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Objective:To observe the changes of the expression level of long non-coding RNA (lncRNA) KCNQ1OT1 and microRNA (miR)-146a-3p in placenta tissues of preeclampsia (PE) patients, as well as their effect and mechanism on the biological functions of trophoblast cells.Methods:A total of 45 cases of hospitalized PE patients in Hainan General Hospital from July 2017 to July 2018 were selected as the PE group, 55 normal pregnant women during the same period were chosed as the control group. The expression level of KCNQ1OT1 mRNA and miR-146a-3p in the placenta tissues between two groups were detected by using quantitative real time (qRT)-PCR. Pearson′s test was furtherly analyzed the correlation between them. Human trophoblast cell line (HTR8/SVneo) were randomly divided into control and lipopolysaccharide (LPS) groups, and then LPS group were divide into four sub-groups,included LPS group, short hairpin RNA (sh)-KCNQ1OT1 (after silencing the expression of KCNQ1OT1), miR-146a-3p inhibitor and sh-KCNQ1OT1+miR-146a-3p inhibitor. The targeting relationship between KCNQ1OT1 and miR-146a-3p were predicted by bioinformatics software and confirmed by luciferase assay. The cell proliferation and invasion capacities were respectively detected by cell counting kit-8 (CCK-8) and transwell assay. The expression level of KCNQ1OT1 mRNA and miR-146a-3p were detected by qRT-PCR and the protein expression level of CXC chemokine ligand 12 （CXCL12） and CXC chemokine receptor type 4 (CXCR4) were tested by western blot.Results:(1) The mRNA expression level of KCNQ1OT1 in the placenta of PE group was lower than that of control group (0.23±0.03 vs 0.51±0.04, P<0.05), and the miR-146a-3p expression level was higher than that of the control group (0.49±0.03 vs 0.31±0.03, P<0.05), there were statistical significant differences between the two groups. (2) Luciferase assay showed that there was a targeting relationship between KCNQ1OT1 and mir-146a-3p. Compared with the control group, the mRNA expression level of KCNQ1OT1 in the LPS group were significantly decreased (0.91±0.03 vs 0.35±0.03, P<0.05), and the expression level of miR-146a-3p were significantly increased (0.22±0.03 vs 0.63±0.04, P<0.05). The cell proliferation, invasion and migration capacities and the protein expression of CXCL12 and CXCR4 significantly reduced in the LPS group compared with control group (all P<0.05). The mRNA expression level of KCNQ1OT1 (0.23±0.03) in the sh-KCNQ1OT1 group were further decreased, the expression of miR-146a-3p (0.85±0.03) were further increased, and the cell proliferation, invasion and migration capacities and the protein expression of CXCL12 and CXCR4 were all further reduced compared with control group，there were significant difference between two groups (all P<0.05). Comparing the miR-146a-3p inhibitor group, and sh-KCNQ1OT1+miR-146a-3p inhibitor group with the sh-KCNQ1OT1 group, respectively, the expression level of KCNQ1OT1 mRNA (0.78±0.04 vs 0.50±0.03) increased, and the expression level of miR-146a-3p (0.42±0.03 vs 0.46±0.03) decreased, the cell proliferation, invasion and migration capacities and the protein expression of CXCL12 and CXCR4 were all increased ,there were statistically significant differences (all P<0.05). Conclusion:KCNQ1OT1 could target the regulation of miR-146a-3p through CXCL12/CXCR4 pathway in the proliferation, invasion an migration of HTR8/SVneo cells, which may be involved in the pathogenesis of PE.
