Bone is an active tissue in which the processes of remodeling are continuous to ensure normal bone integrity and strength. Steroids play an important role in regulating bone growth, development and remodeling. Glucocorticoids excess will induce bone damages especially osteoporosis. Otherwise, estrogen and androgen are bone protective steroids in both female and male. To develop a new selective steroid receptor modulator is one of the targets in future study to treat osteoporosis.
Adult ; Androgens ; physiology ; Bone Development ; Bone Remodeling ; Estrogens ; physiology ; Female ; Glucocorticoids ; physiology ; Humans ; Male ; Middle Aged ; Osteogenesis ; Osteoporosis ; prevention & control

Objective To investigate the distribution of frequency of parathyroid hormone (PTH) gene polymorphisms in healthy adults of Han nationality in Bejing area and relationship between PTH genotypes and bone mineral density(BMD) in young and postmenopausal women. Methods Polymorphisms of PTH gene were detected by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) of restriction enzyme Bst B1 in 270 subjects. If the site of enzyme Bst B1 existed, the PTH genotype was “B”. On the contrary, if base mutation occurred, the genotype was “b”. Some of the PTH genotypes were confirmed by DNA sequences analysis. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiome try. Results Genotype frequencies of BB, Bb, bb were 73.7% , 25.9% , and 0. 4% respectively in adults of Beijing areas. The frequencies of RFLP alleles B, b were 86. 7% and 13.3%. Beijing postmenopausal women frequencies of BB, Bb, bb were 67.1%, 32.2% and 0.7%. B, b alleles frequencies were 83.2% and 16. 8%. We statistically compared bone mineral density at the lumbar 2-4, neck,wards triangle and trochanter major, there was no significant difference between BB and Bb genotype of young women and postmenopausal women groups. Otherwise, no obvious relationship was found between the BMD and PTH genotype in Beijing women. Conclusions PTH gene polymorphisms were not associateds with BMD in Beijing women.

Objective: To evaluate the clinical value of echocardiography in aortic valvuloplasty (AVP) in the low-age pediatric patients with congenital aortic valve stenosis. Methods: We retrospectively studied 39 low-age (at median of 23 months) patients with congenital aortic valve stenosis who received aortic valve repair in our hospital for their echocardiography information, and statistically analyzed the main indicator changes by 4 time points as pre-operation and 1 week, 1-3 months, 6-12 months after the operation respectively. Results: In our study, the bicuspid to tricuspid valve ratio was approximately at 5.5/1 and 2 patients died during peri-operative period. Compared with pre-operative time point, Doppler aortic valve peak velocity (Vmax ) and the mean aortic transvalvular pressure gradient (MPG) were reduced accordingly, for Vmax: (4.30 ± 0.73) m/s vs (2.65 ± 0.78) m/s, (2.93 ± 0.63) m/s, (3.01 ± 0.83) m/s,P<0.01, for MPG: (45.78 ± 15.19) mmHg vs (18.24 ± 10.08) mmHg, (21.01 ± 10.08) mmHg, (22.31 ± 13.41) mmHg. Compared with pre-operative time point, left ventricular ejection fractions (LVEF) were similar in 3 post-operative time points. Compared with 1 week post operative time point, left ventricular end-diastolic anteroposterior diameter (LVEDD) was increased at 6-12 months post-operative time point, the relative wall thickness (RWT) was decreased, bothP<0.05, and aorta valve regurgitation (AR) was increasedP<0.01. Pearson correlation analysis showed that aortic annulus (AA) inner diameter was positively related to LVEDD (r= 0.648,P<0.01), negatively related to Vmax (r= -0.205,P<0.05) and RWT was positively related to Vmax (r= 0.196,P<0.05). There were 6 patients with pre-operatively decreased LVEF, 1 of them died and the rest 5 with elevated LVEF at 6-12 months post-operative period,P<0.05. Conclusion: Echocardiograghy could be used as the ifrst choice of imaging method for diagnosing congenital aortic valve stenosis, it has the important role for in-operative monitoring and post-operative evaluation of AVP in relevant patients.

