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BACKGROUND@#The casein kinase 2-interacting protein-1 (CKIP-1) is important in the development of osteoblasts and cardiomyocytes. However, the effects of CKIP-1 on osteoblast precursor mesenchymal stem cells (MSCs) remain unclear. This study aimed to determine whether CKIP-1 affects osteogenic differentiation in MSCs and explore the relationship of CKIP-1 and inflammation.@*METHODS@#Bone marrow MSCs of CKIP-1 wild type (WT) and knockout (KO) mice were cultivated in vitro. Cell phenotype was analyzed by flow cytometry, colony formation was detected to study the proliferative ability. Osteogenic and adipogenic induction were performed. The osteogenic ability was explored by alizarin red staining, alkaline phosphatase (ALP) staining and ALP activity detection. Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out to determine the mRNA expression levels of osteoblast marker genes. The adipogenic ability was detected by oil red O staining. Content of the bone was analyzed to observe the differences of bone imaging parameters including trabecular bone volume/tissue volume (BV/TV), bone surface area fraction/trabecular BV, trabecular number (Tb.N), and trabecular spacing (Tb.sp). Interleukin (IL)-1β was injected on WT mice of 2 months old and 18 months old, respectively. Difference in CKIP-1 expression was detected by RT-PCR and western blot. The relationship between CKIP-1 and inflammation was explored by RT-PCR and western blot.@*RESULTS@#ALP assays, alizarin red staining, and qRT-PCR showed that MSCs derived from CKIP-1 KO mice exhibited a stronger capability for osteogenesis. Micro-computed tomography detection showed that among 18-month-old mice, CKIP-1 KO mice presented significantly higher bone mass compared with WT mice (P = 0.02). No significant difference was observed in 2-month-old mice. In vivo data showed that expression of CKIP-1 was higher in the bone marrow of aging mice than in young mice (4.3-fold increase at the mRNA level, P = 0.04). Finally, the expression levels of CKIP-1 in bone marrow (3.2-fold increase at the mRNA level, P = 0.03) and cultured MSCs were up-regulated on chronic inflammatory stimulation by IL-1β.@*CONCLUSIONS@#CKIP-1 is responsible for negative regulation of MSC osteogenesis with age-dependent effects. Increasing levels of inflammation with aging may be the primary factor responsible for higher expression levels of CKIP-1 but may not necessarily affect MSC aging.
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OBJECTIVE@#To establish an arterial remodeling model of rats and to investigate the expression and role of Hippo signaling pathway in this model.@*METHODS@#In the model group (n=40), the left common carotid artery was removed through the median incision of the neck. The 6-0 non-absorbable line was used to ligate the carotid artery near the proximal end as far as possible, completely blocking the blood flow. The common carotid artery of rats in control group (n=20) was not ligated using the operative line. After 14 days, the animals were sacrificed and the common carotid arteries were separated through the original surgical pathway and the arteries from the ligature to the distal end were collected. Arterial morphology and fibrosis were observed by HE and MASSON staining. Immunohistochemical staining was used to detect the expressions of anti-α smooth muscle actin (α-MSA) and proliferating cell nuclear antigen (PCNA) in the carotid artery. Western blot was used to detect the expressions of yes associated protein (YAP), transcriptional coactivator with PDZ-binding motif (TAZ), TEAD1, Bcl-2-like protein 4 (Bax), and B-cell lymphoma-2 (Bcl-2).@*RESULTS@#Compared with the control group, the HE staining showed that the vascular remodeling was obvious, the ratio of the neointima/middle membrane was increased significantly, and the MASSON staining indicated that the fibrosis was significantly increased in model group. The immunohistochemical staining suggested that the expressions of α-SMA and PCNA were increased significantly; Western blot suggested that the expressions of YAP, TAZ, TEAD1, and Bcl-2 were increased in carotid artery of the model group. While the expression of Bax and the ratio of Bax/Bcl-2 were decreased.@*CONCLUSION@#A rat model of arterial remodeling mediated by carotid artery ligation was established successfully in this study. Hippo signaling pathway was proved to be activated in the arterial remodeling model induced by carotid artery ligation in rats, and might regulate the change of Bax/Bcl-2 ratio related to proliferation and apoptosis, and subsequently involved in the proliferation of smooth muscle cells to promote vascular remodeling.
