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Objective To observe and study the influence degree of strengthen training for family members for the treatment compliance and unhealthy lifestyle of patients with diabetes mellitus. Methods A total of 64 patients with diabetes mellitus from January 2015 to March 2016 were selected and divided into two groups by the method of random number table,32 patients in control group were taken care with routine nursing of diabetes mellitus,32 patients in observation group were taken care with strengthen training for family members,then the treatment compliance,related knowledge cognition of unhealthy lifestyle and unhealthy lifestyle situation of two groups before the intervention and at 4th, 8th and 12th week after the intervention were compared. Results The treatment compliance, related knowledge cognition of unhealthy lifestyle and unhealthy lifestyle situation of two groups before the intervention all had no obvious differences (all P>0.05). The treatment compliance at 4th, 8th and 12th week after intervention were (6.26 ± 0.32), (6.95 ± 0.36), (7.28 ± 0.40) scores in observation group, and (5.23 ± 0.29), (6.01 ± 0.30), (6.30 ± 0.33) scores in control group, and the differences were significant (t=8.253, 9.201, 10.257, all P<0.05). The related knowledge cognition of unhealthy lifestyle at 4th, 8th and 12th week after intervention were (80.78±5.90), (89.44±6.02), (93.43±6.20) scores in observation group, and (73.38± 5.73), (77.93±5.84), (82.20±6.01) scores in control group, and the differences were significant (t=11.258,12.563, 13.324, all P<0.05 ). The unhealthy lifestyle situation at 4th, 8th and 12th week after intervention were 75.00%(24/32), 90.63%(29/32), 96.88%(31/32) in observation group, and 59.38%(19/32), 68.75%(22/32),78.13%(25/32) in control group, and the differences were significant (χ2=7.251, 7.468, 9.169, all P<0.05). Conclusions The influence of strengthen training for family members for the treatment compliance and unhealthy lifestyle of patients with diabetes mellitus are more active, so the value of strengthen training for family members of patients with diabetes mellitus is higher.
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Objective To investigate the expression of suppressor of cytokine signaling-3 (SOCS-3) gene in placenta,its role in the pathogenesis of pre-eclampsia and its effect on proliferation and migration of HTR-8/SVneo cells.Methods Fifteen women with severe pre-eclampsia hospitalized in the First Affiliated Hospital of Nanjing Medical University from October 2010 to March 2011 and t 5 normal pregnant women during the same time period were investigated.Cultured HTR-8/SVneo cells were transfected with SOCS-3 specific small interfering RNA (siRNA) or negative siRNA as the controls.The expression of SOCS-3 mRNA and protein in placenta and these cells was detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot.Cell proliferation was detected by methyl thiazolyl tetrazolium,cell cycle by flow cytometry and migration by the Transwell test.Two independent t tests were used for statistical analysis.Results The SOCS-3 mRNA and protein levels in the severe pre-eclampsia group were lower than those in the normal group (0.25±0.03 vs 0.71±0.08 and 0.21±0.05 vs 0.75±0.12,t=15.94 and 14.29,respectively,both P＜0.05).SOCS-3 mRNA and protein levels in the transfection group at 24 hours were lower than those in the negative control group (0.39±0.02 vs 1.00±0.04 and 0.003 7±0.001 4 vs 1.514 9±0.035 7,t=27.58 and 73.35,respectively,both P＜0.05).The integral absorbance values of cell proliferation in the transfection group at 48,72 and 96 hours after transfection were 0.23 ± 0.01,0.32±0.02 and 0.37± 0.02,respectively,which were lower than those in the negative control group (0.39± 0.02,0.55 ± 0.04 and 0.86± 0.04,t=2.60,6.64 and 42.44,respectively,all P＜0.05).The cell clonal formation was lower in the transfection group compared with the negative group (116± 15 vs 312±24,t=9.96,P＜0.05).The ratios of G1/G0 and S phase cells in the transfection group were (55.75±2.21) ％ and (31.59±0.83) ％,respectively,and were significantly different from those in the negative control group [(47.88± 1.87) ％ and (37.38± 1.34) ％,t=45.43 and 20.06,respectively,P＜0.05].After 48 hours,cell migration in the transfection group was lower than that in the negative control group (93 ± 11 vs 167± 17,t=21.36,P＜O.05).Conclusion SOCS-3 expression is probably involved in the pathogenesis of pre-eclampsia by being down-regulated and therefore impeding proliferation and migration of the trophoblast.
