Objective To explore the effect and mechanism of metastasis associated gene 1 (MTA1) on epithelial-mesenchymal transition(EMT) of esophageal squamous cell carcinoma(ESCC).Methods Lentivirus infection method was used to establish the MTA1 knocking out cell line (LV3-shMTA1-KYSE410) and the MTA1 overexpressing cell line (LVS-MTA1-KYSE450).Western Blot was used to measure the expression of MTA1 and the proteins associated with EMT process.Furthermore,the expression and localization of E-cadherin and Vimentin were observed by immunofluorescence assay under confocal microscope.Finally,the wound healing method was performed to confirm the changes of migration ability of the established cell lines.Results When KYSE-450 cells were overexpressed MTA1,the expression level of E-cadherin was down-regulated while Vimentin was up-regulated,and the migration ability was enhanced (0.91 ± 0.00 vs.0.23 ± 0.04,P ＜0.05).When MTA1 was knocked out in KYSE-410 cells,the results were the opposite (0.19±0.01 vs 0.53±0.01,P ＜0.05).Conclusion Overexpression of MTA1 may promote the process of epithelial-mesenchymal transition and enhance the migration ability of ESCC.

Objective To explore the correlationship between level of bFGF (basic fibroblast growth factor) in serum and synovia and X-ray severity (Kellgren-Lawrence Grading System) in patients with knee OA. Method 68 patients with knee OA were enrolled into this study. Knee OA grading was evaluated according to the Kellgren-Lawrence classification. bFGF levels in both serum and synovia were examined using enzyme-linked immunosorbent assay. Results The level of bFGF in serum and synovial fluid in patients with knee OA were positively correlated with radiographic severity (r = 0.619, P < 0.001 and r = 0.603, P < 0.001, respectively). Further analysis revealed that there was a positive correlation between the level of bFGF in serum and in synovia fluid (r = 0.688, P < 0.001). Conclusions Levels of bFGF in serum and synovial fluid were significantly increased in patients with OA, and levels of bFGF were positively correlated with radiographic severity. These findings indicate that bFGF levels may be a biomarker of disease severity and could play an crucial part in the pathophysiology of degenerative process in OA.

In this study, ITS2 barcode was used to identify Bupleurum chinense and B. longiradiatum. The ITS2 regions of 48 samples were amplified and sequenced. The sequences obtained above were aligned and the K2P distances were calculated. We used three methods, BLAST1, nearest distance and phylogenetic tree (NJ-tree), to test the identification ability. The results showed that the maximum intraspecific genetic distance of B. chinense was 0.013, and the minimum interspecific genetic distance between B. chinense and B. longiradiatum was 0.049. The NJ-tree can easily identify B. chinense and B. longiradiatum. Therefore, the ITS2 barcode is suitable to identify B. chinense and B. longiradiatum.
Bupleurum ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control

OBJECTIVETo investigate the protective effect of hypothermia combined with dexamethasone on spermatogenesis and the expression of intercellular adhesion molecule 1 (ICAM1) after testicular torsion-detorsion.

METHODSWe made unilateral testicular torsion models in 100 pubertal male Sprague-Dawley rats by 720 degree torsion of the left testis and then randomly divided them into four groups of equal number to be treated with normal temperature + physiological saline (group A), hypothermia + physiological saline (group B), normal temperature + dexamethasone (group C), and hypothermia + dexamethasone (group D). After 48 hours, we collected the testes, observed pathological changes of the testicular tissue by HE staining under the light microscope, detected the apoptosis of spermatogenic cells by TUNEL, and determined the expression of ICAM1 by Western blot.

RESULTSHE staining showed different degrees of testicular tissue injury in the four groups of rats, most obvious in group A, but mild in the other three. The ICAM1 protein expression was significantly higher in group A (0.68 +/-0. 03) than in B (0. 49 +/- 0. 06, P <0. 05) , C (0. 46 +/- 0. 09, P < 0.05) , and D (0.17 +/- 0.08, P <0.01). The nuclei were deep brown or brown. Lots of apoptotic spermatogenic cells were seen in the torsion testis of group A, with a significantly higher apoptosis index ( [33. 13 +/- 3.21 ]%) than in B ( [ 17. 12 +/-5.23 ]%, P < 0.05), C ([14.13 +/- 2.03]%, P <0.05), and D ([9.05 +/- 1.03]%, P <0.01).

CONCLUSIONHypothermia combined with dexamethasone can protect the testis from injury as well as the reproductive function of the testis after testicular torsion-detorsion and reduce the expression of ICAM1.

