Objective To investigate the effects of 1,25(OH)_2D_3 on cell proliferation and cell cycle progression in mouse osteoblasts.Methods Sterile bones of skull of mouse were taken from 30 newborn mouse,and the osteoblast were separated by enzyme digestion methods.After 1,25(OH)_2D_3 in different concentrations were added into culture medium,the effects of 1,25(OH)_2D_3 on cell proliferation of mouse osteoblasts and on cell cycle progression were examined by mono-nuclear celldirect cytotoxicity assay(MTT)reduction assay and flow cytometry respectively.Results After 24,48,72 h of 1,25(OH)_2D_3 incubation,the cell number of osteoblast had significant difference among groups of 1,25(OH)_2D_3 of 10~(-8),10~(-9),10~(-11)mol/L.Significant differences were found in the cell cycle progression in response to 1,25(OH)_2D_3 treatment from the Gl(84.30?1.90)to the G2-M (7.70?0.667)and S(8.00?1.42)phases when compared with those in the control group. Conclusions Cell proliferation of mouse osteoblasts can be inhibited by 1,25(OH)_2D_3 in a concentration-dependent manner.

OBJECTIVETo investigate the collagen constitution of hyperplastic scar (HS) in different ages and the change of relative factors.

METHODSThirty cases with HS were divided into two groups according to patients' age: group 1 (1 - 19 years, A) and group 2 (20 - 50 years, B). The normal skin (NS) from corresponding age of volunteers was employed as control group. The changes in TGFbeta1, collagenase (MMP-1) and tissue inhibitor of metalloproteinase (TIMP-1beta) and the collagen ratio were observed by means of in situ hybridization technique and SABC (Strept-Avidin-Biotin complex) immunohistochemistry and image analysis.

RESULTSThe ratio of type I to type III collagen in A group was 6.48 in average and 3.76 in B group, but there was no evident difference in the ratio during the disease process in both groups. The expression of TGFbeta1 in A group was much higher than that in B group (P < 0.01). The TIMP-1 mRNA expression showed no difference among all age groups in HS patients, but it was much higher than that in NS group. The MMP-1 expression was evidently lower than TIMP-1 expression, and there was no difference in MMP-1 expression compared with NS group.

CONCLUSION(1) The TGFbeta1 expression in HS patients was negatively correlated with age, and the increased expression of TGFbeta1 produced an increase ratio of type I to type III collagen. (2) High level expression of TIMP-1 led to the formation of HS by inhibiting MMP-1 expression, and the expression was not related to age.

Adolescent ; Adult ; Age Factors ; Burns ; metabolism ; pathology ; Child ; Child, Preschool ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagen Type I ; biosynthesis ; Collagen Type III ; biosynthesis ; Female ; Humans ; Infant ; Male ; Matrix Metalloproteinase 1 ; biosynthesis ; Middle Aged ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis ; Transforming Growth Factor beta1 ; biosynthesis ; Young Adult

Objective To determine the correlation of cardiovascular MRI with cardiac biomarkers and electrocardiography(ECG)in acute myocardial infarction(MI).Methods Nineteen patients with first acute MI were selected to undergo MRI on a 1.5 T system within 3-7 days after the onset of symptoms.A first-pass perfusion scan was performed with the administration of Gd-DTPA at a speed after cine MRI for global left ventricle(LV)funotions.Delay-enhanced MRI was performed by using an ECG-gated inversionrecovery fast-gradient echo-pulse sequence 5 to 10 minutes later with second bolus injection at a s peed.Infarct mass(IM),percentage size of infarction(PSI)and LV functions were compared with peak troponin T (peak TnT)and peak ereatine kinase-MB fraction (peak CK-MB).The 12-lead ECG was analysed for STelevation on admission.Pearson and Spearman correlation test and independent-Sample t test wel-e used for statistics.Results The IM (median 6.3 g) was correlated with peak TnT(median 0.8μg/L,r=0.487,P=0.0340)and left ventricle end-systolic volume index(LVESVI)(median 23.4 ml/m2,r=0.480,P=0.038).IM showed a negative correlation with left ventricle ejection fraction(LVEF)(54.1±15.4)/(r=-0.563.P:0.012).The PSI(median 6.0/)was correlated with peak TnT(r=0.583,P=0.009),peak CK.Mn(median 43.0 U/L,r=0.470,P=0.042)and LVSV[(57.6 ±15.0)ml,r=-0.482,P=0.036],peak TnT was also correlated with LVSV(r=-0.524,P=0.021).There were more involved segments(IS)(t=2.972,P=0.009),higher peak TnT(t=2.245,P=0.041)and peak CK-MB(t=2.508,P=0.024)in ST-elevation MI(STEMI)than in non ST-elevation MI(NSTEMI).Conclusions IM directly influences LV functions in acute MI.Peak TnT was a better biomarker reflecting PSI and LV functions.There were more involved segments in STEMI than in NSTEMI.

