Characterizing the skeletal muscle immune microenvironment for sarcopenia: insights from transcriptome analysis and histological validation.

Background: Sarcopenia is a condition characterized by the age-related loss of skeletal muscle mass and function. The pathogenesis of the disease is influenced by chronic low-grade inflammation. However, the specific changes in the immune landscape changes of sarcopenic muscle are not yet fully understood. Methods: To gain insights into the immune cell composition and interactions, we combined single-nucleus RNA sequencing data, bulk RNA sequencing dataset, and comprehensive bioinformatic analyses on the skeletal muscle samples from young, aged, and sarcopenic individuals. Histological staining was then performed on skeletal muscles to validate the distribution of immune cells in clinical samples. Results: We analyzed the transcriptomes of 101,862 single nuclei, revealing a total of 10 major cell types and 6 subclusters of immune cell types within the human skeletal muscle tissues. Notable variations were identified in the immune microenvironment between young and aged skeletal muscle. Among the immune cells from skeletal muscle microenvironment, macrophages constituted the largest fraction. A specific marker gene LYVE1 for skeletal muscle resident macrophages was further identified. Cellular subclasses included four distinct groups of resident macrophages, which play different roles in physiological or non-physiological conditions. Utilizing bulk RNA sequencing data, we observed a significant enrichment of macrophage-rich inflammation in sarcopenia. Conclusions: Our findings demonstrate age-related changes in the composition and cross-talk of immune cells in human skeletal muscle microenvironment, which contribute to chronic inflammation in aged or sarcopenia muscle. Furthermore, macrophages emerge as a potential therapeutic target, thus advancing our understanding of the pathogenesis of sarcopenia.

Identification of key mitochondria-related genes and their relevance to the immune system linking Parkinson's disease and primary Sjögren's syndrome through integrated bioinformatics analyses.

BACKGROUND: Mitochondria are the metabolic hubs of cells, regulating energy production and antigen presentation, which are essential for activation, proliferation, and function of immune cells. Recent evidence indicates that mitochondrial antigen presentation may have an impact on diseases such as Parkinson's disease (PD) and autoimmune diseases. However, there is limited knowledge about the mechanisms that regulate the presentation of mitochondrial antigens in these diseases. METHODS: In the current study, RNA sequencing was performed on labial minor salivary gland (LSG) from 25 patients with primary Sjögren's syndrome (pSS) and 14 non-pSS aged controls. Additionally, we obtained gene expression omnibus datasets associated with PD patients from NCBI Gene Expression Omnibus (GEO) databases. Single-sample Gene Set Enrichment Analysis (ssGSEA), ESTIMATE and Spearman correlations were conducted to explore the association between mitochondrial related genes and the immune system. Furthermore, we applied weighted Gene Co-expression Network Analysis (WGCNA) to identify hub mitochondria-related genes and investigate the correlated networks in both diseases. Single cell transcriptome analysis, immunohistochemical (IHC) staining and quantitative real-time PCR (qRT-PCR) were used to verify the activation of the hub mitochondria-related pathway. Pearson correlations and the CIBERSORT algorithms were employed to further reveal the correlation between hub mitochondria-related pathways and immune infiltration. RESULTS: The transcriptome analysis revealed the presence of overlapping mitochondria-related genes and mitochondrial DNA damage in patients with pSS and PD. Reactive oxygen species (ROS), the senescence marker p53, and the inflammatory markers CD45 and Bcl-2 were found to be regionally distributed in LSGs of pSS patients. WGCNA analysis identified the STING pathway as the central mitochondria-related pathway closely associated with the immune system. Single cell analysis, IHC staining, and qRT-PCR confirmed the activation of the STING pathway. Subsequent, bioinformatic analysis revealed the proportion of infiltrating immune cells in the STING-high and STING-low groups of pSS and PD. Furthermore, the study demonstrated the association of the STING pathway with innate and adaptive immune cells, as well as functional cells, in the immune microenvironment of PD and pSS. CONCLUSION: Our study uncovered a central pathway that connects mitochondrial dysfunction and the immune microenvironment in PD and pSS, potentially offering valuable insights into therapeutic targets for these conditions.

