Activation of cofilin and its relation with distribution of tight junction protein zonula occludens 1 in hypoxic human intestinal epithelial cells / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the effect of hypoxia on cofilin activation in intestinal epithelial cells and its relation with distribution of tight junction protein zonula occludens 1 (ZO-1).</p><p><b>METHODS</b>The human intestinal epithelial cell line Caco-2 was used to reproduce monolayer cells. The monolayer-cell specimens were divided into control group (no treatment), hypoxic group ( exposed to hypoxia), and normoxic group (exposed to normoxia) according to the random number table. Western blotting was used to detect the protein expressions of cofilin and phosphorylatedl cofilin (p-cofilin) of cells in normoxic group and hypoxic group exposed to normoxia or hypoxia for 1, 2, 6, 12, and 24 h and control group, with 9 samples in control group and 9 samples at each time point in the other two groups. The other monolayer-cell specimens were divided into hypoxic group (exposed to hypoxia) and control group (no treatment) according to the random number table. Cells in hypoxic group exposed to hypoxia for 1, 2, 6, 12, and 24 h and control group were obtained. Morphology and distribution of F-actin was observd with laser scanning confocal microscopy, the ratio of F-actin to G-actin was determined by fluorescence method, and distribution of ZO-l and cellular morphology were observed with laser scanning confocal microscopy. The sample number of last 3 experiments was respectively 3, 6, and 3 in both hypoxic group (at each time point) and control group. Data were processed with paired ttest, analysis of variance of repeated measurement, and LSD-t test.</p><p><b>RESULTS</b>The protein expressions of cofilin and p-cofilin of cells between normoxic group exposed to normoxia for 1 to 24 h and control group showed no significant changes (with values from -0.385 to 1.701, t(p-cofilin)values from 0. 040 to 1.538, P values above 0.05). There were no obvious differences in protein expressions of en filmn of cells between hypoxic group exposed to hypoxia for 1 to 24 h and control group ( with values from 1.032 to 2.390, P values above 0.05). Compared with that in control group, the protein expressions of p-cofilin of cells were greatly reduced in hypoxic group exposed to hypoxia for 1 to 24 h (with values from 4.563 to 22.678, P values below 0.01), especially exposed to hypoxia for 24 h. The protein expressions of cofilin of cells between normoxic group and hypoxic group at each time point were close ( with t values from -0.904 to 1.433, P values above 0.05). In hypoxic group, the protein expressions of p-cofilin of cells exposed to hypoxia for 1, 2, 6, 12, and 24 h were 0.87 +/- 08, 0.780 .05, 0.89 +/- 0.07, 0.68+0. 07, and 0.57 +/- 0.06, respectively, significantly lower than those in normoxic group (0.90 +/- 0.07, 0.97 +/- 0.06, 1.00 +/- 0.06, 1.00 +/- 0.05, and 0.99 +/- 0.05, with t values from 3.193 to 16.434, P values below 0.01). In control group, F-actin in the cytoplasm was abundant, most of it was in bunches. The trend of F-actin was disorderly in hypoxic group from being exposed to hypoxia for 1 h, shortened in length or even dissipated. The ratios of F-actin to G-actin of cells in hypoxic group exposed to hypoxia for 12 and 24 h (0.89 +/- 0.12 and 0.84 +/- 0.19) were obviously decreased as compared with that in control group (1. 00, with t values respectively 3. 622 and 3. 577, P values below 0.01). There were no obvious differences in the ratios of F-actin to G-actin of cells between hypoxic group exposed to hypoxia for 1, 2, and 6 h and control group ( with values from 0.447 to 1.526, P values above 0.05). In control group, cells were compact in arrangement, and ZO-1 was distributed continuously along the cytomnembrane. From being exposed to hypoxia for 2 h, cells became irregular in shape in hypoxic group. ZO-1 was distributed in discontinuous fashion along the cytomembrane with breakage in hypoxic group exposed to hypoxia for 24 h.</p><p><b>CONCLUSIONS</b>Hypoxia may cause the disorder of dynamic balance between F-actin and G-actin by inducing cofilin activation, which in turn leads to the changes in distribution of tight junction protein ZO-1 in intestinal epithelial cells.</p>

Detection of plasma cofilin protein for diagnosis of lung cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect serum content of cofilin protein in patients with lung cancer and investigate its clinical value.</p><p><b>METHODS</b>The serum content of cofilin protein was detected in 30 cases of lung cancer and 30 healthy control subjects using enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>The mean serum content of cofilin protein was 0.485∓0.465 ng/ml in patients with lung cancer and 0.203∓0.102 ng/ml in the control subjects, showing a significant difference between them (P<0.05). The content of cofilin protein in patients with stage III and IV lung cancer 0.744∓0.584 ng/ml, significantly higher than that in stage II patients (0.257∓0.126 ng/ml).</p><p><b>CONCLUSION</b>Serum cofilin protein is elevated in patients with lung cancer, especially in cases in advanced stages, suggesting its relation with lung cancer staging.</p>