Animals ; Dexamethasone ; pharmacology ; Disease Models, Animal ; Hypothermia, Induced ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; metabolism ; physiopathology ; Spermatogenesis ; drug effects

ObjectiveTo evaluate the clinical safety and efficacy of SilverHawk directional atherectomy for femoropopliteal occlusive lesions. MethodsEighteen ischemia occlusive lesions in 11 patients of the lower extremity were treated with SilverHawk directional atherectomy.The mean lesion number was 1.6 ± 1. 1 per patient. The mean lesion length was ( 3.4 ± 2. 2 ) cm. The average degree of diameter stenosis was 96％ ± 14％. 9 lesions were totally occlusive. Clinical symptoms included claudication in 4 cases ( Rutherford classes: 3) and critical limb ischemia ( Rutherford classes: 4) in 7 cases. Lesions characteristics were divided by TASC classification: TASC B in 7 cases; TASC C in 1 case (in-stent occlusion); TASC D in 3 cases.Mean ABI was 0. 5± 0.4. Patency was evaluated with color duplex sonography or CTA besides clinical examination during follow-up.ResultsNine totally occlusive lesions were recanalizated successfully via intraluminal approach. 18 lesions achieved technical success (residual stenosis ＜50％ ) leaving 15％ ±7％ mean residual stenosis in mean (8 ±3)min, predilation was needed in one lesion ( in-stent occlusion) prior to atherectomy. Clinical symptoms improved or disappeared with mean ABI 1.07 ±0. 12 and Rutherford grades: 0 (n =9) and 1 (n =2). Patency rate was 100％ with mean 0. 93 ± 0. 14 ABI and Rutherford grades remain unchanged after follow-up of mean ( 9 ± 4 ) monthes.ConclusionsSilverHawk directional atherectomy is safe and effective for the treatment of lower extremity ischemia.

ObjectiveTo evaluate the clinical application of endovascular treatment for severe Takayasu's arteritis (TA).MethodsIn this study,35 target lesions in 32 patients [28 women,mean age (30 ±8) years] with severe Takayasu's arteritis were treated with endovascular merthod.The average length of lesion was 3.1 cm( range 2.7 -5.3).The overall average degree of diameter stenosis was 90％ ± 11％ (range 70- 100)in which 15 lesions were completely occlusive.There were 10 patients whose ESR were higher than 20 mm/h( range 25 -37).Follow-up included physical examination and patency evaluated by color duplex souography/computed tomography angiography/angiography at 6 months and then annually.ResultsRecanalization was unsuccessful in 3 completely occlusive lesions,with a successful rate of 80％(12/15).There was one case in which embolization leading to acute thrombogenesis developed during interventional procedure and resulting in severe stroke.The technical successful rate ( residual stenosis ＜ 50％ ) was 88.6％ ( 31/35 ).The transient cerebral ischemia attack ( TIA ) symptoms disappeared in 31 cases.26 cases were followed up for an average of (19 ± 10) months (range 13 -40).Occipital infarction following severe in-stent restenosis developed 13 months later in one case.Symptomatic in-stent restenosis18monthslaterwasfoundin2cases. Patencyratewas88.5％( 23/26 ).ConclusionsEndovascular treatment is safe and effective for severe TA.Strict indication and accurate targeting the lesions help ensure the success of management.

AIM: To investigate the effects and mechanism of enalapril on nephritis of diabetic mice. METHODS: Diabetes was induced by injection of streptozotocin after uninephrectomy. Rats were randomly divided into three groups: control, diabetes, diabetes treated with enalapril (10 mg?kg~-1 ?d~-1 by gavage). 8 weeks after STZ injection, urine albumin excretion rate (AER) were measured, and glomerular morphology were observed by light microscopy. The levels of malonyldialdehyde (MDA) in renal tissue and urine as well as activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) in renal tissue were determined. Immunohistochemistry for ED-1 (macrophage marker), monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1) were performed by streptavidin-biotin complex (SABC) technique. RESULTS: Increased kidney weight, ratio of kidney weight to body weight, AER and expansion of mesangial as well as tuft areas on histological examination of the kidney were significantly attenuated by the treatment of enalapril (P

