Animals ; Cells, Cultured ; Ethanol ; toxicity ; Hepatocytes ; drug effects ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Transcription Factor RelA ; metabolism

Animals ; Carcinogens ; toxicity ; Female ; Lung Neoplasms ; chemically induced ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Sensitivity and Specificity ; Urethane ; toxicity

Objective To study the influence of melatonin(MT)on the sleep of anxiety and depression in elderly with sleep disorder and elderly patients with non-acute cardiac and/or cerebral vascular diseases.Methods The effect of melatonin on sleep Was assessed in a multi-centers,randomized,double blinded and placebo paralleled comparison clinical study.Two hundreds twenty-four participants aged over 60 years old were cases in sub-health state or with non-acute cardiac and/or cerebral Vascular diseases.They were randomly seperated into two groups,and received ether MT(3-6 mg once daily)or starch for 6 months.AU of the subjects were assessed with Pittsburgh sleep quality index(PSQI).Results The participants'total score of PSQI decreased significantly after 1-6 months use of melatonin,representing the remarkable improvement in sleep quality.After the first month.69.3% showed improvement in sleep quality;through the second,third,fourth,five month,the improvement in total score had been going on,and was significantly better in melatonin group than the control group.At the end of the six month,the sleep quality showed the best improvement,with a 91.2% effective rate.Conclusion Melatonin has an remarkable physical effect of improving the sleep quality in old people.

OBJECTIVETo study whether hematologic malignancy patients with anemia have a lower erythropoietin (EPO) response.

METHODSSerum EPO levels were detected by ELISA in patients with hematologic malignancies and with iron deficiency anemia (IDA). Eighty patients with hematologic malignancies, including 13 multiple myeloma (MM), 7 chronic lymphocytic leukemia (CLL) and 60 non-Hodgkin's lymphoma (NHL) were studied. Thirty of them had anemia(21 NHL,6 MM and 3 CLL). Twenty patients with IDA were studied as the control.

RESULTSHematologic malignancy patients with anemia had higher EPO levels [(97.8 +/- 183.9) IU/L] than those with normal Hb values [(27.8 +/- 85.4) IU/L; P <0.01]. In patients with IDA, serum EPO response was inversely correlated with Hb level (r= -0.5, P <0.05) , but no such inverse correlation was found in the hematologic malignancy patients with anemia (r = -0.14). After corrected for Hb level, the serum EPO levels were significantly lower in anemic patients with hematologic malignancies than in IDA patients (P = 0.032) , indicating a decreased EPO response in the former group.

CONCLUSIONAnemia associated with hematologic malignancy might result from an inappropriately low EPO response. EPO treatment for these patients may be beneficial.

Adolescent ; Adult ; Aged ; Anemia, Iron-Deficiency ; blood ; complications ; Enzyme-Linked Immunosorbent Assay ; Erythropoietin ; blood ; Female ; Hematologic Neoplasms ; blood ; complications ; Hemoglobins ; metabolism ; Humans ; Male ; Middle Aged ; Prospective Studies

BACKGROUNDHypoxia-inducible factor-1alpha (HIF-1alpha) is a transcriptional factor that could improve the stimulation of angiogenesis and the metabolic adaptation of tumor cells to hypoxia. A recent study showed that HIF-1alpha could induce colon cancer cells epithelial-mesenchymal transition (EMT). However, no evidence indicates a similar correlation in human prostate cancer cells. This study was designed to evaluate the effect of HIF-1alpha over-expression on the EMT in human prostate cancer cells.

METHODSWe selected the appropriate cell line for HIF-1alpha induction from those EMT negative prostate cell lines through vimentin gene detection by RT-PCR. As the result, LNCaP cell line is the best one for further experiment. LNCaP cells were transfected with recombinant plasmid pcDNA3.1 (-)/HIF-1alpha and pcDNA3.1 (-) control vector by Lipofectamine 2000 system. The positive cell colonies were confirmed by indirect immunofluorescence labeling. Then Transwell polycarbonate filter was used to analyze the invasive potency. The expression of EMT associated proteins, E-cadherin and vimentin, was detected by Western blotting.

RESULTSAmong four of the EMT negative cell lines, LNCaP was the only one expressed the vimentin gene but not the associated protein. The expression level of HIF-1alpha in LNCaP/HIF-1alpha was distinctly higher than that in LNCaP/pcDNA3.1 and LNCaP. The cell numbers of LNCaP/HIF-1alpha that penetrated through the Transwell filter were higher than that of LNCaP/pcDNA3.1 and LNCaP. Compared with the LNCaP/pcDNA3.1 and LNCaP cells, the expression of vimentin was up-regulated in LNCaP/HIF-1alpha, whereas the expression of E-cadherin was down-regulated.

