Objective In this study, we explored the mechanism of anti-bone resorption of RGD-insulin using osteoclast-like cells (OLCs) from giant cell tumor of bone as an in vitro model. Methods The function of bone resorption was observed and the staining for tartrate-resistant acid phosphatase (TRAPase) was identified. The carbonic anhydrase Ⅱ (CAⅡ) gene expression, apoptosis and the adhesion of OLC that treated by RGD-insulin and echistatin were determined by in situ hybridization, TUNEL staining and adhesion test, respectively. Results OLCs were TRAP positive and had bone resorptive function. After OLCs were treated with 10 -5, 10 -6, 10 -7mol/L RGD-insulin and echistatin respectively, the numbers of TRAP-positive multinucleated cells were (179?3), (238?11), (351?10) per well that treated with echistatin and (229?20), (222?21), (265?17) per well that treat with RGD-insulin.The expression of CAⅡ mRNA decreased, the number of OLCs apoptosis increased in a dose-dependent manner (P

Objective To perform comparative proteomic analysis of human ovarian cancer cell lines for detecting platinum-resistance associated proteins.Methods The total proteins of two sensitive (SKOV3 and A2780)and four resistant(SKOV3/CDDP,SKOV3/CBP,A2780/CDDP and A2780/CBP) human ovarian cancer cell lines were separated by two-dimensional gel electrophoresis(2-DE).The differentially expressed proteins were analyzed using image analysis software,stained with Coomassie Brilliant Blue,then identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS)and database searching.The mRNA and protein levels of the differentially expressed protein which was most significant in all of the four resistant cell lines were validated by RT-PCR and western blotting,respectively.Results Five proteins were found to be significant in four cell lines. Annexin A3 and destrin were up-regulated and nicotinamide-adenine dinucleotide phosphate(NADP)- dependent isocitrate dehydrogenase 1 was down-regulated in all the four resistant samples.Glutathione transferase omega 1 had an increased expression in the other three resistant cell lines except for SKOV3/CBP in which its expression was not changed.However,cofilin 1 represented a different trend.In the two resistant sublines of SKOV3,eofilin 1 had a down-regulation,but it had an up-regulation in the cell lines induced from SKOV3.The expression of annexin A3 was up-regulated by 3-20 fold and the results of RT- PCR and western blotting showed complete consistency with that by 2-DE.Conclusions Proteomic techniques are useful to the identification of the resistance-associated proteins in ovarian cancer platinum- resistant cell lines and five candidates have been found.The five differential proteins might become hopeful candidate biomarkers for resistance.

Objective To investgate the changes of bone mineral density (BMD) and biochemical markers of bone metabolism in elderly male patients with osteoporosis (OP) . Methods BMD and bone mineral content (BMC) of the lumbar spines (L 2～4 ) and the biochemical markers of bone metabolism in serum and urine were measured. Results BMD and BMC in OP group was reduced by 21.6% and 25% respectively vs that in control group. Bone formation indicators of serum levels of ALP and BGP in OP group were increased by 25.4% and 222% respectively vs that in control. At the same time, bone resorption indicators of urine Hop/Cr and NTX/Cr ratios were significantly increased by 22.6% and 223% in OP group than that in control. Serum T level of OP group was lower than that in control group and serum 25 OH D3 was below the normal value in both groups. Conclusions BMD and BMC of the lumbar spines (L 2 4 ) are the main basis for diagnosing the osteoporosis in men. Some elder male patients with osteoporosis are due to high turnover of bone metabolism. Androgen plays an important part in maintaining the bone mass. Vitamin D deficiency in elder men is the main reason occurring osteoporosis.

Objective To explore the mechanism of cerebral infarction caused by hyperhomocystinemia.Methods A hundred and nineteen paitents with acute cerebral infarction were chosen for case group.According to their levels of plasm total homocystine,they were divided into two groups: hyperhomocystinemia group and nonhyperhomocystinemia group.Forty patents without cerebrovascular disease,hepatophy,nephrosis and thyroid gland disease were chosen as control subjects.Plasm levels of total homocystine,serum levels of MDA and IL-8 were measured respectively,their correlations were also studied.Results Plasma levels of tHcy(?mol/L)and serum levels of MDA(nmol/L)and IL-8 (ng/ml)showed a significant increase in case group(19.97,4.41?0.84,0.23?0.08)in comparison with control subjects(9.83,3.24?0.64,0.12?0.08),t values were 8.139,8.021,7.767 respectively(P

