Mechanism of LncRNA DDX11-AS1 in Regulating Proliferation, Migration and Apoptosis of Gastric Cancer Cells by Targeting MiR-497-5p / 胃肠病学

Background: LncRNA expression was up-regulated or down-regulated in gastric cancer, but the mechanism of role of LncRNA in the development and progression of gastric cancer was not been fully clarified. Aims: To investigate the mechanism of LncRNA DDX11-AS1 in regulating the proliferation, migration and apoptosis of gastric cancer cells by targeting the expression of miR-497-5p. Methods: qRT-PCR was used to detect the expressions of DDX11-AS1 and miR-497-5p in gastric cancer tissue and cell lines. Pearson method was used to analyze the correlation between DDX11-AS1 and miR-497-5p in gastric cancer tissue. Gastric cancer HGC-27 cells were randomly divided into NC group, si-con group, si-DDX11-AS1 group, miR-con group, miR-497-5p group, si-DDX11-AS1+anti-miR-con group, si-DDX11-AS1+anti-miR-497-5p group. The cell proliferation was detected by MTT. The cell apoptosis was detected by flow cytometry. Transwell assay was used to detect the migration and invasion ability. The dual luciferase reporter system assay was used to verify the targeted regulatory relationship between DDX11-AS1 and miR-497-5p. The protein expressions of cyclin D1, cleaved caspase-3, MMP-2 and MMP-9 were detected by Western blotting. Results: The expression of DDX11-AS1 in gastric cancer tissue and cell lines was significantly increased (P<0.05), while the expression of miR-497-5p was significantly decreased (P<0.05), DDX11-AS1 was negatively correlated with miR-497-5p (r=-0.754, P<0.05). Interfering the expression of DDX11-AS1 or up-regulating the expression of miR-497-5p significantly inhibited cell proliferation, migration and invasion (P<0.05), apoptosis rate was significantly increased (P<0.05), and the expressions of cyclin D1, MMP-2 and MMP-9 were significantly decreased (P<0.05), and the expression of cleaved caspase-3 was significantly increased (P<0.05). The dual luciferase reporter assay demonstrated that DDX11-AS1 negatively regulated the expression and activity of miR-497-5p. Inhibition of miR-497-5p expression reversed the effect of interference with DDX11-AS1 expression on the biological behavior of gastric cancer cells. Conclusions: Interference with LncRNA DDX11-AS1 expression can inhibit the proliferation, migration, invasion and induce apoptosis of gastric cancer cells by targeting and up-regulating the expression of miR-497-5p.

Evaluation of hypoxic pulmonary vascular remodeling based on fractal theory / 实用放射学杂志

Objective To evaluate the relationship between the spatial structure of pulmonary vascular tree and oxygen partial pressure in patients with chronic obstructive pulmonary disease(COPD) by the fractal dimension method.Methods 106 patients with COPD and 100 healthy people without COPD as controls were included in this study.All of the patients underwent multidetector CT scan and blood gas analysis.The pulmonary vascular trees were generated using post-processing software,and the FD of the pulmonary vascular trees were determined with ImageJ software in a personal computer.The fractal dimension were evaluated in the two groups.The relationship between FD and oxygen partial pressure in patients with COPD was analyzed.Results The FD value of the patients with COPD was lower than that of the patients without COPD (t =5.21,P＜ 0.01).There was a significant correlation between FD and the PaO2 in patients with COPD (r=0.692,P＜ 0.01).Conclusion FD analysis can effectively evaluate the process of pulmonary vascular remodeling induced by hypoxia in patients with COPD,which may be used as an important index for quantitative evaluation of pulmonary vascular remodeling in the course of COPD.

