Bone marrow mesenchymal stem cell-derived exosomal microRNAs target PI3K/Akt signaling pathway to promote the activation of fibroblasts.

BACKGROUND: Fibroblast plays a major role in tendon-bone healing. Exosomes derived from bone marrow mesenchymal stem cells (BMSCs) can activate fibroblasts and promote tendon-bone healing via the contained microRNAs (miRNAs). However, the underlying mechanism is not comprehensively understood. Herein, this study aimed to identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets, and to verify their effects as well as mechanisms on fibroblasts. AIM: To identify overlapped BMSC-derived exosomal miRNAs in three GSE datasets and verify their effects as well as mechanisms on fibroblasts. METHODS: BMSC-derived exosomal miRNAs data (GSE71241, GSE153752, and GSE85341) were downloaded from the Gene Expression Omnibus (GEO) database. The candidate miRNAs were obtained by the intersection of three data sets. TargetScan was used to predict potential target genes for the candidate miRNAs. Functional and pathway analyses were conducted using the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, respectively, by processing data with the Metascape. Highly interconnected genes in the protein-protein interaction (PPI) network were analyzed using Cytoscape software. Bromodeoxyuridine, wound healing assay, collagen contraction assay and the expression of COL I and α-smooth muscle actin positive were applied to investigate the cell proliferation, migration and collagen synthesis. Quantitative real-time reverse transcription polymerase chain reaction was applied to determine the cell fibroblastic, tenogenic, and chondrogenic potential. RESULTS: Bioinformatics analyses found two BMSC-derived exosomal miRNAs, has-miR-144-3p and has-miR-23b-3p, were overlapped in three GSE datasets. PPI network analysis and functional enrichment analyses in the GO and KEGG databases indicated that both miRNAs regulated the PI3K/Akt signaling pathway by targeting phosphatase and tensin homolog (PTEN). In vitro experiments confirmed that miR-144-3p and miR-23b-3p stimulated proliferation, migration and collagen synthesis of NIH3T3 fibroblasts. Interfering with PTEN affected the phosphorylation of Akt and thus activated fibroblasts. Inhibition of PTEN also promoted the fibroblastic, tenogenic, and chondrogenic potential of NIH3T3 fibroblasts. CONCLUSION: BMSC-derived exosomes promote fibroblast activation possibly through the PTEN and PI3K/Akt signaling pathways, which may serve as potential targets to further promote tendon-bone healing.

Comparison of chemical, electrical, and combined activation methods for in vitro matured porcine oocytes.

Factors influencing porcine oocyte activation were systematically studied. This study included (1) the effect of ionomycin plus various chemical agents on activation, (2) comparison of different electrical activation parameters, (3) optimization of combined activation, and (4) evaluation of the optimized protocols. The results showed that (1) blastocyst rates of ionomycin (Ion) + 6-dimethylaminopurine (6-DMAP) (29.7 ± 1.1%), Ion + cytochalasin B (CB) + cycloheximide (CHX) (29.8 ± 1.2%), Ion + CB + 6-DMAP (30.4 ± 1.6%), and Ion + CB + CHX + 6-DMAP (30.2 ± 2.7%) were significantly higher than Ion + CHX (15.8 ± 1.5%, p < 0.05); (2) the parthenogenetic blastocyst formation of electrical activation was optimal when oocytes were activated by three direct current (DC) pulses of 1.00 kV cm(-1) for 80 µs (39.5 ± 1.1%); (3) blastocyst rates of DC + CB + CHX (55.4 ± 1.2%) and DC + CB + 6-DMAP (50.4 ± 2.9%) were significantly higher than DC + 6-DMAP, DC + CB + CHX + 6-DMAP, electrical activation, and chemical activation alone (p < 0.05); and (4) approximately 84% of parthenogenetic blastocysts yielded by the optimized protocol were diploid, which was significantly higher than that of electrical activation blastocysts (40%). Using the optimized electrical and combined activation protocol, high blastocyst rates were generated by intracytoplasmic sperm injection (ICSI) (34.6 ± 1.1%), cytoplasmic microinjection (CI) (52.3 ± 2.2%), and handmade cloning (HMC) (31.2 ± 1.0%), respectively. This study concludes that the optimal activation protocol of in vitro matured porcine oocytes was combined activation with parameter as three DC pulses of 1.00 kV cm(-1) for 80 µs plus CB and CHX treatment.

Epidermal growth factor receptor (EGFR) as a therapeutic target in rheumatoid arthritis.

