRISPR Cas technology, digital PCR technology, sequencing technology, and microfluidic technology, in Salmonella detection. It aims to provide a reference for the development and improvement of Salmonella detection technology and the establishment of infection warning and control systems.

Salmonella is an important foodborne pathogen and one of the main causes of diarrhea. Every year, about 550 million people suffer from diarrhea due to Salmonella infection, of which about 230 000 die. It has become a major global public safety issue. The application fields of Salmonella detection involve food safety, water quality monitoring, animal husbandry, public health monitoring, and medical diagnosis. The detection requirements mainly come from three aspects: pathogen identification, serotype identification, drug resistance and virulence identification. In recent years, the detection technology for Salmonella has made rapid progress, especially the emergence and development of emerging molecular detection technologies, providing new perspectives for Salmonella detection in different scenarios. However, due to the diversity of Salmonella serotypes and the complexity of detection scenarios, existing detection technologies still have some pain points (such as long detection time, cumbersome operation steps, low scene adaptability, etc.). This article will elaborate on the application of several emerging molecular detection technologies with distinct characteristics, such as CRISPR Cas technology, digital PCR technology, sequencing technology, and microfluidic technology, in Salmonella detection. It aims to provide a reference for the development and improvement of Salmonella detection technology and the establishment of infection warning and control systems.

OBJECTIVE: To evaluate the predictive value of plasma heparin-binding protein (HBP) combined with albumin (Alb) for predicting 28-day mortality in patients with sepsis. METHODS: The clinical data of patients with sepsis admitted to the emergency intensive care unit (EICU) of the People's Hospital of Shenzhen Baoan District from March 2020 to March 2024 were retrospectively analyzed. The study began at the time of the first diagnosis of sepsis upon EICU admission and ended upon patient death or at 28 days. The gender, age, length of stay in EICU, underlying diseases, and infection sites were recorded. Within 24 hours of sepsis diagnosis, blood culture results, white blood cell count (WBC), C-reactive protein (CRP), procalcitonin (PCT), blood lactate acid (Lac), HBP, Alb, sequential organ failure assessment (SOFA), acute physiology and chronic health evaluation II (APACHE II), mortality in emergency department sepsis score (MEDS), modified early warning score (MEWS), number of organ failures, use of vasopressors, application of mechanical ventilation, renal replacement therapy, and 28-day prognosis were recorded, the differences in these indicators between two groups were compared. Univariate and multivariate Logistic regression analyses were used to analyze the risk factors of 28-day mortality in patients with sepsis. Receiver operator characteristic curve (ROC curve) was drawn, and the area under the ROC curve (AUC) was calculated to evaluate the early predictive value of various risk factors for 28-day mortality in patients with sepsis. RESULTS: A total of 300 patients with sepsis were included, with 16 excluded, resulting in 284 patients being analyzed. Among them, 191 survived and 93 died within 28 days. There were no statistically significant differences between the two groups in terms of gender, age, underlying diseases, infection sites, blood culture positivity rate, number of organ failures, and length of stay in EICU. Univariate analysis showed that the rate of vasopressor use, the rate of mechanical ventilation, HBP, PCT, CRP, Lac, SOFA score, APACHE II score, MEDS score, and MEWS score were significantly higher in the death group than those in the survival group, while Alb was significantly lower in the death group than that in the survival group. Multivariate Logistic regression analysis showed that HBP and Alb were independent risk factors for predicting 28-day mortality in patients with sepsis [odds ratio (OR) and 95% confidence interval (95%CI) were 1.093 (0.989-1.128) and 1.174 (1.095-1.259), both P < 0.05]. ROC curve analysis showed that both HBP and Alb had certain predictive value for 28-day mortality in patients with sepsis [AUC and 95%CI were 0.820 (0.717-0.923) and 0.786 (0.682-0.890), both P < 0.05]. When the critical value of HBP was 117.50 μg/L, the sensitivity was 85.90%, and the specificity was 70.50%. When the critical value of Alb was 28.30 g/L, the sensitivity was 69.30%, and the specificity was 81.20%. When the two indexes were combined for diagnosis, the AUC was 0.881 (95%CI was 0.817-0.945, P < 0.001), the sensitivity was 92.70%, and the specificity was 76.80%. CONCLUSIONS HBP and Alb are independent risk factors for predicting 28-day mortality in patients with sepsis. The combined prediction efficiency of HBP and Alb for 28-day mortality in patients with sepsis is superior to a single indicator.
Humans ; Sepsis/diagnosis* ; Retrospective Studies ; Predictive Value of Tests ; Intensive Care Units ; Blood Proteins/analysis* ; Prognosis ; Antimicrobial Cationic Peptides/blood* ; APACHE ; Male ; Female ; Organ Dysfunction Scores ; ROC Curve ; Middle Aged ; C-Reactive Protein/analysis* ; Emergency Service, Hospital ; Aged ; Hospital Mortality ; Serum Albumin/analysis*

