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Objective To evaluate the performance of recombinase-aided amplification (RAA) assay in detection of Clonorchis sinensis metacercariae in freshwater fish samples, so as to provide insights into standardization and field application of this assay. Methods Wild freshwater fish samples were collected in the rivers of administrative villages where C. sinensis-infected residents lived in Jiangyan District, Xinghua County and Taixing County of Taizhou City, Jiangsu Province from June to September 2022. Genomic DNA was extracted from six freshwater fish specimens (5 g each) containing 0, 1, 2, 4, 8 and 16 C. sinensis metacercariae for fluorescent RAA assay, and the diagnostic sensitivity was evaluated. Fluorescent RAA assay was performed with genomic DNA from C. sinensis, Metorchis orientalis, Haplorchis pumilio and Centrocestus formosanus metacercariae as templates to evaluate its cross-reactions. In addition, the detection of fluorescent RAA assay and direct compression method for C. sinensis metacercariae was compared in field-collected freshwater fish samples. Results Positive amplification was found in fresh-water fish specimens containing different numbers of C. sinensis metacercariae, and fluorescent RAA assay was effective to detect one C. sinensis metacercaria in 5 g freshwater fish specimens within 20 min. Fluorescent RAA assay tested negative for DNA from M. orientalis, H. pumilio and C. formosanus metacercariae. Fluorescent RAA assay and direct compression method showed 5.36% (93/1 735) and 2.88% (50/1 735) detection rates for C. sinensis metacercariae in 1 735 field-collected freshwater fish samples, with a statistically significant difference seen (χ2 = 478.150, P < 0.001). There was a significant difference in the detection of C. sinensis metacercariae in different species of freshwater fish by both the direct compression method (χ2 = 11.20, P < 0.05) and fluorescent RAA assay (χ2 = 20.26, P < 0.001), and the detection of C. sinensis metacercariae was higher in Pseudorasbora parva than in other fish species by both the direct compression method and fluorescent RAA assay (both P values < 0.05). Conclusions Fluorescent RAA assay has a high sensitivity for detection of C. sinensis metacercariae in freshwater fish samples, and has no cross-reactions with M. orientalis, H. pumilio or C. formosanus metacercariae. Fluorescent RAA assay shows a higher accuracy for detection of C. sinensis infections in field-collected freshwater fish than the direct compression method.
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Objective To investigate the effect of early enteral nutrition combined with probiotics on the nutritional status of patients with severe head injury. Methods Thirty-five patients with severe head injury were randomized into early enteral nutrition group ( control group, n = 18 ) and early enteral nutrition combined with probiotics group (observational group, n = 17). All the patients received enteral nutrition via nasogastric tube within 24-48 hours following admission. Hemoglobin, lymphocyte count,plasma concentrations of total protein, albumin, prealbumin, cholinesterase, and fasting blood glucose were determined before enteral nutrition and at days 8, 15, 21 after enteral nutrition. Arm circumference (AC), triceps skinfold thickness (TSF) and ann muscle circumference ( AMC ) were measured at the same time points. Gastrointestinal reactions as well as the time to first defecation were recorded. Glasgow coma scale (GCS) was assessed. Results There was no difference in terms of hemoglobin, lymphocyte count, plasma concentrations of total protein, prealbumin, and GCS score at each time point between the two groups ( P ＞ 0. 05 ). At day 8 after enteral nutrition, the patients in the observational group exhibited significantly lower level of fasting blood glucose than those in the control group (P ＜ 0.05 ). At day 15,the concentration of plasma cholinesterase in the observational group was significantly higher than that in the control group ( P ＜ 0. 05 ). At day 21, the observational group showed a tendency in increase of plasma albumin level in comparison with the control group( P = 0. 054), and the decrease of AC and AMC was obviously lower than that in the control group ( P ＜0. 05 ) compared with pre-enteral nutrition. The observational group showed less gastro-oesophageal reflux and constipation as well as earlier time to first defecation compared with the control group ( P ＜ 0. 05,0. 01 ). Conclusion Compared with the ordinary enteral nutrition, the enteral nutrition combined with probiotics can attenuate metabolic disorders and improve the nutritional status of the patients with severe head injury.
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Objective To investigate the effects of hyperbaric oxygen (HBO) on the systemic inflammation response in patients with severe traumatic brain injury (TBI), and to explore HBO therapy mechanisms. Methods Seventy patients with severe TBI were randomly divided into a routine treatment group ( RT group, n = 35 ) and an HBO group (n=35). All patients received conventional treatment, but the HBO group received additional early HBO therapy. Twenty age-and sex-matched normal subjects were recruited and served as normal controls. Serum interleukin-6 (IL-6) was measured by ELISA, and C reactive protein (CRP) was also measured on days 1, 7, 14 and 21 after injury. Sequential organ failure assessments (SOFAs) and Glasgow coma scale (GCS) scores were evaluated at the same time points. Systemic inflammatory response syndrome (SIRS) was assessed daily. Results The concentrations of serum IL-6 and CRP increased obviously following TBI, but patients in the HBO group exhibited significantly lower levels at each time point than those in the RT group. In the HBO group fewer cases of SIRS developed,and they had a significantly shorter average duration than those in the RT group. The average SOFA score in the HBO group was significantly lower than that of the RT group at days 14 and 21 after injury, and the GCS scores had improved significantly more by day 21. Compared with the patients who were free of SIRS, the patients with SIRS showed higher levels of IL-6 and CRP, higher SOFA scores as well as lower GCS scores ( all differences statistically significant). Conclusions HBO therapy can attenuate systemic inflammation after TBI, protect the functions of important organs and improve clinical outcomes. Decreasing the level of IL-6 may contribute to the effectiveness of HBO.