Effects of farnesoid X receptor agonist on adiponectin and its receptors / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of GW4064, a farnesoid X receptor (FXR) agonist, on adiponectin and its receptors during the differentiation of 3T3-L1 preadipocytes and on adiponectin receptors in HepG2 cells.</p><p><b>METHODS</b>The mRNA expressions of FXR, PPARγ2, adiponectin, AdipoR1, and AdipoR2 and the protein levels of adiponectin on days 0, 2, 4, 6, and 8 during the differentiation of 3T3-L1 preadipocytes treated with GW4064 were detected by fluorescent real-time PCR and ELISA, respectively. The mRNA expressions of AdipoR1 and AdipoR2 in HepG2 cells were also examined at 0, 12, 24, and 48 h after GW4064 treatment.</p><p><b>RESULTS</b>The mRNA expressions of FXR, PPARγ2, adiponectin, and AdipoR2 in 3T3-L1 preadipocytes and AdipoR2 in HepG2 cells treated with GW4064 was significantly increased compared with the control group (all P<0.05). The protein level of adiponectin was also significantly increased after GW4064 treatment. The expression of AdipoR1 in either 3T3-L1 preadipocytes or HepG2 cells showed no significant changes after GW4064 treatment.</p><p><b>CONCLUSION</b>GW4064 can up-regulate the expressions of FXR, PPARγ2, adiponectin, AdipoR2 in 3T3-L1 preadipocytes and AdipoR2 in HepG2 cells. As adiponectin and its receptors are two important factors in the treatment of non-alcoholic fatty liver disease, FXR agonist may potentially produce therapeutic effect on non-alcoholic fatty liver disease and can regulate adipocytes via up-regulating PPARγ during adipocyte differentiation.</p>

Effects of FXR agonist on leptin and OB-Rb / 实用医学杂志

Objective To investigate the effects of GW4064,one FXR agonist,on the leptin and OB-Rb during the differentiation of 3T3-L1 preadipocytes and on the OB-Rb in the HepG2 cells. Methods The mRNA relative expression of leptin , OB-Rb and the protein of leptin on the day of 0 , 2 , 4 , 6 , 8 during the differentiation of 3T3-L1 preadipocytes after interfered with GW4064 were detected by fluorescent real-time PCR and ELISA , respectively. Meanwhile , the mRNA relative expression of OB-Rb of HepG2 cells after treated with GW4064 0 h , 12 h,24 h,48 h were also examined. Results The mRNA relative expression of leptin in the 3T3-L1 preadipocytes and OB-Rb in the HepG2 cells after treated with GW4064 were significantly increased compared with the controlgroup. Also, the protein level of leptin was similar with the mRNA expression. The all differences were statistically significance (P0.05). Conclusions GW4064 is able to upregulate the expression of leptin in the 3T3-L1 preadipocytes and OB-Rb in the HepG2 cells. Now the role of leptin in the NAFLD is still unkown, however, the low expression of OB-Rb is related with the leptin resistance in the pathogenesis of NAFLD. So we hypothesize that FXR agonist may treat NAFLD through upreglating the expression of OB-Rb and improving leptin resistance.

Role of microparticles in thrombotic diseases / 中华检验医学杂志

Circulating microparticles (MPs) derive from a variety of cells,including platelets,leukocytes,endothelial cells and erythrocytes.In addition,tumor cells release MPs into blood.MPs are formed as a result of membrane lipid remodeling and proteolytic cleavage of cytoskeleton.Exposure of phosphatidylserine (PS) and tissue factor (TF) provide a surface for the assembly of Xase and prothrombianse to activate coagulation and induce thrombosis.Considerable levels of platelet-derived PS+ MPs are detected in healthy subjects.However,the level of PS+ and TF+ MPs,the latter of which predominantly derived from monocytes,greatly increased in thrombotic diseases,such as sepsis,cancer,and myocardial infarction.Therefore,MPs could be a useful biomarker for the evaluation of thrombosis risk.

Application of genome-wide genechip for screening and identifying genes related to CD133(+)CD200(+) colorectal cancer stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen and identity genes related to CD133(+)CD200(+) colorectal cancer stem cells.</p><p><b>METHODS</b>The two subpopulations of colorectal cancer cells, namely CD133(+)CD200(+) and CD133(-)CD200(-) cells, were sorted and verified by flow cytometry. The gene expression profiles of CD133(+)CD200(+)and CD133(-)CD200(-) colorectal cancer cells were examined using Affymetrix Human U133 Plus2.0 genome-wide genechip. The differentially expressed genes between the two cell subpopulations were analyzed to identify the genes responsible for the main effect in association with colorectal cancer stem cells. Real-time quantitative PCR was performed to confirm some of the differentially expressed genes identified by genechip.</p><p><b>RESULTS</b>The genechip result showed that 655 genes were differentially expressed in CD133(+)CD200(+) colorectal cancer stem cells by at least 3 folds, including 290 up-regulated and 365 down-regulated ones. Bioinformatics analysis and gene co-expression network building identified 3 genes (MDM2, PRKACG, and CACNA1G) with specific expression in CD133(+)CD200(+) colorectal cancer stem cells, and this result was confirmed by real-time quantitative PCR analysis.</p><p><b>CONCLUSION</b>A specific gene expression profile of colorectal cancer stem cells has been established through screening and identifying genes related to CD133(+)CD200(+)colorectal cancer stem cells by gene genechip technique, which provides a basis for further study of gene targeting therapy of colorectal cancer.</p>