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BACKGROUND: Pearl has the tranquilizing effect, and it can be applied in the treatment of hypertension. However, there is little report on the prevention and cure effect and mechanism of hydrolyzed pearl liquid on hypertension. OBJECTIVE: To observe the effect of hydrolysis of Hepu pearl (hydrolyzed Nanzhu fluid) on the biological behavior and secretion of human microvascular endothelial cells. METHODS: Human microvascular endothelial cells were cultured and passaged. There were four groups, and the microvascular endothelial cells were incubated in the 200 μL culture medium containing nothing (control group), and 120, 60 and 30 mg/L hydrolysis of Nanzhu fluid. The cell proliferation and migration was detected by cell conuting kit-8 assay and Transwell assay respectively; the cell cycle distribution was tested by flow cytometry; the cell apoptosis was assayed by TUNEL method; the secretion of nitric oxide and reactive oxygen species was tested by nitrale reduetase and chemical fluorescence method, respectively. RESULTS AND CONCLUSION: Compared with the control group, hydrolysis of Nanzhu fluid significantly promoted the proliferation of microvascular endothelial cells in a manner-dependent manner (P ＜ 0.05), suggesting the optimal concentration was 120 mg/L. Compared with the control group, the percentage of cells in S and G2 phase was significantly increased, and the percentage of cells in the G1 phase was significantly reduced in the hydrolysis of Nanzhu fluid group (P＜0.05), indicating hydrolysis of Nanzhu fluid could promote cell cycle progression. Apoptotic cells with green-stained nucleus were invisible in both groups. The number of cell migration in the hydrolysis of Nanzhu fluid group was significantly more than that in the control group (P ＜ 0.05). Compared with the control group, there was a significant increase in the nitric oxide secretion, and a significant decrease in the production of reactive oxygen species in the hydrolysis of Nanzhu fluid group (P＜0.05). To conclude, hydrolysis of Nanzhu fluid can promote the proliferation and migration of human microvascular endothelial cells in vitro, and has the function of promoting the secretion of nitric oxide and inhibiting reactive oxygen species secretion, implying its positive role in the protection of endothelial cell function.
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BACKGROUND: Inadequate sources of islet cells mean that islet cell transplantation for diabetes cannot meet the clinical demand.Therefore,in vitro induction of pancreatic stem cells to differentiate into islets has become a focus of research. OBJECTIVE:To study the effect of Tripterygium wilfordii polysaccharides on the differentiation of pancreatic stem cells from islets in mice, so as to explore the effect of traditional Chinese medicine on the differentiation of pancreatic stem cells into pancreatic beta cells. METHODS:Tripterygium wilfordii polysaccharide was used to induce the differentiation of purified mouse pancreatic stem cells into islets in vitro.The islet-like cell clusters then underwent morphologic observation, dithizone (DTZ) staining, and western blot analysis. RESULTS AND CONCLUSION: Cell morphology, cell growth characteristics and immunocytochemical staining showed that mouse pancreatic stem cells were obtained.They were induced by Tripterygium wilfordii polysaccharide into spherical islet-like structures, which had a spindly pedicle connected with the bottom of the culture flask, and were DTZ-stained to iron red. Western blot assay detected β-cytokine proteins in the islet-like cell clusters. These findings confirm that mouse pancreatic stem cells can be induced to differentiate into islet-like cell clusters containing β cells in vitro by Tripterygium wilfordii polysaccharide.
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Objective To investigate the adjunctive therapeutic effect of recombination growth hormone(rhGH) on the elderly patients with pulmonary infection.Methods 53 elderly patients with pulmonary infection were randomly divided into rhGH group(n=25) and control group(n=28).The rhGH group were received hypodermic introl group were received standard therapy.Clinical efficacy were compared between two groups.The level of albumin (ALB),growth hormone (GH),insulin-llke growth factor-1 (IGF-1),leptin (LP) were measured and the body mass index(BMI) was observed in all cases before and after treatment.Results The overall efficacy of rhGH group and control group was 88.0% versus 60.7%.The results showed significant statistical difference between both groups(P＜0.05).After treating with rhGH for 10 days,the level of BMI,ALB,GH,IGF-1 and LP were increased in the rhGH group,which were(26.1±4.1)kg/m2;(38.4 ±6.6)g/L;(7.0 +0.9) μg/L;(27.3 ±6.1)μg/L;(6.9 ± 1.1)μg/Lrespectively.And the level of BMI,ALB,GH,IGF-1 and LP were (21.8 ± 3.4 ) kg/m2 ;( 29.5 ± 5.1 ) g/L;( 4.0 ±0.4) μg/L;( 22.0±3.8 )μg/L;( 3.8±0.8 )μg/L in the control group after treatment.The results showed significant statistical difference between both groups(P＜0.05).Conclusion There was obvious adjunctive therapeutic effect of rhGH to the elderly patients with pulmonary infection by improving the nutritional state of these patients.