CONCLUSIONSOver-expression of HIF-1alpha stimulates the invasion potency of human prostate carcinoma cells through EMT pathway. The expression of E-cadherin and vimentin, playing established roles in EMT, could be regulated by HIF-1alpha in human prostate cancer cell line.

Cadherins ; genetics ; Cell Line, Tumor ; Epithelial Cells ; chemistry ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; physiology ; Male ; Mesoderm ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; analysis ; Vimentin ; genetics

BACKGROUNDThe cholesterol-lowering statin drugs have some non-lipid-lowering effects, such as inhibiting myocardial remodeling. However, the underlying mechanism is still unclear.

METHODSThe left anterior descending coronary artery was ligated to establish a rat model of heart failure, and the rats were divided into a sham operation (SO) group, myocardial infarction model (MI) group, and MI-atorvastatin group. Changes in hemodynamic parameters were recorded after the final drug administration. Histological diagnosis was made by reviewing hematoxylin and eosin (HE) stained tissue. Real-time quantitative polymerase chain reaction (PCR) was performed to determine the expressions of type I and type III collagen, matrix metalloproteinase-2 (MMP-2), and tissue matrix metalloproteinase inhibitor-2 (TIMP-2). Further, primary rat cardiac fibroblasts were cultured and the MTT assay was performed to determine the effect of atorvastatin on cardiac fibroblast proliferation.

RESULTSThe model of heart failure was established and the results of HE staining and Masson's trichrome staining revealed that the rats in the heart failure group showed obvious hyperplasia of fibrotic tissue, which was significantly reduced in the atorvastatin group. Real-time quantitative PCR showed that the MI group showed a significantly increased expression of type I and type III collagen, MMP-2, and TIMP-2, but a significantly reduced MMP-2/TIMP-2 ratio. Compared with the MI group, the atorvastatin group showed significantly reduced expression of type I and III collagen, unchanged expression of MMP-2, significantly reduced expression of TIMP-2, and an increased MMP-2/TIMP-2 ratio. We further found that atorvastatin significantly inhibited the Ang II-induced fibroblast proliferation and the expression of type I and type III collagen in cardiac fibroblasts while increasing the MMP-2/TIMP-2 ratio.

CONCLUSIONSThese data suggest that atorvastatin can inhibit cardiac fibroblast proliferation and enhance collagen degradation by increasing the MMP-2/TIMP-2 ratio, thereby inhibiting the formation of myocardial fibrosis in rats with heart failure after myocardial infarction.

Animals ; Atorvastatin Calcium ; Collagen ; biosynthesis ; Disease Models, Animal ; Female ; Fibrosis ; Heart Failure ; drug therapy ; pathology ; Heptanoic Acids ; pharmacology ; therapeutic use ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Matrix Metalloproteinase 2 ; genetics ; Myocardial Infarction ; complications ; Myocardium ; pathology ; Pyrroles ; pharmacology ; therapeutic use ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-2 ; genetics ; Ventricular Remodeling ; drug effects

OBJECTIVETo establish immortalized cell line from the urothelium of the urinary bladder and identify the characteristics of the cell line.

METHODSHuman papillomavirus 16 (HPV-16) plasmid was used to transfect urothelium of infant urinary bladder in vitro with the help of Fugene-6, and this plasmid contained E6 and E7 genes of HPV-16. We also identified the existence of HPV-16 E6 and E7 genes and the biological characteristics of the cell line by PCR, immunohistochemistry, and the biology identification.

RESULTSBLTR-4 cell line, produced from the transfection of HPV-16K plasmid, was a cell line from urothelium with the expression of HPV-16 E6 and E7 genes. It had been cultured more than 70 passages, and the characteristics of growth was similar to the immortalized cell line as reported.

CONCLUSIONSBLTR-4 cell line is an immortalized cell line from urothelium of the urinary bladder, which contains HPV-16 E6 and E7 genes. BLTR-4 cell line is a good experimental model to investigate the relationship of the infection of high risk HPV and transitional cell carcinoma (TCC) in vitro.