Objective:To investigate the efficacy and prognostic factors of pars plana vitrectomy (PPV) in the treatment of proliferative diabetic retinopathy (PDR) with chronic renal failure (CRF).Methods:A retrospective study. From January 2016 to June 2021, a total of 82 eyes of 58 patients diagnosed with PDR combined with CRF and treated with PPV in Department of Ophthalmology, The Second Hospital of Hebei Medical University were included in the study. There were 32 cases in males and 26 cases in females. The mean age was (48.45±10.41) years. The course of renal failure was (4.15±3.23) years, and the course of diabetes was (14.45±6.71) years. All patients undergo best-corrected visual acuity (BCVA). The BCVA examination was performed using the international standard Snellen visual acuity chart, which was converted to logarithm of the minimum angle of resolution (logMAR) visual acuity for recording. The mean number of logMAR BCVA was 2.04±0.82 (0.7-2.8). The duration of vitreous hemorrhage averaged (2.65±1.55) months. There were 38 eyes (46.3%, 38/82) with traction retinal detachment; 32 eyes had a history of panretinal photocoagulation (PRP) treatment (39.0%, 32/82). All eyes were treated with 25G PPV. Patients with traction retinal detachment were treated with intravitreal injection of anti-vascular endothelial growth factor (VEGF) 3 days before surgery. Opacification of the lens affected the operation operator combined with phacoemulsification. Biochemical indexes such as hemoglobin, glycosylated hemoglobin, albumin, creatinine, uric acid, and alternative treatment (non-dialysis/hemodialysis/peritoneal dialysis) were collected. Postoperative follow-up time was ≥6 months. χ2 test or Fisher's exact test were used for comparison between groups. A logistic regression model was used for multivariate analysis, and Spearman correlation analysis was used to evaluate the correlations between variables. Results:At 6 months after surgery, the mean logMAR BCVA was 1.16±0.57. Compared with logMAR BCVA before surgery, the difference was statistically significant ( t=-0.837, P<0.001); 44 eyes had BCVA ≥0.1 and 38 eyes had BCVA <0.1. Postoperative vitreous hemorrhage (PVH) was observed in 17 eyes after surgery (20.7%, 17/82). PVH occurred in 15 (46.9%, 15/32), 1 (2.3%, 1/44), and 1 (16.7%, 1/6) eyes in patients without dialysis, hemodialysis and peritoneal dialysis, respectively. There was significant difference between those without dialysis and those on hemodialysis ( χ2=26.506, P<0.05). There was no significant difference between peritoneal dialysis patients and those without dialysis and hemodialysis patients ( χ2=2.694, 2.849; P>0.05). PVH occurred in 3 (10.0%, 3/30) and 14 (27.0%, 14/52) eyes of vitreous cavity filled with silicone oil and perfusion fluid, respectively. The difference was statistically significant ( χ2=3.315, P<0.05); 1 (33.3%, 1/3) and 10 (71.4%, 10/14) eyes were treated with PPV again, respectively, and the difference was statistically significant ( P<0.05). Neovascular glaucoma (NVG) occurred in 12 eyes (14.6%, 12/82). Logistic regression analysis showed that age [odds ratio ( OR) =0.911, P<0.05], diabetic retinopathy (DR) stage ( OR=7.229, P<0.05), renal failure duration ( OR=0.850, P<0.05), operation time ( OR=1.135, P<0.05) was an independent risk factor for poor vision prognosis. Diabetes duration ( OR=1.158, P<0.05), renal failure duration ( OR=1.172, P<0.05) and alternative therapy were independent factors affecting the occurrence of PVH. Diabetes duration ( OR=1.138, P<0.05) and renal failure duration ( OR=1.157, P<0.05) were independent risk factors for postoperative NVG. Spearman correlation analysis showed that PVH was strongly correlated with post-operative NVG ( r=0.469, P<0.01). There was no significant correlation between blood glucose, hemoglobin, creatinine and blood urea nitrogen and prognosis of postoperative vision, PVH and NVG occurrence ( P>0.05). Conclusions:In PDR patients with CRF, DR Stage, age, renal failure course and operation duration are correlated with vision prognosis. Compared with those who do not receive alternative therapy, hemodialysis treatment can reduce the occurrence of PVH and NVG after surgery.