The effect of lysophosphatidic acid (LPA), with a wide range of its different concentrations, upon cultured mouse cortical neurons was assessed by electrophoresis of DNA fragments, HO33342 and TUNEL stainings, and also by ultrastructural examination at times. The results showed that administration of LPA at lower concentrations (0.1-30 micromol/L) dose-dependently protected cortical neurons from apoptosis that was induced by deprivation of serum from the cultural medium, while 50 micromol/L or higher concentrations of LPA failed to show this effect; and moreover, the concentrations higher than 50 micromol/L induced apoptosis in neurons cultured in serum-containing complete medium. These results suggest that a moderate concentration of LPA may play as a survival factor in apoptotic cortical neurons, while an excessive level of LPA induces apoptosis in neurons cultured in complete medium.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Cerebral Cortex ; cytology ; Culture Media, Serum-Free ; Lysophospholipids ; pharmacology ; Mice ; Neurons ; cytology

It has been reported that lysophosphatidic acid (LPA) at its lower concentrations prevents apoptosis induced by serum-deprivation in cultured cortical neurons when LPA is added into the cultural medium with serum withdrawal. The present study was designed to investigate whether LPA could also block the apoptosis induced by beta-amyloid peptide fragment 31-35 (AbetaP31-35) in cultured cortical neurons by using techniques of DNA fragmentation electrophoresis, HO33342 staining, and TUNEL examinations. The results showed that pretreatment of LPA suppressed the AbetaP31-35-induced apoptosis only when LPA was applied to the cultured neurons with lower concentrations (1-10 micromol/L) and especially, with a preceding time of 12-24 h before the AbetaP31-35 exposure. These facts imply that LPA also acts as a neuroprotective factor against AbetaP31-35-induced apoptosis, though the mechanism underlying the protective action in this case may be more complex than that involved in the serum deprivation-induced apoptosis.
Amyloid beta-Peptides ; antagonists & inhibitors ; Animals ; Animals, Newborn ; Apoptosis ; drug effects ; physiology ; Cells, Cultured ; Cerebral Cortex ; pathology ; Lysophospholipids ; pharmacology ; Mice ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; Peptide Fragments ; antagonists & inhibitors

Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.

OBJECTIVETo study the role of apelin in the prevention of pulmonary hypertension induced by hypoxia in rats.

METHODSThe animal model of hypoxic pulmonary hypertension was established by exposing the rats to isobaric hypoxic chamber for 4 weeks (8 h/d, 6 d/ w). Forty male Sprague-Dawley rats were randomly divided into control group (NC), hypoxic group(HH), hypoxic with low-dose apelin (5 nmol/(kg x d) group(LA) and high-dose apelin (10 nmol/(kg x d) (HA). [pGlu]apelin-13 was administered into the rats of apelin groups by mini-osmotic pump subcutaneously. The mean pulmonary arterial pressure(mPAP) and the mean carotid arterial pressure (mCAP) were measured by either right or left cardiac catheterization, and the weight ratio of right ventricule/left ventricule plus septum (RV/(LV + S)) were calculated. The Masson's trichrome stained lung specimens were examined by light microscope to examine the vessel wall area/total area (WA/TA), vessel cavity area/total area (CA/TA) and media thickness of pulmonary arterioles (PAMT). Meanwhile, the lung homogenates were assayed for the activity of supeeroxide dismutase (SOD) and the content of malondialdehyde (MDA).

RESULTS(1) mPAP and RV/(LV + S) of HH group were significantly higher than those of NC group. mPAP of LA and HA groups were lower than those of HH group. The RV/(LV + S) of HA group was significantly lower than that of HH group, but there was no significant difference between HH group and LA group. (2) Masson's trichrome staining revealed that WA/TA and PAMT of HH group were higher than those of NC group. Administration of apelin significantly eliminated WA/TA and PAMT in LA and HA groups. (3) CA/TA of HH group was lower than that of NC group. Administration of apelin significantly elevated CA/TA in LA and HA groups. (4) The activity of SOD and content of MDA in HH group was, respectively, lower and higher than those in NC group. Apelin treatment increased the activity of SOD in LA and HA groups while decreased the content of MDA.

CONCLUSIONSApelin could play an important role in treatment of hypoxic pulmonary hypertension of rats and the mechanisms of protection were associated with vasodilation of pulmonary artery and inhibition of oxidative stress.

Animals ; Cardiotonic Agents ; pharmacology ; therapeutic use ; Hypertension, Pulmonary ; etiology ; physiopathology ; prevention & control ; Hypoxia ; complications ; physiopathology ; Intercellular Signaling Peptides and Proteins ; pharmacology ; therapeutic use ; Male ; Oxidative Stress ; drug effects ; Pulmonary Artery ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Vasodilation ; drug effects

OBJECTIVETo explore the methodology as well as the features, quantificational index, and reference standard of 3.0 Tesla magnetic resonance (MR) diffusion-weighted imaging (DWI) on the normal rabbit's liver.

METHODSTwenty New Zealand white rabbits were enrolled and DWI was performed after anesthetics with multi-b values at 3.0 T MR scanner. Apparent diffusion coefficient (ADC) values as well as the difference between maximum and minimum ADC values, signal strength (SH), noise signal (SD), signal to noise ratio (SNR), and quality index (QI) were recorded and analyzed.

