Objective To evaluate the expression levels of peptidylarginine deiminase 4 (PADI4)and B-cells pecific Moloney leukemia virus insert site-1 (BMI-1 )in esophageal squamous cell carcinoma (ESCC) tissues and pericarcinous tissues.To explore the function and clinical significance in the development of ESCC and their association.Methods The expression levels of PADI4 and BMI-1 were measured by immunohisto-chemistry,Western blotting and quantitative real time PCR in ESCC tissues and pericarcinous tissues from 86 patients.The relationships between the expressions of PADI4 and BMI-1 and the clinicopathologic characte-ristics were analyzed.Results The immunohistochemistry showed that the expressions of PADI4 and BMI-1 in ESCC tissues (68.6% and 73.3%)were significantly higher than those in pericarcinous tissues (37.2% and 30.2%,χ2 =1 7.01 1 ,P =0.000;χ2 =31 .876,P =0.000).Western blotting indicated that the levels of PADI4 and BMI-1 were higher than those in pericarcinous tissues (0.91 9 ±0.098 vs.0.71 8 ±0.1 03,t =2.462,P =0.021 ;0.975 ±0.074 vs.0.71 7 ±0.071 ,t =2.640,P =0.01 4).The expressions of BMI-1 and PADI4 mRNA in ESCC tissues were higher than those in pericarcinous tissues,but the differences were not sta-tistically significant (0.091 ±0.005 vs.0.038 ±0.002,t =1 .701 ,P =0.1 01 ;0.1 1 4 ±0.075 vs.0.048 ± 0.003,t =1 .499,P =0.1 46)by the quantitative real time PCR.The expression of PADI4 was correlated with lymph node metastasis (χ2 =5.771 ,P =0.01 6),depth of invasion (χ2 =6.672,P =0.01 0)and clinical stage (χ2 =5.771 ,P =0.01 6).The BMI-1 gene expression had a correlation with lymph node metastasis (χ2 =7.1 76,P =0.007),the differentiation degree (χ2 =1 3.787,P =0.001 )and clinical stage (χ2 =7.1 76,P =0.007).In addition,there was a positive correlation between PADI4 and BMI-1 expression in ESCC by immunohistochemistry and quantitative real time PCR (r =0.21 4,P =0.047;r =0.534,P =0.005).Conclusion The expression levels of PADI4 and BMI-1 are significantly higher in ESCC compared to pericarcinous tissues.PADI4 and BMI-1 are positively correlated and may contribute to the diagnosis and prog-nosis of the ESCC.

Purpose:To investigate the main prognostic factors of patients with renal cell carcinoma.Methods:104 cases of patients with postoperative renal cell carcinoma were analyzed by Cox regression models.Results:Four factors showed significant relation to prognosis: clinical stage, radical nephrectomy for renal cell carcinoma, hematuria, embolization of renal artery. A predicting equation was established, and the predicting values were calculated according to the individual features of patients. The predicting and actual values were compared, and the sensitivity, specificity and overall concordance were 73.3%, 83.3%, and 80.4% respectively.Conclusions:Radical nephrectomy for renal cell carcinoma and embolization of renal artery were protective factors of renal cell carcinoma and the clinical stage and hematuria were risky factors.

OBJECTIVETo investigate the effects of several vasoactive peptides on the development of arterial restenosis after balloon angioplasty.

METHODSIn rat aortic artery restenosis model produced by denudation of aortic endothelia, we observed changes of endothelin (ET), angiotensin II (AII), calcitonin gene-related peptide (CGRP) and adrenomedullin (Adm) in plasma and aorta with radioimmunoassay and expression of hypertension-related gene (HRG-1) with semi-quantitative RT-PCR, and studied the effects of these peptides on intimal hyperplasia, intima/media ratio and MAPK activities of aortic artery after angioplasty respectively. Furthermore, in cultured cells, we studied the effects of these peptides on vascular smooth muscle cell (VSMC) proliferation and expression of HRG-1 of VSMC from spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats with 3H-TdR incorporation and RT-PCR respectively.

RESULTSAfter angioplasty, the levels of ET and AII in plasma and aorta significantly increased, accompanied with VSMC proliferation and neointima hyperplasia. On day 10 after angioplasty, the levels of ET in plasma and aorta increased by 69% and 124% respectively, compared with sham group (P < 0.01); and the level of aortic AII increased by 80% (P < 0.01). Antiserum against ET or inhibitors of angiotensin converting enzyme (ACE) could significantly inhibit the proliferation of VSMC and neointima formation. Compared with the sham group, on day 3 after angioplasty, the CGRP levels in plasma and aorta increased by 64% and 89% respectively (P < 0.01) and the Adm levels in plasma and tissue increased by 129% and 102% respectively (P < 0.01). On day 10, intravenous administration of CGRP significantly inhibited the proliferation of VSMC and neointima formation induced by balloon aortic injury (by 66% and 79% respectively, P < 0.01). In addition, ET and AII attenuated the expression of HRG-1 in aorta and stimulated mitogen-activated protein kinase (MAPK) activity, while CGRP and Adm potentiated the expression of HRG-1 and inhibited MAPK.

CONCLUSIONSET and AII can stimulate the proliferation of injured intima while CGRP and Adm have an anti-hyperplasia effect after angioplasty. These 4 peptides are involved in the regulation of VSMC proliferation and affect the development of vascular restenosis by regulating the expression of HRG-1 and MAPK activity.