Measurement of three-dimensional changes in lip vermilion in adult female patients after orthodontic extraction: a retrospective longitudinal study.

BACKGROUND: 3D facial scanning has changed the way facial aesthetic is evaluated and has numerous advantages for facial analysis. The specific relationship between lip vermilion morphological changes after orthodontic extraction treatment has not been fully explained. The objective of this study was to evaluate 3D morphological changes after orthodontic extraction treatment in lip vermilion of adult females with dentoalveolar protrusion using a structured light-based scanner. METHODS: Forty-two female subjects (25.2 ± 1.9 years) were recruited as the treatment group; these patients had undergone extraction treatment and achieved better sagittal profiles. Twenty female subjects (25.5 ± 2.1 years) were enrolled in the non-treatment group; these patients did not require any orthodontic treatment. The follow up time for the treatment group was more than 24 months and for the non-treatment group was more than 12 months. 3D facial scans were captured using 3D CaMega. Six landmarks (Ls, Li, R.Chp, L.Chp, R.Ch, and L.Ch), three linear measurements (mouth height, philtrum width, and mouth width), and three area measurements (upper, lower, and total vermilion area) were measured. The spatial deviations of three volumetric measurements (upper, lower, and total vermilion) were constructed for quantitative analysis. Color-coded displacement map were constructed for visualization of the soft-tissue displacement as qualitative evaluation. RESULTS: Mouth height and philtrum width decreased (-0.93 mm and - 1.08 mm, respectively) significantly (p = 0.008 and p = 0.027, respectively), and no significant (p = 0.488) change in mouth width was observed in the treatment group. The lower and total vermilion surface areas decreased (-51.00mm2 and - 69.82mm2, respectively) significantly (p = 0.003 and p = 0.031, respectively) in the treatment group, but no statistically significant (p = 0.752) change was detected in the upper vermilion. In the treatment group, significant retractions were observed in the color-coded displacement map, and three volumetric measurements of vermilion changed significantly (p = 0.012, p = 0.001 and p = 0.004, respectively). Significant differences were found between the treatment group and the non-treatment group in the linear, area and volumetric measurements. CONCLUSIONS: This study established a method for qualitative and quantitative evaluation of the lip vermilion. Significant 3D retraction of the lip vermilion after the extraction treatment was found, with morphological variation between upper and lower vermilion.

Thickness change of masseter muscles and the surrounding soft tissues in female patients during orthodontic treatment: a retrospective study.

BACKGROUND: Facial esthetics is a major concern of orthodontic patients. This study aims to evaluate orthodontic treatment-related thickness changes of the masseter muscles and surrounding soft tissues and the potential factors that would influence these changes during orthodontic treatment in female adults. METHODS: Forty-two female adult patients were included in this retrospective study and were divided into extraction (n = 22) and nonextraction (n = 20) groups. Pretreatment and posttreatment cone-beam computed tomography (CBCT) images were superimposed and reconstructed. The thickness changes of the masseter area of facial soft tissue (MAS), masseter muscles (MM) and surrounding fat tissue (FT) were measured. Pretreatment age, treatment duration, sagittal relationship (ANB), and vertical relationship (Frankfort-mandibular plane angle, FMA)-related MAS, MM and FT changes were compared between extraction and nonextraction groups. Spearman's correlation coefficient was calculated between the above variables. Regression analysis was conducted to confirm the causal relations of the variables. RESULTS: The thickness of MAS and MM significantly decreased in both groups, with larger decreases (> 1 mm) in the extraction group. There were strong correlations (r > 0.7) between the thickness decrease in MAS and MM in both groups and moderate correlations (r > 0.4) between MAS and FT in the nonextraction group. A significantly greater decrease of MAS and MM were found to be moderately correlated with a smaller FMA (r > 0.4) in the extraction group. Scatter plots and regression analysis confirmed these correlations. CONCLUSIONS: Masseter muscles and the surrounding soft tissue exhibited a significant decrease in thickness during orthodontic treatment in female adults. Low-angle patients experienced a greater decrease in soft tissue thickness in the masseter area in the extraction case. But the thickness changes were clinically very small in most patients.