OBJECTIVETo investigate the role and significance of Wnt/beta-catenin signaling pathway regulating GSK-3beta, STAT3, Smad3 and TERT in hepatocellular carcinoma (HCC).

METHODSThe HCC cell line HepG2 was transfected with small interfering RNA (siRNA) directed against beta-catenin. Proteins were extracted and the expressions of beta-catenin, GSK-3beta, p-GSK-3beta, STAT3, Smad3 and TERT were detected by Western blot at 72 h and 96 h respectively after transfection.

RESULTSbeta-catenin expression was inhibited at both time points and the expression at 96 h was higher than that at 72 h (t = 4.43, P < 0.05). Interestingly, GSK-3beta and p-GSK-3beta expressions increased gradually at 72 and 96 h (tGSK-3beta= 4.98, tp-GSK-3beta= 29.83, P < 0.05) respectively, and STAT3 expression showed no alteration after transfection (F = 0.49, P > 0.05). Smad3 expression was increased at 72 h (t = 10.67, P < 0.05) and decreased to normal at 96 h (t = 1.26, P < 0.05), while TERT expression decreased at 72 h (t = 4.18, P is less than 0.05) and increased to normal at 96 h (t = 1.26, P > 0.05).

CONCLUSIONSWnt/beta-catenin signaling pathway is related to the expressions of GSK-3beta, Smad3 and TERT, but perhaps not related to STAT3 protein expression in HCC. It suggested that Wnt/beta-catenin signaling pathway might participate in HCC genesis and development through regulating the above three factors.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Small Interfering ; Signal Transduction ; Wnt Proteins ; metabolism ; beta Catenin ; metabolism

OBJECTIVETo compare the sonographic findings with pathological features of ovarian thecoma, and to analyze the relationship between them.

METHODSThe sonograms of 45 ovarian thecoma cases were reviewed retrospectively and categorized into three subtypes as sound attenuation pattern, homogeneous hypoechoic pattern and solid and cystic mixed pattern. The pathological findings were classified as theca cell-predominant, fibroblast-predominant and mixed thecoma according to the cellular composition of the tumors. Hyaline degeneration and luteinization of the tumors were recorded. The pathologic findings of each subtype based on sonography were compared.

RESULTSOf the 45 patients, there were 34 (75.6%) solid ovarian lesions, 15 (33.3%) of those showed a sound-attenuation pattern with an anterior hypoechoic zone and posterior acoustic attenuation in sonography, the other 19 (42.2%) cases had homogeneous hypoechoic pattern with no posterior acoustic attenuation, and the remaining 11 (24.4%) cases presented as a solid and cystic mixed pattern. There were no significant differences in pathological cellular composition among the three sonographic subtypes. Five solid tumors containing hyaline degeneration and one with luteinization were found to have posterior acoustic attenuation. The solid and cystic mixed thecomas showed cystic degeneration and hemorrhage.

CONCLUSIONSolid ovarian thecomas usually have typical sonographic features, which may be associated with degeneration but not with cellular composition within the tumor.

Adult ; Aged ; Aged, 80 and over ; CA-125 Antigen ; blood ; Female ; Humans ; Middle Aged ; Ovarian Neoplasms ; blood ; classification ; diagnostic imaging ; pathology ; Retrospective Studies ; Thecoma ; blood ; classification ; diagnostic imaging ; pathology ; Ultrasonography, Doppler, Color ; methods ; Young Adult

OBJECTIVETo study the human myxovirus resistant protein A (MxA), a specifically induced peptide by interferon I, and to use its level as a diagnostic criterion for viral infections.

METHODSAnti-MxA antisera from immunized mice were prepared with the expressed MxA protein of pET32a-MxA in E. coli BL-21(DE3). To confirm the antiserum activity and specificity, the expression product of BL21, wild type MxA pEGFP-C1-wMxA and site-directed mutant MxA pEGFP-C1-mMxA(N589S) stably transfected 3T3 cells and induced A549 cells were detected by Western blot with the antisera using non-MxA transfected or non-IFN-beta induced cells, intact A549, NIH 3T3 cells transfected with pEGFP-C1 and pET32a (+)-transformed BL-21 as controls.

