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Background and Objectives@#Osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) plays a critical role in the success of lumbar spinal fusion with autogenous bone graft. This study aims to explore the role and specific mechanism of miR-34c-5p in osteogenic differentiation of BMSCs. @*Methods@#and Results: Rabbit model of lumbar fusion was established by surgery. The osteogenic differentiation dataset of mesenchymal stem cells was obtained from the Gene Expression Omnibus (GEO) database, and differentially expressed miRNAs were analyzed using R language (limma package). The expressions of miR-34c-5p, miR-199a-5p, miR-324-5p, miR-361-5p, RUNX2, OCN and Bcl-2 were determined by qRT-PCR and Western blot. ELISA, Alizarin red staining and CCK-8 were used to detect the ALP content, calcium deposition and proliferation of BMSCs. The targeted binding sites between miR-34c-5p and Bcl-2 were predicted by the Target database and verified using dual-luciferase reporter assay. MiR-34c-5p expression was higher in rabbit lumbar fusion model and differentiated BMSCs than normal rabbit or BMSCs. The content of ALP and the deposition of calcium increased with the osteogenic differentiation of BMSCs. Upregulation of miR-34c-5p reduced cell proliferation and promoted ALP content, calcium deposition, RUNX2 and OCN expression compared with the control group. The effects of miR-34c-5p inhibitor were the opposite. In addition, miR-34c-5p negatively correlated with Bcl-2. Upregulation of Bcl-2 reversed the effects of miR-34c-5p on ALP content, calcium deposition, and the expressions of RUNX2 and OCN. @*Conclusions@#miR-34c-5p could promote osteogenic differentiation and suppress proliferation of BMSCs by inhibiting Bcl-2.
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Background and Objectives@#Osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) plays a critical role in the success of lumbar spinal fusion with autogenous bone graft. This study aims to explore the role and specific mechanism of miR-34c-5p in osteogenic differentiation of BMSCs. @*Methods@#and Results: Rabbit model of lumbar fusion was established by surgery. The osteogenic differentiation dataset of mesenchymal stem cells was obtained from the Gene Expression Omnibus (GEO) database, and differentially expressed miRNAs were analyzed using R language (limma package). The expressions of miR-34c-5p, miR-199a-5p, miR-324-5p, miR-361-5p, RUNX2, OCN and Bcl-2 were determined by qRT-PCR and Western blot. ELISA, Alizarin red staining and CCK-8 were used to detect the ALP content, calcium deposition and proliferation of BMSCs. The targeted binding sites between miR-34c-5p and Bcl-2 were predicted by the Target database and verified using dual-luciferase reporter assay. MiR-34c-5p expression was higher in rabbit lumbar fusion model and differentiated BMSCs than normal rabbit or BMSCs. The content of ALP and the deposition of calcium increased with the osteogenic differentiation of BMSCs. Upregulation of miR-34c-5p reduced cell proliferation and promoted ALP content, calcium deposition, RUNX2 and OCN expression compared with the control group. The effects of miR-34c-5p inhibitor were the opposite. In addition, miR-34c-5p negatively correlated with Bcl-2. Upregulation of Bcl-2 reversed the effects of miR-34c-5p on ALP content, calcium deposition, and the expressions of RUNX2 and OCN. @*Conclusions@#miR-34c-5p could promote osteogenic differentiation and suppress proliferation of BMSCs by inhibiting Bcl-2.
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Objective To investigate vitamin D ( VD) level and its influencing factors in 4-15-year-old children in southwest of Zhejiang province, so as to guide intervention strategy for improving VD level. Methods A total of 2 620 4 -15-year-old healthy children who visited Lishui People's Hospital for regular physical check-up between January and December 2014 were divided into 4 age groups, namely 4 -6 years (kindergarten), 7 -9 years (lower grades in primary school), 10 -12 years (higher grades in primary school), and 13-15 years (junior middle school).We randomly selected 200 children from each age group using a random number table.A survey using self-developed questionnaire was conducted to obtain information regarding diet, exposure to sunlight, and VD supplement.Serum 25-( OH) D level was measured with electro-chemiluminescence.VD nutritional status and composition, diet, sunlight exposure, VD supplement were com-pared among the groups.Results In southwest Zhejiang, the mean VD level in primary and middle school students was ( 24.76 ±8.39 ) μg/L, clinical deficiency rate was 31.5%, subclinical deficiency rate was 45.0%, and only 23.5% reached the appropriate level, which was even lower ( 14.5%) in junior middle school students.The VD deficiency rate elevated significantly along with the growth of age (P=0.036).The level of VD was positively correlated with sunlight exposure and VD supplementation, and showed significant in-ter-group differences ( all P7 years should be given more attention to in the intervention of increasing sunlight exposure. Students with difficulties in getting adequate sunlight exposure need VD supplements.
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Objective: To observe the effect of Baihe Zhimu Decoction on serum sex hormone and uterus index of ovariectomized rats with kidney-yin deficiency. Methods: 3-month years old femal SD rats which were ovariectomized were divided randomLy into 6 groups: ovariectomized, Baihe Zhimu DecoctionⅠ, Baihe Zhimu DecoctionⅡ, Baihe Zhimu Decoction Ⅲ, Baihe Zhimu Decoction Ⅳ, then the normal group was set. The rats were induced to kidney-yin deficiency by using etraiodothyronine, then ig Baihe Zhimu Decoction for 4 weeks. The levels of sex hormone E2, FSH, LH in serum were detected and the indexes of uterus were caculated to observe the effect of Baihe Zhimu Decoction on sexual hormone and sexual organ in ovariectomized rats with kidney-yin deficiency. Results: Compared with normal group, the index of uterus and E2 level declined, and the level of FSH and LH increased (P
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Objective To observe the effect of serum containing Baihe Zhimu Decoction(BZD) on ovarian granular cells(GC) injury induced by cadmium chloride(CdCl2) in rats.Methods Totally fifteen female SD rats aged 23 days,3 months and 12 months(5 rats in each age group) were enrolled into the study.The number of ovary GC was counted.The growth of GC within 10 days was observed continuously for the selection of the well-growth ovarian GC.On the third day of the culturing,the screened ovary GC was cultured for another 48 hours in 10,20,40,60 and 80?mol/L CdCl2 solution respectively at 37℃ with 5% CO2 added.The optical density(OD) at 490nm was determined by methyl thiazolyl tetrazolium(MTT) assay for the screen of the optimal concentration of CdCl2 solution.Then the ovary GC were cultured in 96-well plate for 72 hours with serum containing BZD(at the final concentration of 1.25%,2.5%,5%,10% and 20% respectively) added.Meanwhile,the blank control group(cultured with 10% DMEM culture medium),model group(cultured with 10% DMEM culture medium and optimal concentration of CdCl2 solution),and blank serum group(cultured with blank serum and optimal concentration of CdCl2 solution)were set up.After the culturing for 48 hours,the OD value of all the groups was detected to evaluate the effect of BZD-containing serum on CdCl2-induced ovarian GC injury in rats.Results The number of GC in 23-day rats was the biggest and their growth was the best in the manner of exponential growth,indicating that GC in 23-day rats was suitable for the modeling.Ovarian injury was increased with the concentration of CdCl2,and 40?mol/L was the optimal concentration for CdCl2,which induced the shrinkage of GC edge.Serum containing BZD promoted the growth of GC,and counteracted the CdCl2-induced GC injury.Conclusion The protective mechanism of BZD-containing serum on CdCl2-induced GC injury is probably related with the counteraction of CdCl2-induced injury and with the increase of ovarian GC activity and quantity.