Impact of increased serum 8-hydroxy-2'-deoxyguanosine levels on extent of coronary artery lesions in elderly patients with type 2 diabetes.

OBJECTIVE: Patients with type 2 diabetes (T2DM) are prone to cardiovascular disease, and both conditions are linked to oxidative DNA damage, which produces 8-hydroxy-2'-deoxyguanosine (8-OHdG). We investigated the impact of 8-OHdG on coronary heart disease (CHD) in elderly patients with T2DM. METHODS: We assessed the demographic, clinical, and biochemical characteristics of 147 patients with T2DM (mean age 73.29 ± 8.19 years) with or without CHD. Serum 8-OHdG was detected by enzyme-linked immunosorbent assay. CHD was diagnosed as ≥50% stenosis in at least one main branch of the coronary arteries determined by coronarography, evaluated by Gensini score. RESULTS: Serum 8-OHdG, number of stenotic branches, and Gensini score were all significantly increased in the CHD group. After adjustment for various factors, the number of stenotic branches and Gensini score remained positively correlated with 8-OHdG levels in the CHD group. Coronary artery lesions were significantly more severe in the CHD compared with the non-CHD group when 8-OHdG levels were >0.523 ng/mL. The number of stenotic branches and Gensini score were significantly independently associated with 8-OHdG levels in patients with T2DM. CONCLUSIONS: 8-OHdG is a marker of oxidative DNA damage, and is highly associated with the extent of coronary artery lesions in ageing patients with T2DM.Trial registration: Registration number: 1.0/20170720; date of registration 26/07/2016 (retrospectively registered).

Posteroanterior cephalometric analysis of White-American and Chinese adolescents: a cross-sectional study.

Objective: To characterize ethnic differences between Chinese and White-Americans between 8.5 and 17.5 years of age, with respect to transverse cephalometric characteristics and to establish transverse craniofacial normative values for Chinese adolescents. Methods: Two-hundred fifty-seven and 547 posteroanterior cephalograms were selected from 35 White-Americans and 157 Chinese with individual normal occlusions. Transverse measurements were obtained and compared between ethnicities to guide determination of normative values for Chinese adolescents. Student's t-test or one-way analysis of variance was used, as appropriate. Results: Chinese girls demonstrated significantly larger measurements than White-American girls at all ages, with the exception of nasal width. Chinese boys exhibited larger measurements than White-American boys at different ages. Chinese boys had larger measurements than girls for most measurements. These data established normative values for Chinese adolescents. Discussion: Ethnic differences existed between Chinese and White-American adolescents with respect to transverse craniofacial measurements. Transverse normative values were established for Chinese adolescents.

Association of C1q/TNF-related protein-1 (CTRP1) serum levels with coronary artery disease.

OBJECTIVE: Complement C1q tumor necrosis factor-related proteins (CTRPs), belonging to the CTRP superfamily, are extensively involved in regulating metabolism and the immune-inflammatory response. The inflammatory process is linked to the pathogenesis of coronary artery disease (CAD). Here, we investigated the association of serum levels of CTRP1 with CAD. METHODS: Study participants were divided into two groups according to the results of coronary angiography: a control group (n = 63) and a CAD group (n = 76). The concentrations of serum CTRP1 and inflammatory cytokines were determined by enzyme-linked immunosorbent assay. Further analysis of CTRP1 levels in individuals with different severities of CAD was conducted. The CAD severity was assessed by Gensini score. RESULTS: Serum levels of CTRP1 were significantly higher in CAD patients than in controls (17.24 ± 1.07 versus 9.31 ± 0.56 ng/mL), and CTRP1 levels increased with increasing severity of CAD. CTRP1 levels were positively correlated with concentrations of tumor necrosis factor-α and interleukin-6. Multiple logistic regression analysis showed that CTRP1 was significantly associated with CAD. CONCLUSIONS: Our data showed close associations of serum CTRP1 levels with the prevalence and severity of CAD, indicating that CTRP1 can be regarded as a novel and valuable biomarker for CAD.