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Objective To investigate the effect of transforming growth factor β1 (TGF-β1) on the expression of matrix metalloproteinase 9 (MMP-9),tissue inhibitor of metalloproteinase 1 (TIMP-1),nuclear factor kappa B(NF-κB) and the possible signalling pathways in human amniotic cells WISH.Methods The WISH cell line was cultured.WISH cells were added with TGF-β1 of different concentrations (0,2,10 and 20 ng/ml,respectively) for 24 hours.Then,reverse transcription (RT) PCR and western blotting were used to analyze the protein and mRNA expression of TIMP-1 and MMP-9; and the expression of NF-κB was analyzed by western blot.Results (1) The profile of TIMP-1 mRNA (0.413 ±0.036,0.623 ±0.058,1.392 ±0.124,1.387 ±0.102) in WISH cells elevated when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).In accordance with TIMP-1 mRNA,the expression of TIMP-1 also elevated with the increase of TGF-β1 (0.357 ± 0.031,0.596 ± 0.048,1.243 ± 0.097 and 1.359 ± 0.121,respectively).And when 2,10 or 20 ng/ml of TGF-β1 was added,the TIMP-1 mRNA and protein were significantly higher than the TIMP-1 mRNA and protein when no TGF-β1 was added(P ＜ 0.05).(2)In contrast with TIMP-1,MMP-9 mRNA (1.325 ±0.056,0.987 ±0.081,0.610 ±0.034,0.347 ±0.023) in WISH cells decreased when the concentration of TGF-β1 increased (0,2,10,20 ng/ml).The MMP-9 protein (1.119 ±0.064,1.008 ±0.052,0.578 ±0.041,0.401 ±0.015) also decreased with the increase of TGF-β1.And when 2,10 or 20 ng/ml of TGF-β1 was added,the MMP-9 mRNA and protein were significantly lower than the MMP-9 mRNA and protein when no TGF-β1 was added (P ＜ 0.05).(3) The NF-κB protein (1.423 ±0.065,1.116 ± 0.045,0.796 ± 0.041,0.359 ± 0.021) was significandy reduced with the increase of TGF-β1 (0,2,10,20 ng/ml; P ＜ 0.05).Conclusions The mRNA and protein expression of TIMP-1 decreased when TGF-β1 was low in WISH cells,whereas those of MMP-9 elevated when TGF-β1 was low.The unbalance of TIMP-1 and MMP-9 was related to the pathology of the premature rupture of membrane.And the NF-κB singalling pathway might be an important mechanism in the regulation of TIMP-1 and MMP-9 system.
