1.Effect of microRNA-106b on the invasion and proliferation of trophoblasts through targeting MMP-2
Jing LI ; Jinming WANG ; Yuehua LIU ; Zhan ZHANG ; Ning HAN ; Jingyan WANG ; Shanhui XUE ; Ping WANG
Chinese Journal of Obstetrics and Gynecology 2017;52(5):327-332
Objective To investigate the expression of microRNA-106b(miR-106b)in the placentas of patients with pre-eclampsia and its relationship with matrix metallopeptidase(MMP)-2,and its effect on the invasion and proliferation of trophoblasts. Methods (1) Placental tissues were collected from patients with mild pre-eclampsia (mPE, n=30) , severe pre-eclampsia (sPE, n=30) and normal pregnant women (n=40). Human choriocarcinoma cell lines JAR and JEG3 were assigned to the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group, respectively. (2) The target gene of miR-106b(such as MMP-2) was predicted by bioinformatics. Dual-luciferase reporting system was used to verify the regulation of miR-106b on the expression of MMP-2. (3) The expressions of miR-106b and MMP-2 were measured by quantitative real-time PCR (qRT-PCR) and western blot. (4) Cell proliferation was determined by MTT assay. (5) Invasive activities in each group were assessed by cell transwell invasion assays. Results (1) Predicting result of bioinformatics indicated that MMP-2 was one of the target genes of miR-106b. Dual-luciferase activity assay demonstrated that MMP-2 was the direct target of miR-106b(P<0.01).(2) The results of qRT-PCR.①The expression of miR-106b in the placentas of mPE, sPE, normal pregnant women were 2.89±0.04, 1.96±0.03, 1.01±0.03, respectively (P<0.05). And the expression of MMP-2 mRNA in the placentas of mPE, sPE, normal pregnant women were 1.87±0.05, 0.69±0.03, 2.78±0.03, respectively (P<0.05).②The expression of miR-106b in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 2.39 ± 0.03, 1.03 ± 0.04, 0.73 ± 0.03, 1.11 ± 0.04, respectively (P<0.05). And its expression in the JEG3 cell line were 2.17±0.04, 1.18±0.04, 0.61±0.03 and 1.22±0.03, respectively (P<0.05). ③The expression of MMP-2 mRNA in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 0.45±0.15, 1.02±0.03, 2.28±0.03, 1.11±0.03, respectively (P<0.05). And its expression in the JEG3 cell line were 0.58±0.03, 1.25±0.15, 2.25±0.03, 1.21±0.03, respectively (P<0.05). (3) The results of western blot.①The expression of MMP-2 protein in the placentas of mPE, sPE, normal pregnant women were 1.63 ± 0.04, 0.55±0.03, 2.82±0.03, respectively (P<0.05).②The expression of MMP-2 protein in the JAR cell line in the miR-106b mimics group, the mimics negative control group, the miR-106b inhibitor group and the inhibitor negative control group were 0.41 ± 0.03, 0.97 ± 0.03, 2.25 ± 0.03, 1.01 ± 0.03, respectively (P<0.05). And its expression in the JEG3 cell line were 0.53±0.03, 1.20±0.03, 2.31±0.04, 1.19±0.03, respectively (P<0.05). (4) miR-106b could inhibit the proliferation of JAR and JEG3 cells, cell proliferation rates in the miR-106b mimics group were lower than that in the mimics negative control group (P<0.05). And cell proliferation rate in the miR-106b inhibitor group was higher than the inhibitor negative control group (P<0.05) . (5) The numbers of JAR cell that passed the membrane in the miR-106b mimics group, the mimics negative control group. The miR-106b inhibitor group and the inhibitor negative control group were 61±15, 79±13, 134±13, 80±12, respectively( P<0.05). And the numbers of JEG3 cell that passed were 57±12, 71±15, 128±15, 70± 14, respectively (P<0.05). Conclusion The miR-106b could inhibit the invasion and proliferation of JAR and JEG3 cells through targeting MMP-2, and have a relationship with the pathogenesis of pre-eclampsia.
2.Mechanism Underlying the Role of HIF-1αduring Hepatocyte Apoptosis in Intermittent Hypoxia with Pulmonary Emphysema in Rats
Jingyan ZHAN ; Huaiyong CHEN ; Qi WU ; Xin SUN ; Jing FENG ; Xue LI ; Long XU ; Cuiping BAO
Tianjin Medical Journal 2014;(2):156-159
Objective To investigate the effect of intermittent hypoxia (IH) with pulmonary emphysema on the ex-pression of hypoxia inducible factor-1α(HIF-1α),Bax and Bcl-2, and the mechanism underlying the role of HIF-1αin he-patocyte apoptosis thereof. Methods Sixty rats were randomly divided into four groups: normal control group, rats were treated normally;IH group, rats were treated by 30 s nitrogen and then 90 s air, and rats were treated by from 9:00-17:00 daily;pulmonary emphysema group, rats were treated by smudging for half an hour, twice a day (8:00 and 18:00);IH with pul-monary emphysema group, rats were treated by 30 s nitrogen and then 90 s air from 9:00-17:00 daily. After exposure four-teen weeks, rats were killed. qRT-PCR assay was conducted to detect the expression of HIF-1α mRNA, Bax mRNA and Bcl-2 mRNA in live tissues. Results The expressions of HIF-1αmRNA, Bax mRNA and Bax/Bcl-2 were significantly higher in IH with pulmonary emphysema group than those in control group,IH group and pulmonary emphysema group (P<0.05). The expression level of Bcl-2 mRNA was significantly lower in IH with pulmonary emphysema group than that of con-trol group and pulmonary emphysema group (P < 0.05), but no significant difference compared with that of IH group (P >0.05). The levels of HIF-1αand Bax were positively correlated with the level of Bax/Bcl-2 (r=0.732 and 0.699),but the lev-els of HIF-1αand Bax were negatively correlated with the level of Bcl-2 (r=-0.705). Conclusion The expression levels of HIF-1αmRNA, Bax mRNA and Bcl-2 mRNA were over-regulated in hepatocytes induced by intermittent hypoxia with pul-monary emphysema. The HIF-1αexpression was correlated with Bax and Bcl-2, suggesting that HIF-1αmay promote the hepatocyte apoptosis through transcriptional co-activators, Bax and Bcl-2.
