Apigenin accelerates lipopolysaccharide induced apoptosis in mesenchymal stem cells through suppressing vitamin D receptor expression / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Transplantation of mensenchymal stem cells (MSCs) has been proposed as a promising way for tissue engineering. However, the application of MSCs for transplantation will undergo apoptosis due to the extremely harsh microenvironment such as excessive inflammation. Apigenin (API) has been reported to protect cells against inflammatory damage and cell death by exhibiting anti-inflammatory and anti-oxidative capacity. Here we investigated the modulatory effects of API in lipopolysaccharide (LPS)-mediated inflammation and apoptosis of MSCs, and further defined the underlying mechanism.</p><p><b>METHODS</b>Effects of different concentrations of API (0, 5, 10, 20, 40 and 80 µmol/L) for 24 hours, and LPS (0, 0.5 and 5.0 µg/ml) for 6 hours and 24 hours on MSCs viability were assayed by MTT. Based on this, MSCs were pretreated with different concentrations of API (0 - 40 µmol/L) at the indicated times (6, 12 and 24 hours) followed by exposure to 5 µg/ml LPS for 24 hours. MTT, phase-contrast microscopy, annexinV/propidium iodide (PI) double stain flow cytometry (FCM) and Hoechst staining were applied to explore the effects of API on MSCs induced by 5 µg/ml LPS for 24 hours. In addition, reverse-transcription polymerase chain reaction (RT-PCR) was applied to detect the mRNA expression of pro-inflammatory factors including cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-κB), pro-apoptotic gene caspase-3, Bad, and anti-apoptotic gene Bcl-2. Moreover, AutoDock software was used to imitate the docking score of API and vitamin D receptor (VDR). In parallel, Western blotting and RT-PCR were used to investigate protein and mRNA expression of VDR.</p><p><b>RESULTS</b>MSCs stimulated with LPS 5 µg/ml for 24 hours was used as a model of apoptosis induced by over inflammatory stimulus. API (0 - 40 µmol/L) had non-toxic effect on MSCs; however, it could decrease mRNA expression of COX-2, iNOS and NF-κB at different time points in MSCs induced by LPS, except for API at the concentration of 5 µmol/L.</p><p><b>RESULTS</b>from phase-contrast microscopy, MTT, Hoechst staining and AnnexinV/PI double stain FCM demonstrated that with the increasing concentrations of API and extension of administrating time, significant morphological changes of MSCs occurred, viability of cells was strongly inhibited, and meanwhile, apoptosis of LPS-administrated MSCs was exacerbated, compared with LPS individual group. In addition, API promoted caspase-3, Bad mRNA expression and inhibited Bcl-2 mRNA expression in a time-dependent and concentration- dependent manner. Further study found that pro-apoptosis effect of API was related to suppress VDR expression.</p><p><b>CONCLUSIONS</b>API could inhibit the expression of inducible inflammatory factors, therefore exert the strong anti-inflammatory function. However, API could not protect MSC apoptosis induced by LPS but amplified the apoptosis. The apoptosis is related to Bad/Bcl-2 increasing and caspase-3 activation, which is mediated through suppressing VDR expression.</p>

Experimental study of icariin in inducing bone marrow mesenchymal stem cell differentiation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To research the effect of icariin (ICA) on the proliferation and differentiation of bone mesenchymal stem cells (BMSCs) and to study its influence on the expressions of transforming growth factor-beta1 (TGF-beta1) and bone morphogenetic protein-2 (BMP-2) in the progress of BMSCs differentiating into osteoblast, for providing an experimental evidence to explain the mechanism of ICA, also for exploring the feasibility of establishing a platform upon TGF-beta, and BMP-2 to screen out the medicine in preventing and treating osteoporosis.</p><p><b>METHODS</b>After the most effective concentration of ICA for promoting the differentiation of BMSCs into osteoblast was judged with the indices like alkaline phosphatase (ALP), etc., a grouped experiment was conducted for the sake of studying the effect and mechanism of ICA in its process of inducing BMSCs differentiation into osteoblast through detecting expression of ALP and calcium nodes, as well as the expressions of TGF-beta1, and BMP-2 with ELISA.</p><p><b>RESULTS</b>The most effective concentration of the ICA on the BMSCs differentiation was judged as 1 x 10(-9) mol/L, ICA of that concentration showed effects in enhancing the expressions of osteoblast-indices and increasing the secretion of TGF-beta1, and BMP-2. Besides, the increase of TGF-beta1, and BMP-2 was revealed in all the groups, in which ICA showed its influence visibly.</p><p><b>CONCLUSION</b>ICA could promote the differentiation of BMSCs into osteoblast; the up-regulation of TGF-beta1, and BMP-2 expressions is possibly one of the action mechanisms of various interventional drugs in their differentiation promoting progress.</p>

The methods of shorting proximal femoral and total hip arthroplasty for Crowe IV dysplastic hip of adults / 中华外科杂志

<p><b>OBJECTIVES</b>To discuss the methods and outcome of shorting proximal femoral and total hip arthroplasty for Crowe IV dysplastic hip of adults.</p><p><b>METHODS</b>From July 2000 to February 2006, 13 cases of osteoarthritis secondary to severe development dysplastic hip were treated by total hip replacement and the shorting proximal femoral.</p><p><b>RESULTS</b>The duration of follow-up ranged from 4 months to 55 months. The average score increased from 36.9 to 84.1 points after the surgery according to Harris. All the patients could walk independently. Their paces were improved obviously and the function of their hips was satisfactory.</p><p><b>CONCLUSIONS</b>The treatment by total hip arthroplasty and the shorting of posterior femoral is effective and efficient for osteoarthritis secondary to Crowe IV development dysplastic hip in adults. The long-term followup is necessary for further study.</p>

The methods of shorting of proximal femoral and total hip arthroplasty for old femoral neck fracture with severe hip joint dislocation / 中华外科杂志

<p><b>OBJECTIVE</b>To discuss the effective method and outcome to old femoral neck fracture with severe hip joint dislocation.</p><p><b>METHODS</b>From April 1996, 7 cases of old femoral neck fracture with severe hip joint dislocation were treated by the shorting of posterior femoral and total hip arthroplasty.</p><p><b>RESULTS</b>The average age of 7 patients was 51 years old and the mean follow-up was 27.3 months. The mean Harris score was improved to 84.3 from 36.7 before operation in the latest follow-up. The prosthesis position of acetabular and femoral was excellent, there was no sinking or softening of artificial joint, nonunion of femoral osteotomy.</p><p><b>CONCLUSION</b>The preliminary clinical results are quite satisfactory of the shorting of posterior femoral and total hip arthroplasty to old femoral neck fracture with severe hip joint dislocation, the follow-up is necessary for further long-term outcome.</p>