Subject(s)
Animals , Rats , Carotid Arteries , Metabolism , Carotid Artery, Common , Cell Proliferation , Myocytes, Smooth Muscle , Protein Serine-Threonine Kinases , Metabolism , Signal Transduction , Vascular Remodeling , PhysiologyABSTRACT
Objective To investigate the correlation between the expression level of high temperature requirement serine protease A1 (HtrA1) in nucleus pulposus and the degree of intervertebral disc degeneration (Pfirrmann grade).Methods Thirty-six patients who underwent excision of nucleus pulposus were examined by magnetic resonance imaging (MRI) before operation,and the Pfirrmann grading of all patients was performed according to the sagittal T2 weighted MRI.The expression of HtrA1 in nucleus pulposus tissue was detected by reverse transcription polymerase chain reaction and Western blot.The correlation between the expression level of HtrA1 in nucleus pulposus tissue and Pfirrmann grade was analyzed.Results MRI in all 36 patients showed that there were 3 cases of Pfirrmann grade Ⅰ,10 cases of grade Ⅱ,11 cases of grade Ⅲ,7 cases of grade Ⅲ,and 5 cases of grade Ⅴ.The mRNA and protein expressions of HtrA1 in nucleus pulposus tissue increased with the increase of Pfirrmann grade.There were significant differences in the expression level of HtrA1 among different Pfirrmann grade groups (P<0.05) except for the difference between grade Ⅲ and Ⅴ (P>0.05).Spearman rank correlation analysis revealed that there was a rank correlation between expression level of HtrA1 and Pfirrmann grade (P<0.000 1).Conclusion The mRNA and protein expressions of HtrA1 in nucleus pulposus tissue increase with the increase of Pfirrmann grade,indicating HtrA1 is correlated with the degree of intervertebral disc degeneration.
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The large defect of oral and maxillofacial region doesn't only affect the function and aesthetics but also has an adverse impact on patients' psychology. The traditional way to restore the defects are limited by donor site and secondary trauma. In recent years,the oral mucosal tissue engineering has developed rapidly and provides a new solution for craniofacial reconstruction. Tissue-engineered oral mucosa is an ideal substitute of oral mucosa. It can be used in clinical settings and in vitro experiments. This articles review the recent advances in tissue-engineered oral mucosa and its applications.
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Objective To compare the control effect on postoperative GER disease between tubular EC stomach esophagus anastomosis and the traditional full stomach esophagus anastomosis.Methods From September 2010 to October 2015 in Bozhou People's Hospital,85 patients diagnosed with esophageal cancer undergoing elective resection were randomly divided into a tubular stomach EC group (45 cases) and total gastrectomy group (40 cases),two patients underwent esophageal resection,wherein the tubular stomach set of rows of tubular esophagus stomach anastomosis,total gastrectomy group underwent conventional full stomach esophagus anastomosis.After the surgery until the patient to return to normal gastrointestinal function uses dynamic monitor its pH 24h esophageal pH monitoring chamber,the other respectively after 1 March using RDQ Scale GER-related symptoms in patients with score,at the same time Statistics after 1 March of the occurrence of GER.Results There were no deaths occurred,and no occurrence of postoperative anastomotic fistula and thoracic gastric emptying dysfunction,etc;the two groups were almost reached full monitoring 24 h,and between groups while monitoring the total time,Li position monitoring time,there was no significant supine monitoring time (P＞0.05);24 h reflux episodes long tubular gastric reflux group and significantly less than the number of total gastrectomy group,the longest duration of reflux and pH value ＜4.00 The cumulative time was significantly shorter in total gastrectomy group,DeMeester scores were significantly lower than the total gastrectomy group,between groups were statistically significant (P＜0.01);postoperative gastric tube 1,March RDQ score and incidence of GER significantly lower than the total gastrectomy group,between groups were statistically significant (P＜0.01 or P＜0.05).Conclusion Tubular stomach esophagus anastomosis compared with conventional full stomach esophagus anastomosis resection of esophageal cancer has a more ideal GER disease control effect,and can provide a reference for the choice of nastomosis ways for patients with esophageal cancer surgery.
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Objective To observe the expression of β-arrestin2 in cardiac tissues of ovariectomzied (OVX) rats, and to explore the role of β-arrestin2 in estrogen-mediated cardiovascular protection in OVX rats. Methods OVX rats were evaluated by monitoring the serum levels of estrogen and the ratio of' uterine weight to body weight. The cardiovascular function of rats was assessed by measuring the ratio of heart weight to body weight, blood pressure and heart rate after estrogen replacement treatment for 4 weeks, Western blotting analysis was- performed to detect the protein expression of β-arrestin1, β-arrestin2 and angiotensin receptor type 1 (AT1R) in the cardiac tissues. Results Compared with rats in the sham group, the serum estrogen level and the ratio of uterine weight to body weight in OVX rats were decreased significantly (P<0. 05); meanwhile, the cardiovascular parameters including blood pressure, heart rate and the ratio of heart weight to body weight were significantly increased (P<0.05) in OVX rats, and they were attenuated after treatment with estrogen replacement. Compared with the sham controls, the cardiac expression of β-arrestin2 was decreased significantly in OVX rats (P<0.05), with up-regulated estrogen expression and unchanged expression of β-arrestin1 and AT1R. Conclusion β-Arrestin2 expression is decreased in the cardiac tissues of OVX rats, which can be increased by estrogen replacement therapy.
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<p><b>OBJECTIVE</b>To investigate the chemical constituents of Epimedium brevicornum.</p><p><b>METHOD</b>The chemical constituents were isolated by using silica gel column chromatography and preparative TLC. The structures were identified on the basis of physical-chemical constants and spectral data.</p><p><b>RESULT</b>Five compounds were isolated and identified as hyperoside, icariin, epimedin B, epimedin C, inositol.</p><p><b>CONCLUSION</b>Compound I and III - V were isolated from the plant for the first time.</p>