Coronary Occlusion ; surgery ; Drug-Eluting Stents ; adverse effects ; Humans ; Male ; Middle Aged ; Sirolimus ; administration & dosage ; Thrombosis ; complications

Carcinoma, Renal Cell ; secondary ; Cystadenocarcinoma, Serous ; pathology ; Female ; Humans ; Neoplasms, Glandular and Epithelial ; pathology ; Ovarian Neoplasms ; pathology

Objective To study the expressions of metadherin (MTDH) and cyclinD1 in esophageal squamous cell carcinoma (ESCC) and their clinical significances. Methods The protein expressions of MTDH and cyclinD1 were detected by immunohistochemistry in 78 cases of ESCC. Results The positive expression rate of MTDH in ESCC was 71.79%(56/78) and the positive expression rate of cyclinD1 in ESCC was 74.36%(58/78). The expressions of MTDH and cyclinD1 were significantly correlated with the degree of differentiation and lymph node metastasis (both P< 0.05), but not with the age, gender of patients and depth of tumor invasion (all P> 0.05). Conclusion The over expressions of MTDH and cyclinD1 protein may involve in the occurrence and development of esophageal carcinoma, which play important roles in the invasion and metastasis of esophageal cancer.

Objective:To investigate the effect of miRNA-618 (miR-618) on the cell proliferation and apoptosis of acute monocyte leukemia THP-1 cells.Methods:Real-time polymerase chain reaction (PCR) was used to detect the relative expression level of miR-618 in THP-1 cells and monocytes isolated from peripheral blood of the healthy people. Overexpression of miR-618 plasimid vector was constructed and empty vector was treated as the negative control; and then the two vectors were transfected with THP-1 cells; finally, miR-618 overexpression group and negative control group were set. THP-1 cell proliferation and apoptosis of both groups were detected by using CCK-8 method and flow cytometry, respectively. TargetScan was used to predict the target gene of miR-618 and it was verified by using luciferase reporter assay.Western blot was used to detect the protein levels of THP-1 cells in miR-618 overexpression group and negative control group, and predicted miR-618 target gene in peripheral blood monocytes of the healthy people.Results:PCR showed that the expression level of miR-618 was lower in THP-1 cells compared with that in monocytes isolated from peripheral blood of the healthy people ( P < 0.05). CCK-8 assay showed that compared with the negative control group, the proliferation ability of THP-1 cells in miR-618 overexpression group was decreased (the absorbance values at 0, 24, 48 and 72 h after transfection: 0.20±0.03 vs. 0.20±0.03, 0.28±0.02 vs. 0.35±0.03, 0.34±0.03 vs. 0.43±0.04, 0.39±0.02 vs. 0.53±0.05, all P < 0.05), and the late apoptosis rate was increased [(27.1±0.1)% vs. (14.9±0.1)%, t=2.13, P=0.03]. The target gene of miR-618 was ARPP19 predicted by using TargetScan software. Luciferase reporter assay showed that the relative luciferase activity of THP-1 cells in group transfected with wild-type ARPP19 gene plasmid+miR-618 gene plasmid was higher than that in the blank control group and group transfected with wild-type ARPP19 gene plasmid+miR-618 empty vector (0.170±0.003 vs. 0.100±0.004, 0.100±0.001, all P < 0.05). Western blot indicated the expression level of ARPP19 protein in THP-1 cells of miR-618 overexpression group was lower than that of the negative control group, while the expression levels of ARPP19 protein of peripheral blood monocytes of the healthy people in both groups were similar. Conclusion:miR-618 can inhibit the cell proliferation and promote apoptosis of THP-1 cells by inhibiting the expression of of THP-1 cells ARPP19 in acute monocyte leukemia.

OBJECTIVETo investigate the impact of the Artemisia annua plant-derived drug, artesunate, on proliferation of primary rat hepatic stellate cells (HSCs), and to analyze the underlying molecular mechanisms of its anti-fibrogenic effects involving the inhibition of transforming growth factor-beta 1 (TGF-b1) expression and secretion in liver.

METHODIsolated, cultured, and activated primary rat HSCs were divided into sixteen groups, including one untreated control group and fifteen artesunate-treated experimental groups with 125, 150, 175, 200 or 225 mumol/L for 24, 48 or 72 hours. The rate of cellular proliferation was measured using the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. TGF-b1 mRNA expression was evaluated by reverse transcription-polymerase chain reaction and protein expression was evaluated by Western blotting. Enzyme-linked immunosorbent assay was used to evaluate secreted levels of TGF-b1 protein.

RESULTSArtesunate significantly inhibited proliferation of cultured HSCs in a dose- and time-dependent manner (all, P less than 0.01). After 24 hours of exposure, the inhibition ratios of the various artesunate concentrations were: 6.06%+/-1.44% (125 mumol/L), 21.47%+/-5.57% (150 mumol/L), 42.00%+/-7.36% (175 mumol/L), 67.12%+/-4.55% (200 mumol/L), and 79.83%+/-3.67% (225 mumol/L). Artesunate significantly inhibited the TGF-b1 mRNA expression in HSCs, and the higher the drug concentration, the higher the degree of inhibition (all, P less than 0.01). In addition, artesunate significantly inhibited the expression of intracellular and secreted TGF-b1 protein (all, P less than 0.01). In response to artesunate (mumol/L concentrations), the TGF-b1 levels were (164.24+/-6.88) pg/ml (0μmol/L), (102.68+/-4.45) pg/ml (150μmol/L), (86.54+/-5.56) pg/ml (175μmol/L), and (56.55+/-5.66) pg/ml (200μmol/L).

