Exosomes secreted from osteocalcin-overexpressing endothelial progenitor cells promote endothelial cell angiogenesis.

Exosome secretion is an important paracrine way of endothelial progenitor cells (EPCs) to modulate resident endothelial cells. The osteocalcin (OCN)-expressing EPCs have been found to be increased in cardiovascular disease patients and are considered to be involved in the process of coronary atherosclerosis. Since OCN has been proven to prevent endothelial dysfunction, this study aimed to evaluate the effect of exosomes derived from OCN-overexpressed EPCs on endothelial cells. Exosomes derived from EPCs (Exos) and OCN-overexpressed EPCs (OCN-Exos) were isolated and incubated with rat aorta endothelial cells (RAOECs) with or without the inhibition of OCN receptor G protein-coupled receptor family C group 6 member A (GPRC6A). The effects of exosomes on the proliferation activity of endothelial cells were evaluated by CCK-8 assay, and the migration of endothelial cells was detected by wound healing assay. A tube formation assay was used to test the influence of exosomes on the angiogenesis performance of endothelial cells. Here, we presented that OCN was packed into Exos and was able to be transferred to the RAOECs via exosome incorporation, which was increased in OCN-Exos groups. Compared with Exos, OCN-Exos had better efficiency in promoting RAOEC proliferation and migration and tube formation. The promoting effects were impeded after the inhibition of GPRC6A expression in RAOECs. These data suggest that exosomes from OCN-overexpressed EPCs have a beneficial regulating effect on endothelial cells, which involved enhanced OCN-GPRC6A signaling.

Emodin azide methyl anthraquinone derivative induced G0/ G1 arrest in HER2/neu-overexpressing MDA-MB-453 breast cancer cells.

PURPOSE: Our previous data have shown that emodin azide methyl anthraquinone derivative (AMAD) triggered mitochondrial- dependent cell apoptosis involving caspase-8-mediated Bid cleavage, and induced proteasomal degradation of HER2/neu by blocking Her2/neu binding to Hsp90. In the present study, we futher investigated the effect of this compound on the cell cycle and related molecular mechanisms in HER2/neu-overexpressing MDA-MB-453 breast cancer cells. METHODS: The cell cycle distribution was tested by flow cytometry. The expression of cell cycle-related proteins was determined by Western blot analysis; DNA agarose gel electrophoresis was used to examine the apoptosis of MDAMB- 453 cells induced by emodin AMAD. RESULTS: After MDA-MB-453 cells were treated with different concentrations of emodin AMAD for 24 hrs, cells were arrested in G0/G1 phase, and the expression of G0/G1 related proteins c/Myc, Cyclin D1, CDK4 and p-Rb changed. DNA fragmentation appeared on the agarose gel in a concentration- dependent manner. CONCLUSION: Emodin AMAD induced G0/G1 arrest in Her2/ neu-overexpressing MDA-MB-453 cancer cells. This G0/G1 arrest was associated with decreasing protein expression of c-Myc, Cyclin D1, CDK4, and p-Rb.

Chlorogenic Acid Inhibits Proliferation, Migration and Invasion of Pancreatic Cancer Cells via AKT/GSK-3ß/ß-catenin Signaling Pathway.

BACKGROUND: Chlorogenic acid (CA, United States Patent No. 10772340), a natural biologically active food ingredient, displays potent antitumor activity against a variety of cancer cells. However, the mechanism underlying its anticancer effect is not well elucidated. OBJECTIVE: In the present study, we hope to dissect the mechanism underlying the anticancer effects of CA in pancreatic cancer cells. METHODS: The cytotoxicity of CA in pancreatic cancer cells was determined by MTT assay. Flow cytometry was performed to evaluate the cells apoptosis, while a clonogenic assay was carried out to check the colony formation of cancer cells. Transwell assay was performed to assess the cells migration and invasion. The protein expression of AKT/GSK-3ß/ß-catenin signaling pathway was detected by Western Blot. RESULTS: Our data indicated that CA inhibited the proliferation of PANC-28 and PANC-1 cells in a dose and time-dependent manner. CA was able to inhibit colony formation, migration, and invasion ability and trigger apoptosis in PANC-28 and PANC-1 cells. Further study showed that CA down-regulated the expression of AKT, p-AKT(Thr308), p-GSK-3ß(Ser9), ß-catenin, N-cadherin, and vimentin while enhancing the expression of cleaved-caspase 3 and cleaved-caspase 7 in PANC-28 and PANC-1 cells. CONCLUSION: Our study provides significant evidence that CA is able to inhibit the growth of pancreatic cancer via the AKT/GSK-3ß/ß-catenin signaling pathway.

The Deficiency of the ASD-Related Gene CHD8 Disrupts Behavioral Patterns and Inhibits Hippocampal Neurogenesis in Mice.