The epidermal growth factor receptor (EGFR) has an important role in the hyperplastic growth of tumor. Similar to tumor growth, rheumatoid arthritis (RA) synovium is hyperplastic, invasive, and expresses EGFR and its ligands. Activation of EGFR signaling is responsible for synovial fibroblast proliferation in RA. Furthermore, in addition to its role in proliferation, EGFR and its ligands can induce cytokine production of synovial fibroblasts during the pathogenesis of RA. Agents that target EGFR have yielded promising results in animal experiments involving RA, pharmacologic modulations targeting EGFR, or its ligands may give rise to new therapeutic approaches for RA. In this review article, we will discuss the biological features of EGFR and summarize recent advances regarding the role of EGFR in the pathogenesis and treatment of RA.

Type 17 T-helper cells might be a promising therapeutic target for osteoporosis.

Osteoporosis, a disease characterized by low bone mass and deterioration of bone tissue, is a pressing public health problem. Recent studies have suggested a possible role of T-helper (Th) cells in the pathogenesis of bone loss which occurs in systemic inflammatory diseases. However, there are contradictions in the published literature regarding the functional role of Th1/Th2 cells in the regulation of the differentiation of osteoclasts. These paradoxes have now been clarified by the recent discovery of Th17 cells, a novel subset of Th cells that selectively secrete several proinflammatory cytokines, mainly IL-17. It has been confirmed that Th17 cells have stimulatory effects on osteoclastogenesis and accelerate bone loss in animal models with inflammatory disorders. Targeting Th17 cells or IL-17 may inhibit the bone resorption with RA. Thus, we are led to suppose that Th17 cells might be promising therapeutic targets in osteoporosis.

[Changes of leptin resistance, blood lipids and inflammatory response before and after the exercise therapy in children with obesity].

OBJECTIVE: Some research has shown that C-reactive protein (CRP), leptin, soluble leptin receptor (sLR) and blood lipids are involved in the development of obesity. This study aimed to investigate the changes of leptin resistance, blood lipids and inflammatory response before and after the exercise therapy in children with obesity. METHODS: Fifty-one obese children at ages of 12 years received an exercise therapy for 2 months. The levels of serum leptin, sLR, triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C) and high-sensitivity C-reactive protein (hs-CRP) were measured before and after the exercise therapy. Forty normal children served as the control group. RESULTS: Compared with the control group, serum levels of leptin, TG, TC, LDL-C and hs-CRP and the body mass index (BMI) in the obese group increased (p<0.01), while the serum level of sLR decreased significantly (p<0.05). The levels of hs-CRP, leptin, TC, TG, LDL-C and BMI in the obese group were significantly reduced after the exercise therapy (p<0.05). In the obese group, the serum leptin level was positively correlated with the levels of blood lipids and hs-CRP (p<0.05); serum levels of leptin and hs-CRP were negatively correlated with the sLR level (p<0.05); the hs-CRP level was positively correlated with the levels of blood lipids (p<0.01). CONCLUSIONS: Leptin resistance and the changes of blood lipids and inflammatory response are found in children with obesity. Exercise therapy can partially improve these changes.

[Rapid determination of fatty acids in soybeans [Glycine max (L.) Merr.] by FT-near-infrared reflectance spectroscopy].

Current breeding programs dealing with fatty acid (FA) concentrations in soybean [Glycine max (L. ) Merr.] require large numbers for gas chromatographic analyses, thus it is important to develop a method for rapid determination of fatty acid by Near-Infrared Reflectance spectroscopy (NIRS) in soybeans. The objective of this work was to study the potential of fourier-transform near-infrared reflectance spectroscopy (FT-NIRS) to estimate the fatty acid concentrations in Chinese soybean varieties. One hundred and eight of soybean cultivars or lines (the calibration set: 64; the external validation set: 44) were scanned within 4000-12500 cm(-1) of wavenumbers using a standard sample cup by NIRS machinery, and analyzed the fatty acids by gas chromatograph (GC) methods. Equations were developed using partial least squares (PLS) regression and cross validation for multivariate calibration in this study. The optimal spectral region was selected from 6101.9 to 5446.5 cm(-1) based on the OPUS 5.0 software. Cross validation results showed that major FA components such as oleic acid (R2(CV) = 0.94), linoleic acid (R2(CV) = 0.87), linolenic acid (R2(CV) = 0.85), and total saturates (R2(CV) = 0.88) were accurately determined by the proposed equations as compared with the reference data obtained by the GC method. External validation results also demonstrated that equation for oleic acid had the highest predictive ability R(2)val = 0.91), root mean square error of predication (RMSEP) value was 2.47 g x kg(-1) dry weight, the ratios of RMSEP to the standard deviation (SD) was 0.29, which was usable for quality assurance application. Moreover, equations for palmitic acid, stearic acid, linoleic acid, linolenic acid, and total saturates were predicted with the determination coefficients ranging from 0.66 to 0.76, RMSEP values from 0.37 to 2.74 g x kg(-1) dry weight, and RMSEP/SD values from 0.47 to 0.53, which could be used for sample screening. Therefore, we confirmed that a reliable estimation of the major fatty acid components is possible by using NIRS technique in soybeans.

