Yin-Yang1 (YY1),as a ubiquitous nuclear transcription factor with double transcriptional activity in eukaryotic cells,regulates many genes transcription and plays an important role in the cellule biological processes.Overexpression of YY1 is found in several tumor types recently,and may play a role in tumor development and progression by regulating oncogenes,tumor suppressor genes,angiogenesis related factors,as well as apoptosis.YY1 may become a new kind of tumor markers,is conducive to estimate the prognosis of patients with tumor and gain important breakthrough in cancer targeted therapy.

Objective To investigate the effects of high glucose and insulin on the expression of glucose transporter 4 (GLUT4), Cbl-associated protein (CAP) and cytoskeleton protein F-actin of glomerular mesangial cells (GMCs), in order to explore the function of GLUT4, Cbl-associated protein and F-actin in the pathogenesis and development of diabetic nephropathy (DN). Methods Cultured 1097 rat glomerular mesangial cells were divided into 8 groups: control, 10-9 mol/L insulin, 10-8 mol/L insulin, 10-6 mol/L insulin, high glucose (30 mmol/L), mannitol (25 mmol/L mannitol+5 mmol/L glucose), high glucose plus 10-6 mol/L insulin, high glucose plus 10-9 mol/L insulin. Expression of CAP mRNA and GLUT4 was measured by RT-PCR and immunohistochemistry method. F-actin was stained by rhodamine-pholloidin and the fluorescent intensity was calculated by image analysis system. Results The expression of GLUT4 mRNA and protein, CAP mRNA was found in normal giomerular mesangial cells (control), and there was no significant difference in 10-9 mol/L insulin group. The expression of GLUT4 mRNA (P<0.05) and protein (P<0.01), CAP mRNA (P<0.01) level was decreased in high glucose group compared with that of control group, but there was no significant difference in mannitol group. The expression of GLUT4 and CAP mRNA up-regulated with the increase of concentration of insulin. The expressions of GLUT4 mRNA in 10-8 mol/L insulin and 10-6 mol/L insulin groups were 2.06-fold and 2.66-fold of 10-9 mol/L insulin group, of GLUT4 protein were 1.93-fold and 2.83-fold of control, and of CAP mRNA were 1.91-fold and 2.15-fold of control, respectively. The expressions of GLUT4 mRNA, GLUT4 protein, CAP mRNA in high glucose plus insulin group were 2.15-fold, 2.08-fold, 2.14-fold of high glucose group respectively. High glucose decreased the fluorescent intensity of F-actin to 44.5% (P<0.01). 10-8 mol/L insulin and 10-6 mol/L insulin groups increased to 1.224-fold (P<0.05), 1.296-fold (P<0.01) in a concentration-dependent manner. The spearman correlation coefficient between GLUT4 and F-actin was 0.929 (P=0.001), between GLUT4 mRNA and CAP mRNA was 0.905 (P=0.002). Conclusions (1) A certain expression of GLUT4 mRNA and protein, CAP mRNA from GMC is found in normal glomerular mesangial cells. (2) High glucose can inhibit the expression of GLUT4 and CAP mRNA significantly, and facilitate the depolymerization of F-aetin. (3) Insulin can reverse down-regnlation of GLUT4 and CAP mRNA caused by high glucose. (4) GLUT4, CAP and F-actin are important factors in the development of DN.

[Objective] To investigate the roles of Nrf2-Keap1 system in the protection of skin from ultraviolet B (UVB)-induced damage.[Methods] The dorsal surface of ears of 8 wild-type and 8 nrf2-null mutant 8-week-old female mice was exposed to a single dose of UVB irradiation (200 mJ/cm2) by using a FL120SE UV lamp source.Then,the morphology of ears was observed with the measurement of thickness before,as well as on day 1,2,4,7,9,11 and 14 after,the irradiation.Biopsy specimens were taken from the ears 36hours after the irradiation and subjected to hematoxylin and eosin staining as well as terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay.The test Results were recorded and statistically analyzed by rank sum test and t test.[Results] A significant increase was observed in the thickness of mouse ears and number of sunburn cells per high-power field (400 ×) in nrf2-null mutant mice compared with wild-type mice ((0.49 ± 0.22) cm vs.(0.25 ± 0.03) cm,P＜ 0.01; 17.0 ± 3.9 vs.5.0 ± 1.7,t=13.8,P＜ 0.01).The number of TUNEL positive cells in the nrf2-null mutant mice was about 5 times that in the wild mice.The sunburn reaction appeared more intense and persistent in nrf2-null mutant mice than in the wild mice.[Conclusion] Nrf2-Keap1 pathway may protect skin against acute UVB damage,including cell apoptosis and oxidative damage.

