Endotoxins are common exogenous pyrogens. Excessive endotoxins in medical devices and injections can lead to serious consequences such as sepsis, septic shock, and even death. Therefore, endotoxin detection plays a crucial role in medical, pharmaceutical, and food sectors. The wide application of Limulus amebocyte lysate (LAL) has led to a sharp decline in the number of horseshoe crabs. Moreover, the LAL assay has limitations such as interbatch variations and difficulty in quantification. The recombinant factor C (rFC) assay is stable between batches, highly sensitive, and capable of quantitation, and thus it can be used as an alternative for the LAL assay. However, the high cost and complex procedures involved in producing recombinant factor C have limited the widespread application of this method. In order to simplify the preparation and reduce the production cost of recombinant factor C, this study focuses on the production of recombinant factor C based on the baculovirus expression system. Multiple measures such as a high-yield and anti-apoptotic vector qBac-IIIG, the optimal signal peptide, and the optimized codon were used to reach the goal of endotoxin detection with cell supernatant. This method simplifies the steps of protein purification. The sensitivity of the supernatant reached 0.05 EU/mL in a 1-L fermentation system, and 500 000 detecting reactions can be supported per liter of fermentation broth. This study increases the yield and activity of recombinant factor C, simplifies the procedures of protein purification, and reduces the cost, laying a foundation for the promotion and application of recombinant factor C in endotoxin detection.
Animals ; Recombinant Proteins/genetics* ; Horseshoe Crabs/chemistry* ; Baculoviridae/metabolism* ; Endotoxins/analysis* ; Protein C/biosynthesis* ; Genetic Vectors/genetics* ; Arthropod Proteins/genetics* ; Enzyme Precursors ; Serine Endopeptidases

Objective To analyze the fluid resistance characteristics of different drafting formations in marathon swimming using computational fluid dynamics(CFD)method,and provide theoretical guidance for selecting optimal drafting strategies in competitions and training.Methods Multi-swimmer models were established via three-dimensional body scanning technology,and various formation models(I-,A-,V-,L-,H-type)were created by adjusting lateral and longitudinal distances between swimmers.The ANSYS Discovery Live software was used to simulate the overall resistance of different models and the resistance of individual swimmers within formations.Results The I3-type formation exhibited an overall drag reduction effect,reducing total resistance by 55.21％,whereas other formations increased overall resistance.The V-type formation showed the most significant resistance increase(31.88％).During drafting,the lowest resistance position was the rear position in the I3-type formation,while the highest resistance position was the middle position in the L-type formation.When leading,the fluid resistance of the leading swimmer in the A-type formation was significantly greater than that of an individual swimmer(P＜0.05).Conclusions Longitudinal drafting formations demonstrated superior drag reduction effects,with the rear position in a three-person longitudinal arrangement showing the optimal drag reduction.Considering both tactical considerations and drag reduction effects,swimmers are advised to avoid the middle position in lateral formations.

Objective To analyze the fluid resistance characteristics of different drafting formations in marathon swimming using computational fluid dynamics(CFD)method,and provide theoretical guidance for selecting optimal drafting strategies in competitions and training.Methods Multi-swimmer models were established via three-dimensional body scanning technology,and various formation models(I-,A-,V-,L-,H-type)were created by adjusting lateral and longitudinal distances between swimmers.The ANSYS Discovery Live software was used to simulate the overall resistance of different models and the resistance of individual swimmers within formations.Results The I3-type formation exhibited an overall drag reduction effect,reducing total resistance by 55.21％,whereas other formations increased overall resistance.The V-type formation showed the most significant resistance increase(31.88％).During drafting,the lowest resistance position was the rear position in the I3-type formation,while the highest resistance position was the middle position in the L-type formation.When leading,the fluid resistance of the leading swimmer in the A-type formation was significantly greater than that of an individual swimmer(P＜0.05).Conclusions Longitudinal drafting formations demonstrated superior drag reduction effects,with the rear position in a three-person longitudinal arrangement showing the optimal drag reduction.Considering both tactical considerations and drag reduction effects,swimmers are advised to avoid the middle position in lateral formations.

Objective:To investigate the independent predictors of the long-term clinical outcomes in patients with branch atheromatous disease (BAD) in lenticulostriate artery (LSA) territory.Methods:Patients with LSA-BAD admitted to the Department of Neurology, Zhongnan Hospital of Wuhan University from January 1, 2016 to June 1, 2019 were enrolled retrospectively. Their demography, vascular risk factor, and baseline clinical data were collected. The National Institutes of Health Stroke Scale (NIHSS) was used to evaluate the severity of stroke. The clinical outcomes were evaluated by the modified Rankin Scale at 6 months. 0-2 was defined as good outcome, and >2 was defined as poor outcome. Multivariate logistic regression analysis was used to determine the independent influencing factors of clinical outcomes in patients with LSA-BAD. Results:A total of 81 patients with LSA-BAD were enrolled. Their age 59.20±11.75 years (range, 39-81 years), 53 were male (65.4%), and median baseline NIHSS score was 1.0 (interquartile range, 0-4.0). Forty-one patients (50.6%) received intravenous thrombolysis. At 6-month follow-up after the onset, 63 patients (77.8%) had a good outcome, and 18 (22.2%) had a poor outcome. The baseline NIHSS score of the poor outcome group was significantly higher than that of the good outcome group (6.5 [0-9.0] vs. 1.0 [0-3.0]; Z=2.395, P=0.017), while the proportion of mild stroke (61.6% vs. 98.4%; χ2=17.595, P<0.001) and patients receiving intravenous thrombolysis (38.9% vs. 54.0%; χ2=4.450, P=0.035) were significantly lower than those of the good outcome group. Multivariate logistic regression analysis showed that after adjusting for other confounding factors, venous thrombolysis was independently correlated with the good outcome (odds ratio 0.099, 95% confidence interval 0.011-0.924; P=0.042), while the high baseline NIHSS score was independently associated with the poor outcome (odds ratio 1.736, 95% confidence interval 1.262-2.388; P=0.001). Conclusion:Intravenous thrombolysis is helpful to improve the outcomes of patients with LSA-BAD, and a higher baseline NIHSS score is an independent predictor of the poor outcome.

The authors described the hospital group reform made by Jiangsu Kangfu Medical Group,and its main actions taken and initial success in public hospital organizational structure.Based on such studies, they recommend further clarification of the rights and responsibilities, for sectional integration;appropriate adjustment to update the supervision mechanism;building of the appointment system to speed up personnel system reform.

This paper introduced overseas experiences and theories of health systems in view of the ongoing medical and health system reform in China.It proposed the super health ministry reform,justified its necessity,and clarified key issues of such a reform,in addition to discussing how to improve the system of macro-health.