Integrated investigation of DNA methylation, gene expression and immune cell population revealed immune cell infiltration associated with atherosclerotic plaque formation.

BACKGROUND: The clinical consequences of atherosclerosis are significant source of morbidity and mortality throughout the world, while the molecular mechanisms of the pathogenesis of atherosclerosis are largely unknown. METHODS: In this study, we integrated the DNA methylation and gene expression data in atherosclerotic plaque samples to decipher the underlying association between epigenetic and transcriptional regulation. Immune cell classification was performed on the basis of the expression pattern of detected genes. Finally, we selected ten genes with dysregulated methylation and expression levels for RT-qPCR validation. RESULTS: Global DNA methylation profile showed obvious changes between normal aortic and atherosclerotic lesion tissues. We found that differentially methylated genes (DMGs) and differentially expressed genes (DEGs) were highly associated with atherosclerosis by being enriched in atherosclerotic plaque formation-related pathways, including cell adhesion and extracellular matrix organization. Immune cell fraction analysis revealed that a large number of immune cells, especially macrophages, activated mast cells, NK cells, and Tfh cells, were specifically enriched in the plaque. DEGs associated with immune cell fraction change showed that they were mainly related to the level of macrophages, monocytes, resting NK cells, activated CD4 memory T cells, and gamma delta T cells. These genes were highly enriched in multiple pathways of atherosclerotic plaque formation, including blood vessel remodeling, collagen fiber organization, cell adhesion, collagen catalogic process, extractable matrix assembly, and platelet activation. We also validated the expression alteration of ten genes associated with infiltrating immune cells in atherosclerosis. CONCLUSIONS: In conclusion, these findings provide new evidence for understanding the mechanisms of atherosclerotic plaque formation, and provide a new and valuable research direction based on immune cell infiltration.

Neutrophil-to-Lymphocyte Ratios Are Closely Associated With the Severity and Course of Non-mild COVID-19.

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection is spreading worldwide. Measuring the prevention and control of the disease has become a matter requiring urgent focus. Objective: Based on coronavirus disease 2019 (COVID-19) clinical data from Wuhan, we conducted an in-depth analysis to clarify some of the pathological mechanisms of the disease and identify simple measures to predict its severity early on. Methods: A total of 230 patients with non-mild COVID-19 were recruited, and information on their clinical characteristics, inflammatory cytokines, and T lymphocyte subsets was collected. Risk factors for severity were analyzed by binary logistic regression, and the associations of neutrophil-to-lymphocyte ratios (N/LRs) with illness severity, disease course, CT grading, inflammatory cytokines, and T lymphocyte subsets were evaluated. Results: Our results showed that the N/LRs were closely related to interleukin (IL)-6 and IL-10 (P < 0.001, P = 0.024) and to CD3+ and CD8+ T lymphocytes (P < 0.001, P = 0.046). In particular, the N/LRs were positively correlated with the severity and course of the disease (P = 0.021, P < 0.001). Compared to the values at the first test after admission, IL-6 and IL-10 were significantly decreased and increased, respectively, as of the last test before discharge (P = 0.006, P < 0.001). More importantly, through binary logistic regression, we found that male sex, underlying diseases (such as cardiovascular disease), pulse, and N/LRs were all closely related to the severity of the disease (P = 0.004, P = 0.012, P = 0.013, P = 0.028). Conclusions: As a quick and convenient marker of inflammation, N/LRs may predict the disease course and severity level of non-mild COVID-19; male sex, cardiovascular disease, and pulse are also risk factors for the severity of non-mild COVID-19.

T-plate fixation for unstable proximal clavicula fractures.

OBJECTIVE: The aim of this study was to evaluate the clinical results of T-plate fixation and anterior sternoclavicular ligament repair in proximal clavicle fractures. METHODS: Between August 2013 and August 2016, a total 12 patients (10 men and 2 women; mean age: 44.1 ± 9.1 years (range, 25-59 years)) with unstable proximal clavicle fractures (Throckmorton, type D) were treated with T-type plate fixation, bridging the sternoclavicular joint, and anterior sternoclavicular ligament repair. Average operative time, associated injuries, postoperative complications, postoperative fracture healing time and follow-up time were recorded. The outcomes were evaluated with radiographic assessment, visual analog scale (VAS) pain score and Rockwood SCJ scoring system. All the patients were evaluated on postoperative 3rd, 6th, and 12th months. RESULTS: The average surgery time was 78.0 ± 8.47 minutes while fracture healing time was 4.51 ± 0.95 months. According to Rockwood SCJ scoring system, 9 cases (75%) were in excellent, 2 cases (16.7%) in good and 1 case (8.3%) in fair condition at 12 months follow-up. The average Rockwood SCJ score was 7.7 ± 0.75 preoperatively, 12.7 ± 0.86 by 3 months, 13.0 ± 0.73 by 6 months and 13.3 ± 0.49 by 12 months. The VAS pain score was 7.9 ± 1.15 (preoperative score), 3.4 ± 1.52 (3 months follow-up), 3.0 ± 1.32 (6 months follow-up) and 2.1 ± 1.07 (12 months follow-up). The VAS and Rockwood SCJ scores were significantly improved postoperatively (p < 0.05). There was no intraoperative complication, while one patient had redislocation of the sternoclavicular joint after implant removal. CONCLUSION: T-type plate fixation with anterior sternoclavicular ligament repair might be a reliable and effective treatment method in unstable proximal clavicle fractures (type D) with few complications and satisfactory clinical results after 12 months follow-up. LEVEL OF EVIDENCE: Level IV, therapeutic study.

[Correlation between Gan Depression Qi Stagnation Syndrome and Thyroid Stimulating Hormone Level in Adolescent Polycystic Ovary Syndrome Patients].

Objective To observe the distribution features of Gan depression qi stagnation syndrome (GDQSS) , and to analyze the correlation between GDQSS scores and thyroid stimulating hormone (TSH) level in adolescent polycystic ovary syndrome (PCOS) patients. Methods Disease location and disease nature were syndrome identified in PCOS outpatients confirmed at Department of Gynecology, Ningbo Municipal Hospital of TCM. Totally 110 patients in line with GDQSS were graded by GDQSS score from low to high. GDQSS score ranging 70 -100 belonged to grade I (mild) ; 101 -150 to grade II (moderate) ; >150 to grade III (severe). The correlation between the severity of GDQSS and TSH level was an- alyzed using Pearson and Partial correlation analyses. Results Levels of TSH, testosterone (T) , pro- lactin (PRL) were the lowest in grade I GDQSS patients, and they were the highest in grade lIl GDQSS patients (P <0. 01). Pearson correlation analysis showed that GDQSS score was positively correlated with TSH level (r =0. 676, P <0. 01). They were still positively correlated by using Partial correlation after controlling body mass index (BMI) , waist hip ratio (WHR) , homeostasis model of assessment-insulin resistance (HOMR-IR) factors (r =0. 663, P <0. 01). Conclusion Adolescent PCOS patients with obvi- ous GDQSS symptoms might be associated with increased TSH level.