Efficacy and initial clinical evaluation of optical genome mapping in the diagnosis of structural variations / 中华预防医学杂志

To investigate the efficacy and value of optical genome mapping (OGM) in detecting chromosomal structural variations. In a clinical study about high-precision analysis of genomic structural variation for complex genetic diseases, a retrospective study was performed on the cases with karyotyping at the department of Obstetrics and Gynecology, and Endocrinology of Peking Union Medical College Hospital from January to December 2021. Ten cases with abnormal karyotype was detected by OGM. Partial cases were verified by fluorescence in situ hybridization (FISH), SNP array or CNV-seq. Results of ten cases, nine were detected with abnormality by OGM, including unbalanced chromosomal rearrangements (n=3), translocation (n=5) and paracentric inversion (n=1), and the results were in concordance with other standard assays. However, one case with breakpoint and reconnected at centromere has not been detected. In conclusion, ten samples were comprehensively analyzed by karyotyping, FISH, SNP array or CNV-seq, and OGM, and results demonstrated that optical genome mapping as a new technology can not only detect unbalanced rearrangements such as copy number variants as well as balanced translocations and inversions, but more importantly, it can refine breakpoints and orientation of duplicated segments or insertions. So it can contribute to the diagnosis of genetic diseases and prevent birth defect. However, the current technology is not yet capable of detecting breakpoints of balanced structural variations lying within unmapped regions.

Unfractionated heparin attenuates endothelial barrier dysfunction via the phosphatidylinositol-3 kinase/serine/threonine kinase/nuclear factor kappa-B pathway / 中华医学杂志(英文版)

BACKGROUND@#Vascular endothelial dysfunction is considered a key pathophysiologic process for the development of acute lung injury. In this study, we aimed at investigating the effects of unfractionated heparin (UFH) on the lipopolysaccharide (LPS)-induced changes of vascular endothelial-cadherin (VE-cadherin) and the potential underlying mechanisms.@*METHODS@#Male C57BL/6 J mice were randomized into three groups: vehicle, LPS, and LPS + UFH groups. Intraperitoneal injection of 30 mg/kg LPS was used to induce sepsis. Mice in the LPS + UFH group received subcutaneous injection of 8 U UFH 0.5 h before LPS injection. The lung tissue of the mice was collected for assessing lung injury by measuring the lung wet/dry (W/D) weight ratio and observing histological changes. Human pulmonary microvascular endothelial cells (HPMECs) were cultured and used to analyze the effects of UFH on LPS- or tumor necrosis factor-alpha (TNF-α)-induced vascular hyperpermeability, membrane expression of VE-cadherin, p120-catenin, and phosphorylated myosin light chain (p-MLC), and F-actin remodeling, and on the LPS-induced activation of the phosphatidylinositol-3 kinase (PI3K)/serine/threonine kinase (Akt)/nuclear factor kappa-B (NF-κB) signaling pathway.@*RESULTS@#In vivo, UFH pretreatment significantly attenuated LPS-induced pulmonary histopathological changes (neutrophil infiltration and erythrocyte effusion, alveolus pulmonis collapse, and thicker septum), decreased the lung W/D, and increased protein concentration (LPS vs. LPS + UFH: 0.57 ± 0.04 vs. 0.32 ± 0.04 mg/mL, P = 0.0092), total cell count (LPS vs. LPS + UFH: 9.57 ± 1.23 vs. 3.65 ± 0.78 × 10/mL, P = 0.0155), polymorphonuclear neutrophil percentage (LPS vs. LPS + UFH: 88.05% ± 2.88% vs. 22.20% ± 3.92%, P = 0.0002), and TNF-α (460.33 ± 23.48 vs. 189.33 ± 14.19 pg/mL, P = 0.0006) in the bronchoalveolar lavage fluid. In vitro, UFH pre-treatment prevented the LPS-induced decrease in the membrane expression of VE-cadherin (LPS vs. LPS + UFH: 0.368 ± 0.044 vs. 0.716 ± 0.064, P = 0.0114) and p120-catenin (LPS vs. LPS + UFH: 0.208 ± 0.018 vs. 0.924 ± 0.092, P = 0.0016), and the LPS-induced increase in the expression of p-MLC (LPS vs. LPS + UFH: 0.972 ± 0.092 vs. 0.293 ± 0.025, P = 0.0021). Furthermore, UFH attenuated LPS- and TNF-α-induced hyperpermeability of HPMECs (LPS vs. LPS + UFH: 8.90 ± 0.66 vs. 15.84 ± 1.09 Ω·cm, P = 0.0056; TNF-α vs. TNF-α + UFH: 11.28 ± 0.64 vs. 18.15 ± 0.98 Ω·cm, P = 0.0042) and F-actin remodeling (LPS vs. LPS + UFH: 56.25 ± 1.51 vs. 39.70 ± 1.98, P = 0.0027; TNF-α vs. TNF-α + UFH: 55.42 ± 1.42 vs. 36.51 ± 1.20, P = 0.0005) in vitro. Additionally, UFH decreased the phosphorylation of Akt (LPS vs. LPS + UFH: 0.977 ± 0.081 vs. 0.466 ± 0.035, P = 0.0045) and I kappa B Kinase (IKK) (LPS vs. LPS + UFH: 1.023 ± 0.070 vs. 0.578 ± 0.044, P = 0.0060), and the nuclear translocation of NF-κB (LPS vs. LPS + UFH: 1.003 ± 0.077 vs. 0.503 ± 0.065, P = 0.0078) in HPMECs, which was similar to the effect of the PI3K inhibitor, wortmannin.@*CONCLUSIONS@#The protective effect of UFH against LPS-induced pulmonary endothelial barrier dysfunction involves VE-cadherin stabilization and PI3K/Akt/NF-κB signaling.