The study of the relationship between influenza virus infection and the dysfunction of vascular smooth muscle cells / 中华微生物学和免疫学杂志

Objective To research the influenza virus infection on rat vascular smooth cells number,proliferation,apoptosis,the amount of IL-6,sFas,platelet-derived growth factor(PDGF) and the mechanism of atherosclerosis.Methods Flow cytometry,enzyme-linked immunosorbent assay and cell count experiments were used to detect these indicators at 0 h,6 h,12 h,24 h,48 h.Results After influenza virus infected at 0 h,proliferation,apoptosis condition were 10.39％,0.44％,respectively; at 6 h,proliferation,apoptosis respectively increased to 12.68％,0.73％ ; proliferation reached the peak at 12 h (18.01％),instead apoptosis decreased to 0.14％ ; at 24 h,proliferation decreased to 12.89％ and apoptosis markedly increased to 1.09％ ; at 48 h,proliferation further reduced to 7.07％ and apoptosis reached the peak(4.61％).The number of cells and the cytokine secretion were statistically significant to control group (P＜0.05).Conclusion Influenza virus infection might lead to change of cell proliferation and apoptosis and involve the atherosclerosis form and development,and cytokines played an important role in them.

Study on the acetoacetate/β hydroxybutyrate determination in classification of type 1 and type 2 diabetes mellitus / 中华内分泌代谢杂志

The clinical values of acetoacetate ( AcAA ) and β hydroxybutyrate ( βHBA ) determination in classification of type 1 and2 diabetes were explored. 102 normal control subjects,33 cases of type 1 diabetes, and 104cases of type 2 diabetes were enrolled. Serum AcAA, βHBA, fasting plasma glucose ( FPG), C-peptide, and insulin levels were measured. The results showed that serum AcAA, βHBA, total ketone tody (TKB) levels in the diabetic groups were significantly higher than those of the normal group( P＜0. 01 ). AcAA, βHBA, TKB levels in type 1diabetes were higher as compared with those of type 2 diabetes( P＜0.01 ). The AcAA, βHBA, and TKB levels were negatively related with C-peptide and insulin in diabetic patients( P＜0. 01 ). All the type 1 diabetic patient were found to have TKB and lower C-peptide levels. TKB positive and lower C-peptide in type 2 diabetes were found in 47% and 26% respectively. Receiver operating characteristic (ROC) curve suggested that the area under the ROC curve of type 1 and type 2 diabetes was 0.926. The optimal operating point of the total ketone body was 0. 532 mmol/L with higher sensitivity and specificity. Enzymatic determination of acetoacetate and β hydroxybutyrate seems to have important clinical values for classification of type 1 and 2 diabetes.

The relation of aquaporinl gene expression and kidney injury in rats with disseminated intravascular coagulation / 中华急诊医学杂志

Objective To characterize the effects of AQP1 expression on kidney damage in rat disseminated intravascular coagulation(DIC) caused by lipopolysaccharide(LPS) dosing. Method Fifty male Wistar rats (clean grade) were randomly assigned into 5 groups of 10 rats. The 10 control rats were dosed with 10 ml of 0.9%NaCl solution by a drip via the vena caudalis within 4 h, and blood and tissues were obtained after treatment completion. In the DIC groups, the rats were dosed with LPS (30 mg/kg body weight in 10 ml of 0.9% NaCl solution) by a drip via the vena caudalis within 4 h, and blood and tissues were obtained at 4, 6, 8 and 10 h. The blood platelet(PLT) count, prothrombin time(PT) , activated partial thromboplastin time(APTT), fibrin(FIB) and D-dimer(D-D) were detected. Hematoxylin and eosin(HE) staining was used to examine the pathologic changes in the lung and kidney tissues of each group (both hematologic parameters and tissue pathologic changes were used to judge the course of DIC). The AQP1 gene expression levels in the kidney tissues from the groups were evaluated by the mRNA levels using RT-PCR. Statistical analyses were performed by the SNK- q method. Results The PLT count, PT, APTT, FIB and D-D examinations revealed remarkable changes in all DIC groups compared with the control group (P < 0.01). The AQP1 mRNA level was significantly decreased in the DIC group at 4 h compared with the control group (P < 0.01) , and further decreased to the minimum level in the DIC group at 6 h. Moreover, cloudy swelling of renal tubular cells was observed at 6 h and cell degeneration and necrosis were observed at 8 h among the DIC groups. Conclusions Downregulation of AQP1 mRNA expression occurred before damage to the renal tubular cells in DIC, indicating that AQP1 expression may be involved in the kidney damage observed in rat DIC.