ObjectiveThe contamination of foodborne pathogens in ready-to-eat fruits and vegetables in Shanghai was analyzed to provide a scientific basis for food safety， risk assessment and related supervision. MethodsFrom June to September 2021， a total of 143 batches of12 kinds of ready-to-eat fruits and vegetables such as lettuce， chicory， and cherry tomatoes were collected from farmers’ markets， supermarkets， and e-commerce platforms. The total number of bacterial colonies， Salmonella， Listeria monocytogenes， Staphylococcus aureus， Cronobacter spp. and Shiga toxin-producing Escherichia coli in the samples were tested according to National Food Contamination and Harmfulness Risk Monitoring Manual. ResultsAmong the 143 batches， foodborne pathogens were detected in 68 batches， with a total detection rate of 47.55% （68/143）. A total of 79 strains of foodborne pathogens were detected. The detection rate of Staphylococcus aureus was the highest （32.87%， 47/143）， followed by Cronobacter spp. （20.98%， 30/143）， Salmonella （0.70%， 1/143）， Listeria monocytogenes （0.70%， 1/143）， Shiga toxin-producing Escherichia coli （0.00%）. Furthermore， the detection rate was higher in different ready-to-eat fruits and vegetables： chicory （17.33%）， cucumber （17.14%）， cherry tomatoes （16.00%）， and honeydew melon （15.38%）， respectively. Meanwhile， the contamination rate of pathogens in ready-to-eat fruits and vegetables from farmers’ markets， supermarkets， and e-commerce platforms was relatively high. ConclusionReady-to-eat fruits and vegetables in Shanghai are contaminated by foodborne pathogens. The prevention and control of the contamination of post-harvest fruits and vegetables should be strengthened to reduce the risk of foodborne disease outbreaks.

Humans ; Aged ; Acute Kidney Injury ; Risk Factors ; Creatinine

Behçet’s syndrome is a kind of chronic systemic vasculitis with involvement of multiple organs. Intestinal involvement of Behçet’s syndrome is presently named as intestinal Behçet’s syndrome (disease). Recently, there is considering another kind of disease type with only typical intestinal ulcers. Since it is difficult to differentiate intestinal Behçet’s syndrome from Crohn’s disease, intestinal tuberculosis, intestinal lymphoma, as well as intestinal manifestations of many other autoimmune diseases, and there is limited evidence for the therapy of intestinal Behçet’s syndrome, proposing diagnosis and treatment recommendations for intestinal Behçet’s syndrome through evidence-based judgment will be of great significance for clinical practice.