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Recombinase polymerase amplification (RPA) is a newly developed isothermal amplification technology with high sensitivity and specificity. The combination of RPA and lateral flow strips (LFS) enables rapid identification of target genes. This technique has been widely used in medicine, food, botany, and other fields. This review generalizes the use of RPA-LFS technology for the diagnosing pathogenic microorganisms, providing a reference for point-of-care diagnosis of pathogenic microorganisms.
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Objective:To explore the changes of T follicular regulatory (T FR) cells/T follicular helper (T FH) cells and their related cytokines in peripheral blood of children with dust mite allergic asthma, and their clinical significance. Methods:A total of 25 children with acute dust mite allergic asthma (the asthma group) in Affiliated Hospital of Jiangnan University from January to December 2021 and 16 age- and sex-matched healthy volunteers (the healthy control group) at the same time were enrolled in the retrospective study.The percentages of peripheral T FR cells and T FH cells of the 2 groups were measured by flow cytometry.The plasma levels of cytokines[interleukin (IL)-10, IL-21] of the 2 groups were assessed by the flow cytometric microsphere-based array technology.The specific IgE (sIgE) levels of dust mites in 2 groups were detected by fluorescence enzyme immunoassay.The percentage of eosinophils in peripheral blood detected by blood cell analyzer.Data between groups were compared by t-test, and the correlation among indicators was analyzed by Spearman rank correlation analysis. Results:The asthma group had evident T FR cells/T FH cells immune imbalance.Compared with the healthy control group, the asthma group had a significantly lower T FR cells level[(0.11±0.03)% vs.(0.13±0.03)%], a significantly higher T FH cells level[(5.07±1.75)% vs.(3.80 ± 1.60)%], and a significantly lower ratio of T FR cells /T FH cells(0.02±0.01 vs.0.05±0.03) ( t=2.29, 2.30, 3.71; all P<0.05). Compared with the healthy control group, the asthma group had a significantly higher IL-21 level[(547.85±195.13) ng/L vs.(404.94±110.41) ng/L], and a significantly lower IL-10 level[(10.18±3.49) ng/L vs.(14.79±5.65) ng/L] ( t=2.60, 3.15; all P<0.05). The ratio of T FR cells/T FH cells in asthma group was negatively correlated with sIgE ( r=-0.444 2, P=0.026 1), but not related to the eosinophil percentage ( r=-0.135 2, P=0.519 3). Conclusions:Children with dust mite allergic asthma suffer from T FR cells/T FH cells subset imbalance.The imbalanced T FR cells, T FH cells and their related cytokines IL-10 and IL-21 may play a role in regulating the production of asthma sIgE.
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Objective:To evaluate the effect of artesunate on intestinal ischemia/reperfusion (I/R)-induced lung injury in mice and relationship with heme oxygenase-1 (HO-1).Methods:Twenty-four healthy SPF male C57BL/6 mice, aged 8-9 weeks, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (group Sham), intestinal I/R group (group I/R), artesunate group (group A), and artesunate plus HO-1 inhibitor Zinc protoporphyrin Ⅸ(ZnPP) group (group AS). The model of intestinal I/R injury was established by occluding the superior mesenteric artery for 45 min followed by 2 h reperfusion in anesthetized animals.Artesunate 40 mg/kg was injected via the tail vein at 1 h before ischemia in group A. ZnPP 7.5 mg/kg was injected via the tail vein at 12 h before ischemia, and artesunate 40 mg/kg was injected via the tail vein at 1 h before ischemia in group AS.The animals were sacrificed at the end of reperfusion, and the lung tissues were obtained for microscopic examination of the pathologic changes and for determination of the wet/dry lung weight (W/D) ratio, myeloperoxidase (MPO) activity, malondialdehyde (MDA) content, expression of interleukin-6 (IL-6) mRNA (by fluorescence quantitative polymerase chain reaction) and apoptotic index (AI) (by TUNEL). The lung injury score was assessed. Results:Compared with group Sham, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly increased, and the expression of IL-6 mRNA was up-regulated in group I/R ( P<0.05). Compared with group I/R, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly decreased, and the expression of IL-6 mRNA was down-regulated in group A ( P<0.05). Compared with group A, the lung injury score, W/D ratio, MPO activity, MDA content and AI were significantly increased, and the expression of IL-6 mRNA was up-regulated in group AS ( P<0.05). Conclusions:Artesunate can alleviate intestinal I/R-induced lung injury, and the mechanism may be related to activation of HO-1 in mice.