RESULTSThe antisera had specific positive immunoreactivity to the NIH3T3 cells transformed with pEGFP-C1-wMxA and pEGFP-C1-mMxA, INF-beta induced A549 cells and BL21 proteins expressed with pET32a (+)-MxA. The hybridization signals from IFN-beta induced A549 cells depended on the IFN-beta inducing concentrations. Meanwhile, immunohistochemical assay showed that NIH 3T3 cells with pEGFP-C1-wMxA and pEGFP-C1-mMxA had > 98% of positive cells at 1:50 dilution of the serum and A549 cells induced by 20 ng/mL IFN-beta for 48 h showed 95% positive cells. pEGFP-C1-transfected NIH 3T3 cells were all negative.

CONCLUSIONAnti-sera are highly specific to diversified MxAs. The antibody is detectable by Western blot, immunocytochemistry and immunofluorescence assay.

Animals ; Antibody Specificity ; Cell Line, Tumor ; GTP-Binding Proteins ; genetics ; immunology ; metabolism ; Gene Expression Regulation ; Humans ; Mice ; Myxovirus Resistance Proteins ; NIH 3T3 Cells ; Species Specificity

OBJECTIVETo observe the effects of different human parathyroid hormone 1-34 (hPTH1-34) administration on SaoS-2 cells, and explore the mechanism of bone formation improvement.

METHODSEach cycle covered 48 h. SaoS-2 cells were continuously or intermittently stimulated by 50 ng/ml hPTH1-34 for 1, 3, 6, 12, and 24 h in each cycle. Total RNA was extracted by Trizol kit. Alkaline phosphatase (ALP), osteocalcin or bone Gla-containing protein (BGP) and cyclic adenosine monophosphate (cAMP) levels were measured by chemical method, radioimmunoassay and competitive protein binding method, respectively. c-fos gene expression was semi-quantified by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSALP level was time-dependently increased in 1, 3 and 6 h stimulation, especially in 3 and 6 h (compared with control, P < 0.01; P < 0.05 or P < 0.01 compared with continuous stimulation). The cAMP level was time-dependently increased in 3 and 6 h incubation (P < 0.05 compared with control and continuous stimulation). Intermittent hPTH1-34 stimulation had more effects on cAMP level than continous action (P < 0.001). hPTH1-34 intermittent stimulation of 1, 3, and 6 h enhanced c-fos gene expression time-dependently.

CONCLUSIONSIntermittent hPTH1-34 stimulation has a stronger effect on osteoblast than continuous action, especially in 3, 6 h in each cycle intermittent stimulation. The synchronous responses of c-fos, ALP and cAMP to hPTH1-34 suggest that hPTH1-34 affect Saos-2 cells through cAMP dependent protein kinase A (PKA) pathway and c-fos gene paly an important role.

Alkaline Phosphatase ; analysis ; Cells, Cultured ; Humans ; Osteoblasts ; cytology ; Osteocalcin ; analysis ; Osteogenesis ; drug effects ; Osteosarcoma ; genetics ; pathology ; Parathyroid Hormone ; pharmacology ; Parathyroid Hormone-Related Protein ; pharmacology ; Peptide Fragments ; pharmacology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo observe the changes of urinary deoxypyridinoline crosslink/creatinine (UDpd/Cr) in rats after OVX and intervention by estrogen and bisphosphonate and investigate the possible application of deoxypyridinoline in osteoporosis diagnosis and treatment.