Adiponectin modulates the function of endothelial progenitor cells via AMPK/eNOS signaling pathway.

Endothelial progenitor cells have been shown to differentiate into endothelial cells and to play a pivotal role in vascular homeostasis. Adiponectin has anti-atherogenic and anti-inflammatory properties via directly acting on vascular cells. The aim of the present study is to explore the effect of adiponectin on major functions involved in survival, migration, and tube formation of endothelial progenitor cells and to explore the underlying mechanism. In this study, we transferred adiponectin gene into endothelial progenitor cells via lentiviral vectors and investigated the proliferation, migration and tube formation of these transfected cells. We found that adiponectin is highly expressed in endothelial progenitor cells and promotes their proliferation, migration and tube formation. Western blot data showed that the former two processes were mediated through the AMPK/Akt/eNOS pathway, the latter via the AMPK/eNOS pathway. Use of the AMPK inhibitor (Compound C) or Akt inhibitor (MK-2206) reduced eNOS phosphorylation and attenuated adiponectin-induced endothelial progenitor cell proliferation, migration and tube formation compared to the controls (p < 0.05). Taken together, these data indicated that adiponectin promotes endothelial progenitor cell proliferation and migration via AMPK/Akt/eNOS signaling pathway and promotes tube formation through AMPK/eNOS, suggesting that adiponectin-transduced endothelial progenitor cell transplantation is a potential therapeutic target for vascular disease.

Association of serum C1q/TNF-related protein-3 (CTRP-3) in patients with coronary artery disease.

BACKGROUND: Recent studies have demonstrated that complement C1q tumor necrosis factor related proteins (CTRPs) have diverse biological influences on the cardiovascular system. CTRP 3 is a member of the CTRP superfamily, which may play a pivotal role in the pathogenesis of coronary artery disease (CAD). Here, we investigated whether serum levels of CTRP 3 are associated with the prevalence and the severity of CAD. METHODS: In this study, 145 eligible participants were included who underwent coronary angiography. According to the result of the coronary angiography, all participants were divided into two groups: non-CAD group (n = 66) and CAD group (n = 79). The CAD group was further divided into single-vessel (n = 25), double-vessel (n = 30) and triple-vessel (n = 24) disease groups in line with different lesioned vessels of CAD. Plasma CTRP 3 concentration was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Serum levels of CTRP 3 were significantly higher in CAD patients than in non-CAD patients (CAD: 56.68 ± 3.63 ng/ml, non-CAD: 44.10 ± 3.20 ng/ml, p < 0.01). Significant differences of CTRP 3 levels were also found between single-vessel group and triple-vessel group (single-vessel group: 44.80 ± 3.14 ng/ml, triple-vessel group: 75.07 ± 9.41 ng/ml, p < 0.005). Multiple logistic regression analysis revealed that CTRP 3 levels, together with HDL cholesterol and glucose, correlated with CAD. CONCLUSIONS: Elevated serum CTRP 3 levels were closely related to the prevalence and severity of CAD, suggesting that it might be regarded as a novel biomarker for CAD.

Effects of antioxidant system on coronary artery lesions in patients with abnormal glucose metabolism.

