Aortic valve calcification (AVC) is a pathological process correlated with multiple disease causes and actively regulated by cardiac valve cells. In this study, porcine aortic valve myofibroblasts cultured in vitro were treated with 50 μg z L(-1) of pathological factor tumor necrosis factor α (TNF-α). Tanshinone II A (TSN) with the concentration of 50 mg x L(-1) and TNF-α were combined in incubating cells for 72 h (3 d) and 120 h (5 d). The Western blotting and Real-time PCR were adopted to detect the changes in smooth muscle α actin (α-SMA), bone morphogenetic protein 2 ( BMP2), alkaline phosphatase (ALP) in cells, and expressions of key effect proteins GSK-3β and β-catenin on Wnt/β-catenin signal pathway. According to the findings, TNF-α can significantly increase the expression of myofibroblasts α-SMA and add the transformation activity to them, with nearly no expression of BMP2, ALP and mRNA in the control group and the TSN group but significant increase in their expressions in the TNF-α group (P < 0.01), which showed osteoblast-like phenotype. Moreover, TNF-α down-regulated the expression of up-streaming regulator GSK-3β and mRNA expression (P < 0. 01) , notably increased the expression of key effect protein β-catenin, but with no significant difference in mRNA with the control group and the TSN group. The result demonstrated that TSN showed a certain inhibitory effect on TNF-α's pathological impact (P < 0.05) in a time-dependent manner. Inflammatory factor TNF-α may promote the transformation of aortic valvular myofibroblasts to osteoblast-like phenotype by activating Wnt/β-catenin signal pathway in aortic valvular myofibroblasts, so as to cause AVC. Tanshinone II A can have a preventive effect in AVC by activating GSK-3β proteins and regulating signal transduction of Wnt/β-catenin signal pathway.
Animals ; Aortic Valve ; cytology ; drug effects ; metabolism ; Cells, Cultured ; Diterpenes, Abietane ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Glycogen Synthase Kinase 3 ; genetics ; metabolism ; Glycogen Synthase Kinase 3 beta ; Myofibroblasts ; cytology ; drug effects ; metabolism ; Osteoblasts ; cytology ; drug effects ; metabolism ; Swine ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; beta Catenin ; genetics ; metabolism

Objectivc To observe the short-term changes of anterior corneal biometry after discontinuation of orthokeratology in patients with 2-year wearing.Methods Retrospective study.Sixty myopic patients aged from 8-14 years old during October 2012 and October 2014 were wearing orthokeratology for 2 years in Tianjin Medical University Eye Hospital.According to the degree of myopia,they are divided into three groups(SE≤-2.00 D for group A,-2.00 D ＜ SE ≤-4.00 D for group B and -4.00 D ＜ SE ≤-6.00 D for group C).The recovery of anterior corneal curvature,including flat K(FK),steep K(SK),average K (AVEK),changes of axial length and central corneal thickness (CCT) at 1 week,2 weeks,1 month after discontinuation of orthokeratology were observed.Results There was no statistical differences in FK,SK,AVEK before and 2 weeks after wearing orthokeratology in group A (all P ＞ 0.05).While in group B,there was no significant difference in FK,SK,AVEK before and 1 month after wearing orthokeratology;There were statistically differences in FK,SK,AVEK at 1 month after discontinuation in group C compared with the baseline (all P ＜0.05).As for CCT,there was no statistical differences among group A,B,C after discontinuation of orthokeratology for 2 weeks (all P ＞ 0.05).There were statistical differences in the axial length between 1 week and 2 weeks after discontinuation of orthokeratology in group B and C (all P ＜ 0.05);There were statistical differences in the axial length between 1 month and 2 weeks after discontinuation of orthokeratology in three groups (all P ＜ 0.05);Compared with the state before wearing orthokeratology,the increase of axial length in group A,B,C were (0.43 ± 0.36) mm,(0.35 ± 0.21)mm and (0.36 ± 0.29) ram,respectively.Conclusion The time course of returning to the original corneal parameter varies among different degree of myopia,and the axial length has no significant growth after short-term discontinuation.

