ObjectiveTo explore the light sensitivity and kinetic of the new optogenetics tools Channelrhodopsin-XXM2.0 (XXM2.0) and Channelrhodopsin-PsCatCh2.0 (PsCatCh2.0), and analyze whether they could be used to restore the visual function by optogenetics.MethodsMolecular biology techniques were used to link the gene fragments of XXM2.0 and PsCatCh2.0 to the vector pCIG(c)-msFoxn3 containing ampicillin resistant screening gene and reporter gene to form new plasmid pCIG(c)-msFoxn3-XXM2.0 and pCIG(c)-msFoxn3-PsCatCh2.0. The constructed plasmids were transfected into HEK 293T cells, and light responses were recorded in the whole cell mode with the HEKA patch clamp system. The photocurrent was recorded under three light intensity included 2.7×1016, 4.7×1015, and 6.4×1014 photons/(cm2·s). And then, XXM2.0 and PsCatCh2.0 were stimulated with 2.7×1016 photons/(cm2·s) and fully recovered. The opening and closing time constants were analyzed with Clampfit 10.6 software. At the same light intensity, photocurrents of XXM2.0 and PsCatCh2.0 were recorded by the light pulse stimulating of 2-32 Hz. The current attenuation was analyzed at long intervals of 4000 ms and short intervals of 200 ms after repeated stimulation. Comparisons between groups were performed by independent samples t test.ResultsRestriction endonuclease sites of EcoRⅠ and EcoRⅤ were successfully introduced at XXM2.0 and PsCatCh2.0 sequences. When the digestion was completed, they were ligated by T4 DNA ligase to construct new plasmids pCIG(c)-msFoxn3-XXM2.0 and pCIG (c)-msFoxn3-PsCatCh2.0, and then transfected on HEK 293T cells. The light intensity dependence was showed in XXM2.0 and PsCatCh2.0. The greater light intensity was accompanied by the greater photocurrent. Under the light intensity 6.4×1014 photons/(cm2·s) below the retinal safety threshold, large photocurrent was still generated in XXM2.0 and PsCatCh2.0 with 92.8±142.0 and 13.9±5.6 pA (t=1.24, 1.24; P=0.28, 0.29). The opening time constants of XXM2.0 and PsCatCh2.0 were 23.9±6.7 and 2.4±0.8 ms, and the closing time constants were 5803.0±568.2 and 219.9±25.6 ms. Compared with PsCatCh2.0, the opening and closing time constant of XXM2.0 were both larger than PsCatCh2.0. The differences were statistically significant (t=7.10, 31.60; P=0.00, 0.00). In terms of response frequency, XXM2.0 and PsCatCh2.0 could follow to 32 Hz high-frequency pulsed light stimulation, and all could respond to repeated light stimulation at a long (4000 ms) and a short time (200 ms) interval with the small current decay rate.ConclusionXXM2.0 and PsCatCh2.0 could be activated under light intensity with safety for the retina, and could respond to high frequency (at least 32 Hz) pulsed light stimuli with low current attenuation, which could meet the characteristics of opsins required to restore the visual function by optogenetics.
[Abstract]Acute type A aortic dissection is a lethal disease that requires immediate surgical intervention and lifesaving measures. The treatment of this condition primarily involves addressing the complex structure and vital role of the aortic root. Since 1968, surgical techniques for Aortic dissection type A have rapidly advanced, resulting in significantly improved patient outcomes. In recent years, various approaches to aortic root management have emerged. This article provides a comprehensive overview of these approaches.
From June 2002 to December 2023, there were 5 patients with criss-cross admitted to the General Hospital of the Northern Theater Command, including 3 males and 2 females, aged 18 months to 25 years, and weighing 13-49 kg. There were 5 patients of atrioventricular position, 3 patients of right ventricular loop, 2 patients of left ventricular loop, 3 patients of normal atrioventricular connection, and 2 patients of inconsistent connection. Combined intracardiac malformations: 1 patient of simple ventricular septal defect combined with pulmonary hypertension, 1 patient of corrected transposition of the great arteries combined with ventricular septal defect, atrial septal defect, and pulmonary artery stenosis, 1 patient of corrected transposition of the great arteries combined with ventricular septal defect, atrial septal defect, and left atrioventricular valve insufficiency, and 2 patients of right ventricular double outlet combined with ventricular septal defect and pulmonary artery stenosis. The surgical methods included 2 patients of intracardiac anatomical correction, 1 patient of bidirectional vena cava pulmonary artery anastomosis, and 2 patients of total extracardiac ductal cava pulmonary artery anastomosis. All 5 patients were discharged smoothly.
ObjectiveTo explore the clinical efficacy of single-branch intraoperative stent combined with modified bilateral cerebral perfusion in type A aortic dissection. MethodsA retrospective analysis was conducted on the clinical data of patients who underwent surgery for type A aortic dissection at the First Affiliated Hospital of Anhui Medical University from January 2021 to May 2024. Patients were divided into the experimental group (single branch stent combined with modified bilateral cerebral perfusion) and the control group (traditional surgical method, straight stent+unilateral cerebral perfusion) according to the surgical method. Propensity score matching analysis was used to match the two groups of patients with a ratio of 1:1, and the perioperative data and clinical efficacy after matching were compared. ResultsA total of 14 patients were included in the experimental group, including 13 males and 1 female, with an average age of (47.4±16.1) years. There were 56 patients in the control group, including 38 males and 18 females, with an average age of (52.1±11.7) years. After propensity score matching, 14 patients were included in each group. Compared with the control group, the ventilator support time [(27.4±24.3) h vs. (93.4±88.0) h, P=0.018], length of stay in the intensive care unit [(2.8±1.8) d vs. (8.7±6.5) d, P=0.009], and postoperative awakening time [(5.4±2.2) h vs. (8.8±4.8) h, P=0.047] in the experimental group were shorter than those in the control group, with statistically significant differences. ConclusionSingle-branch intraoperative stent combined with modified bilateral cerebral perfusion can effectively shorten ventilator support time, length of stay in the intensive care unit, and postoperative awakening time, and is safe and effective for the treatment of type A aortic dissection, which is worth further promotion.