Effects of cofilin phosphorylation on the actin cytoskeleton reorganization induced by shear stress / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To explore the effects of cofilin on the actin cytoskeleton reorganization in osteoblasts induced by fluid shear stress.</p><p><b>METHODS</b>Fluid shear stress (1.2 Pa) was applied to osteoblasts for 0 (control group), 15, 30, 45, 60, 120 min in vitro. Cells were stained with fluorescein isothiocyanate (FITC)-phalloidin for fiber-actin, and confocal laser scanning microscope(CLSM) was used to observe the fluorescence of fiber-actin. Western blotting was used to detect the expression of the cofilin and the phospho-cofilin.</p><p><b>RESULTS</b>Actin filaments became organized into stress fibers that were thicker and more abundant than those in non-flowed cells. The fluorescence intensity (38.00 ± 6.88) of fiber-actin after 120 min (42.93 ± 6.41) loading it was 2.8 times as much as that in control group (15.41 ± 3.60, P < 0.05). Additionally, the level of phospho-cofilin protein was dramatically elevated after loading. Fluid shear stress induced an initial decrease of cofilin at 60 min. However, at 120 min cofilin (0.254 ± 0.026) increased to 1.5 times as much as that at 60 min (0.162 ± 0.004).</p><p><b>CONCLUSIONS</b>The results indicate that cofilin phosphorylation mediates fiber-actin reorganization in the osteoblasts induced by fluid shear stress.</p>

Effects of Siwu decoction on bone marrow protein expression of blood deficiency mice induced by cyclophosphamide / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of Siwu decoction on protein expression of blood deficiency mice induced by cyclophosphamide (CIX) and discuss the possible molecular mechanism on blood enriching function of Siwu decoction.</p><p><b>METHOD</b>Blood deficiency mice were established by injecting ip with 250 mg x kg(-1) CTX. Proteomic technologies were applied to identify the different protein.</p><p><b>RESULT</b>Siwu decoction could restore the changes of 12 up-regulated and 3 down-regulated proteins in bone marrow of blood deficiency mice induced by cyclosphosphamide.</p><p><b>CONCLUSION</b>Siwu decoction could effect expression of proteins which functions including apoptosis, proliferation and differentiation of the haematopoietic stem/progenitor cell. The regulation in the molecular level might be the mechanism of stimulating hematopoiesis in bone marrow fo siwu decocetion.</p>

Role of Slingshot-1L in peripheral eosinophils of asthmatic patients / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of Slingshot-1L (SSH-1L) in peripheral blood eosinophils of asthmatic patients.</p><p><b>METHODS</b>Peripheral vein blood sample of 30 ml was collected from 15 healthy volunteers, 15 asthmatic patients with glucocorticoid treatment and 15 asthmatic patients without the treatment. The eosinophils were isolated, purified and counted for each sample, and SSH-1L/beta-actin gene fragments were amplified simultaneously by RT-PCR for the total RNA. SSH-1L protein was detected by Western blotting from the total protein of the peripheral eosinophils. The expressions of SSH-1L at both mRNA and protein levels are compared between different groups.</p><p><b>RESULTS</b>SSH-1L/beta-actin ratio significantly increased in untreated patients with asthma attacks in comparison with healthy volunteers (P<0.05), which did not occur in patients treated with glucocorticoids (P>0.05). The optical density of SSH-1L protein significantly increased in untreated asthmatic patients (P<0.05), but not in patients treated with glucocorticoids (P>0.05), as compared with the healthy volunteers.</p><p><b>CONCLUSION</b>Significantly increased SSH-1L expression in peripheral eosinophils may play an important role in the activation and migration of eosinophils.</p>

Nano-ESI-MS/MS identification on differentiation-associated proteins in M1 mouse myeloid leukemia cells induced by IL-6 / 中国医学科学院学报

<p><b>OBJECTIVE</b>To identify two differentiation-associated proteins induced by rhIL-6 in M1 mouse myeloid leukemia cells.</p><p><b>METHODS</b>Protein spots were excised from 2-D gels and digested in-gel with trypsin. The trypsin lysis products were first analyzed by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) through peptide mass fingerprinting and then performed peptide sequencing by nano-electrospray ionization mass spectrometry/mass spectrometry (nano-ESI-MS/MS). The database search was finished with the Mascot search engine (http://www.matrixscience.co.uk) using the data processed through MaxEnt3 and MasSeq.</p><p><b>RESULTS</b>The two proteins were not revealed by peptide mass fingerprint using MALDI-TOF-MS, while they were respectively identified as Destrin and Putative protein after the sequence of their trypic peptides were obtained by the nano-ESI-MS/MS techniques.</p><p><b>CONCLUSION</b>Nano-ESI-MS/MS technique can successfully identify the two differentiation-associated proteins induced by rhIL-6 and has great advantage in protein analysis.</p>