NOD mice were treated intraperitoneally with tripterygium wilfordii ployglycosidium (TWP) for 4 weeks to observe the incidence of cyclophosphamide accelerated diabetes.The apoptosis of?-cells was detected by TUNEL,the expression of caspase-3 in islets of the NOD mice was detected by immunohistochemistry and the expression of caspase-8 mRNA in pancreas by RT-PCR.The results revealed that the incidence of diabetes in TWP group was lower than that in control group.The apoptosis index of?-cells was decreased in TWP group. The expression of caspase-3 in islets and the expression of caspase-8 mRNA in pancreas in TWP group were lower than those in control group.

Adult ; Aged ; Aged, 80 and over ; Epistaxis ; therapy ; Female ; Gloves, Protective ; Hemostatic Techniques ; Humans ; Male ; Middle Aged

OBJECTIVETo compare the clinical effects of the circumcision stapler, circumcision cerclage, and traditional circumcision in the treatment of phimosis and redundant prepuce.

METHODSUsing the circumcision stapler (group A), foreskin cerclage (group B), and traditional circumcision (group C), we treated 276 patients with phimosis or redundant prepuce. We made comparisons among the three groups in the operation time, intraoperative blood loss, intraoperative and 24-hour postoperative pain scores, and incidence of postoperative complications. Results: The operation time, intraoperative blood loss, and intraoperative pain score were (6.52 ± 2.45) min, (1.93 ± 0.82) ml, and 1.37 ± 0.68 in group A and (7.24 ± 1.86) min, (1.51 ± 0.72) ml, and 1.20 ± 0.79 in group B, all significantly lower than (28. 36 ± 4.22) min, (9.52 ± 3.29) ml, and 3.06 ± 0.75 in group C (P <0.05). The 24-hour postoperative pain score was remarkably higher in group B than in A and C (3. 18 ± 0. 82 vs 1. 85 ± 0. 63 and 1. 82 ± 0. 75, P <0. 05). The incidence rate of postoperative complications was markedly lower in group A than in B (5. 43% vs 14. 13%, P < 0.05), but with no significant differences between either A and C or B and C (P >0.05).

CONCLUSIONThe circumcision stapler, with its advantages of simple operation, minimal invasiveness, fewer complications, and better cosmetic result, deserves a wider clinical application.

Blood Loss, Surgical ; Circumcision, Male ; adverse effects ; instrumentation ; methods ; Foreskin ; Humans ; Incidence ; Male ; Pain Measurement ; Pain, Postoperative ; diagnosis ; Penis ; abnormalities ; Phimosis ; therapy ; Postoperative Complications ; Postoperative Period

OBJECTIVETo investigate the effect of adenovirus expressing human bone morphogenetic protein-7 (hBMP-7) on proliferation and differentiation of human dental pulp cells.

METHODSThe replication-deficient adenoviral vector encoding hBMP-7 gene was constructed by using homologous recombinant modality. The efficiency of transfection was evaluated by fluorescent microscopy and flow cytometry. The expression of hBMP-7 protein in adenovirus-infected dental pulp cells was determined by Western blot. The proliferation of cells was tested by MTT method, the activity of alkaline phosphatase was assayed, von Kossa staining was used to detect mineralized nodule formation, and the expression of DSPPmRNA in cells was detected using semi-quantitative RT-PCR.

RESULTSGreen fluorescent protein was visible under fluorescent microscopy. Higher transfection efficiency (91.1 +/- 1.0)% could be obtained at MOI of 75. Western blot from dental pulp cells infected with Ad-hBMP-7 for 48h detected protein expression of a hBMP-7 gene. The activity of alkaline phosphatase in cells was significantly higher than those of the control groups (P < 0.05). The cells infected with Ad-hBMP-7 had the ability of mineralization. DSPP mRNA expression of cells was in a time- and dose- dependent manner.

CONCLUSIONSAd-hBMP-7 can induce human pulp cells into odontoblasts, but has no obvious effect on their proliferation.

Adenoviridae ; genetics ; Bone Morphogenetic Protein 7 ; genetics ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Dental Pulp ; cytology ; Humans ; Odontoblasts ; cytology ; Transfection

UNLABELLEDTo study the infected root of Panax quinquefolium on the contents of ginsenosides.