ObjectiveTo explore the value of virtual touch tissue quantification (VTQ) in the evaluation of clinical pathological typing of advanced gastric cancer.MethodsFifty six patients who had been diagnosed as gastric cancer were examined using acoustic radiation force impulse.According to clinical pathological typing,all cases were divided into highly-moderately differentiated adenocarcinoma (14 cases) and non-highly-moderately differentiated adenocarcinoma (42 cases).A comparison with clinical pathologic results was made after surgery.The correlation of VTQ results and clinical pathological typing of gastric cancer was analyzed.ResultsThe VTQ value of highly-moderately differentiated adenocarcinoma was lower than that of non-highly-moderately differentiated adenocarcinoma [(1.49 ± 0.44) m/s vs (2.12 ± 0.45) m/s],with statistical significance( t =-4.53,P ＜0.05).According to the maximum area under the ROC curve,the cutoff value of VTQ was 1.795 m/s,the sensitivity and specificity were 78％ and 86％,respectively,the Youden's index was 0.64,and with high reproducibility(Kappa =0.81).ConclusionsVTQ could initial estimate the clinical pathological typing of advanced gastric cancer before operation.

ObjectiveTo evaluate the consistency of HER2 gene status before and after neoadjuvant chemotherapy in breast cancer by FISH (fluorescence in situ hybridization) and IHC (immunohistochemistry) techniques,and analyze the factor of the difference in the result and the feasibility of HER2 gene tested by FISH in the neoadjuvant chemotherapy breast cancer patients.Methods FISH and IHC for HER2 gene expression status was performed on the archival paraffin-embedded sections of breast cancer tissues before and after neoadjuvant chemotherapy from 135 Chinese female patients,x2 test of paired comparison of enumeration data and Kappa analysis were used to compare the difference and consistency of this two techniques.ResultsThe detection rate of HER2 status in punctured cancer tissues before neoadjuvant chemotherapy by FISH and HER2 did not show statistical difference in our research while the opposite result were showed in cancer tissues after neoadjuvant chemotherapy.Moreover,the two techniques of HER2 test were less concordant in patients accepted taxanes neoadjuvant chemotherapy than CAF treatment.ConclusionsThe consistency of FISH and IHC techniques of cancer tissues before neoadjuvant chemotherapy gained advantage compared to the ones after neoadjuvant chemotherapy.Patients received neoadjuvant chemotherapy especially taxanes should take the test of HER2 gene status by FISH technique.

ObjectiveTo explore the value of virtual touch tissue quantification (VTQ) in evaluating Lauren classification of advanced gastric carcinoma.MethodsForty-one patients with gastric cancer proved by endoscopic biopsy performed preoperative VTQ examination,and the findings were compared with postoperative pathologic results via hematoxylin -eosin and Alcian Blue-Periodic Acid Schiff (AB-PAS) staining.ResultsIn 41 patients,26 cases were diagnosed as diffuse type and 15 cases as intestinal type by pathology after operation.The shear wave velocity(SWV) of diffuse type was higher than that of intestinal type [(1.72± 0.83)m/s vs (1.05± 0.66)m/s,t =2.819,P=0.008] measured by virtual touch tissue quantification (VTQ).According to the area under the ROC curve,the cut- off value of SWV in gastric cancer tissues for assessing the diffuse type was 1.045 m/s with a sensitivity of 80.8％ and specificity of 73.3 ％ respectively.ConclusionsVTQ could be considered as a promising method to distinguish the Lauren classification in patients with advanced gastric carcinoma.

Objective To study the inhibitory effects of δ-opioid receptor activation in serumdeprivation induced apoptosis of human liver cells and the proposed protein kinase C(PKC)pathway mechanism.Methods MTT assay was used to detect the survival rate of human liver cells in vitro and Annexin V-FITC/PI double staining was used to detect the cell apoptosis rate.Flow cytometry was used to analyze cell cycle,RT PCR used to analyze the PKC mRNA and Western Blot analysis was used for detecting the protein expression of PKC and Caspase-3.Results After serum-deprivation for 48h of cultured human liver cells in vitro,significant liver cell apoptosis occurred.The apoptosis was suppressed by δ-opioid receptor activation,which manifested as a slower rate of apoptosis,decreased expression of Caspase-3and increased expression of PKC.After GF109203X was added,the inhibitory effects of DADLE decreased markedly.Conclusion Activation of δ-opioid receptor on the membrane of human liver cells has inhibitory effects on serum-deprivation induced apoptosis of liver cells.The underlying mechanism may be associated with PKC pathway activation.