Chromodomain helicase DNA-binding 8 (CHD8) is a gene that poses a high risk for autism spectrum disorder (ASD) and neurological development delay. Nevertheless, the impact of CHD8 haploinsufficiency on both hippocampus neurogenesis and behavior remains uncertain. Here, we performed behavioral assessments on male and female CHD8 heterozygous mice. The study discovered that both male and female CHD8 heterozygous mice displayed an impairment in preference for social novelty. Concurrently, CHD8 heterozygous mice exhibited anxiety-like behavior. However, its cognitive capacity for learning and memory is within the expected range. Furthermore, we discovered a reduction in the number of both immature and mature new neurons in mice with CHD8 heterozygous, resulting in an impeded neurogenesis process in the hippocampus. Taken together, our findings indicate that CHD8 plays a crucial role in the regulation of hippocampal neurogenesis, and further suggest that ASD-like behaviors observed in CHD8 heterozygous mice may be associated with disruptions in hippocampal neurogenesis.

Plaque characteristics in patients with ST-segment elevation myocardial infarction and early spontaneous reperfusion.

BACKGROUND: Early spontaneous reperfusion (ESR) is not an uncommon phenomenon in clinical settings. AIMS: The aim of this study was to detect potential mechanisms of ESR in patients with STEMI. METHODS: This prospective study enrolled a total of 241 consecutive patients with STEMI undergoing optical coherence tomography (OCT) from July 2016 to August 2019. Forty-five patients (18.7%) met angiographic ESR criteria (TIMI 3 flow on the initial angiogram). Among those without ESR (TIMI 0 flow on initial angiogram), 45 patients were assigned to the control group according to propensity score matching with the ESR group. RESULTS: Although the baseline characteristics of the groups were comparable, non-ruptured plaque (62.2% vs 35.6%) predominated and plaque rupture (37.8% vs 64.4%) was less common in the ESR group (p=0.011). Red thrombus (44.4% vs 77.8%) was also less common in the ESR group (p=0.001). Lastly, compared to the control group, the ESR group underwent fewer emergent stent placements (68.9% vs 91.1%, p=0.008). CONCLUSIONS: Relief of coronary occlusion induced by a non-ruptured plaque may contribute to ESR in patients with STEMI.

The Social Utility and Health Benefits for Older Adults of Amenity Buildings in China's Urban Parks: A Nanjing Case Study.

As China's population rapidly ages, research and discussion on how to better optimize public spaces for the elderly's health and benefit continue to deepen. This study uses observational surveys and questionnaires to investigate the elderly visitors of Nanjing's urban parks and explore the impact the parks' amenity buildings (structures built to provide visitors with conveniences, e.g., shelters and pavilions) has on their health and associated socialization tendencies. Data were collected from ten amenity buildings in ten separate parks to compose a total dataset of 728 activity statistics and 270 valid questionnaires. The study's results indicate that amenity buildings significantly increase opportunities for older adults to socialize and thereby can increase this demographic's associated health benefits. The social activities formed around amenity buildings are found to improve social interactions and connectedness among older adults more compared to other age groups. Elderly participation in social activities is also found to positively correlate with environmental characteristics. High-quality landscapes ensure healthy development of social activities within amenity buildings and promote the occurrence and continuation of social interactions. In order of highest to lowest impact on elderly activities, the following factors were identified and scored: amenity building scale, lighting, comprehensive surrounding environment, surrounding amenities, water features, and vegetation. This research also reveals that among existing amenity buildings, there is insufficient support for certain activities and therefore, parks need to be improved to address this deficiency. Overall, this study indicates that under China's current aging trends, amenity buildings have become an especially important infrastructure within urban public space, and their design trend is to incorporate the dual characteristics of "recreation + society".

Ganoderic acid A from Ganoderma lucidum ameliorates lipid metabolism and alters gut microbiota composition in hyperlipidemic mice fed a high-fat diet.

Ganoderic acid A (GA) is one of the most abundant triterpenoids in Ganoderma lucidum, and has been proved to possess a wide range of beneficial health effects. The aim of the current study is to investigate the amelioration effects and mechanism of GA on improving hyperlipidemia in mice fed a high-fat diet (HFD). The results showed that GA intervention significantly inhibited the abnormal growth of body weight and epididymal white adipose tissue (eWAT), prevented the hypertrophy of epididymal adipocytes, and ameliorated the biochemical parameters of serum and liver related to lipid metabolism in HFD-fed mice. Histological analysis also showed that the excessive accumulation of lipid droplets in the liver induced by HFD-feeding was greatly alleviated by GA intervention. In addition, GA intervention also increased the level of short chain fatty acids (SCFAs) in the intestine and promoted the excretion of bile acids (BAs) through feces. High-throughput sequencing of bacterial full-length 16S rDNA revealed that daily supplementation with GA made significant structural changes in the gut microbial population of mice fed with HFD, in particular modulating the relative abundance of some function related microbial phylotypes. The relationships between lipid metabolic parameters and gut microbial phylotypes were also revealed by correlation analysis based on a heatmap and network. The result showed that 46 key gut microbial phylotypes (OTUs) were markedly correlated with at least one lipid metabolic parameter. Moreover, UPLC-QTOF/MS-based liver metabolomics showed that 111 biomarkers (47 up-regulated metabolites and 64 down-regulated metabolites) were significantly changed after high-dose GA intervention (75 mg kg-1 day-1), compared with the HFD-fed hyperlipidemic mice. Metabolic pathway enrichment analysis of the differential hepatic metabolites demonstrated that GA intervention had significant regulatory effects on primary bile acid biosynthesis, fatty acid biosynthesis, amino sugar and nucleotide sugar metabolism, inositol phosphate metabolism, and so on. In addition, GA intervention regulated the mRNA levels of hepatic genes involved in fatty acid metabolism and bile acid homeostasis. These findings present new evidence supporting that GA from G. lucidum has the potential to alleviate lipid metabolic disorders and ameliorate the imbalance of gut microflora in a positive way.

