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Objective:To explore the dynamic phenotype of type Ⅱ alveolar epithelial cells(AEC Ⅱ)in radiation-induced lung fibrosisand its role in the formation of fibrosis.Methods:Totally 90 C57BL/6J female mice were divided into 2 groups: irradiation group (50, thoracic irradiation with a single dose of 20 Gy X-rays), control group (40, sham irradiation). At 24 h, 4 and 12 weeks after irradiation, 5 mice were euthanized and the lungs were collected for pathological observation. The other lungtissues were collected for the isolation of primary AEC Ⅱ cells with microbeadssorting.The mRNA expressions of proSP-C, HOPX, vimentin, β-catenin and TGF-β1 in AEC II cells were detected by RT-PCR.Results:Acute pneumonitis was observed in the lungs at 24 h after irradiation and alleviated in accompany with partial alveolar septal thickening and a small amount of collagen deposition at 4 weeks after irradiation. The collagen deposition became more pronounced at 12 weeks after irradiation, together with collapsed and fused alveolar cavities, alveolar septal hyperplasia, and pulmonary fibrosis formation.The mRNAexpression levels of proSP-C and HOPX in primary AEC Ⅱ cells increased at 24 hours after irradiation and then approached to a peak value at 4 weeks after irradiation ( F=8.441, 3.586, P=0.036). The mRNA expression levels of vimentin, a biomarker of EMT, was increased significantly at 4 weeks and continued up to 12 weeks after irradiation( F=8.358, P=0.001). The mRNA expression levels of profibrotic factors β-catenin and TGF-β1 were both significant increased at 12 weeks after irradiation( F=4.62, 3.279, P=0.044). Conclusions:The phenotypeof AECⅡ cells could not only be transformed from proSP-C+ to HOPX+ /proSP-C+ , HOPX+ /proSP-C+ /vimentin+ , and vimentin+ /proSP-C, but also differentiated into mesenchymal cells with highly expressed profibrotic factors, thereby inducing EMT process, which either played a role in the repair of radiation-induced lung injury or triggered radiation-induced fibrosis.
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Objective@#To investigate the mechanism of cleft palate in mice induced by excessive all-trans retinoic acid (atRA).@*Methods@#The pregnant mice were randomly divided into atRA-treated group (n=27) and control group (n=27). atRA-treated group was given by gavage a single dose of atRA (100 mg/kg) at 8: 00 AM on gestation day 10 (GD10) and the control group was given by gavage the isopyknic corn oil. At GD13-GD15, the fetal mice palate development was observed by HE staining. The mouse embryonic palatal mesenchymal cell proliferation was detected by 5-bromo-2-deoxyuridine (BrdU) immunohistochemistry. The expressions of Smad2, phospho-Smad2 (p-Smad2), Smad4 and Smad7 in mouse embryonic palatal mesenchyme were analyzed by Western blotting.@*Results@#At GD13-GD15, compared with the control, the ratio of BrdU-positive cells in the palatal mesenchyme of atRA-treated fetuses decreased significantly (P<0.05), especially at GD14, atRA inhibition rate was (65.4±1.7)%. Moreover, atRA decreased the levels of p-Smad2 and Smad4 in embryonic palate mesenchymal cells, whereas the expression of Smad7 was significantly increased at GD14 and GD15.@*Conclusions@#atRA may lead to cleft palate by inhibiting the activation of Smad signaling pathway and affecting the proliferation of palatal mesenchymal cells.