Effect of high epidural anesthesia on interleukin-6 and soluble interleukin-2 receptor in patients with dilated cardiomyopathy / 中国组织工程研究

BACKGROUND: Serum cytokines in patients with dilated cardiomyopathy are increased obviously, and the expression of interleukin-6mRNA is also observed in myocardial tissues. High epidural anesthesia can block the vicious cycle involving cytokines and improve cardiac function. OBJECTIVE: To observe the changes of interleukin-6 and soluble interleukin-2 receptor in patients with dilated cardiomyopathy after high epidural anesthesia treatment. DESIGN: A case-controlled analysis. PARTICIPANTS: Thirty-five inpatients with dilated cardiomyopathy were selected from the Department of Cardiology, First Hospital Affiliated to Harbin Medical University, from October 2001 to May 2002. All the patients were randomly divided into high epidural anesthesia group and conventianal treatment group. High epidural anesthesia group consisted of 22patients, 15 males and 7 females, whose cardiac function was classified into grade Ⅱ in 4 patients, grade Ⅲ in 9 and grade Ⅳ in 9. Conventional treatment group consisted of 13 patients, 11 males and 2 females, whose cardiac function was grade Ⅱ in 1 patient, grade Ⅲ in 5 and grade Ⅳ in 7. Healthy control group comprised 21 people, 13 males and 8 females,who received physical examination at the same period. INTERVENTIONS: Patients with dilated cardiomyopathy were treated with high epidural anesthesia and conventional method, whereas those in conventional treatment group were treated with conventional method only.Elbow venous blood of 3 Ml was sampled from all patients on empty stomach in the morning before treatment and after 4-week treatment. The level of serum interleukin-6 and soluble interleukin-2 receptor was measured with enzyme-linked immunoadsorbent assay. MAIN OUTCOME MEASURES: The level of serum interleukin-6 and soluble interleukin-2 receptor of patients in eachgroup. RESULTS: Blood samples of the 56 subjects were qualified and entered the final analysis.①The level of serum interleukin-6 was higher in dilated car diomyopathy group (13.9ng/L) than in healthy control group (11.22 ng/L) (z= -3.072, P ＜ 0.05). ② The level of serum interleukin-6 in high epidural anesthesia group was obviously decreased aftertreatment (11.42 ng/L) as compared with that before treatment (20.42ng/L) (z =2.582 9, P ＜ 0.05).The le vel of serum interleukin-6 in conventional treatment group was similar before and after treatmant(12.16 ng/L and 12.80 mg/L, z = -1.89,P ＞ 0.05). The difference of interleukin-6 level in high epidural anesthesiagroup (-2.04 ng/L)was obviously higher than that in conventional treat ment group (0.28 ng/L) (P ＜ 0.01) before and after treatment.③The level of serum soluble interleukin-2 receptor was higher in dilated car diomyopathy group[(1 306.17±1.46)ng/L]than in healthy control group [(1078.95±1.23) ng/L] (t =2.51, P ＜ 0.05). ④ The level of soluble intedeukin-2 receptor in high epidural anesthesia group was decreased after treatment [(1 086.68±1.34)ng/L]as compared with thatbeforetreatment [(1 328.01±1.51) ng/L], (t =2.145, P ＜ 0.05). The level of soluble interleukin-2 receptor in conventional treatment group was similar before and after treatment [(1 473.33±1.66) ng/L and (1 331.07±1.52) ng/L,t=-1.06, P ＞ 0.05]. CONCLUSION:The level of serum interleukin-6 and soluble inter leukin-2 receptor is significantly decreased after high epidural anesthesia, suggesting that high epidural anesthesia can regulate cytokines better than conventional treatment.The regulation of high epidural anesthesia is related to inhibition of sympathetic nervous system and humoral-im mune system.