Immunomodulatory and neuroprotective mechanisms of Huangqi glycoprotein treatment in experimental autoimmune encephalomyelitis.

Previous studies have shown that Huangqi glycoprotein (HQGP) has an anti-inflammatory effect in vitro, and suppressed experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis; however, the mechanism underlying its effect is largely unknown. In this manuscript we investigated the mechanisms by which HQGP protect mice from EAE. HQGP was extracted from Astragalus membranaceus and purified by anion-exchange and gel filtration chromatography. HQGP delayed disease onset, reduced disease severity and alleviated inflammation and demyelination in the central nervous system (CNS). Moreover, HQGP reduced the infiltration of pathogenic immune cells and increased the expression of microtubule-associated protein 2 (MAP-2) and neuronal nuclei (NeuN) in the CNS. HQGP treatment also reduced the expression of chemokines such as CCL2 and CCL5 and the production of tumor necrosis factor α (TNF-α), interleukin (IL)-1ß, IL-6, but increased the level of IL-10. These results demonstrate that HQGP suppressed EAE development by modulating the immune system and the infiltration of leukocytes to the CNS as well as promoting axon and neural repair.

[The influence of zi-hua burn cream on the survival of random skin flaps in rats].

OBJECTIVE: To observe the effect of zi-hua burn cream on the survival of skin flaps in rats, and its mechanisms. METHODS: 72 Wistar rats, were randomly divided into four groups as zi-hua group(n = 18, external application of alfalfa burn cream), control group (n = 18, external application of heparin sodium cream), model group (n = 18, external application of vaseline) , negative control (n = 18, no operation). 8 cm x 2 cm random skin flaps with pedicle on the side of head were designed on the back of Wistar rats. The drug was applied on the flap surface, 2 times a day. The survival of skin flaps was observed. The change of serum superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO), turner necrosis factor-alpha(TNF-alpha)and interleukin-6(IL-6)were compared at 1,2,3,7 d after operation, and histologic examination was performed. RESULTS: The survival rate of zi-hua group (73.58 - 10. 74)% was significantly higher than that of model group (33.40 - 16.05) %, showing a statistical difference (Q = 10.63, P <0.01). There was no significant difference between the zi-hua group and control group (71.65 +/- 11. 92) %. The level of serum SOD, NO in zi-hua group and control group was higher than that in model group, while the level of serum MDA, TNF-alpha and IL-6 was lower than that in model group(P <0.01). On 7 day after operation, skin flaps tissue edema,necrosis and inflammatory cell infiltration in zi-hua group and control group was less obvious than that in model group. There was significant proliferation of granuloma and fibroblast and formation of neonatal capillary in zi-hua group and control group. The vascular density in zi-hua group was obviously higher than that in the model group and control group(P <0. 01). CONCLUSIONS: Zi-hua burn cream could significantly improve the blood supply of skin flaps, increase the survival rate of skin flaps in rats. Its mechanism may be associated with the anti-free-radical-damage action, improve local microcirculation, improve the NO content, reduce the TNF-alpha and IL-6 level, reduce inflammation factor release, improve oxidative stress state, and reduce inflammation reaction.

[Study of MSCs in vitro cultured on demineralized bone matrix of mongrel].

PURPOSE: To manufacture demineralized bone matrix(DBM) of mongrel and to explore the feasibility of DBM as scaffold of bone tissue engineering. METHODS: Thigh bones of mongrel were degreased, demineralized, deproteined, freezed, dried and sterilized to form DBM. Mesenchymal stem cells(MSCs) were seeded onto the scaffold and their growth were examined by inverted phase contrast microscope and scanning electron microscope. RESULTS: DBM had a three-dimensional mesh structure.The mean pore diameter of DBM was (254.39+/-88.71)microm and the pore rate was about 70%.MSCs could adhere to the surface and inner walls of DBM, proliferated well and secreted a large amount of extracellular matrix. CONCLUSION: DBM has satisfactory biocompatibility.