1.Construction of adenoviral vector encoding Calponin-1 siRNA and its effect on human myometrium cells in vitro
Yonghong GU ; Changju ZHOU ; Lingyu HU ; Qian CHEN ; Weishe ZHANG
Journal of Central South University(Medical Sciences) 2009;34(12):1231-1237
Objective To investigate the effect of Calponin-1 suppression on human myometrium cells through adenovirus mediated siRNA. Methods Human uterine smooth muscle tissues were digested with enzymes, cultured and confirmed with immunocytochemistry. Aadenovirus siRNA-Calponin-1 plasmid was transfected into primary cultured uterine smooth muscle cells in vitro. The expressions of Calponin-1 mRNA and protein were analyzed by RT-PCR and Western blot, respectively.Results The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid was successfully constructed, and Calponin-1 siRNA mediated by recombinant adenovirus resulted in markedly reduced expression of Calponin-1 mRNA and protein in human myometrium cells. The gray values of Calponin-1 mRNA in the uterine smooth muscle cells in the experimental, blank control, and empty vector groups were 316.3±39.2, 1048.5±126.4 and 1027.2±127.5, respectively. The gray values of Calponin-1 protein were 323.3±43.2, 1021.5±143.4, and 1019.2±144.5,respectively. The difference between the experimental group and the blank control group as well as the empty vector group was significant (P< 0.05). There was no significant difference between the empty vector group and the blank control group (P>0.05).Conclusion The pAdEasy-pShuttle-U6-Calponin-1 siRNA plasmid can inhibit the expression of Calponin-1 in human myometrium cells in vitro,which may be a useful approach to determine the role of Calponin-1 in delivery.
2.Clinical value of angiogenin in predicting the prognosis of patients with idiopathic pulmonary fibrosis
Yanling BAI ; Haiyan ZHU ; Qiyu SUN ; Guozhong GU ; Lingyu ZHANG ; Ying LI ; Baofeng YANG
Chinese Critical Care Medicine 2017;29(9):789-793
Objective To explore the relationship between angiogenin-1/2 (Ang-1/2) and clinical parameters of idiopathic pulmonary fibrosis (IPF), and to assess the value of Ang-1/2 in predicting the prognosis of patients with IPF.Methods A retrospective analysis was conducted. Ninety-one patients diagnosed as IPF by high resolution CT (HRCT) and lung biopsy admitted to Daqing Oil Field General Hospitalfrom March 2014 to January 2015 were enrolled. The general data, serum parameters and pulmonary function parameters of all patients were collected. After treatment, all of the 91 patients were followed-up to 2 years. The patients were divided into favorable prognosis group and unfavorable prognosis group according to follow-up results. The differences in all parameters between the two groups werecompared. The relationship between Ang-1, Ang-2 and lung function parameters was analyzed by Pearson correlation analysis. Cox proportional hazard regression model was used to evaluate the effect of clinical parameters on the prognosis of patients with IPF. The effect of Ang-2 in predicting prognosis of patients with IPF was analyzed by receiver operating characteristic (ROC) curve.Results During the 2-year follow-up period, 30 of 91 patients showed a favorable prognosis, and 55 showed an unfavorable prognosis with a poor prognosis rate of 64.71%, and 6 patients withdrew from the study due to loss of follow-up and death. Compared with the favorable prognosis group, Ang-2 level in the unfavorable prognosis group was significantly increased (μg/L: 2.88±1.63 vs. 1.89±1.22,t = 2.909,P= 0.005), but Ang-1 only showed a slight increase (μg/L: 28.70±14.26 vs. 25.62±11.95,t = 1.005,P = 0.318). The results of Pearson correlation analysis showed that Ang-2 level was negatively correlated with forced expiratory volume in 1 second (FVC1) and the percentage of carbon monoxide diffusing capacity accounting for the expected value (DLCO%;r value was -0.227 and -0.206, andP value was 0.147 and 0.253, respectively), but no significant correlation between the level of Ang-1 and FVC1 as well as DLCO% was found (r value was -0.153 and -0.121, andP value was 0.147 and 0.253, respectively). Cox proportional hazard regression model analysis showed that the prognosis of patients with IPF was significantly affected by smoking time and Ang-2 (bothP< 0.05), and the influence of Ang-2 was greater [relative risk (RR): 1.236 vs. 1.006, P= 0.037]. Age, gender, smoking and the levels of FVC1, DLCO% and Ang-1 had no significant effect on the prognosis of IPF patients (allP> 0.05). Prognostic analysis showed that the area under ROC curve (AUC) of Ang-2 for predicting prognosis of patients with IPF was 0.692, and the best diagnostic point was 0.35μg/L, the sensitivity was 61.8%, the specificity was 73.3%, the positive predictive value was 69.8%, and the negative predictive value was 65.7% which indicated that Ang-2 could predict the prognosis of patients with IPF.Conclusion Ang-2 could assess the prognosis of patients with IPF, which is expected to be used as an indicator of predicting the prognosis of patients with IPF.
4.Expression of Calponin-1 and Transgelin in human uterine smooth muscles in non-labor and labor situation.
Qian CHEN ; Yonghong GU ; Changju ZHOU ; Lingyu HU ; Changying PENG
Journal of Central South University(Medical Sciences) 2010;35(10):1073-1079
OBJECTIVE:
To investigate the expression of Calponin-1 and Transgenlin in the uterine smooth muscles during normal labor on-sets, and to evaluate their effect on initiating the normal labor.
METHODS:
A total of 14 uterine bodies and lower segments of human pregnancy were divided to a non-labor group (NIL) and a labor group(IL). Immunohistochemical technology and Western blot were used to determine the expression of Calponin-1 and Transgelin in the 2 groups.
RESULTS:
Immunohistochemical detection and Western blot showed that Calponin-1 protein in the uterine smooth muscle tissue of the body and the lower uterine segment of smooth muscle tissues had significant difference (P<0.05). The expression of Transgelin in the uterine body smooth muscle tissue in the IL was higher than that in the NIL(P<0.05). In the lower uterine segments of the smooth muscle, the expression of Transgelin was not significantly different in the 2 groups (P>0.05).
CONCLUSION
Calponin-1 of the uterine smooth muscle and Transgelin of the uterine body smooth muscle may involve in the regulation of uterine smooth muscle contractility, which is closely related to child birth launch.
Adult
;
Calcium-Binding Proteins
;
genetics
;
metabolism
;
Female
;
Humans
;
Labor Onset
;
metabolism
;
Microfilament Proteins
;
genetics
;
metabolism
;
Muscle Proteins
;
genetics
;
metabolism
;
Myometrium
;
metabolism
;
Pregnancy
;
Uterine Contraction
;
metabolism