Adrenomedullin ; Angioplasty, Balloon ; adverse effects ; Angiotensin II ; pharmacology ; therapeutic use ; Animals ; Aorta, Thoracic ; cytology ; metabolism ; Calcitonin Gene-Related Peptide ; pharmacology ; therapeutic use ; Cell Division ; drug effects ; Cells, Cultured ; Coronary Restenosis ; etiology ; metabolism ; Endothelin-1 ; pharmacology ; therapeutic use ; Male ; Mitogen-Activated Protein Kinases ; metabolism ; Muscle, Smooth, Vascular ; cytology ; Peptides ; pharmacology ; therapeutic use ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Rats, Wistar ; Vasodilator Agents ; pharmacology ; therapeutic use

Objective To investigate the effects of microRNA-486 (miR-486) expression on the cardiac function in rats with myocardial infarction (MI),through direct injection of an adenovirus carrying the miR-486 gene into the myocardium.Methods Totally 120 male rots were divided into 4 groups.Different target materials were infused into the surrounding areas of MI after building a successful disease model.Expression of miR-486 in the surrounding areas of left ventricular MI was detected using real-time PCR at different time points.Cardiac function was measured by cardiac ultrasound at 4 weeks.Using TUNEL,TTC,Masson,and CD31 staining to measure the organic changes in heart tissues.Results The expression of miR-486 in miR-486 group was higher than the other three groups.The expression of miR-486 in the three groups decreased when the time of MI in rats was prolonged.The cardiac function of the miR-486 group was better than that of the MI and AD groups.The MI area in the miR-486 group was smaller than that in the other two groups.Compared to the Sham group,the CVF at the infarct border zone was increased in the MI,Ad,and miR-486 groups.The AI in the MI and Ad groups was significantly increased compared to that in the miR-486 group.MVD was increased in the miR-486 group compared to that in the Sham,MI,and Ad groups.Conclusion miR-486 improved the cardiac function after MI in rats,through lightening collagen deposition,inhibiting apoptosis,and inducing angiogenesis in ischemic regions.

Objective To study the effects of chronic exposure to inorganic arsenic (iAs) in drinking water on bone mineral density (BMD) in mice and its underlying mechanisms.Methods Five-month-old female C57BL/6 mice were randomly divided into sham groups and ovarectomy (OVX) groups (n =19 mice each group),which were further randomly assigned into control group (distilled water) and iAs exposure groups [5 mg/L and 20 mg/L,inorganic arsenite (iAsⅢ):inorganic arsenate (iAsv) =1 ∶ 1].Following 3 months of exposure to iAs,BMD of the mice were determined by the dual energy X-ray detector.RAW 264.7 cell line and bone marrow hematopoietic stem cells (BMHSC) primarily isolated from C57BL/6 mice were used to study the in vitro effects of iAs on osteoclast differentiation and underlying mechanisms.During differentiation induced by receptor activator of nuclear factor-κ B ligand (RANKL,50 μg/L) and macrophage colony-stimulating factor (M-CSF,30 μg/L),RAW 264.7 cell line were treated with 0.00,0.25,0.50,0.75,1.00,1.50 μmol/L iAsⅢ,while BMHSC with 0.0,0.2,0.4,0.6,0.8,1.0 μmol/L iAsⅢ for 6 days.Based on the effect of iAsⅢ on the differentiation of RAW cells,RAW 264.7 cell line were treated by 0.6 μmol/L iAsⅢ combined with 0,5,10 mmol/L of N-acetyl-cysteine (NAC).Tartrate resistant acid phosphatase (TRAP)-positive red-colored cells with 3 or more nuclei were considered mature osteoclast.Results The femoral BMD of the mice [(80.04 ± 4.06) mg/cm2] that had been exposed to 20 mg/L of iAs for 3 months was substantially decreased compared to that of sham control mice [(84.44 ± 4.40) mg/cm2].As expected,the BMD of the OVX group [(76.36 ± 3.36) mg/cm2] was significant decreased compared to that of the sham control group (P ＜ 0.05).However,the BMD among the OVX groups showed no significant difference [5 mg/L:(77.74 ± 4.91) mg/cm2;20 mg/L:(75.56 ± 3.71) mg/cm2,P ＞ 0.05].In vitro studies,the iAsⅢ evidently affected the osteoclast differentiation in a concentration-dependent fashion.Low concentrations of iAs Ⅲ exposure significantly augmented osteoclast differentiation in the two cell models while high concentrations showed inhibitory effect.In RAW 264.7 cells,the number of osteoclasts in different groups was significantly different (F =1 522,P ＜ 0.05),in the 0.50 μmol/L iAs Ⅲ group the number of osteoclasts reached the peak.In the BMHSC,the nmnber of osteoclasts in different groups was also significantly different (F =1 781,P ＜ 0.05),in the 0.6 μmol/L iAsⅢ group the number of osteoclasts reached the peak.NAC pretreatment significantly abolished low-level iAsⅢ(0.6 μmol/L)-induced augmentation of osteoclast differentiation in a concentration-dependent fashion (0 mmol/L:109.33 ± 3.06;5 mmol/L:56.00 ± 2.65;10 mmol/L:22.67 ± 0.58,F =1 940,P ＜ 0.05).Conclusions The inhibitory effect of iAs on bone metabolism is dependent on the availability of ovary function,suggesting that iAs may interfere with estrogen metabolism and/or function to disturb bone metabolism.Oxidative stress induced by iAs exposure stimulates osteoclast differentiation,and the increased osteoclast differentiation may be involved in the reduction of BMD caused by chronic iAs exposure.These preliminary findings suggest that antioxidant intervention may be an effective approach to prevent osteoporosis induced by chronic iAs exposure.