Cell Line, Transformed ; Humans ; Oncogene Proteins, Viral ; genetics ; Papillomaviridae ; genetics ; Papillomavirus E7 Proteins ; Papillomavirus Infections ; virology ; Plasmids ; genetics ; Repressor Proteins ; genetics ; Transcription, Genetic ; Transfection ; Tumor Virus Infections ; virology ; Urinary Bladder ; cytology ; Urinary Bladder Neoplasms ; virology

OBJECTIVESTo evaluate the application of two-dimensional electrophoresis in the research of differentially expressed proteins in human spermatozoa.

METHODSTwo-dimensional gel electrophoresis was performed on 4 sperm samples from normal healthy men and another 4 from asthenospermia patients. After silver staining, the differential expression proteins were analyzed by PDQuest 2D analysis software.

RESULTSSeven differential protein spots were identified, 2 expressed highly in the asthenospermia sperm but lowly in the normal spermatozoa, while the other 5 expressed just the opposite way.

CONCLUSIONThe protein profiles of differential expression between the normal spermatozoa and idiopathic asthenospermia were established and some differential proteins were found. The data obtained in this study may help prepare the ground for further studies on the isolation and identification of differentially expressed proteins in human asthenospermia sperm.

Case-Control Studies ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Male ; Oligospermia ; metabolism ; Seminal Plasma Proteins ; analysis ; biosynthesis ; Spermatozoa ; chemistry

AIMTo investigate the effect of puerarin on expressions of MMP-2 and TIMP-2 in the kidney of diabetic rats.

METHODSUninephrectomized male Wistar rats were used to induce diabetes by intraperitoneal injection of streptozocin (65 mg x kg(-1)). Puerarin was given daily by intraperitoneal injection from the third day of induction of diabetes for 16 weeks. Using in situ hybridization and immunohistochemistry to detect MMP-2, TIMP-2 mRNA expressions and MMP-2, TIMP-2, collagen IV and Laminin expressions in diabetic kidneys with image analysis system, Flow cytometry was used to detect the expressions of TGFbeta1, MMP-2 and TIMP-2.

RESULTSCompared with those in kidneys of control group, expressions of MMP-2 mRNA and proteins were lower, while the expressions of both TGFbeta1 and TIMP-2 were higher in the diabetic kidney (P < 0.05). The level of MMP-2 expression was advanced, while expression of TIMP-2 was reduced by puerarin treatment (P < 0.05).

CONCLUSIONPuerarin showed some renal protective effect on diabetic nephropathy, partly through inhibition of excessive deposition of glomeruli extracellular matrix by up-regulating MMP-2 and down-regulating TIMP-2 expressions besides reducing the blood glucose.

Animals ; Collagen Type IV ; metabolism ; Diabetic Nephropathies ; chemically induced ; metabolism ; physiopathology ; Isoflavones ; pharmacology ; Kidney ; enzymology ; metabolism ; pathology ; Laminin ; metabolism ; Male ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; Peptide Fragments ; metabolism ; Protective Agents ; pharmacology ; RNA, Messenger ; biosynthesis ; Rats ; Rats, Wistar ; Streptozocin ; Tissue Inhibitor of Metalloproteinase-2 ; biosynthesis ; genetics ; Transforming Growth Factor beta ; metabolism ; Transforming Growth Factor beta1

OBJECTIVETo observe the expression profiles between two metastasis-associated proteins in different prostate cancer cell lines and explore the molecular mechanisms of bone metastatic potentials.

METHODSExpressions of E-cadherin and vimentin in two prostate cancer cell lines (LNCaP and IA8) with different metastatic potentials were detected by Western blotting.

RESULTSThere was remarkable difference in the expressions of E-cadherin and vimentin between the highly metastatic cell line and the lowly one. As one of the adhesion associated proteins, E-cadherin was detected with high level of expression in LNCaP cell line, which was well known as low metastatic potential. However, E-cadherin did not expressed in IA8 with high metastatic potential. And as one of the cytoskeleton proteins, vimentin expression was high in IA8, but not in LNCaP.

CONCLUSIONThere is definitely difference in the metastatic phenotypes (E-cadherin and vimentin) among cell lines with different metastatic potentials. The expressions of E-cadherin and vimentin proteins may play important roles in promoting and inhibiting the metastasis of prostate cancer respectively, and may be considered to be valuable in evaluating the malignant degree, predictable metastasis and prognosis of prostate cancers.

Cadherins ; biosynthesis ; Cell Line, Tumor ; Humans ; Male ; Neoplasm Metastasis ; Prognosis ; Prostatic Neoplasms ; metabolism ; pathology ; Vimentin ; biosynthesis