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Objective To investigate the effect of epidermal growth factor (EGF) on the expression of Matrix metalloproteinase-9 (MMP-9) and the signalling pathways involved in the trophoblast cell line JEG-3. Methods The JEG-3 trophoblast cell line was used in this study. (1) JEG-3 cells were cultured with various concentrations of EGF (0, 1, 10,20 ng/ml) for 24 hours and the expression of MMP-9 was tested by western blotting and reverse transcription PCR (RT-PCR). (2) Western blotting and RT-PCR were also used to investigate the expression of MMP-9 expression after incubation for 0,4,12 and 24 hours with EGF treatment (10 ng/ml) in JEG-3 cells. (3) According to the different added ingredients, JEG-3 cells were divided into some groups: control group (without EGF), EGF group (exposure to l0ng/ml EGF),EGF + inhibitors group (exposure to 10 ng/ml EGF +20 ng/ml SB203580 or exposure to 10 ng/ml EGF + 10ng/ml U0126) inhibitors group (exposure to 20 ng/ml SB203580 or exposure to 10 ng/ml U0126). Western blotting were used to investigate the expression levels of MMP-9, nuclear factor kappa B (NF-kB) ,p38MAPK, phospho-p38MAPK (p-p38MAPK) , extracellular -signal regulated kinase (ERK) and phospho-ERK (p-ERK) protein in JEG-3 cells after incubation for 24 hours. Results (1) The profiles of MMP-9mRNA were increased by various concentrations of EGF (0, 1 , 10, 20 ng/ml) in JEG-3 cells after 24hculture. The expression of MMP-9 mRNA in JEG-3 cells exposure at 1 ng/ml of EGF (0. 567 ±0. 056) , 10ng/ml of EGF (1. 392 ±0. 133) , 20 ng/ml of EGF (1. 971 ±0. 067) were significantly higher respectively (P <0. 05) , compared with 0 ng/ml of EGF treatment (0. 166 ±0. 015). Similarly, MMP-9 mRNAs were also increased with the increasing incubation time. Compared to EGF (10 ng/ml) stimulation for 0 h (0.253 ±0.044), the MMP-9 mRNA profiles were 0. 470 ±0. 026, 1.061 ±0. 115, 1. 453 ±0. 180 for 4,12 and 24 hours, respectively (P < 0. 05). (2) In accordance to the mRNA profiles, the expression of MMP-9 protein was also increased by different concentrations of EGF (0,1, 10, 20 ng/ml) in JEG-3 cells after 24 h-culture. The abundance of MMP-9 protein in the three groups was 0. 043 ±0. 012, 0. 085 ±0. 008, 0. 142 ±0. 015, with a significantly higher expression, compared with 0 ng/ml of EGF treatment (0. 004 ±0.001, P < 0.05) respectively. Similarly, MMP-9 proteins were also increased with the increasing incubation time. Compared to EGF(10 ng/ml) stimulation for 0 h (0. 030 ±0. 009) , the profiles of MMP-9 protein were 0. 137 ± 0. 010, 0. 240 ± 0. 010, 1.240 ±0.061 for 4, 12 and 24 hours, respectively (P < 0.05). (3) Both p38MAPK and ERK signalling pathways were activated by EGF in JEG-3 cells. The expression of p-p38MAPK was significantly higher (without or with 10 ng/ml EGF, 234. 1 ± 4. 1 vs.260. 9 ± 2. 5 , P < 0. 05) , however, the p38 MAPK inhibitor SB203580 markedly suppressed the increase in p-p38MAPK content induced by EGF(227. 9 ±2. 4 vs. 260. 9 ±2. 5, P<0. 05). Similarly, the expression of p-ERK was significantly higher with EGF treatment (812. 2 ±3. 5) vs. without EGF group (453.4±5.8) (P <0. 05) , while the ERK inhibitor U0126 significantly inhibited the increased p-ERK content in response to EGF treatment (71. 0 ± 1. 2 vs. 812. 2 ± 3. 5, P < 0. 05) . (4) The p38MAPK inhibitor SB203580 significantly reduced the expression of EGF-induced MMP-9 (0. 645 ± 0. 270 vs. 1. 476 ± 0. 452, P < 0. 05)and NF-kB (0.530 ± 0.026 vs. 0.959 ± 0. 017, P < 0. 05) . (5) The ERK inhibitor U0126 also significantly reduced the expression of EGF-induced MMP-9 (0. 623 ±0. 030 vs. 2. 112 ±0. 056, P <0. 05)and NF-kB (0. 325 ± 0. 082 vs. 0. 939 ± 0. 153, P < 0. 05). Conclusion EGF induced the expression of MMP-9 in a time and dose-dependant manner in JEG-3 cells. EGF enhanced MMP-9 expression through the activation of p38MAPK and ERK signalling pathways in JEG-3 cells.