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<p><b>OBJECTIVE</b>To investigate the possibility of differentiation of human mesenchymal stem cells (hMSC) into epidemic cells in vitro.</p><p><b>METHODS</b>hMSCs were segregated from normal adult human bone marrow by Percoll solution (1.073 g/ml) , and were cultured, purified, and amplified to 3th passage in vitro. Then the hMSCs were randomly divided into control group ( with treatment of normal L-DMEM medium) and experimental group (with treatment of L-DMEM medium containing epidermal growth factor,insulin,tretinoin, calcium chloride). After 7 days of culture, the morphologic changes of hMSCs in the 2 groups were observed with inverted phase contrast microscope. The expressions of P63 and PCK of hMSCs were assessed with immunohistochemical methods.</p><p><b>RESULTS</b>The shape of hMSCs in experimental group became irregular or oblong in shape, while that in control group were still in spindle shape. Immunohistochemical results showed that hMSCs were P63 and PCK positive in the experimental group, while those in control group were negative.</p><p><b>CONCLUSION</b>Human mesenchymal stem cells can differentiate into epidemic cell in vitro.</p>
Subject(s)
Humans , Bone Marrow Cells , Cell Biology , Metabolism , Cell Differentiation , Cells, Cultured , Epithelial Cells , Cell Biology , Keratins , Metabolism , Membrane Proteins , Metabolism , Mesenchymal Stem Cells , Cell BiologyABSTRACT
OBJECTIVE:To investigate the therapeutic efficacy of recombination human growth hormone(rhGH)on the patients with chronic obstructive pulmonary disease(COPD)complicated with spontaneous pneumothorax(SP).METHODS:58 patients with COPD complicated with SP were randomly divided into two groups:29 cases in the control group received conventional therapy including anti-infection therapy,oxygen therapy and nutritional support,and another 29 in the trial group received low dose of rhGH(0.1 U?kg-1?d-1)hypo quaque nocte for 7 days started from the second day after admission in addition to the conventional therapy as employed for control group.The time of chest tube stay and the length of hospital stay were compared between the two groups.The body mass index(BMI)and serum levels of growth hormone(GH),insulin-like growth factor-1(IGF-1)and albumin(ALB)were measured in all cases before and after treatment.RESULTS:The time of chest tube stay and the length of hospital stay in the trial group vs.the control group were(5.5?1.8)d vs.(9.3?2.2)d(P0.05);however,after treatment,all indexes(BMI,ALB,GH and IGF-1)were higher in the trial group than in the control group(P
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<p><b>OBJECTIVE</b>To explore the activation of the signal transduction pathways related with the carcinogenesis of sporadic colon cancers.</p><p><b>METHODS</b>A gene microarray monitoring activation of 8 signal transduction pathways (PathwayFinder GEArray) was used to screen the differentially expressed genes between colorectal cancer and normal colon tissue. The differentially expressed genes were further analyzed by RT-PCR, using RNA extracted from cancer tissue and matched normal colon mucosa of 35 patients with colorectal cancer.</p><p><b>RESULTS</b>The expression of hsf1, hsf27 and inos was increased in colon cancer compared with normal colon mucosa using PathwayFinder GEArray. The RT-PCR results showed that the expression of hsf1 was detected in 86% of patients(30/35)and the expression of inos detected in 63% patients(22/35).</p><p><b>CONCLUSION</b>Hsf1 induces heat shock stress signaling pathway, which might play a role in the carcinogenesis of sporadic colorectal cancer.</p>
Subject(s)
Humans , Colorectal Neoplasms , Genetics , Metabolism , DNA-Binding Proteins , Genetics , Gene Expression Regulation, Neoplastic , Heat Shock Transcription Factors , Intracellular Signaling Peptides and Proteins , Genetics , Nerve Tissue Proteins , Genetics , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction , Physiology , Transcription Factors , Tumor Suppressor Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To isolate MSCs from adult human bone marrow cells and to induce them into adipocytes.</p><p><b>METHODS</b>MSCs were isolated from adult human bone marrow aspirated by Percoll and expanded in L-DMEM. The surface antigen of MSCs, CD14, CD34, CD45, CD44, VLA-1, HLA-DR and cell cycle were analysed on a FACScan flow cytometer. MSCs were cultured in adipogenisis inducing medium including insulin, 1-methyl-3-isobutylxanthine, indomethine and dexamethasone for 7 days and stained with Oil Red O.</p><p><b>RESULTS</b>MSCs grew as adherent cells and expanded more than 10 passages. They were positive for CD44 and negative for CD14, CD34, CD45, HLA-DR. The expression of VLA-1 was weak. After 7 days of adipocyte inducing, about 85%of the cells displayed accumulation of lipid vacuoles, as detected by Red Oil O.</p><p><b>CONCLUSION</b>MSCs isolated and cultured from adult human bone marrow can be induced to adipogenisis committed differentiation.</p>