Background and purpose:MicroRNA(miRNA) is a class of small non-coding RNA playing an important regulatory role in many tumors. This study investigated which miRNA might negatively regulate the expression of Aurora-B in osteosarcoma cells, and to lay the foundation for the further investigation of the effort and regulation of Aurora-B in osteosarcoma malignant phenotype.Methods:Bioinformatics prediction software (http://www.targetscan.org) and luciferase assays were used to investigate which miRNA might target to modulate the Aurora-B. Real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot assay were used to further verify which miRNA could negative regulate the expression ofAurora-B gene.Results:Bioinformatics prediction showed let-7 family have the possibility to modulate the expression of Aurora-B; Luciferase assays showed thatAurora-B might be the target gene of let-7a/b/c/d/e/f/g/i; RTFQ-PCR and Western blot analysis testiifed that both the expression levels of Aurora-B mRNA and Aurora-B protein were signiifcantly decreased in Let-7a/g/i up-regulated U2-OS and HOS cells, compared to the cells in the negative control group; but in Let-7b/c/d/e/f up-regulated U2-OS and HOS cells, the expression levels of Aurora-B mRNA and Aurora-B protein have no signiifcant difference, compared to the cells in the negative control group.Conclusion:Let-7a/g/i may downregulate the expression of Aurora-B in human osteosarcoma cells.

Potential target proteins binding to VirB4 of type Ⅳ secretion system were screened during Brucella infected bovine embryonic trophoblast cells .Brucella VirB4 genes were amplified by PCR with species-specific primers .Expression vector pGBKT7-virB4 was constructed and analysed by sequencing and restriction enzymes ,transforming to the yeast strain Y187 and testing self-activation and toxicity .The cells model and cDNA library of bovine embryonic trophoblast cells infected with Brucella abortus strain were constructed respectively .Utilizing yeast two-hybrid system was employed to screen the target proteins of bovine embryo trophoblastic cells which was conjunctive with virB4 .These proteins were detected by real-time fluo-rescence quantitative PCR .The results suggested that bait plasmid pGBKT7-virB4 was successfully transformed into the Y187 and there was no toxicity and self-activation;the cDNA library of bovine embryonic trophoblast cells infected with Brucella abortus strain was constructed .There screened 13 positive plasmids in which Q10 and SLC3A2 were up-regulated at the mRNA level .In this paper ,we reported the interactions between the VirB4 protein of Brucella and the bovine embryo trophoblastic cells ,which provide an upstream work for further elucidating the pathogenesis of Brucella infection of the host cell .

Objective:To investigate the effect of sakubatril/valsartan on left ventricular systolic dyssynchrony in young patients with dilated cardiomyopathy (DCM).Methods:Forty-nine patients with left ventricular systolic asynchronism DCM from July 2017 to July 2018 in Guangdong Medical University Affiliated Hospital were randomly divided into two groups. The experimental group was treated with sakubatra valsartan and the control group was treated with lotensin for 12 months. Left ventricular end-diastolic diameter (LVEDd), left ventricular end-systolic diameter (LVESd) and ejection fraction (EF) were measured by two-dimensional color Doppler echocardiography. The left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV), ejection fraction (EF) and systolic dyssynchrony index (SDI) of 16 segments were measured by three-dimensional color Doppler ultrasound. Plasma NT-proBNP was tested before and after treatment.Results:After treated for 12 months, three-dimensional color Doppler ultrasonography showed the LVEDV in the experimental group decreased significantly: (180.5 ± 42.7) ml vs. (160.5 ± 45.6) ml, P<0.05. The left ventricular systolic asynchronism (SDI) of 16 segments in the experimental group significantly improved, and the value of left ventricular SDI significantly decreased compared with that of the control group: (8.2 ± 3.6)% vs. (10.8 ± 4.1)%, P<0.05. Plasma NT-proBNP decreased more significantly in the experimental group compared with that in the control group: (105.54 ± 13.25) ng/L vs. (137.27 ± 14.36) ng/L, P<0.05. Conclusions:Sakubatril/valsartan can effectively improve left ventricular systolic dyssynchrony in young patients with dilated cardiomyopathy.

OBJECTIVETo investigate the effect of co-expression of bone morphogenetic protein 2 (BMP2) and Sox9 on chondrogenic differentiation of mesenchymal stem cells (MSCs) in vitro and provide experimental evidence for tissue engineering of cartilage.

METHODSMouse embryonic bone marrow MSC C3H10T1/2 cells were infected with recombinant adenovirus expressing BMP2, Sox9 and green fluorescent protein (GFP) for 3-14 days, with cells infected with the adenovirus carrying GFP gene as the control. The mRNA expression of the markers of chondrogenic differentiation, including collagen type II (Col2a1), aggrecan (ACAN), and collagen type X (Col10a1), were determined by real-time PCR. Alcian blue staining was used for quantitative analysis of sulfated glycosaminoglycan in the cellular matrix. The expression of Col2a1 protein was assayed by immunohistochemical staining and Western blot analysis.