Objective To observe temporal expression of matrix metalloproteinases(MMP-13) and tissue inhibitor of metalloproteina-ses-1 (TIMP-1) in lung of newborn rats with hyperoxia induced chronic lung disease (CLD),and to explore the relationship of CLD with MMPs.Methods The neonatal rats within 24 hours after birth were randomly divided into hyperoxia-exposed group(n=40) and control group(n=40).On postnatal 1,3,7,14 and 21 days,lung tissue of rats in 2 groups were collected.Lung histological changes were evaluated by hematoxylin and eosin and Masson stain;Collagen Ⅰ was detected by enzyme linked immunoadsorbent assay;MMP-13 and TIMP-1 were identifide by immunohistochemistry.Results Exposured to hyperoxia enviroment for 21 days,the number of alveolar decreased,terminal air space enlarged,inter-alveolar septa thickened,and deposition of interstitial collagen fibers.On 14 and 21 days,collagen Ⅰ in the lung of hyperoxia-exposed group increased significantly compared with that of control group(P0.05),obviously decreased on 21 day(P

AIM: To study the clinical characteristics of corneal perforation ( CP) .
METHODS:A retrospective analysis in July 1995 to July 2010 the First Affiliated Hospital of Fujian Medical University diagnosed CP 72 patients ( 72 eyes ) , clinical characteristics of all the patients were analyzed.
RESULTS: The incidence of corneal ulcer perforation rised year by year, the morbidity of male and female was 17:7, the onset age focused on 48 years old. Of 23 industrial workers ( 32%) with clear history of trauma, pathogeny identification results:top two:fungal infection and Acanthamoeba keratitis. A using history of glucocorticoid was found in 10 cases.
CONCLUSION:There are plenty of primary causes of CP such as traumas, fungal infection, Acanthamoeba keratitis, eroded keratitis, etc. CP happens in middle-aged males in Fujian province, most traumas are the causes, the main pathogenic bacteria is fungal infection.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Medicine, Chinese Traditional ; Mice ; Microscopy, Electron ; Sputum ; drug effects

Objective To discuss the effects of extract of Ginkgo BilobaLeaves(EGb) on expression of cytokine of renal interstitial fibrosis induced by transforming growth factor-?1 (TGF-?1) and extracellular matrix. Methods Cultured human kidney cells(HKC) were divided into three groups: control group,TGF-?1(8 ng/mL) group,and TGF-?1(8 ng/mL) added EGb(25,50,100,150 mg/L)group.After 72 h,expression of ?-SMA was detected by cell immunochemistry ABC,and collagen type I by Real-time PCR and Western blotting. Results Treated with TGF-?1(8 ng/mL) for 72 h,expression of ?-SMA and collagen type Ⅰ were up-regulated markedly compared with control group.Treated with EGb(25,50,100,150 mg/L)and TGF-?1(8 ng/mL)concomitantly for 72 h,expression of ?-SMA and collagen typeⅠ were down-regulated in dosage dependent manner compared with TGF-?1 group. Conclusion EGb can inhibit expression of ?-SMA and collagen type I in HKC induced by TGF-?1,and the possible mechanism might be related to the inhibition of EGb on renal tubular epithelial-myofibroblast transdifferentiation.

Objective To investigate the distribution of ompA gnne in 15 Chinese reference standard strains belonging to 15 serogroups of Leptospira interrogate, and to express recombinant OmpA ( rOmpA ) and to identify immunogenicity and immunoprotection of rOmpA. Methods Genomic DNAs from different leptospiral strains were extracted by phenol-chloroform method. Entire ompA gene fragments from the strains were amplified by PCR and then sequenced after T-A cloning. A prokaryotic expression system of ompA gene from L. interrogans strain 56601 was constructed, and the expression and yield of rOmpA were determined by SOS-PAGE plus Bio-Rad Agarose Image Analyser. Rabbits were immunized with rOmpA for obtaining antiserum, and immunodiffusion test was used to measure the antiserum's titer. Western blot assay was performed to determine the immunoreaetivity of rOmpA with the antiserum against rOmpA and antiserum against whole cell of L. interrogans strain 56601, while mi-croscopic agglutination test (MAT) was applied to detect the cross agglutination to the 15 L. interrogans strains. A leptospire adhering cell model and a leptospire infecting guinea pigs model were used to determine the adhesion-bloc-king effect of rOmpA antiserum and immunoprotection of rOmpA. Results All the 15 L. interrogans strains, but not L. biflexa strain Patoe Ⅰ , had sequence conserved ompA genes. The yield of rOmpA was approximate 20% of the total bacterial proteins, rOmpA could induce rabbits to produce antibody and immunodiffusion titer of the anti-serum was 1:4. Both antisera against rOmpA and against whole cell of L. interrogans strain 56601 were able to pro-duce positive Western blot signs to rOmpA, and the former offered 1 : 20-1 : 320 MAT titers to the 15 L. interrogans strains. 1: 10-1:160 dilutions of rOmpA antiserum could efficiently block L. interrogans strain 56601 adhering to J774A. 1 cells, and 100 μg and 200 μg rOmpA displayed 50.0% and 75.0% immunoprotective rates in the infee-ted guinea pigs. Conclusion ompA gene only exists in genomes of different pathogenic L. interrogans serogroups. rOmpA has relatively stronger antigenicity, cross immunoreactivity and certain immunoprotection, implying that this recombinant protein may be used as a candidate antigen for developing universal genetic engineering vaccine of L. interrogans.