METHODS40 female 6-month-old virginal Wistar rats were divided into 5 groups, ovariectomized or sham ovariectomized. (1) Ovxb (n = 8): sacrificed at 6 weeks after OVX; (2) Sham (n = 8): sham ovariectomized; (3) Ovxe (n = 8): sacrificed at 14 weeks after OVX; (4) O + E (n = 9):OVX + 17 beta estradiol [20 micrograms/(kg.d) ih]; (5) O + C (n = 7):OVX + cimadronate [0.2 mg/(kg.d)]; Treatment started 6 weeks after OVX and lasted 8 weeks. Rats in group 2-5 were sacrificed at 14 weeks after OVX. Urinary and serum biochemical parameters were measured, pQCT scanning of femur, bone biomechanical test in femur were determined.

RESULTSOVX resulted in increasing of UDpd/Cr 133.3% (P < 0.01). The ratio of UCa/Cr also increased in OVX groups but without any significant compared with Sham (P > 0.05). UDpd/Cr were reduced by 54.6% and 51.8% (P < 0.01) in O + E, O + C group respectively compared with Ovxe. The significant negative correlationships were found between UDpd/Cr and bone mass, BMD and biomechanic characteristics.

CONCLUSIONSUDpd/Cr ratio is a sensitive bone resorption marker, a marked changes were observed when the rats ovariectomized or treated with estradiol and cimadronate. There were best correlation between UDpd/Cr and bone mineral density and bone biomechanic characteristics. It is fair to apply UDpd/Cr ratio for osteoporosis diagnosis and treatment.

Amino Acids ; urine ; Animals ; Bone Density ; Creatinine ; urine ; Diphosphonates ; therapeutic use ; Estradiol ; therapeutic use ; Female ; Osteoporosis ; drug therapy ; urine ; Ovariectomy ; Rats ; Rats, Wistar

BACKGROUNDFibroblast growth factor 9 (FGF9), expressed in brain, kidney and developing skeletal tissues, can physiologically inhibit endochondral ossification; but little is known about how FGF9 affects osteoblasts and its detailed regulatory mechanism. Here we examined the effect of FGF9 on the activity of the murine Runt-related transcription factor 2 (Runx2) gene promoter in preosteoblast MC3T3-E1 and premyoblast C2C12 cells.

METHODSPlasmids containing the Runx2 promoter region were transfected into MC3T3-E1 and C2C12 cells and stably transfected cell lines were established. The method of luciferase reporter gene activation was used to examine the effects of FGF9 on the promoter activity.

RESULTSFGF9 (10 ng/ml) increased Runx2 promoter activity in MC3T3-E1 cells. When MC3T3-E1 cells were treated with FGF9 plus the various inhibitors or activator of the intracellular signaling transducation pathways, including 10 micromol/L U0126 (the inhibitor of mitogen-activated protein kinase kinase), 10 micromol/L SB203580 (the inhibitor of p38/mitogen activated protein kinase), or 1 micromol/L C6 ceramide (an activator of mitogen activated protein kinase), the luciferase expression did not change significantly compared with that of the cells treated with FGF9 only. However, when C2C12 cells were treated with 10 ng/ml FGF9, Runx2 gene promoter activity first decreased and then increased over a period of 1 to 5 days. Among the above inhibitors, only U0126 (10 micromol/L) completely blocked the effects of FGF9 on Runx2 gene promoter activity.

CONCLUSIONSOur data showed that FGF9 can affect Runx2 gene promoter activity in MC3T3-E1 and C2C12 cells. The action of FGF9 appears to depend partly on the mitogen-activated protein kinase kinase/mitogen-activated protein kinase pathways in C2C12 cells.

Animals ; Cells, Cultured ; Core Binding Factor Alpha 1 Subunit ; genetics ; Fibroblast Growth Factor 9 ; pharmacology ; MAP Kinase Signaling System ; Mice ; Myoblasts ; drug effects ; metabolism ; Osteoblasts ; drug effects ; metabolism ; Promoter Regions, Genetic