RESULTSAdenovirus-mediated BMP2 expression induced chondrogenic differentiation of C3H10T1/2 cells. Overexpression of Sox9 effectively enhanced BMP2-induced expression of the chondrogenic markers Col2a1, aggrecan and Col10a1 mRNAs, and promoted the synthesis of sulfated glycosaminoglycan and Col2a1 protein in C3H10T1/2 cells.

CONCLUSIONCo-expression of BMP2 and Sox9 can promote chondrogenic differentiation of MSCs in vitro, which provides a new strategy for tissue engineering of cartilage.

Animals ; Bone Morphogenetic Protein 2 ; genetics ; metabolism ; Cartilage ; cytology ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; SOX9 Transcription Factor ; genetics ; metabolism ; Tissue Engineering

Organ transplantation is the most effective method to treat end-stage organ failure. As the increase of transmission risk of donor-derived diseases, the quality, safety and selection criteria of transplanted organs become more and more important. Chapter 7 of the European Union's Guide to the Quality and Safety of Organs for Transplantation (6th Edition) proposed basic requirements in terms of donor and organ quality assessment, selection criteria and procedures, which were worthy of study and practice in clinical practice.

Objective To explore the characteristics and significance of functional connectivity (FC) of affective network (AN) in patients with postpartum depression (PPD) under resting state. Methods A total of 23 patients with PPD (PPD group) and 28 healthy postpartum women (control group) were examined using resting-state fMRI. As two critical nodes of AN, amygdala (AMYG) and subgenual anterior cingulate cortex (sgACC) were selected as the regions of interest (ROI) to analyze the differences of functional connectivity strength (FCS) of two regions from other brain regions between two groups, followed by Pearson correlation analysis on the abnormal FCS and the Edinburgh postnatal depression scale (EPDS) score in PPD group. Results Compared to the control group, the patients in PPD group showed the extensively reduced FCS (P<0.05, Alphasim correction) between AMYG and frontal cortex, temporal cortex, hippocampus, cerebellum and orbitofrontal cortex, while there were enhanced FCS (P<0.05, Alphasim correction) between sgACC and parietal cortex, occipital cortex, thalamus, superior temporal gyrus and cingulate cortex. Moreover, in PPD group, the reduced FCS between left AMYG and left medial orbitofrontal cortex was negatively correlated with EPDS scores (r=-0.62, P=0.02). Conclusion Patients with PPD have dysfunctional connectivity of AN in multiple brain regions. The weaker FCS between left amygdala and left medial orbitofrontal cortex is, the more severe depression. The dysfunctional connectivity of AN may provide an effective mechanism-based biomarker underlying PPD.

This paper is aimed to investigate the effect of titanium (Ti) particles and tumor necrosis factor alpha (TNF-alpha) on the expressions of MMP-1, 2, 3 in human synovial cells, so as to explore the possible mechanism of osteolysis post-operation of metal-on-metal total joint arthroplasty in human synovial cells induced by Ti particles. In vitro cell cultures, human synovial cells were treated by Ti particles and/or TNF-alpha. The total RNA was isolated at 2 hours after the treatment. The gene expression of MMP-1, 2, 3 was analyzed by Semi-quantitative Reverse-transcriptional PCR and quantitative real-time PCR. Cell supernatant was collected at 12, 24, 48 hours after the treatment and Gelatin zymography was performed to detect the activity of MMP-2. Compared to those in the control group (untreated), Ti particles and TNF-alpha increased the gene expression of MMP-1, 2, 3 respectively (P < 0.05), and the effect of combination of the two was even more significant (P < 0.01). The trend of activities of MMP-2 is similar with gene expression. Ti particles and TNF-alpha increased MMP-2 activities by 1.3 times and 1.5 times respectively (P < 0.05), and the combination of the two increased by 1.7 times (P < 0.01). Ti particles and TNF-alpha-induced the stimulation of MMP-1, 2, 3 expressions and MMP-2 activities in human knee joint synovial cells may be involved in aseptic loosening after metal-on-metal arthroplasty through increasing the degradation of bone matrix and declining of osseous support structure mechanics.
Cells, Cultured ; Humans ; Joint Prosthesis ; Knee Joint ; cytology ; Matrix Metalloproteinase 1 ; genetics ; metabolism ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 3 ; genetics ; metabolism ; Matrix Metalloproteinases ; genetics ; metabolism ; Particle Size ; Prosthesis Failure ; adverse effects ; RNA ; genetics ; metabolism ; Synovial Membrane ; cytology ; enzymology ; Titanium ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology