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Objective@#To investigate the effect of Toll-like receptor 9 (TLR9) on high glucose-induced retinal ganglion cells-5 (RGC-5) apoptosis and the inhibitory effects of small interfering RNA-TLR9 (si-TLR9) on apoptosis.@*Methods@#RGC-5 cells were divided into normal control group, high glucose group, high glucose+ negative control group and high glucose+ si-TLR9 group which cells were respectively dealt with normal culture medium, high glucose medium, transfection of non-specific siRNA under high glucose and transfection of siRNA-TLR9 under high glucose.The expression of TLR9 mRNA was detected by real time PCR; the survival rate of the cells was evaluated by MTT assay; the apoptotic rate of the cells was detected by flow cytometry; the caspase-3 activity in the cells was detected by related kit, and the expressions of TLR9, B cell lymphoma (bcl-2), bcl-2 associated X protein (bax), p38 mitogen-activated protein kinases (p38MAPK) and phosphorylated (p)-p38MAPK proteins were detected by Western blot.@*Results@#The expressions of TLR9 mRNA and protein in the high glucose+ si-TLR9 group were significantly decreased in comparison with the high glucose+ negative control group (both at P<0.05). The cell survival rate of the high glucose group and high glucose+ negative control group was (78.36±5.13)% and (75.12±4.25)%, respectively, which was significantly lower than (95.48±7.25)% in the normal control group, and that in the high glucose+ si-TLR9 group was (86.58±5.32)%, which was significantly reduced in comparison with the high glucose+ negative control group (all at P<0.05). The apoptotic rate of the cells in the high glucose group and high glucose+ negative control group was (13.23±1.22)% and (12.52±1.38)%, respectively, which was significantly higher than (2.26±0.15)% of the normal control group, and apoptotic rate in the high glucose+ si-TLR9 group was (7.15±0.24)%, which was significantly lower than that in high glucose+ negative control group (all at P<0.05). The expression of bax in the cells of the high glucose+ si-TLR9 group was significantly decreased, and the expression of bcl-2 in the cells of the high glucose+ si-TLR9 group was significantly increased in comparison with the high glucose+ negative control group (both at P<0.05). Caspase-3 activity in the cells was significantly decreased in the high glucose+ si-TLR9 group compared with the high glucose+ negative control group (P<0.05). The relative expression of p-p38MAPK in the cells was significantly decreased in the high glucose+ si-TLR9 group compared with the high glucose+ negative control group (P<0.05).@*Conclusions@#Down-regulation of TLR9 expression in the RGC-5 cells can inhibit the apoptosis induced by high glucose probablely by suppressing p38MAPK signaling pathway.
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Objective To investigate the effect of Toll-like receptor 9 (TLR9) on high glucose-induced retinal ganglion cells-5 (RGC-5) apoptosis and the inhibitory effects of small interfering RNA-TLR9 (si-TLR9) on apoptosis.Methods RGC-5 cells were divided into normal control group,high glucose group,high glucose+ negative control group and high glucose + si-TLR9 group which cells were respectively dealt with normal culture medium,high glucose medium,transfection of non-specific siRNA under high glucose and transfection of siRNA-TLR9 under high glucose.The expression of TLR9 mRNA was detected by real time PCR;the survival rate of the cells was evaluated by MTT assay;the apoptotic rate of the cells was detected by flow cytometry;the caspase-3 activity in the cells was detected by related kit,and the expressions of TLR9,B cell lymphoma (bcl-2),bcl-2 associated X protein (bax),p38 mitogen-activated protein kinases (p38MAPK) and phosphorylated (p)-p38MAPK proteins were detected by Western blot.Results The expressions of TLR9 mRNA and protein in the high glucose+si-TLR9 group were significantly decreased in comparison with the high glucose+negative control group (both at P＜0.05).The cell survival rate of the high glucose group and high glucose+negative control group was (78.36±5.13)％ and (75.12± 4.25) ％,respectively,which was significantly lower than (95.48± 7.25) ％ in the normal control group,and that in the high glucose+si-TLR9 group was (86.58±5.32)％,which was significantly reduced in comparison with the high glucose+negative control group (all at P＜0.05).The apoptotic rate of the cells in the high glucose group and high glucose+negative control group was (13.23 ± 1.22) ％ and (12.52± 1.38) ％,respectively,which was significantly higher than (2.26±0.15)％ of the normal control group,and apoptotic rate in the high glucose+si-TLR9 group was (7.15±0.24) ％,which was significantly lower than that in high glucose+negative control group (all at P＜0.05).The expression of bax in the cells of the high glucose+si-TLR9 group was significantly decreased,and the expression of bcl-2 in the cells of the high glucose+si-TLR9 group was significantly increased in comparison with the high glucose+ negative control group (both at P＜ 0.05).Caspase-3 activity in the cells was significantly decreased in the high glucose+si-TLR9 group compared with the high glucose+negative control group (P＜0.05).The relative expression of p-p38MAPK in the cells was significantly decreased in the high glucose+ si-TLR9 group compared with the high glucose+negative control group (P＜0.05).Conclusions Down-regulation of TLR9 expression in the RGC-5 cells can inhibit the apoptosis induced by high glucose probablely by suppressing p38MAPK signaling pathway.
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Objective To investigate the relationship between serum 25-OH vitamin D3 (25-OHD3)levels and carpal tunnel syndrome(CTS)in type 2 diabetes.Methods Data from 102 patients with type 2 diabetes were collected,and patients were divided into a CTS group(n=29)and a non-CTS group(n=73)according to clinical symptoms,signs,and electrophysiological testing.Fasting venous blood was collected to assay serum 25-OHD3 levels that were compared between the two groups.The correlation between 25-OHD3 and clinical parameters for type 2 diabetes was analyzed.Results The incidence of vitamin D3 deficiency was 65.5％ (n=19)in the CTS group and 46.6％(n=34)in the non CTS group,and the difference was statistically significant(x2 =16.85,P＜0.01).Levels of 25-OHD3 were lower in the CTS group(14.13±4.91) μg/L than in the non-CTS group(17.08±3.35)μg/L(t =4.067,P ＜ 0.01).Levels of 25 OHD3 were negatively correlated with those of glycosylated hemoglobin(HbA1c)(r =-0.176,P =0.041)and the duration of diabetes(r=-0.29,P=0.01) Conclusions Low 25-OHD3 is a risk factor for carpal tunnel syndrome in type 2 diabetes and is negatively associated with the duration of diabetes and HbA1c in patients with type 2 diabetes.
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Objective To explore the correlation of bone mineral density (BMD) with levels of N-terminal osteocalcin (N-MID),total collagen type 1 amino-terminal extension peptide,β-collagen special sequence (β-CTX),25-hydroxy vitamin D [25 (OH) D3] and insulin-like growth factor-1 (IGF-1) in postmenopausal women with type 2 diabetes.Methods 116 elderly postmenopausal women with type 2 diabetes were divided into 3 groups according to BMD level:normal group (n=29),osteopenic group (n=49) and osteoporosis group (n=38).Body height,weight,body mass index (BMI) were measured.Fasting blood glucose (FBG),2-hour postprandial glucose (PPG),glycated hemoglobin (HbAlc),24-hour urinary protein (UPro) and levels of N-MID,T-P1NP,β-CTX,25(OH)D3 and IGF-1 were determined.General informations and bone metabolic markers were statistically analyzed and compared between the 3 groups.Results There were no significant differences in levels of FBG,PPG,HbA1c,25(OH)D3 and duration between the 3 groups (all P＞0.05).For the comparison of normal group with osteopenic and osteoporosis groups,BMI and IGF-1 levels were decreased (both P＜0.05) ; T-P1NP and N-MID levels were decreased [(19.85 ± 2.83)μg/Lvs.(12.23±2.75) μg/L,(9.16±2.86) μg/L; (7.12±2.05) μg/Lvs.(5.06±2.38) μg/L,(4.92±2.12) μg/L,all P＜0.05]; 24 h urine protein was increased (P＜0.05); β-CTX level was increased [(1.32±0.38) μg/Lvs.(1.97±0.58) μg/L,(2.14±0.73) μg/L,P＜0.05].Logistic regression analysis showed that BMD was positively correlated with total collagen type 1 aminoterminal extension peptide,IGF-1,25(OH)D3,N MID and BMI (r=0.351,0.217,0.213,0.273,0.203,respectively,P＜0.01 or 0.05),and BMD was negatively correlated with β-CTX and urinary protein levels (r =-0.236 and-0.216,P ＜ 0.05).Conclusions Osteopenia or osteoporosis occurs more often in postmenopausal women with type 2 diabetes.The change of BMD is closely related with bone metabolism markers and IGF 1 level.Routine tests and early prevention can avoid fracture in postmenopausal women with type 2 diabetes.
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Objective To understand the features of cognitive impairment in patients with subclinical hypothyroidism. Methods The cognitive function of the patients with subclinical hypothyroidism and the healthy controls was tested using Mini-Mental State Examination (MMSE) and Wechsler Memory Scale (WMS). The change in cognitive function was observed and analyzed before and after L-T4 treatment. Results Fifty-seven patients with subclinical hypothyroidism and the same number of healthy controls were enrolled in this study. There were no significant differences between two groups in biochemical indexes and MMSE score (all P＞0.05). But there were significant differences between two groups in WMS score, including pictures, memories, learning, touch, understanding memory and back several test scores (all P＜0.05). There existed certain degree of cognitive dysfunction mainly displayed in memory, attention, etc. After three months of L-T4 treatment, all had significant improvement (all P＜0.05). Conclusions Patients with subclinical hypothyroidism have cognitive dysfunction, when the thyroid stimulating hormone is restored to normal level, cognitive function is also improved.