BACKGROUND:Bone marrow stem cell transplantation can improve heart function and prevent ventricle remodeling.At present,the adult bone marrow stem cells used for transplantation primarily included bone marrow mononuclear cells (BM-MNCs) and mesenchymal stem cells (MSCs),and endothelial progenitor cells.The curative effects and precise mechanisms of transplantation of various bone marrow stem cells remain unknown.OBJECTIVE:To compare the effects of transplantation of autologous BM-MNCs and MSCs via the coronary artery on ventricle remodeling subsequent to acute myocardial infarction (AMI). DESIGN,TIME AND SETTING:Randomized controlled animal experiment performed at the Center for Clinical Research,Hebei Provincial People's Hospital,Electron Microscope Room,Hebei Medical University between March 2005 and December 2006.MATERIALS:Thirty-six male Jizhong pigs,were randomly divided into 4 groups:control group (n = 6),infarct model group (n = 10),BM-MNC group (n = 10),and MSC group (n = 10).METHODS:Porcine autologous BM-MNCs were isolated by gradient density centrifugation,and MSCs were obtained by adherence method.Prior to transplantation,both BM-MNCs and MSCs were colloidal gold labeled.Except the infract model group,pigs in the other 3 groups were developed into AMI models by oppressing the left anterior descending branch with balloon catheter.Ninety minutes after modeling,(6.0±1.3)×107 autologous BM-MNCs and (4.5±2.1)x 107 MSCs were respectively transplanted into pigs in the BM-MNC group and the MSC group via the coronary artery and cultured for 28 days.MAIN OUTCOME MEASURES:Observation of pathological changes of cardiac muscle tissue by light and electron microscope;Examination of cardiac function by ultrasonograph;Detection of the number of blood vessels and apoptotic myocardial cells,and expression of nuclear factor-κB (NF-κB) and troponin Ⅰ and its correlation to cardiac function by immunohistochemistry;Detection of mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in the cardiac tissue as well as its correlation to cardiac function by reverse transcription-polymerase chain reaction (RT-PCR).RESULTS:In the MSC group,there was proliferation of a great deal of blood vessels as well as growth of abnormal cell masses around the coronary vessels,while the BM-MNC group exhibited the "budding" of many capillary vessels.Prior to transplantation,cardiac function indices were basically similar among each group (F = 1.550,P＞0.05).Twenty-eight days after transplantation,left ventricular ejection fraction was significantly lower in the control,BM-MNC,and MSC groups than in the infarct model group (F = 5.30,P＜0.05),while endocardial fractional shortening was significantly higher (F = 10.67,P＜0.01).Compared with the infarct model group,the number of blood vessels in the infarct zone and infarct border zone was increased in the BM-MNC group (F=29.56-34.87,P＜0.01) and had no apparent change in the MSC group.In the BM-MNC and MSC groups,apoptotic myocardial cells in the infarct zone and infarct border zone were significantly reduced (F=14.31-35.34,P＜0.01 ) and troponin I expression rate was significantly increased (F=19.05,P＜0.01 ),as compared with the infarct model group.In addition,NF-κB positive rate in the infarct border zone was significantly lower in the BM-MNC and MSC groups than in the infarct model group (F=19.05,P＜0.01).VEGF gene expression level in the infarct border zone was significandy higher in the BM-MNC group than in the infarct model group and MSC group (F = 49.41,P＜0.01).bFGF gene expression level in the infarct border zone was significantly higher in the MSC group than in the infarct model and BM-MNC groups (F=4.71,P＜0.01).LVEF was negatively correlated to myocardial cell apoptosis rate and NF-κB level (r=-0.441 1,P＜0.05;r=-0.579 6,P＜0.01 ).LVEF was positively correlated to number of blood vessels,VEGF and bFGF expression (r=0.775,P＜0.01;r=0.565 1,P＜0.05;r=0.573 5,P＜0.05).CONCLUSION:Transplantation of both autologous BM-MNC and MSC via coronary artery can improve the condition of left ventricular remodeling subsequent to myocardial infarction.The improvement of cardiac functions is related to the increase of blood vessels,VEGF and bFGF expression,the decrease of myocardial cell apoptosis and NF-κ B level in cardiac muscle tissues after stem cell transplantation.BM-MNC transplantation better promotes blood vessel proliferation and VEGF expression in the cardiac tissue but produces worse effects on bFGF gene expression than MSC transplantation.