Comparison of two methods for hemodynamic assessment in children with tetralogy of Fallot after radical surgery / 中国胸心血管外科临床杂志

@#Objective To explore the hemodynamic assessment after radical surgery in children with tetralogy of Fallot (TOF) by both echocardiography and Mostcare monitor. Methods Clinical data of 63 children with TOF who underwent radical surgery in our hospital from February 2016 to June 2018 were retrospectively analyzed, including 34 males and 29 females, aged 6-24 (9.82±5.77) months. There were 19 patients undergoing transannular patch reconstruction of the right ventricular outflow tract (a transannular patch group) while 44 patients retained the pulmonary valve annulus (a non-transannular patch group) . The echocardiography and Mostcare monitor parameters were recorded and brain natriuretic peptide was tested at the time points of 0, 8, 12, 24 and 48 hours after operation (T 0, T 1, T 2, T 4) to analyze their correlations and the change trend at different time points after radical surgery. Results The left ventricular ejection fraction at T 1 (43.49%±3.82%) was lower than that at T 0 (48.29%±4.55%), T 2 (45.83%±3.69%), T 3 (53.76%±4.43%) and T 4 (60.54%±3.23%, P<0.05). The cardiac index at T 1 (1.85±0.35 L·min−1·m−2) was lower than that at T 0 (2.11±0.38 L·min−1·m−2), T 2 (2.07±0.36 L·min−1·m−2), T 3 (2.42±0.37 L·min−1·m−2) and T 4 (2.82±0.42 L·min−1·m−2, P<0.05). The cardiac circulation efficiency at T1 (0.19±0.05) was lower than that at T 0 (0.22±0.06), T 2 (0.22±0.05), T 3 (0.28±0.06) and T 4 (0.34±0.06, P<0.05). The right ventricular two-chambers view fraction area change at T 1 (23.17%±3.11%) was lower than that at T 0 (25.81%±3.74%), T 2 (25.38%±3.43%), T 3 (30.60%±4.50%) and T 4 (36.94%±5.85%, P<0.05). The pulse pressure variability was the highest at T 0 (18.76%±3.58%), followed by T 1 (14.81%±3.32%), T 2 (12.44%±2.94%), T 3 (10.39%±2.96%) and T 4 (9.18%±1.92%, P<0.05). The blood brain natriuretic peptide was higher at T 1 (846.67±362.95 pg/ml) than that at T 0 (42.60±18.06 pg/ml), T 2 (730.95±351.09 pg/ml), T 3 (510.98±290.39 pg/ml) and T 4 (364.41±243.56 pg/ml, P<0.05). There was no significant difference in left ventricular ejection fraction, cardiac circulation efficiency and heart index between the two groups (P>0.05). The right ventricular two-chambers view fraction area change of the transannular patch group was significantly lower than that of the nontransannular patch group at each time point (P<0.05). The blood brain natriuretic peptide and pulse pressure variability of the transannular patch group were significantly higher than those of the non-transannular patch group (P<0.05). Left ventricular ejection fraction was positively correlated with cardiac index (r=0.637, P=0.001) and cardiac circulation efficiency (r=0.462, P=0.001) while was significantly negatively correlated with blood brain natriuretic peptide (r=–0.419, P=0.001). Conclusion Both methods can accurately reflect the state of cardiac function. Mostcare monitor has a good consistency with echocardiography. Using transannular patch to recontribute right ventricular outflow tract in operation has more influence on right ventricular systolic function. The Mostcare monitor can guide the hemodynamic management after surgery in real time, continuously and accurately.