Cardiac three-dimensional electrophysiological labeling technology is the prerequisite and foundation of atrial fibrillation (AF) ablation surgery, and invasive labeling is the current clinical method, but there are many shortcomings such as large trauma, long procedure duration, and low success rate. In recent years, because of its non-invasive and convenient characteristics, ex vivo labeling has become a new direction for the development of electrophysiological labeling technology. With the rapid development of computer hardware and software as well as the accumulation of clinical database, the application of deep learning technology in electrocardiogram (ECG) data is becoming more extensive and has made great progress, which provides new ideas for the research of ex vivo cardiac mapping and intelligent labeling of AF substrates. This paper reviewed the research progress in the fields of ECG forward problem, ECG inverse problem, and the application of deep learning in AF labeling, discussed the problems of ex vivo intelligent labeling of AF substrates and the possible approaches to solve them, prospected the challenges and future directions for ex vivo cardiac electrophysiology labeling.
Objective To construct the lentiviral vector containing homo sapiens forkhead box C2 (Foxc2) gene and to detect its expression in bone marrow mesenchymal stem cells (BMSCs) of rabbits. Methods Human Foxc2 gene coding region fragment was obtained by RT-PCR and then cloned into the plasmid of LV-green fluorescent protein (GFP) to prepare Foxc2 lentiviral plasmid. Foxc2 lentiviral plasmid, pGC-LV, pHelper1.0, and pHelper2.0 were co-transfected into 293T cells to obtain recombinant virus containing Foxc2 gene. The lentiviral titer was detected. BMSCs were isolated from bone marrow of rabbit and infected with Foxc2 recombined lentiviral, then the optimum multiplicity of infection (MOI) was determined by detecting the intensity of fluorescence expression. The expression of Foxc2 in the infected BMSCs was determined at 1, 3, and 7 days after transfection by inverted fluorescence microscope and Western blot. After osteogenic induction, Alizarin red staining was done to observe the formation of mineralized nodule. Results The Foxc2 recombinant lentiviral vector was constructed and was confirmed by restriction enzyme digestion and sequencing analysis. It could efficiently transfect 293T cells and express in 293T cells. The lentiviral titer was 2 × 108 TU/mL. The optimum MOI was 200. The inverted fluorescence microscope observation showed that the Foxc2 gene expressed in 84.5% ± 4.8% of infected BMSCs at 3 days after transfection. The expression of Foxc2 in infected BMSCs was stable and high, and increased gradually within 7 days after transfection by Western blot. At 2 weeks after osteogenic induction, Alizarin red staining showed that there were a large number of red calcified matrix deposition in the cytoplasm. Conclusion Foxc2 recombined lentivirus with high viral titer is successfully constructed and packaged, and the Foxc2 gene can be transfected into BMSCs with stable and high expression of Foxc2 in infected cells, and these cells may be applied for gene therapy of avascular necrosis of the femoral head.
Objective To study the biological activity of recombinant adeno-associated virus vector (rAAV) coexpressing human vascular endothel ial growth factor165 (hVEGF165) and human bone morphogenetic protein 7 (hBMP-7) genes in vitro so as to provide a new method for the therapeutics of osteonecrosis. Methods The 3rd passage rabbit bone marrow mesenchymal stem cells (BMSCs) were transfected with rAAV-hVEGF165-internal ribosome entry site (IRES)-hBMP-7(experimental group) and green fluorescent protein (GFP) labeled rAAV-IRES-GFP (control group). The expressions ofhVEGF165 and hBMP-7 were detected by ELISA assay at the 1st, 2nd, 3rd, 7th, 14th days and Western blot assay at the14th day after transfection. The expression consistencies of hVEGF165 and hBMP-7 were observed by immunofluorescence assay at the 14th day after transfection. The biological activity of hVEGF165 was assessed by angiopoiesis experiment of the 3rd passage human umbil ical vein endothel ial cells (HUVEC). The biological activity of hBMP-7 was assessed by mineral ization of BMSCs detected by ALP staining and al izarin red staining. Results With infecting time, the hVEGF165 and hBMP-7 expressions increased gradually in two groups, showing significant difference between two groups (P lt; 0.05). The expressions of hVEGF165 and hBMP-7 were positive in experimental group and negative in control group, respectively. Immunofluorescence assay showed positive expressions of hVEGF165 and hBMP-7 in the exprimental group and negative expression in the control group, the expression of hVEGF165 and hBMP-7 had good consistencies. hVEGF165 secreted from BMSCs enhanced HUVEC migration, prol iferation and tube formation in experimental group. There was significant difference in the number of blood vessel between two groups (P lt; 0.05). The ALP staining showed more bly stained granules in experimental group than in control group. There was significant difference in the number of the mineral ized nodules between two groups (P lt; 0.05). Conclusion The rAAV-hVEGF165-IRES-hBMP-7 has good biological activity in vitro.