CONCLUSIONArtesunate exerts anti-fibrogenic effects on HSCs in vitro, possibly by reducing the expression, translation and secretion of TGF-b1.

Animals ; Artemisinins ; pharmacology ; Cells, Cultured ; Hepatic Stellate Cells ; drug effects ; secretion ; Rats ; Rats, Wistar ; Transforming Growth Factor beta1 ; metabolism

Objective To evaluate the quality of domestic hepatitis C diagnostic reagents objectively,and to build up the quality control systems for assessment of hepatitis Cdiagnostic reagents.Methods4080 serum samples from blood donors were collected and detected with EIA kits.146anti-HCV positive and negative samples were selected and tested repeatedly by two different imported ( Murex and Ortho) and domestic anti-HCV EIA kits(InTec,ZHONGSHAN BIO-TECH,WANTAI and KHB),then confirmed by CHIRON RIBA HCV 3.0 and PCR qualitative reagents.The samples were tested by nucleic acid quantitative assay and the RNA positive samples were detected by genotyping reagents.ResultsThe quality control systems of diagnostic reagents of anti-HCV and HCV RNA were constructed.Each quality control system was consisted of 50 samples,including 20 anti-HCV/HCV RNA positive,20 anti-HCV/HCV RNA negative and 10 diluted specimens for sensitivity evaluation.The positive samples with dominant HCV genotypes in China contained strong,moderate and weak positive samples.The negative samples involved those S/CO value ( signal-to-cutoff ratios ) close to threshold.Conclusion The quality control systems established in this study are suitable for assessment of the new and improved domestic hepatitis C diagnostic reagents.

Purpose　To investigate the significance of tumor suppressor gene p15 and p16 expression in soft tissue leiomyosarcoma(LMS) and leiomyoma(LM). Methods　Expressions of p15 and p16 was studied in 38 cases of LMS and 20 LM with immunohistochemical method. Results　The abnormal expression rates of p15 and p16 in LMS were 18.4% and 42.1% respectively, and were significantly different. p15 abnormal expression rate in LM was 60%, significantly higher than in LMS. The total abnormal expression rate of p15 and p16 was 52.6% in LMS studied. Grade Ⅰ tumor had the significantly higher abnormal expression rate than that of grade Ⅱ or Ⅲ tumor. Fifteen cases of LMS were followed. The abnormal expression rate of p15 and p16 in recurrent group was 75%, significantly higher than that in non recurrent group. Conclusions　Loss of expression of p16 protein is more frequent in LMS than that of p15, whereas deletion of p15 protein is more frequent in LM than in LMS. Loss of functions of p15 together with p16 participates in the tumorigenesis and progression of LMS, and is correlated with pathologic grading and prognosis of LMS.

Using a bioassay-guided fractionation technique, two compounds were isolated from the roots of Incarvillea younghusbandii Sprague through silica gel, reverse-phase C18 column chromatography and reverse-phase HPLC. Their structures were identified as acteoside (1) and isoacteoside (2) by ESI-MS, GC-MS, 1D- and 2D-NMR. 1 and 2 showed *OH scavenging capacity similar with benzoic acid, higher O2*- (or *OH) scavenging capacity than ascorbic acid, far higher hepatic LPO inhibitory activities than 2, 6-di-tert-butyl-4-methylphenol (BHT) or ascorbic acid, and more powerful effect on protecting erythrocytes from oxidative damage than ascorbic acid. The *OH scavenging capacity was positively proportional to the concentrations of 1 and 2 ranging from 0.015 6 to 0.500 0 mg x mL(-1). The hepatic LPO inhibitory activities increased with the increasing concentrations of 1 and 2 from 0.001 9 to 0.250 0 mg x mL(-1), but decreased slightly with the increasing concentration from 0.250 0 to 1.0000 mg x L(-1).

Animals ; Aquaporin 4 ; metabolism ; Blood-Brain Barrier ; physiopathology ; Brain Edema ; physiopathology ; Brain Ischemia ; metabolism ; physiopathology ; Capillary Permeability ; physiology ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; physiopathology

Objective To construct tissue-engineered skin via in vitro inoculation of epidermal stem cells(ESCS)onto de-epidermized dermis.Methods Skin tissue was obtained from the foreskin of a healthy 6-year-old child.and keratinocytes were isolated by two-step trypsinization method followed by the collection of ESCS via rapid adhesion by collagen Ⅳ.The ESCS were identified by morphological observation and immunohistochemical staining with K19 and integrin β1.To construct tissue-engineered skin,selected ESCS were seeded onto the surface of de-epidermized dermis followed by a one-week culture immersed in the medium and a subsequent 4-week culture at the air-medium interface.The tissue-engqneered skin was evaluated with haematoxylin & eosin(HE)staining as well as keratin immunohistochemistry.Results Micro scopically,cultured ESCs showed a paving stone-like appearance and grew into colonies.Immunohistochemistry revealed that the ESCs were positive for integrin-β1 and keratin 19.After 5 weeks of culture,3-6 layers of epidermal cell were observed on the dermis with the formation of stratum corneum.Keratin protein was observed in the artificial epidermal skin.Conclusion Tissue-engineered skin is successfully constructed with epidermal stem cells and de-epidermized dermis in vitro.