METHODThe contents of three major ginsenosides Rg1, Re and Rb1 were determined by HPLC compared quantitatively between the different degree infected roots and normal root in the phloem and xylem.

RESULTRg1 in phloem and xylem of varying degrees infected root showed no significant difference, but Rb1 decreased 26.3% and 28.3% respectively in medium and serious infected roots comparing to normal root. Re in phloem of seriously infected roots decreased in xylem significantly.

CONCLUSIONThe results indicate that the variation of ginsenosides in different degrees infected roots exists and the proportion of Rg1, Re and Rb1 in the total ginsenosides changes.

Ginsenosides ; chemistry ; Panax ; chemistry ; microbiology ; Plant Diseases ; microbiology ; Plant Roots ; chemistry ; microbiology

AIMTo elucidate the effect of CIHH on cellular immunity and humoral immunity in rat by using flow cytometry method, immunohistochemistry method and electron microscopy techniques.

METHODSForty-eight male adult Sprague-Dawley rats were randomly divided into 4 groups: control(CON) group, 14 days CIHH (CIHH14) group, 28 days CIHH (CIHH28) group, 42 days CIHH (CIHH42) group. The animals in CIHH groups were exposed to 14, 28 and 42 days hypobaric hypoxia(simulated 3 000 m altitude, 5 h per day), respectively. Half of the animals in each group was treated with normaxia and the other half animals were treated with acute hypoxia for 1 h. CD3, CD4, CD8 T lymphocytes, natural killer (NK) cells, IgG, cortisol, epirenamine and C-reactive protein were examined. The weight and ultrastructure of thymus and spleen were observed.

RESULTS(1) Compared with CON, both indexes of thymus and spleen in CIHH14 rats were increased significantly. Spleen index, but not thymus index, was increased in CIHH28 and CIHH42 rats. The thymocytes and spleen cytes in rat were injuryed during acute hypoxia, but the damage in CIHH rats was significant slighter than that in CON rats. (2) Compared with CON, CIHH28 and CIHH42, CD8 in CIHH14 rats were decreased, ratios of CD4/CD8 was increased and NK was decreased. (3) The rats of CON during acute hypoxia showed that CD4 was increased, CD8 was decreased, ratio of CD4/CD8 was elevated, and NK was increased. But there were no significant changes of CD3, CD4, CD8 and NK in CIHH28 and CIHH42 animals during acute hypoxia. (4) Compared with CON, CIHH28 and CIHH42, cortisol in CIHH14 rats was increased obviously, Epirenamine, cortisol and C-reactive protein in CON rats were increased, but there were no obvious changes in CIHH rats before and after acute hypoxia.

CONCLUSIONCIHH protects the immune function of rat against acute hypoxia, which is related with the regulation of neuroendocrine.

Altitude Sickness ; physiopathology ; Animals ; Atmospheric Pressure ; Hypoxia ; physiopathology ; Immunity, Cellular ; physiology ; Immunity, Humoral ; physiology ; Male ; Neuroimmunomodulation ; physiology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spleen ; immunology ; T-Lymphocytes ; immunology ; Thymus Gland ; immunology

OBJECTIVETo investigate the interaction between p38 mitogen-activated protein kinase signal transduction pathway and nuclear factor (NF)-kappaB/IkappaB system on the proinflammatory cytokines release after burn trauma.

METHODSHuman monocyte line THP-1 were incubated with serum from eight healthy controls, burn sera, burn sera pretreatment with SB203580, and burn sera pretreatment with pyrrolidine dithiocarbamate (PDTC). After 24 hours incubation with serum, tumor necrosis factor (TNF)-alpha and interleukin-1beta (IL-1beta) levels in THP-1 culture supernatants were measured by ELISA. The activities of p38 MAPK and expressions of IkappaBalpha in THP-1 were measured by Western blot analysis. The EMSA method was used to characterize the binding activities of NF-kappaB and activating protein (AP)-1 in THP-1.

