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Objective:To investigate the relationship between long non-coding RNA (lncRNA) DHRS4-AS1 and disease-free survival in osteosarcoma patients and the mechanisms of its effect on proliferation and migration of osteosarcoma cells in vitro.Methods:The data of DHRS4-AS1 transcriptome levels and survival status of osteosarcoma patients in GEPIA database were collected since the database was established, and the patients were divided into high DHRS4-AS1 expression group and low DHRS4-AS1 expression group based on the median DHRS4-AS1 transcriptome level, with 59 cases in each group, and the Kaplan-Meier method was used to analyze the disease-free survival of the two groups. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of DHRS4-AS1 in osteosarcoma cell lines MG-63, HOS, 143B, U-2OS, Saos2 and normal osteoblast cell line hFOB1.19, and the osteosarcoma cell line with the lowest DHRS4-AS1 expression level was selected for subsequent experiments. The plasmid carrying DHRS4-AS1 sequence and the plasmid carrying negative control sequence were transfected into the selected osteosarcoma cells as DHRS4-AS1 group and control group. CCK-8 method was used to detect the proliferation of each group of cells, and the absorbance value was used as the cell proliferation ability; cell scratch assay was used to detect the migration of each group of cells. The bioinformatics website starBase V2.0 was used to predict the target genes of DHRS4-AS1, and the dual luciferase reporter gene assay was used to verify the targeting relationship between DHRS4-AS1 and the target genes. The expression levels of target genes and downstream genes of osteosarcoma cells in control group and DHRS4-AS1 group were detected by qRT-PCR and Western blotting.Results:Survival analysis showed that the disease-free survival of osteosarcoma patients in the high DHRS4-AS1 expression group in GEPIA database was superior to that of the low DHRS4-AS1 expression group ( P < 0.001). Compared with normal osteoblastic hFOB1.19 cells, the expression level of DHRS4-AS1 was low in all osteosarcoma cells (all P < 0.01), with the lowest expression level of DHRS4-AS1 in U-2OS cells ( P < 0.001). Cell proliferation ability was reduced in U-2OS cells of the DHRS4-AS1 group after 1, 2, 3 and 4 d of culture compared with the control group (all P < 0.05). The migration rate of U-2OS cells in the DHRS4-AS1 group was lower than that in the control group [(31±6)% vs. (63±4)%, t = 4.38, P = 0.005]. starBase V2.0 website predicted that DHRS4-AS1 complementarily bound to miRNA-411-3p (miR-411-3p); dual luciferase reporter gene assay showed that miR-411-3p overexpression reduced the luciferase activity of the wild-type DHRS4-AS1 reporter gene ( P < 0.001), but had no effect on the luciferase activity of the mutant DHRS4-AS1 reporter gene ( P > 0.05). qRT-PCR showed that the relative expression of miR-411-3p in U-2OS cells of the DHRS4-AS1 group was low (0.22±0.06 vs. 1.06±0.23, t = 3.55, P = 0.012) and the relative expression of metastasis suppressor MTSS1 mRNA was high (5.58±1.03 vs. 1.06±0.22, t = 4.28, P = 0.005) compared with the control group; Western blotting showed that MTSS1 expression was elevated, and the expression levels of cell proliferation phenotype proteins CDK3 and cyclin C and cell migration phenotype proteins ZEB2 and KLF8 were low. Conclusions:Osteosarcoma patients with high expression of lncRNA DHRS4-AS1 have better disease-free survival, and its expression is low in osteosarcoma cell lines. DHRS4-AS1 may promote MTSS1 gene expression and inhibit cell proliferation and migration by targeting and down-regulating miR-411-3p expression in osteosarcoma cells.
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Objective:To investigate the level of plasma Betatrophin in pregnant women with gestational diabetes mellitus (GDM) and its correlation with the control of blood glucose.Methods:Forty-five pregnant women with GDM(GDM group) who received regular obstetric examinations in the Huaihua First People′s Hospital from July 2019 to January 2021 and 50 pregnant women with normal glucose tolerance (NGT) (NGT group) during the same period were enrolled in this study. Blood glucose and blood lipid indicators were collected, plasma Betatrophin level was detected, Logistic regression analysis was used to screen the influencing factors of blood glucose control effect, the pregnancy outcome was followed up, the predictive value of Betatrophin level in blood glucose control and pregnancy outcome was evaluated by receiver operating characteristic (ROC) curve.Results:The levels of systolic blood pressure, diastolic blood pressure, fasting plasma glucose (FPG), 2 h postpartum blood glucose (2 h PG), glycosylated hemoglobin (HbA 1c), fasting insulin (FINS), 2 h postprandial insulin (2 h FINS), insulin resistance index (HOMA-IR), low density lipoprotein cholesterin (LDL-C) and plasma Betatrophin in the GDM group were higher than those in the NGT group, and insulin function index (HOMA-β) and high density lipoprotein cholesterin (HDL-C) were lower than those in the NGT group ( P<0.05). Pearson correlation analysis showed that plasma Betatrophin level was positively correlated with HbA 1c and HOMA-IR in pregnant women and the GDM group ( r = 0.310, 0.314, 0.341, 0.333; P<0.05). In the GDM group, 12 patients with poor glucose control, 33 patients with good glucose control, the FPG, HbA 1c, HOMA-IR and plasma Betatrophin levels in poor glucose control patients were higher than those in good glucose control patients, HOMA-β was lower than that in the good glucose control patients: (5.82 ± 0.98)mmol/L vs. (5.04 ± 1.11) mmol/L, (9.78 ± 2.15)% vs. (8.22 ± 1.41)%, 2.71 ± 0.56 vs. 2.24 ± 0.48, (1 345.12 ± 256.32) ng/L vs. (1 165.10 ± 217.41) ng/L, 144.15 ± 22.71 vs. 158.63 ± 20.26, there were statistical differences ( P<0.05). The area under the curve of plasma Betatrophin level to predict the effect of blood glucose control was 0.775. A total of 8 pregnant women with GDM had poor pregnancy outcome, and the area under the curve predicted pregnancy outcome by plasma Betatrophin level was 0.728. Conclusions:The level of plasma Betatrophin in patients with GDM is closely related to the degree of insulin resistance and the effect of blood glucose control, and can provide some reference for clinical evaluation and therapeutic effect prediction.
