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 study the expression of Survivin and Caspase-3 in uterine smooth muscle tumour (USMT) and it' s significance. Methods Expression of Survivin and Caspase-3 protein were determined by immunohistochemistry Two-step method on 30 uterine Cellular Uterine Leiomyoma (CUL), 10 normal uterine smooth muscle (NSM), 10 Ordinary Uterine Leiomyoma (OUL), 15 Leiomysarcoma (LMS). Results Survivin level in normal uterine smooth muscle are very low, and in OUL, CUL, LMS was on increasing trend, the OL group and the LCA group have statistical significant difference(P<0.05). Caspase-3 level in NSM, OUL, CUL, LMS was on decreasing trend. The NSM group and the CL group, the NSM group and the LCA group both have statistical significant difference (P<0.05). Conclusion Maybe Survivin can inhibit apoptosis and extend cells lives by inhibit the activity of Caspase-3, so it played an important role in the development progress from benign uterine tumour to malignant uterine tumour. The detect of Survivin and Caspase-3 may be useful in the differential diagnosis of uterine smooth muscle tumours.

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.

Objective To analyze nucleophosmin (NPM1) gene mutations in exon 12 in patients with acute myeloid leukemia (AML) and evaluate the clinical appliance of three methods which are frequently used for detecting gene mutation. Methods Genomic DNA from bone marrow of 54 AML patients was detected by PCR for NPM1 exon 12 and screened by PCR-capillary electrophoresis, denature high-performance liquid chromatography (DHPLC) and direct sequencing separately. FLT3-ITD (FMS-like tyrosine kinease internal tandem duplication) was detected by agarose gel electrophoresis and PCR-capillary electrophoresis. Results Seven AML sample harbored NPM1 gene mutations. Five of them were the most common mutation, known as type A (an insertion of a TCTG tetranucleotide at position 960 bp). One of them was type D (an insertion of a CCTG tetranuclectide at position 960 bp). The new variant was a deletion of a TGGCAGTG sequence at 958 bp and insertion of a GCCCGCGGTTTA sequence instead. The detection ratio of the three methods was all 100% and capillary electrophoresis was more rapid, reliable and easier than the other two methods. Moreover it could detect FLT3-ITD simultaneously. The resolving power of DHPLC was affected by many factors. The direct sequencing method was tedious and the heterozygous sequence might be misread. Conclusions There is a new mutation at position 958 bp with a 12-nucleotide insertion and substitution. PCR-capillary electrophoresis is convenient to screen NPM1 mutations of AML in clinical practice.

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).