[Development of new soybean germplasm with null lipoxygenase and Kunitz trypsin inhibitor genes].

Soybean is one of the most important sources of plant protein for human. Soybean protein is a kind of high-quality protein composed of balanced amino acids, which contains all kinds of amino acids, especially 8 amino acids necessary for human. But it also contains some components that are not good for human and affect food quality, such as lipoxygenase (Lox) and trypsin inhibitor (Ti). Those are important anti-nutritious factors. Nutritional value and processing quality of soybean can be improved by means of development of new variety with null Lox and Ti. In this paper, new soybean germplasms that pyramided multiple genes of high quality, null lipoxygenase and trypsin inhibitor genes (Ix1, Ix2, Ix3 and ti) were developed by means of cross and biochemical marker-assisted selection of progenies for null lipoxygenase and trypsin inhibitor genes using known Lox and Ti markers (protein markers). Female parents were soybean varieties Ludou 4, Zhongpin 661, Yudou 8,91D15, wei8640 popularized in Huanghuaihai Plain. Male parents were varieties introduced from US, trypsin-inhibitor (Ti)-deficient varieties P. I. L83-4387 and near isogenic lines of varieties Century for lipoxygenase (Lox)-deficient genes, Century-2 (Ix2), Century-2.3 (Ix2Ix3) and Century-1.3 (Ix1 Ix3). These new germplasms will promote soybean breeding for improved quality production,and utilization.

[Development of a new type soybean germplasm with null lipoxygenase isozymes].

Soybean protein is a kind of high-quality protein composed of balanced amino acids. It contains all kinds of amino acids, especially eight amino acids necessary for human, but also contains some components that are not good for human and affect food quality, such as Lipoxygenase (Lox) and Trypsin inhibitor (Ti). Nutritional value and processing quality of soybean can be improved by means of development of new variety with null Lox and Ti. In this paper, a new type soybean germplasm with null lipoxygenase isozymes was developed by Institute of Crop Sciences, Chinese Academy of Agricultural Sciences through years of biochemical marker assistant selection for null lipoxygenase by means of isoelectric focusing-polyacrylamid gel electrophoresis (IEF-PAGE) in the hybrid progenies of "96P17" (Female parent, a null lipoxygenase 2.3 line) and "93704" (Male parent, a null lipoxygenase 1.3 line). It is the first new soybean germplasm with null Lox1.2.3 genes in our country, which will contribute to soybean breeding for high quality, soybean production and utilization. In this paper, the development process of new type soybean germplasm is described.

[Development of a new soybean variety with null trypsin inhibitor and lipoxygenase 2.3 genes--zhonghuang 16 and its cultivation practices].

Soybean protein is a kind of high-quality protein composed of balanced amino acids, which contains all kinds of amino acid, especially 8 amino acids necessary for human, but also contains some components that are not good for human and affect food quality, such as Trypsin inhibitor and Lipoxygenase. Nutritional value and processing quality of soybean can be improved by means of development of new variety with null Lox and Ti. In this paper, a new soybean variety Zhonghuang 16 (originally name as Zhongzuo 96-952) was developed by Institute of Crop Breeding and Cultivation Chinese Academy of Agricultural Sciences through years of biochemical marker assistant selection for null trypsin inhibitor by Native-polyacrylamid gel electrophoresis (Native-PAGE) and null lipoxygenase by means of isoelectric focusing-polyacrylamid gel electrophoresis (IEF-PAGE) in the hybrid progenies of "ti15176" (Female parent)--a high-yielding, mosaic virus resistant and null trypsin inhibitor line and "Century-2.3" (Male parent)--a null lipoxygenase near isogene line of an elite American variety "Century". This variety was subjected to Beijing regional trial for summer-sowing soybean during 1999-2000, and to Beijing demonstration test in 2001. In 2002, it was passed the examination and approval by the Beijing Committee of Crop Variety Examination and Approval because of its outstanding characteristics such as high and stable yielding, good quality (high protein and fat content, high protein content and good protein quality-null Ti and Lox2.3), disease resistant and good general character. It is the first new soybean variety with null Ti and Lox2.3 genes in our country. In this paper, the development process and cultivation of Zhonghuang 16 were described.