OBJECTIVEArrested lung development and lung fibrosis are characteristic pathological changes in chronic lung disease (CLD). Therefore, the role of extracellular matrix (ECM) remodeling in lung fibrosis has been emphasized recently. Plasmin system is also an important factor to modulate ECM degradation and matrix metalloproteinase (MMP) system. In this study, the authors established an animal model of CLD induced by inhaling high concentration oxygen (hyperoxia) to investigate the changes and functions of urokinase-plasminogen activator (u-PA) and plasminogen activator inhibitor-1 (PAI-1) in CLD.

METHODSFull-term newborn rats were continuously exposed to oxygen (0.90 - 0.95 O(2)) or room air within 12 h after birth. On day 1, 3, 7, 14, 21 in hyperoxia groups and air controls, lung pathology in newborn rats were observed. The changes of u-PA and PAI-1 protein and mRNA expression were measured by Western blotting and RT-PCR.

RESULTS(1) The pathological findings of the lung tissue: on day 3, there was a few inflammatory cells exuded out, bleeding, edema, and interstitial cells increased in hyperoxia group. On day 7 and thereafter, the terminal air space size of the oxygen-exposed rat became large, there was inflammatory response and more interstitial cells, interstitium was thicker, and collagen deposited. (2) u-PA expression: On day 3, the u-PA protein expression increased in hyperoxia group compared with controls (115.52 +/- 7.10 vs. 96.51 +/- 6.33), P < 0.01. On day 7 to day 21, u-PA protein expression (97.66 +/- 7.98, 99.91 +/- 7.60, 103.23 +/- 6.24) was lower than in the control groups (112.43 +/- 6.01, 123.25 +/- 8.35, 103.23 +/- 6.24), P < 0.05, < 0.01 and < 0.01, respectively. u-PA mRNA increased on d 3 in hyperoxia group compared with controls (1.18 +/- 0.07 vs. 0.99 +/- 0.05), P < 0.01. On d 7 to 21, mRNA expression (1.01 +/- 0.06, 1.10 +/- 0.12, 1.27 +/- 0.06) was lower than that in the controls (1.15 +/- 0.08, 1.51 +/- 0.32, 1.60 +/- 0.24) too, P < 0.01. (3) PAI-1 expression: From d 7 to 21 of oxygen exposure, PAI-1 protein expression (147.83 +/- 12.27, 149.07 +/- 11.17, 161.42 +/- 13.08) increased compared with the controls (116.18 +/- 10.67, 113.73 +/- 15.58, 126.60 +/- 8.59), P < 0.01, < 0.05 and < 0.01, respectively. mRNA expression (1.49 +/- 0.28, 1.46 +/- 0.31, 1.51 +/- 0.33) increased compared with the control group (0.94 +/- 0.01, 0.94 +/- 0.03, 0.98 +/- 0.03), P < 0.05, < 0.01 and < 0.05, respectively.

CONCLUSIONSIn the early stage of hyperoxic exposure, the balance of u-PA/PAI-1 mRNA and protein increased, plasmin and degradation activity increased, which may increase the degradation of ECM in lung base membrane. During the middle and late stage, the expression of u-PA/PAI-1 mRNA and protein decreased, plasmin and degradation activity were lower, in parallel to thicker interstitium, suggesting that the imbalance of u-PA/PAI-1 may also play a role in lung fibrosis in CLD induced by hyperoxia.

Animals ; Female ; Hyperoxia ; complications ; Lung ; metabolism ; pathology ; Lung Diseases, Interstitial ; etiology ; metabolism ; pathology ; Male ; Plasminogen Activator Inhibitor 1 ; genetics ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Urokinase-Type Plasminogen Activator ; genetics ; metabolism