RESULTSIn comparison with normal controls, burn sera resulted in a significant higher level release of TNF-alpha and IL-1beta in THP-1 [(7.30 +/- 0.84) ng/ml vs (2.20 +/- 0.28) ng/ml, P < 0.05; (2.88 +/- 0.38) ng/ml vs (0.81 +/- 0.14) ng/ml, P < 0.05], which were significantly inhibited by pretreatment with SB203580 or PDTC. Burn sera showed increased activities of p38 MAPK and AP-1 in THP-1 (4728 +/- 582 vs 1291 +/- 163, P < 0.05; 946 +/- 137 vs 361 +/- 40, P < 0.05), which were abolished by pretreatment with SB203580 but not PDTC. The expression of IkappaBalpha in THP-1 incubated with burn sera was significantly decreased than those incubated with control sera (1211 +/- 115 vs 2658 +/- 318, P < 0.05), which were abolished by pretreatment with PDTC but not SB203580. Burn sera also leaded to an increased activity of NF-kappaB in THP-1 (1636 +/- 170 vs 317 +/- 32, P < 0.05), which were abolished by pretreatment with PDTC but not SB203580.

CONCLUSIONSThere are no direct interaction between p38 MAPK signal transduction pathway and NF-kappaB/IkappaB pathway. These two pathways, which regulate the production of TNF-alpha and IL-1beta in monocyte following burn trauma, are parallel and independent.

Adolescent ; Adult ; Burns ; immunology ; physiopathology ; Female ; Humans ; I-kappa B Proteins ; physiology ; Immune Sera ; pharmacology ; In Vitro Techniques ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Monocytes ; drug effects ; physiology ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; physiology ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism ; physiology

OBJECTIVETo investigate the effect of cadmium (Cd) on estrogen receptor and to assess its endocrine disrupting action.

METHODSThe estrogen receptor rich supernatant was prepared from the ovariectomized Sprague-Dawley rats. The effects of cadmium on estrogen binding were performed using a sing-dose ligand-binding assay. Extract from uterus were treated with various concentrations of cadmium (0, 10(-3), 10(-5) or 10(-7) mol/L) for various pre-incubation time (0, 30, 60, 90 min) by means of orthogonal experimental design with orthogonal layout of L16(4(5)) (the experiment was repeated for 5 times). In addition to the radioinert competitor, each assay included a zero tube and a DES standard curve for quality control purpose. Data for cadmium and the DES standard curve were plotted as percent [3H]-E2 bound versus log (molar concentration), and the IC50 for cadmium was determined. The RBA for cadmium was calculated by dividing the IC50 of DES in terms of the IC50 of cadmium.

RESULTSCadmium could not block the binding of estradiol to the receptor because hormone binding did not change with increasing cadmium concentration or increasing preincubation time. The results showed that the binding of [3H]-estradiol to uterine cytosols was not significant (P > 0.05). The Bmax (its unit is pmol/mg protein) of various concentrations of cadmium (0, 10(-3), 10(-5) or 10(-7) mol/L) for pre-incubating 0 min is 203.15 +/- 75.16, 203.41 +/- 22.78, 220.82 +/- 45.35, 209.10 +/- 49.66 respectively; The Bmax of them for pre-incubating 30 min is 215.67 +/- 92.97, 139.79 +/- 53.78, 205.27 +/- 23.60, 172.63 +/- 55.09 respectively. The Bmax of them for pre-incubating 60 min is 197.11 +/- 50.68, 203.24 +/- 66.33, 183.92 +/- 31.89, 183.33 +/- 32.70, respectively. The Bmax of them for pre-incubating 90 min is 229.69 +/- 76.88, 175.70 +/- 70.28, 164.26 +/- 24.46, 150.78 +/- 65.97 respectively. Mean IC50 for cadmium is 10(-4) - 10(-3) M. If the affinities of DES binding to estrogen receptors was taken to be 100%, the relative binding affinities of cadmium was 10(-6) - 10(-7). The results indicated that cadmium had only a very poor affinity with estrogen receptor.

CONCLUSIONIn vitro assay cadmium did not have distinct disrupting effect on binding of estradiol to estrogen receptors from rat uterine.

Animals ; Cadmium ; toxicity ; Female ; In Vitro Techniques ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; drug effects ; metabolism ; Uterus ; drug effects ; metabolism