RESULTSWith b value increased, the ADC values decreased accordingly (P < 0.001). The difference between maximum and minimum ADC values with b = 1000 s/mm2 was the least (good stability), b = 600 s/mm2 was the second least, and b = 300 s/mm2 was greatest (bad stability). The SH decreased at the same time (P < 0.001), but the difference among DWI with b =600, 800, and 1000 s/mm2 was not statistically significant (P > 0.05). The SD decreased at the same time (P < 0.001), but the difference between DWI with b = 800 s/mm2 and b = 1000 s/mm2 was not statistically significant (P > 0.05). The SNR decreased at the same time (P < 0.001), but there were no significant differences between DWI with b = 600 s/mm2 and b = 800 s/mm2 or b = 600 s/mm2 and b = 1000 s/mm2 (P > 0.05). The SNR of DWI with b = 800 s/mm2 and b = 1000 s/mm2 was lower. The QI decreased at the same time (P < 0. 001) , but the difference between DWI with b = 800 s/mm2 and b = 1000 s/mm was not statistically significant (P > 0.05).

CONCLUSIONWhen 3.0 T MR DWI is applied for rabbit liver, it is better to use b = 600 s/mm2 for reducing scanning time and assuring better diffusion weights, quantity of images, and stability of ADC measurement.

Animals ; Diffusion Magnetic Resonance Imaging ; methods ; Liver ; Male ; Rabbits

OBJECTIVETo investigate whether S. aureus could activate NF-κB signaling pathway in human osteoblasts.

METHODSImmunoblot and electrophoretic mobility shift assay were used to detect the degradation of I-κBα and activation of NF-κB in human osteoblasts following infection with S.aureus, respectively, and there were investigated the activated state of NF-κB signaling pathway in human osteoblasts. In addition, enzyme-linked immunosorbent assay was used to measure the secretion of IL-6 in culture supernatants, which was represented as one of important cytokines in osteomyelitis, and an inhibitor of NF-κB, SN50, which was added to human osteoblasts culture prior to 1 hour at 50 µmol/L before the infection of S.aureus, was used to determine whether S.aureus-activated NF-κB signaling pathway regulates IL-6 secretion of human osteoblasts.

RESULTSS.aureus could induce the degradation of I-κBα (I-κBα(15 min)/I-κBα(0 min) = 0.409 ± 0.245 and I-κBα(30 min)/I-κBα(0 min) = 0.061 ± 0.010) and activation of NF-κB in human osteoblasts in a time and dose-dependent manner following infection. In addition, the secretion of IL-6 in the supernatants of human osteoblasts ((2.17 ± 0.11) µg/L) was suppressed by 50 µmol/L SN50 compared to without the addition of SN50 ((3.58 ± 0.31) µg/L) (F = 174.25, P < 0.05).

CONCLUSIONSS.aureus could activate NF-κB signaling pathway in human osteoblasts, which could regulate cytokines secretions of human osteoblasts.

Cells, Cultured ; Humans ; Interleukin-6 ; secretion ; NF-kappa B ; metabolism ; Osteoblasts ; metabolism ; Signal Transduction ; Staphylococcal Infections ; metabolism

OBJECTIVETo evaluate the effect of intrajejunal nutrition on uptake of amino acid and enzyme-protein synthesis in pancreatic acinar cell and subcellular fractionation and zymogen granules in dogs with acute pancreatitis.

METHODSFifteen dogs were induced acute pancreatitis by retrograde injection of 5% sodium-taurocholate into the pancreatic duct. Radioactive tracing and electron microscope were used to evaluate the change of amino acid uptake, enzyme-protein synthesis in acinar cell, subcellular fractionation, the quantitative analysis of mean zymogen granule number and mean zymogen granule area after injection L-(3)H-phenylalanine 30, 60, 120 1nd 180 min on the 7(th) day.

RESULTSThe radioactivity of L-(3)H phenylalanine uptake by pancreatic acinar cells and incorporations of L-(3)H phenylalanine into newly synthesized enzyme-protein peaked at 60 min. In enteral nutrition (EN) group it was higher that that in parenteral nutrition (PN) group (P < 0.05), and then gradually declined. The radioactivity peaked at 60 min in zymogen granule, lysosomal-mitochondria and microsomal subcellular fractionation. The latter two decreased, bat there was no significant difference (P > 0.05). The change of the mean number and mean area of zymogen granules were not significant different between the EN group and PN group (P > 0.05).

CONCLUSIONEN or PN do not stimulate pancreatic acinar uptake amino acid and enzyme-protein synthesis in acinar cell and subcellular fractionation.

Acute Disease ; Amino Acids ; metabolism ; Animals ; Disease Models, Animal ; Dogs ; Enteral Nutrition ; Enzyme Precursors ; biosynthesis ; Female ; Male ; Pancreas, Exocrine ; metabolism ; Pancreatitis ; pathology ; physiopathology ; therapy ; Parenteral Nutrition ; Random Allocation ; Treatment Outcome