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Objective:To evaluate the role of cannabinoid type 2 receptor (CB2R) in sevoflurane postconditioning-induced reduction of intestinal ischemia-reperfusion (I/R) injury in rats.Methods:Twenty-four clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 220-270 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (group Sham), intestinal I/R group (group I/R), intestinal I/R+ sevoflurane postconditioning group (group Sevo) and intestinal I/R+ sevoflurane postconditioning+ CB2R antagonist AM630 group (group AM). The model of intestinal I/R injury was established by occluding the superior mesenteric artery for 45 min followed by 2 h reperfusion.In the group Sevo, 2% sevoflurane was inhaled immediately at the beginning of reperfusion for 30 min.In the group AM, CB2R antagonist AM630 3 mg/kg was intraperitoneally injected at 1 h before ischemia, and the other treatments were similar to those previously described in group Sevo.At 2 h of reperfusion, the animals were sacrificed after anesthesia, and small intestinal tissues were obtained for microscopic examination of the pathologic changes which was scored according to Chiu and for determination of wet/dry weight ratio (W/D ratio), for detection of malondialdehyde (MDA) content (by thiobarbituric acid colorimetry), for determination of myeloperoxidase (MPO) activity (by MPO assay) and cleaved caspase-3 protein expression (by Western blot). Results:Compared with group Sham, the Chui score, W/D ratio, MDA content and MPO activity in the intestinal tissues were significantly increased, cleaved caspase-3 expression was up-regulated in group I/R ( P<0.05). Compared with group I/R, the Chui score, W/D ratio, MDA content and MPO activity in the intestinal tissues were significantly decreased, cleaved caspase-3 expression was down-regulated in group Sevo ( P<0.05). Compared with group Sevo, the Chui score, W/D ratio, MDA content and MPO activity were significantly increased, cleaved caspase-3 expression was up-regulated in group AM ( P<0.05). Conclusion:CB2R is involved in the process of sevoflurane postconditioning-induced reduction of intestinal I/R injury in rats.
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Gamma-aminobutyric acid (GABA) is an inhibitory neurotransmitter in the adult central nervous system (CNS), however, it causes excitation in the immature CNS neurons. The shift from GABA-induced depolarization to hyperpolarization in postnatal brain is primarily due to progressive decrease in the expression of the Na-K-2Cl symporter 1 (NKCC1) and increased expression of the K-Cl cotransporter 2 (KCC2). Unlike CNS neurons, both immature and mature neurons in the enteric nervous system (ENS) are depolarized by GABA. Molecular mechanisms by which GABA excites ENS neurons are unclear. It is understood, however, that the excitatory action depends on elevated intraneuronal Cl. We aimed to test a hypothesis that high intracellular Cl in ENS neurons is maintained by activity of the NKCCs. We found that NKCC2 immunoreactivity (IR) was expressed in the ENS of the rat colon on postnatal day 1 (P1). The expression level of NKCC2 continuously increased and reached a steady high level on P14 and maintained at that level in adulthood. NKCC1 IR appeared in ENS on P14 and maintained through adulthood. KCC2 IR was not detectable in the ENS in any of the developmental stages. Both NKCC1 IR and NKCC2 IR were co-expressed with GABA receptors in ENS neurons. Exogenous GABA (1 mmol/L) caused membrane depolarization in the ENS neurons. The reversal potential of GABA-induced depolarization was about -16 mV. Blockade of NKCC by bumetanide (50 μmol/L) or furosemide (300 μmol/L) suppressed the depolarizing responses to GABA. Bumetanide (50 μmol/L) shifted the reversal potential of GABA-induced depolarization in the hyperpolarizing direction. Neither the KCC blocker DIOA (20 μmol/L) nor the Cl/HCO exchanger inhibitor DIDS (200 μmol/L) suppressed GABA-evoked depolarization. The results suggest that ENS neurons continuously express NKCC2 since P1 and NKCC1 since P14, which contribute to the accumulation of Cl in ENS neurons and GABA-evoked depolarization in neonate and adult ENS neurons. These results provide the first direct evidence for the contribution of both NKCC2 and NKCC1 to the GABA-mediated depolarization.
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Animais , Ratos , Bumetanida , Neurônios , Receptores de GABA-A , Simportadores , Ácido gama-AminobutíricoRESUMO
Objective To analyze the occurrence and prognosis of congenital heart disease interventional therapy related arrhythmias, and to discuss the prevention and treatment. Methods We retrospectively analyzed the occurrence and prognosis of congenital heart disease interventional therapy related arrhythmias among a total of 223 cases admitted for in terventional therapy between February 2014 to January 2015. Resu1ts 8 cases developed different degree and nature of arrhythmias after the interventional therapy. Among these cases, 3 of them had arrhythmias after ASD occlusion, including one was frequent atrial contraction, one was paroxysmal atrial tachycardia, and one was sinus bradycardia with accelerated junctional rhythm. All of them converted back to sinus rhythmm spontaneously from 2-3 hours to maximum 1 week after operation. 4 cases had arrhythmias after VSD occlusion, including one case of ventricular tachycardia, one case of Ⅰ° degree atrioventricular block who recovered spontaneously after surgery, one case of Ⅲ° degree atrioventricular block and one case of intermittent complete left bundle branch block who retruned to sinus rhythmn after 1 week of symptomatic treatment. One case had ventricular fibrillation after pulmonary valve balloon dilatation and was treated by defibrillation and temperany pacing to convent to sinus. Conc1usions Arrhythmias is a common complication of congenital heart disease after interventional therapy, and most of them are temporary and transient changes. However, once serious arrhythmias happened, the success rate of surgery and postoperative curative effect can be directly effected, or may even lead to mortality.