Animals ; Cytoskeletal Proteins ; biosynthesis ; genetics ; Drugs, Chinese Herbal ; pharmacology ; Hepatocytes ; metabolism ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the changes of the mRNA expressions of myocardial cytoskeletal proteins in endotoxemic rats.

METHODSThirty-seven Wistar rats were randomized into two groups with injection of 10 mg/kg lipopolysaccharide (LPS) or normal saline through the femoral vein. The cardiac function of the rats was monitored continuously for 24 h, and the morphological changes of the cardiac myocytes were observed with HE staining and electron microscope. The mRNA levels of myocardial cytoskeletal proteins including actin, tubulin and desmin were determined by RT-PCR.

RESULTSNo significant difference was found in the number of CD3(+)T lymphocytes in the TILs between different groups. After the immunotherapy, the peLPS injection resulted in significant impairment of the cardiac function and myocardial microstructure of the rats with reduced heart rate and left ventricular systolic pressure (LVSP). The mRNA expression of actin in the cardiac myocytes measured by fluorescence optical density was reduced significantly 8 h after LPS injection, and that of tubulin was decreased significantly 24 h after LPS treatment; desmin mRNA expression showed no significant variation after LPS injection.

CONCLUSIONLPS can significantly impair the cardiac function of the rats possibly by inducing damages of the myocardial cytoarchitecture and causing changes in the mRNA expressions of such cytoskeletal proteins as actin and tubulin.

Actins ; genetics ; metabolism ; Animals ; Cytoskeletal Proteins ; genetics ; metabolism ; Endotoxemia ; chemically induced ; metabolism ; Female ; Lipopolysaccharides ; Myocardium ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Tubulin ; genetics ; metabolism

OBJECTIVETo investigate the role of the TEKT4 protein in the pathogenesis of idiopathic asthenozoospermia.

METHODSWe separated and purified the ejaculated sperm from idiopathic asthenozoospermia patients and normozoospermic men by Percoll discontinuous density gradients, and detected the distribution and the expressions of TEKT4 mRNA and TEKT4 protein by RT-PCR and Western blot.

RESULTSRT-PCR revealed that the expression of TEKT4 mRNA was significantly lower in the sperm of the idiopathic asthenozoospermia patients than in those of the normozoospermic men (0.59 +/- 0.13 vs 0.75 +/- 0.15, t = 4.325, P < 0.05), and Western blot confirmed the results of RT-PCR (0.48 +/- 0.14 vs 0.69 +/- 0.13, t = 5.939, P < 0.05).

CONCLUSIONThe expression of TEKT4 is significantly decreased in the ejaculated sperm of idiopathic asthenozoospermia patients, which might be one of the causes of idiopathic asthenozoospermia.

Adult ; Asthenozoospermia ; metabolism ; Blotting, Western ; Case-Control Studies ; Cytoskeletal Proteins ; metabolism ; Humans ; Male ; RNA, Messenger ; genetics ; Sperm Motility ; Spermatozoa ; metabolism

Bacterial cell division is strictly regulated in the formation of equal daughter cells. This process is governed by a series of spatial and temporal regulators, and several new factors of interest to the field have recently been identified. Here, we report the requirement of gluconate 5-dehydrogenase (Ga5DH) in cell division of the zoonotic pathogen Streptococcus suis. Ga5DH catalyzes the reversible reduction of 5-ketogluconate to D-gluconate and was localized to the site of cell division. The deletion of Ga5DH in S. suis resulted in a plump morphology with aberrant septa joining the progeny. A significant increase was also observed in cell length. These defects were determined to be the consequence of Ga5DH deprivation in S. suis causing FtsZ delocalization. In addition, the interaction of FtsZ with Ga5DH in vitro was confirmed by protein interaction assays. These results indicate that Ga5DH may function to prevent the formation of ectopic Z rings during S. suis cell division.
Bacterial Proteins ; chemistry ; genetics ; metabolism ; Cell Division ; Cell Shape ; Cytoskeletal Proteins ; chemistry ; genetics ; metabolism ; Mutation ; Oxidoreductases ; deficiency ; genetics ; metabolism ; Protein Binding ; Streptococcus suis ; enzymology

One of the most common causes of male infertility is asthenospermia, whose pathogenesis, however, is not yet clear. Recent researches have found that some genes (such as tektin-2, DNAI1, DNAH5, DNAH11, AKAP4, SEPT4 and Smcp) and proteins (such as sperm proteins ACTB, ANXA5, PRM1, PRM2 and SABP and seminal proteins Tf, PSA, PAP and Fractalkine) are associated with asthenospermia. The finding of these molecular markers has provided a base for the explanation of the molecular mechanism of asthenospermia, and these markers may become the diagnostic and therapeutic targets of the disease.
A Kinase Anchor Proteins ; genetics ; Animals ; Asthenozoospermia ; genetics ; metabolism ; Cytoskeletal Proteins ; genetics ; DNA Methylation ; genetics ; GTP Phosphohydrolases ; genetics ; Humans ; Male ; Mutation ; Septins

Progressive tumor growth is dependent on angiogenesis. The mechanisms by which endothelial cells (ECs) are incorporated to develop new blood vessels are not well understood. Recent studies reveal that the ezrin radixin moesin (ERM) family members are key regulators of cellular activities such as adhesion, morphogenetic change, and migration. We hypothesized that ezrin, one of the ERM family members, may play important roles in ECs organization during angiogenesis, and new vessels formation in preexisting tissues. To test this hypothesis, in this study, we investigated the effects of ezrin gene silencing on the migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) in vitro. HUVECs were transfected with plasmids with ezrin-targeting short hairpin RNA by using the lipofectamine-2000 system. Wound assay in vitro and three-dimensional culture were used to detect the migration and angiogenesis capacity of HUVECs. The morphological changes of transfected cells were observed by confocal and phase contrast microscopy. Our results demonstrated that the decreased expression of ezrin in HUVECs significantly induced the morphogenetic changes and cytoskeletal reorganization of the transfected cells, and also reduced cell migration and angiogenesis capacity in vitro, suggesting that ezrin play an important role in the process of HUVECs migration and angiogenesis.
Cell Movement ; genetics ; Cytoskeletal Proteins ; genetics ; metabolism ; Cytoskeleton ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; metabolism ; physiology ; Humans ; Neovascularization, Physiologic ; genetics