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Objective To investigate the changes of coagulation function and blood rheology in acute exacerbation of chronic obstructive pulmonary disease (AECOPD) and effects of depside salt from salvia miltiorrhiza.Methods The patients with AECOPD were randomly divided into depside salt from salvia miltiorrhiza anticoagulation group (45 patients) and conventional treatment group (45 patients).At the same time 50 normal were choosed as control group (50 patients).On the 14th day after treatment,4 indexes of coagulation,D-dimer and of blood rheology in depside salt from salvia miltiorrhiza anticoagulation group and conventional treatment group were determine and compared with those of normal control group.Results The levels of Fbg (4.6 ± 0.7) g/L and D-dimer (1.58 ± 1.13) mg/L were significantly different between depside salt from salvia miltiorrhiza anticoagulation group and normal controls.After depside salt from salvia miltiorrhiza anticoagulation,The levels of FbgFbg (3.3 ± 1.2) g/L and D-dimer (0.48 ± 0.36)mg/L were decreased significantly (P ＜ 0.01).The 1 evels of Fbg (3.3 ± 1.2) g/L and D-dimer (0.48 ± 0.36)mg/L were decreased significantly in depside salt from salvia miltiorrhiza anticoagulation group compared with those in Conventional treatment group (P ＜ 0.01).But PT,APTT and TT level were not significantly different in three groups (P ＜0.05).After treatment,hemodynamic indexes:whole blood high shearing viscosity (4.76 ± 1.35)mPa.S,low shearing viscosity (8.69 ± 2.36) mPa.S plasma viscosity (2.32 ± 0.26) mPa.S and hematocrit (40.3 ± 2.38)％ in depside salt from salvia miltiorrhiza anticoagulation group indicated significant differences compared to those in Conventional treatment group[(5.50.3 ± 1.34) mPa.S,(12.30 ± 2.30) mPa.S,(2.32 ± 0.26) mPa.S,(47.89 ± 3.13)％)] (P ＜ 0.01).Conclusion Patients with in AECOPD have hypercoagulable state.depside salt from salvia miltiorrhiza could improved hypercoagulable state in acute exacerbation of chronic obstructive pulmonary disease and decrease blood viscosity.
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Objective To study the effect of adaptive support ventilation (ASV) in treatment of severe asthma.Methods Forty-nine cases of severe asthma were divided into ASV group (25 cases) and control group (24 cases,tradition mechanical ventilation).The arterial blood gas,respiratory dynamics,mechanical ventilation time,hospital stay and thorax barotrauma was compared between two groups.Results The arterial blood gas and respiratory dynamics was improved after mechanical ventilation compared with that before mechanical ventilation in two groups,and there was significant difference (P ＜ 0.05 or ＜ 0.01).The airway peak voltage,lung dynamic compliance and platform pressure after mechanical ventilation of 2,12 and 24 h in ASV group was better than that in control group[2 h:(33 ± 12) cm H2O(1 cm H2O =0.098kPa) vs.(37 ± 11) cm H2O,(16 ± 9) ml/cm H2O vs.(17 ± 10) ml/cm H2O,(27 ± 6) cm H2O vs.(30 ±12) cm H2O; 12 h:(23 ± 12) cm H2O vs.(25 ± 11) cm H2O,(28 ± 6) ml/cm H2O vs.(23 ± 10) ml/cm H2O,(20 ±6) cm H2O vs.(25 ±4) cm H2O; 24 h:(18 ± 12) cm H2O vs.(20 ± 11) cm H2O,(32 ±9)ml/cm H2O vs.(28 ± 10) ml/cm H2O,(12 ±7) cm H2O vs.(16 ±7) cm H2O],and there was significant difference(P＜ 0.05 or ＜ 0.01).The mechanical ventilation time and hospital stay in ASV group was shorter than that in control group [(46 ± 8) h vs.(56 ± 6) h,(7 ± 2) d vs.(10 ± 3) d],and there was significant difference (P＜ 0.01).The thorax barotrauma was not observed in ASV group; 3 cases showed subcutaneous emphysema and 2 cases showed pneumothorax in control group.Conclusions ASV mode could decrease airway peak voltage and platform pressure,improve arterial blood gas and lung dynamic compliance,shorten mechanical ventilation time and hospital stay.It is safe and effective for patients with severe asthma.