Cognitive impairment is a common complication of diabetes. Hippocampus plays an important role in cognitive function. In hyperglycemia, synaptophysin, a crucial synaptic vesicle membrane protein in hippocampus neuron is found to be down-regulated. Recent evidences have shown that angiotensin IV can facilitate memory acquisition and recovery. However, whether it can also improve cognitive functions of diabetic rats with cognitive disorder, and the possible mechanisms are uncertain. Hence, the objectives of this study. Forty fi ve Sprague Dawley male rats were randomly divided into three groups: Control, diabetic group and diabetes with angiotensin IV treatment group. The cognitive functions, mainly learning and memory of the rats were evaluated using Morris water maze task. The synapses ultrastructure, relative mRNA concentrations and protein expression levels of synaptophysin in hippocampus CA1 area were estimated using transmission electron microscope, RT-PCR, immunohistochemistry and western blotting, respectively. Our study showed that in the diabetic rats with angiotensin IV treatment, the cognitive impairment as measured by Morris water maze task improved, the ultrastructure of synapses in hippocampus reversed, the relative mRNA concentrations and protein levels of synaptophysin in hippocampus signifi cantly increased, when compared with diabetic rats. We conclude that angiotensin IV plays an important role in improving cognitive function of diabetic rats. The possible mechanisms are up-regulating the expression of synaptophysin and normalizing the ultrastructure of synapses in hippocampus.

To investigate the high-risk factors for newborn hearing loss and to provide information for preventing the development of hearing loss and delaying its progression, from May 2003 to June 2006, neonates who failed to pass the universal newborn hearing screening (UNHS) were referred to Jinan Newborn Hearing Screening and Rehabilitation Center from 7 newborn hearing screening centers in seven cities of Shandong province. One-to-one pair-matched case-control method was employed for statistical analysis of the basic features of definitely identified cases. High-risk factors relating to the bilateral hearing loss were evaluated by univariate and multivariate Logistic regression analysis. Our results revealed that 721 transferred newborns who didn't pass the hearing screening received audiological and medical evaluation and 367 were confirmed to have hearing loss. Of them, 177 neonates with hearing loss who met the matching requirements were included in the study as subjects. Univariate analysis showed that high-risk factors related to hearing loss incuded age of father, education backgrounds of parents, parity, birth weight, gestational weeks, craniofacial deformity, history of receiving treatment in neonatal intensive care unit (NICU), neonatal disease, family history of otopathy and family history of congenital hearing loss. Multivariate Logistic regression analysis revealed that 4 independent risk factors were related to bilateral hearing loss, including parity (OR=16.285, 95% CI 3.379-78.481), neonatal disease (OR=34.968, 95% CI 2.720-449.534), family history of congenital hearing loss (OR=69.488, 95% CI 4.417-1093.300) and birth weight (OR=0.241, 95% CI 0.090-0.648). It is concluded that parity, neonatal disease and family history of hearing loss are the promoting factors of bilateral hearing loss in neonates and appropriate intervention measures should be taken to deal with the risk factors.

LncRNAH19 has been implicated as having both oncogenic and tumor suppression properties in cancer. LncRNAH19 transcripts also serve as a precursor for miR-675. However, it is unknown whether LncRNAH19 and miR-675 are involved in the migration and invasion of hepatocellular carcinoma (HCC) cells. The purpose of this study was to investigate the effect and mechanism of LncRNAH19 and miR-675 on migration and invasion of HCC cells. The migration and invasion of HCC cells were measured by Transwell migration and invasion assays after transfection of HCC cells with miR-675 inhibitors and LncRNAH19siRNA. The levels of LncRNAH19 and miR-675 were detected by quantitative reverse transcriptase real-time polymerase chain reaction (qRT-PCR), and the protein expression of AKT, GSK-3β and Cdc25A by Western blotting analysis. The expression levels of LncRNAH19 and miR-675 were higher in MHCC-97H cells than in L02, Huh-7 and HepG2 cells. Transwell migration assay revealed that the miR-675 inhibitor and LncRNAH19siRNA could significantly increase the migration of HCC cells (P<0.01) as compared with the control group. Transwell invasion assay demonstrated that the miR-675 inhibitor and LncRNAH19siRNA could significantly increase the invasion of HCC cells (P<0.01) as compared with the control group. Western blotting analysis showed that the expression levels of AKT and Cdc25A were significantly increased (P<0.05), and the expression level of GSK-3β was significantly decreased (P<0.05) after treatment with miR-675 inhibitors and LncRNAH19siRNA as compared with the control group. These findings suggested that inhibition of LncRNAH19 and miR-675 expression can promote migration and invasion of HCC cells via AKT/GSK-3β/Cdc25A signaling pathway.