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Objective To develop and validate a simple and reliable HPLC method for the analysis of flee and total carnitine in human serum and to investigate its clinical significance. Methods After proteins in serum were precipitated with a precipitating reagent, carnitine in serum was derivatized to form its ester. HPLC separation of the sample solution was performed on a Lichrospher SiO2 column and detected by ultraviolet absorbance at 260 nm. A mobile phase composed of acetonitrile-citric acid-triethanolamine was found to be the most suitable for this separation at a flow rate of 1.2 ml/min and enabled the baseline separation of the carnitine from interferences with isocratic elution. The free and total carnitine levels in serum were studied in 347 subjects. Results Under the chromatographic conditions described, the carnitine derivative had a retention time of approximately 10 min. Good separation and detectability of carnitine in human serum sample were obtained. The method proved to be linear in the range of carnitine from 0 μmol/L to 400 μmol/L The relative standard deviations of within-assay ( n = 5 ) for free and total carnitine analysis were 3.36% and 1.97% , respectively, and between-assay (n =7) for free and total carnitine analysis were 3.04% and 1.77%, respectively. The average recovery was 98. 2% for free camitine and 96.3% for total carnitine, respectively. The average L-carnitine concentrations in the 347 subjects were as follows: total carnitine (52. 2 ± 8. 6 ) μmol/L, free carnitine ( 42. 3 ± 8. 3 ) μmol/L and acylcarnitine ( 9. 9 ± 2. 9 )μmol/L in the male group ( n = 182), and total carnitine (48.2 ± 9. 9 ) μmol/L, free camitine ( 37.9 ±8. 7) μmol/L and acylcarnitine ( 10. 3 ± 3.5 ) μmol/L in the female group ( n = 165 ). Statistical analysis showed that total and free carnitine levels of male were higher than that of female ( P <0. 01 ) while there was not statistical difference of acylcarnitine levels between two groups ( P > 0. 05). Conclusion The method has been successfully applied to the simultaneous determination of free and total carnitine in serum with good sensitivity, specificity and repeatability, and this is a useful guidance for the clinic therapy and the mechanism study on the diseases associated with carnitine.
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Objective: Carnitine,the only carrier for the fatty acid crossing into mitochondria for oxidization,has a significant effect on the metabolism serum lipids.The purpose of this study is to investigate the effects of different doses of carnitine supplement on serum lipids in hyperlipemia rabbits. Methods: Twenty-eight rabbits were equally randomized into a control and three experimental groups,the former fed on high-fat diet,and the latter on carnitine at the dose of 100,200 and 400 mg/kg respectively every day in addition to high-fat diet.Blood samples were collected from the central artery of the rabbits for the measurement of the total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),apolipoprotein B(ApoB) and free carnitine in the serum,their body weight obtained at 0,1,2,3,4 and 5 weeks,and the effects of different doses of carnitine supplement were observed on serum lipids.Results: All the 28 rabbits were included in the result analysis.The hyperlipemia model was successfully established after 4 weeks of high-fat diet.Compared with the control group,the experimental groups showed a remarkable decrease in the levels of TC,TG,LDL-C and AapoB and an increase in that of HDL-C in the serum,positively correlated with the dose and time of carnitine supplement.With the lengthening of time,the effects of different doses of carnitine supplement on the serum lipids was gradually decreased.Conclusion: Carnitine supplement can inhibit the increase of the levels of TC,TG,LDL-C and AapoB in the serum of the rabbits with hyperlipemia induced by high-fat diet and the effect is positively correlated with the dose and time of carnitine supplement.
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Objective: Oxidative modification of low density lipoprotein(LDL) plays an important role in the development of premature atherosclerosis in autoimmune disorders.Oxidized LDL(ox-LDL) reportedly forms stable complexes with ?2-glycoprotein I(?2-GPI),a major autoantigen for anticardiolipin antibodies,in circulation and the intima of atherosclerotic lesions. This study aims to investigate the serum ?2-GPI/ox-LDL concentration in Chinese patients with systemic lupus erythematosus(SLE),and clinical diagnostic value.Methods: The concentrations of ?2-GPI/ox-LDL complexes were analyzed in 47 SLE patients and 42 healthy controls by ELISA.Results: The serum ?2-GPI/ox-LDL concentrations were significantly higher in the SLE patients than in the controls(\U/ml vs\U/ml,P