Objective To evaluate the effectiveness of open wedge high tibial osteotomy (OWHTO) in treatment of medial unicompartmental knee osteoarthritis (MUKOA). Methods A clinical data of 61 cases with MUKOA who were treated with OWHTO between January 2015 and January 2017 were retrospectively analyzed. There are 14 males and 47 females with an average age of 52.8 years (mean, 44-60 years). The body mass index ranged from 19.1 to 34.7 kg/m2 (mean, 25.3 kg/m2). Twenty-seven cases were left side and 34 cases were right side. The disease duration was 1-9 years (mean, 5.3 years). The MUKOA was rated as stage Ⅱ in 33 cases and stage Ⅲ in 28 cases. Preoperative Hospital for Special Surgery (HSS) score was 56.0±3.7. Walking visual analogue scale (VAS) score was 4.6±1.0. Results The operation time was 49-85 minutes (mean, 66.5 minutes). The length of incision was 10-13 cm (mean, 11.0 cm). The total overt blood loss was 80-210 mL (mean, 139.1 mL). The postoperative bed-rest time was 1-10 days (mean, 4.7 days). All patients were followed up 12-24 months (mean, 17.3 months). The bearing area of tibial platform at 3 months after operation was 60.3%-66.8%, with an average of 63.4%. At 3 and 6 months after operation, the HSS score was 79.1±4.2 and 85.3±3.1 respectively, and the VAS score was 1.7±0.7 and 0.6±0.5 respectively, all showing significant differences (P<0.05). Conclusion OWHTO is an ideal choice for treating MUKOA with less postoperative complications. The force line could be corrected by OWHTO. However, the preoperative preparations are very important, especially that the open angle should be measured accurately.
ObjectiveTo compare the short-term effectiveness between arthroscopic cystectomy and internal drainage combined with cystectomy in popliteal cyst.MethodsBetween March 2014 and March 2017, 56 patients with symptomatic popliteal cyst were enrolled in the study, randomized block design was used to divided the patients into trial group (arthroscopic cystectomy combined with internal drainage group, n=28) and control group (arthroscopic internal drainage group, n=28). Excluding those who had incomplete follow-up and received surgery for other diseases postoperatively, 26 patients in the experimental group and 27 patients in the control group were finally enrolled in the study. There was no significant difference in gender, age, side, course of disease, maximum diameter and grade of popliteal cyst, and associated diseases between two groups (P>0.05). The operation time, duration of popliteal ecchymosis and the middle back of calf tenderness were observed postoperatively. The circumference of calf at 1 day, 1 week, and 2 weeks after operation were measured and the differences were calculated with the measurement before operation. Lower extremity venous thrombosis was observed by color doppler ultrasonography at 1 week after operation. The effectiveness was evaluated by Rauschning and Lindgren grading criteria. And MRI was used to observe whether the popliteal cyst disappeared or decreased and measured its maximum diameter at 1 year after operation.ResultsPatients in both groups were followed up 12-14 months, with an average of 12.5 months. The operation time, duration of popliteal ecchymosis, and the middle back of calf tenderness of the trial group were all longer than those in the control group (P<0.05), the differences of circumference of calf at 1 day, 1 week, and 2 weeks after operation of the trial group were greater than those in the control group (P<0.05). Color doppler ultrasonography of the lower extremity at 1 week after operation found that the intermuscular venous thrombosis occurred in 2 cases of the trial group, while no lower extremity thrombosis was found in the control group; and the difference between two groups was not significant (P=0.236). According to the Rauschning and Lindgren grading criteria, there were 16 cases of grade 0, 6 cases of grade 1, and 4 cases of grade 2 in the trial group, and 17 cases of grade 0, 4 cases of grade 1, and 6 cases of grade 2 in the control group at 1 year after operation. There was no significant difference between 2 groups (Z=–1.872, P=0.078). Nine cases (34.62%) of the trial group and 13 cases (48.15%) of the control group still have residual cysts by MRI, the maximum diameter of which was less than 2 cm. The cysts disappeared in the remaining patients in both groups, and there was no recurrence during the follow-up. There was no significant difference in cyst residual rate between 2 groups (χ2=2.293, P=0.852).ConclusionCompared with arthroscopic internal drainage, the short-term effectiveness of the arthroscopic internal drainage combined with cystectomy had no significant improvement, and the operation time was prolonged, the postoperative complications were obviously increased.