OBJECTIVE: To evaluate the effects of catalase (CAT), superoxide dismutase (SOD), and glutathione (GSH) system on the range and severity of coronary artery lesions in patients with abnormal glucose metabolism. METHODS: This study included 766 patients with abnormal glucose metabolism. Levels of fasting blood lipids, blood glucose, CAT, SOD, GSH, glutathione reductase (GR), and glutathione peroxidase (GSH-Px) were measured in all the patients. Coronarography was performed, and the degree of the lesions in each coronary artery was quantitatively analyzed using the Gensini scoring system. RESULTS: In patients with impaired glucose regulation and diabetes, the number of coronary artery branches with stenosis and the Gensini scores were inversely correlated with the plasma levels of CAT, SOD, GSH, GH, and GSH-Px (P < 0.001). Patients were grouped according to the Gensini scores. As the scores increased, the levels of CAT, SOD, GSH, GSH-Px, and GR gradually decreased. Logistic gradual regression analysis showed that GSH-Px and CAT were independent risk factors associated with the coronary artery lesions in three or more branches. CONCLUSIONS: Decreased plasma levels of CAT, SOD, GSH, GR, and GSH-Px were inversely correlated, at least to some extent, with the extent of coronary artery lesions. Particularly, GSH-Px and CAT were independent risk factors associated with coronary artery lesions involving three or more branches. This suggests that long-term hyperglycemia leads to enhancement of oxidative stress.

Expression of brain adiponectin in a murine model of transient cerebral ischemia.

OBJECTIVE: Adiponectin is a hormone that is mainly secreted by fat cells. Adiponectin has anti-inflammatory and anti-atherosclerotic effects, and a protective effect against ischemic brain injury, but the level of expression of adiponectin in brain tissue is unknown. In the current study, a mouse model of transient cerebral ischemia was used to determine the level of expression of adiponectin in ischemic brain tissue. METHODS: Sixty CD-1 mice underwent transient middle cerebral artery occlusion. The level of expression of adiponectin in mouse brain tissues 1 hour, 4 hours, 1 day, 3 days, and 7 days, after cerebral ischemia/reperfusion injury were determined using a real-time quantitative polymerase chain reaction, Western blot, and immunohistochemistry. RESULTS: The level of expression of adiponectin in mouse ischemic brain tissues increased after cerebral ischemia/reperfusion injury and was higher in the central area of ischemia than in the peripheral area. The level of expression of adiponectin occurred only in vascular endothelial cells. There was no significant change in the level of expression of adiponectin mRNA in brain tissue pre- and post-ischemia/reperfusion injury. CONCLUSION: After cerebral ischemia/reperfusion injury, adiponectin accumulated in the vascular endothelial cells of ischemic brain tissues, and non-endogenous adiponectin was generated. Circulating adiponectin accumulated in ischemic brain tissues through its role in adhering to damaged vascular endothelial cells.

Overexpression of adiponectin improves neurobehavioral outcomes after focal cerebral ischemia in aged mice.

AIMS: To study whether adiponectin (APN) could improve neurological outcomes in aged mice after ischemic stroke. METHODS: Adeno-associated virus carrying APN gene was injected into aged and young adult mice 7 days before transient middle cerebral artery occlusion (tMCAO). Atrophic volumes and neurobehavioral deficiencies were determined up to 28 days after tMCAO. Focal angiogenesis was determined based on blood vessel number in the ischemic regions. RESULTS: Increased atrophic volume and more sever neurobehavioral deficits were found in the aged mice compared with young adult mice (P < 0.05). AAV-APN gene transfer attenuated atrophic volume and improved neurobehavioral outcomes, along with increased focal angiogenesis in both aged and young adult mice, compared with control animals (P < 0.05). In addition, the attenuation of atrophic volume and the improvement in neurobehavioral outcomes were much more significant in aged mice than in young adult mice after AAV-APN administration (P < 0.05). The number of microvessels in aged AAV-APN mouse ischemic brain was higher than in young adult AAV-APN treated mouse brain (P < 0.05). CONCLUSIONS: Our results demonstrate that APN overexpression reduces ischemic brain injury and improves neurobehavioral function recovery in aged mice than in young mice, suggesting APN is more beneficial in aged animals after ischemic stroke.