Animals ; Apoptosis ; drug effects ; genetics ; COS Cells ; Cercopithecus aethiops ; Cytoskeletal Proteins ; genetics ; metabolism ; Eye Proteins ; genetics ; metabolism ; Glaucoma, Open-Angle ; genetics ; metabolism ; Glycoproteins ; genetics ; metabolism ; Humans ; Mutation ; Ubiquinone ; analogs & derivatives ; pharmacology

OBJECTIVETo investigate the expression of microRNA-183 (miR-183) and Ezrin protein in stage II( gastric cancer (GC).

METHODSSpecimens of stage II( GC and paracancer tissues (5 cm away from the tumor tissues) were collected from 72 patients. Real-time PCR was used to detect the miR-183 expression. Immunohistochemistry was used to examine the Ezrin protein expression in the tumor tissue. The associations of miR-183 expression with the clinicopathologic features of stage II( GC and Ezrin expression were analyzed.

RESULTSmiR-183 expression was lower in stage II( gastric cancer tissues compared with the paracancer tissues samples(median relative expression, 0.676 vs. 1.000, P<0.05). Low expression of miR-183 was significantly associated with histological differentiation(0.429 vs. 0.907, P<0.05), lymph node metastasis(0.507 vs. 0.908, P<0.05). The survival was shorter in patient with low expression of miR-183(63.0±4.0) as compared to those with high expression of miR-183(75.2±3.8)(P<0.05). There was a negative correlation between the expression of miR-183 and Ezrin(r=-0.272, P<0.05).

CONCLUSIONSmiR-183 is down-regulated in stage II( GC, and associated with the differentiation, metastasis, and prognosis. Ezrin is a potential regulatory protein of miR-183.

Adult ; Aged ; Aged, 80 and over ; Cytoskeletal Proteins ; metabolism ; Female ; Humans ; Male ; MicroRNAs ; genetics ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Staging ; Prognosis ; Stomach Neoplasms ; genetics ; metabolism ; pathology

OBJECTIVETo investigate the effects of 8-methoxypsoralen on human melanocytes [Ca(2+)]i and cytoskeleton actin organization in vitro.

METHODSHuman melanocytes were obtained from normal foreskins. Laser confocal microscope was employed to measure [Ca(2+)]i and rhodamine-conjugated phalloidin was used to visualize the cytoskeleton actin.

RESULTS8-methoxypsoralen increased [Ca(2+)]i and induced organization of actin stress fiber cytoskeleton.

CONCLUSION8-methoxypsoralen might influence the migration of melanocytes by increasing the intracellular free Ca(2+) concentration and cytoskeleton actin reorganization.

Actins ; biosynthesis ; genetics ; Calcium ; metabolism ; Cell Movement ; drug effects ; Cells, Cultured ; Cytoskeletal Proteins ; biosynthesis ; genetics ; Humans ; Melanocytes ; cytology ; drug effects ; metabolism ; Methoxsalen ; pharmacology ; Skin ; cytology

BACKGROUNDBrain acid soluble protein 1 (BASP1) is identified as a novel potential tumor suppressor in several cancers. However, its role in thyroid cancer has not been investigated yet. In the present study, the antitumor activities of BASP1 against the growth and migration of thyroid cancer cells were evaluated.

METHODSBASP1 expression in thyroid cancer tissues and normal tissues were examined by immunohistochemical staining and the association between its expression and prognosis was analyzed. pcDNA-BASP1 carrying full length of BASP1 cDNA was constructed to restore the expression of BASP1 in thyroid cancer cell lines (BHT-101 and KMH-2). The cell proliferation in vitro and in vivo was evaluated by WST-1 assay and xenograft tumor models, respectively. Cell cycle distribution after transfection was analyzed using flow cytometry. Cell apoptosis after transfection was examined by annexin V/propidium iodide assay. The migration was examined using transwell assay.

RESULTSBASP1 expression was abundant in normal tissues while it is significantly decreased in cancer tissues (P = 0.000). pcDNA-BASP1 restored the expression of BASP1 and significantly inhibited the growth of BHT-101 and KMH-2 cells as well as xenograft tumors in nude mice (P = 0.000). pcDNA-BASP1 induced G1 arrest and apoptosis in BHT-101 and KMH-2 cells. In addition, pcDNA-BASP1 significantly inhibited the cell migration.

CONCLUSIONSDownregulation of BASP1 expression may play a role in the tumorigenesis of thyroid cancer. Restoration of BASP1 expression exerted extensive antitumor activities against growth and migration of thyroid cancer cells, which suggested that BASP1 gene might act as a potential therapeutic agent for the treatment of thyroid cancer.

Aged ; Animals ; Apoptosis ; genetics ; physiology ; Calmodulin-Binding Proteins ; genetics ; metabolism ; Cell Cycle ; genetics ; physiology ; Cell Line, Tumor ; Cell Movement ; genetics ; physiology ; Cell Proliferation ; genetics ; physiology ; Cytoskeletal Proteins ; genetics ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mice, Nude ; Middle Aged ; Nerve Tissue Proteins ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology ; Xenograft Model Antitumor Assays