Objective To observe the change of intima/media thickness ratio and expression of inflammatory factors in renal small artery of diabetic rats, and to explore the correlations of intim/media ratio with inflammatory factors and vascular lesions of diabetic nephropathy (DN) rats. Methods Seventy healthy SD rats were randomly divided into diabetic nephropathy group (DN, n=40) and normal control group (N, n=30). DN rat model was induced by intraperitoneal injection of streptozotocin (STZ). Thirty-five DN rats were successfully established. N group received same dose of citrate buffer. Rats were sacrificed after 4, 12, 24 weeks respectively.The intima/media thickness ratio in renal small artery was detected by immunofluorescence. The monocyte chemoattractant protein-1 (MCP-1) protein and mRNA expression of renal small artery were detected by immunohistochemistry and in-situ hybridization at each time point. Results Blood glucose and urine protein excretion (24 h) at different time points in DN group were significantly higher than those of N group (P＜0.05). From the 12th week, Scr, BUN, serum phosphorus were significantly higher than those of N group (P＜0.05). At the 4th week, renal small artery had the expression of MCP-1 protein and mRNA. The expression increased gradually with time, reached the highest at the 24th week, and was significantly higher than that of N group at each time point (P＜0.05). Immunofluorescence results showed that as compared to N group, in the first 4 weeks, intima/media thickness ratio in DN group was not different, at the 12th week the ratio was higher but without significant difference, at the 24th week the ratio was significantly higher (P＜0.05). Small artery intima/media thickness ratio of DN group was positively correlated with MCP-1, cholesterol, triglyceride, serum phosphate (r=0.742, P＜0.01; r=0.740, P＜0.01; r=0.829, P＜0.01; r=0.580, P＜0.01). Conclusions The arterioles intima/media thickness ratio of early DN is significantly correlated with MCP-1, lipids and phosphorus. MCP-1 may be involved in the DN vascular disease.

Objective To observe the expression of bone matrix proteins and the change of intima-tunica media thickness ratio in diabetic rat small renal artery and to explore their correlation and effects on diabetic nephropathy. Methods Seventy healthy SD rats were randomly divided into diabetic group(DN,n=40)and normal control group(N,n=30).DN rat model was induced by streptozotocin(STZ)intraperitoneal injection and the N group rats were given the same dose of citrate buffer.Thirty-five rats were successfully induced in DN group.The rats were sacrificed at week 4,12 and 24,respectively.The protein and mRNA expression of core-bind factor alpha 1(Cbfcd).bone morphogenetic protein 2(BMP-2)and matrix Gla protein(MGP)in smallrenal artery were detected by immunohistochemistry,in-situ hybridization and real-time PCR at each time point. Results Cbfctl and BMP-2 were expressed obviously in small renal artery of DN group by immunohistochemistry stain and in-situ hybridization from 4 to 24 weeks compared with N group at each time point,reaching the peak at week 24.Real-time PCR showed that the MGP mRNA was evidently increased at week 4,slightly decreased at week 12,lowest at week 24in DN group.The BMP-2 mRNA began to increase from week 4 onward,being peak at week 24in DN group.The ratio of intima to tunica media thickness had no significant difference in DN group compared with N group at week 4,but at week 12 and 24 there were significant difference between them.There was a positive correlation between Cbfα1 and BMP-2 expression,but they were negatively correlated with the expression of MGP.The ratio of intima to tunica media thickness was significantly-correlated with the expression of Cbfα1 and BMP-2. Conclusions The ratio of intima to tunica media thickness is positively correlated with Cbfα1 and BMP-2 in small renal artery of early DN.Cbfα1,BMP-2 and MGP may be involved in the progression of vascular lesions in DN.

Objective To explore possible mechanisms of hepatic fibrosis by investigating the ultrastructural dynamic changes of liver tissue, especially several kinds of cells related to hepatic fibrosis.. Methods. Murine schistosomal hepatic fibrosis model was established by infecting mice with Schistosoma japonicum cercariae. Routine transmission electron microscopy was used to observe the liver tissue. H.E. staining was used for examining the pathological changes. . Results . H.E. staining showed that the model was established successfully. Ultrastructural observation showed that at the 6th week after infection, the necrosis of hepatocytes around the acute granulomas occurred; the number of sinusoidal endothelial fenestrae and vitamin A droplets in fat-storing cells decreased; large phagosomes and rough endoplasmic reticulum could be seen in the cytoplasm of Kupffer′s cells. At the 8th week, steatosis was found in some hepatocytes, some microvilli emerged on a few inter-hepatocytic surfaces and the inter-hepatocytic spaces were enlarged. Large collagen fibrillar bundles filled in the perisinusoidal spaces, and capillarization of hepatic sinusoids was observed. Secretory vesicles filled with collagen fibrils appeared in the cytoplasm of fat-storing cells with large amount of collagenous fiber bundles surround the cells. Rough endoplasmic reticulum increased in Kupffer′s cells. At the 10th week, fat-storing cells were activated and transformed into myofibroblasts. At the 12th week, the number of myofibroblasts decreased but that of fibroblasts and fiber cells increased. . Conclusion . Activation of fat-storing cells and transformation from fat-storing cells into myofibroblasts are the critical link in the development of hepatic fibrogenesis following schistosome infection. Kupffer′s cells, necrotic hepatocytes and sinusoidal endothelial cells may relate to the activation of fat-storing cells. Capillarization of hepatic sinusoids possibly accelerates the development of hepatic fibrosis.