3.Human leukocyte antigen-G 14 bp deletion polymorphism in severe pre-eclampsia
Zhan ZHANG ; Jingyan WANG ; Linlin ZHANG ; Shang GUO ; Liting JIA ; Hui LI ; Jing LI ; Juan JU ; Shouhua RONG
Chinese Journal of Obstetrics and Gynecology 2010;45(5):348-352
Objective To investigate the relationship between human leukocyte antigen-G( HLA-G) gene Exon 8 14 bp deletion polymorphism and the pathogenesis of severe pre-eclampsia.Methods Forty-two pregnant women with severe pre-eclampsia,who admitted to the Third Affiliated Hospital of Zhengzhou University from October 2008 to February 2009,and their newborns were chosen as the severe pre-eclampsia group.Another 45 healthy gravidas at the third trimester and their newborns were chosen as the control.All gravidas in both groups were Han Nationality.HLA-G Exon 8 genotyping was detected by PCR in both groups and the allele frequencies and genotype frequencies were compared between the two groups.The genotype frequencies of maternal-neonatal pairs were also analyzed.Results ( 1 ) In the severe pre-eclampsia group,14% of the maternal-neonatal pairs were homozygote of 14 bp deletion,and significantly higher frequency 33% (15/45) was found in the control group (P =0.038).(2) No significant difference was found in the allele frequencies and genotype frequencies of HLA-G 14 bp deletion polymorphism among all the mothers between the two groups (P >0.05).(3) The + 14 bp and-14 bp allele frequencies of HLA-G 14 bp deletion polymorphism in newborns in the severe pre-eclampsia group were 44% (37/84) and56% (47/84),respectively,and 30% (27/90) and 70% (63/90) in the control group.Although there was no significant difference between the two groups,but differences in trends was identified (χ2= 3.678 P = 0.055) ; The genotype (-14 bp/-14 bp) frequency of neonates in the severe pre-eclampsia group showed no difference compared with that in the control group[29% (12/42) vs 49% (22/45)],but differences in trends was also found (P =0.052).Conclusions HLA-G 14 bp deletion polymorphism is associated with the susceptibility of severe pre-eclampsia in Chinese Han nationality.Maternal-fetal genotype pairs of-14 bp/-14 bp may have reduced risk of severe pre-eclampsia.
4.Effect of health education based on outcome-based concept on self-management ability and glycemic control in hospitalized patients with type 2 diabetes
Xiaoyan CHEN ; Jiawei WANG ; Xiujing WANG ; Jing ZHAN ; Jingyan TAN ; Jia LYU ; Qingying TAN ; Lingyu ZHENG
Chinese Journal of General Practitioners 2024;23(6):641-646
Objective:To explore the effect of health education based on outcome-based concept on self-management ability and glycemic control in hospitalized patients with type 2 diabetes melittus (T2DM).Methods:A total of 103 T2DM patients admitted in the Endocrinology Department of 903 Hospital of PLA from March 2022 to September 2022 were recruited and randomly divided into study group ( n=52) and control group ( n=51). Routine health education was given to all patients, while additional outcome-based health education was provided by diabetes specialist nurses for the study group during hospitalization. At 12 weeks after discharge, the self-management ability and glycemic control were compared between the two groups. Results:At 12 weeks after discharge, patients in the study group had significantly better self-management ability (dietary control: (4.06±0.75) vs. (3.70±0.88), t=2.50, regular exercise: (3.88±0.62) vs. (3.52±0.94), t=2.30, medical compliance: (4.47±0.51) vs. (4.12±0.64), t=3.14, self blood glucose monitoring: (3.43±0.87) vs. (2.94±0.95), t=2.71, foot care: (3.56±0.57) vs. (2.77±0.87), t=5.42, and management of hyper or hypoglycemia:(3.65±0.72) vs. (3.24±0.96), t=2.48); glycemic control (fasting blood glucose: (6.31±0.90) vs. (6.88±1.37)mmol/L, t=-2.46, 2-h postprandial blood glucose: (8.39±1.64) vs. (9.27±2.50)mmol/L, t=-2.11, HbA1c: (6.76±0.98)% vs. (7.17±0.93)%, t=-2.22). The control rate of HbA1c in the study group was significantly higher and the incidence of hypoglycemia was significantly lower than those in the control group (78.8%(41/52) vs. 58.8%(30/51), χ 2=4.82 and 11.5%(6/52) vs. 29.4%(15/51), χ 2=5.07, respectively). Conclusions:The outcome-based health education can effectively enhance self-management ability and glycemic control in hospitalized type 2 diabetic patients.