Objective: To investigate the effects of ribosomal protein 16 (RPS16) on tumorigenesis and development of prostate cancer. Methods: Western blotting (WB) was used to detect the differences of RPS16 levels in 25 cases of prostate cancer tissues and 33 prostate hyperplasia, and immunohistochemistry (IHC) was performed to detect RPS16 levels in 48 prostate cancer tissues and 42 benign tissues. The relationship between RPS16 level and clinical pathological parameters of the patients was analyzed. The RPS16 small interfering RNA (siRNA) was transiently transfected into DU145 and LNCaP cells by liposome method, including RPS16-siRNA1, RPS16-siRNA2 and RPS16-siRNA3. Random disturbance RPS16-siRNANC was used as negative control, cells without transfection were blank control. The efficiency of RNA interference was detected by WB 48-72 h after transfection. RPS16-siRNA with highest efficiency was chosen for subsequent cell proliferation assay, flow cytometry (FCM) and transwell assay in order to detect the effects of RPS16 on cell proliferation, cell cycle and invasion ability of DU145 and LNCaP cells. Results: WB results showed that the level of RPS16 protein in the tissues of prostate cancer was higher than that of the benign group (P=0.008). IHC results showed RPS16 protein level was significantly higher in tumor tissues than benign tissues (P=0.009). RPS16 expression was not correlated with age and metastasis, but significantly correlated with clinical stage (P=0.044) and pathological grade of the tumor (P=0.004). RPS16 siRNA can not only significantly reduce the expression of RPS16 protein in DU145 and LNCaP cells, but also inhibit the proliferation and invasion of the cancer cells, so that the cell cycle arrested in G2/M phase. Conclusion: The high expression of RPS16 protein could enhance the proliferation and invasive ability of prostate cancer cells.

CD4-Positive T-Lymphocytes ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Interleukin-2 Receptor alpha Subunit ; Sequence Analysis

Trichinella spiralis infection in rodents is a well-known model of intestinal inflammation associated with hypermotility. The aim of the study was to use this experimental model to elucidate if Th17 cells are involved in the development of gastrointestinal hypermotility. Colonic smooth muscle contractility was investigated in response to acetylcholine. The levels of IL-17, IL-23 and TGF-beta1 in colon were measured by Western blotting. Flow cytometric detection of intracellular IFN-gamma/IL-4/IL-17 cytokine production was used to analyze the proportions of CD4+ T cells subsets in colon. Our results showed that colonic muscle contractility was increased 2 weeks post infection (PI) and stayed high 12 weeks PI when no discernible inflammation was present in the gut. The proportion of Th17 cells and the expression of IL-17 were up-regulated in colon 2 weeks PI and returned to normal 8 weeks PI. The content of IL-17 was correlated with the colonic smooth muscle hypercontracility 2 weeks PI. Meanwhile, TGF-beta1 was increased 2 weeks PI, while IL-23 was normal. Our results suggest that Th17 cells affect the colonic muscle contractility in mice infected with Trichinella spiralis at intestine stage but not at muscle stage and the effect of Th17 cells on muscle contractility might be induced by TGF-beta1. Other cytokines might be involved in the hypercontracility of colonic smooth muscle at muscle stage.

Objective To find an ideal animal model of acute respiratory distress syndrome (ARDS) through investigating the characteristics of three two-hit animal models of ARDS.Methods Forty-eight SD rats were randomly divided into 4 groups: Control group [2.5 mL/kg normal saline (NS) i.v.given at 0 min and 30 min];OA+OA group [0.5 mL/kg oleic acid (OA) i.v.given at 0 min and 30 min];LPS+LPS group [2.5 mg/kg lipopolysaccharide (LPS) i.v.given at 0 min and 30 min];and OA+LPS group [0.5 mL/kg OA i.v.given at 0 min and 2.5 mg/kg LPS,i.v.given at 30 min].The samples were collected at 5 h after the second drug injection.White blood cells count (WBC),polymorphonuclear leukocyte ratio (PMN%),total protein concentration,tumor necrosis factor α (TNF-α) level in bronchoalveolar lavage fluid (BALF),arterial blood gas analysis and lung wet-dry weight ratio (W/D) were measured,respectively.Pathological changes in the lung tissues were observed and histological scores were evaluated.Results Compared with those in the control group,PaCO2,WBC,PMN%,total protein concentration and TNF-α levels in BALF were significantly increased,while PaO2 was dramatically decreased (P<0.01) in the OA+OA,LPS+LPS and OA+LPS groups.The levels of protein concentration in BALF and lung W/D ratio in the OA+LPS group were significantly higher than these in the LPS+LPS group (P<0.05 for all),but had no statistically significant difference compared with these in the OA+OA group.The levels of WBC,PMN% and TNF-α in BALF in the OA+LPS group were significantly higher than those in the OA+OA group (P<0.05),but not significantly different from those in the LPS+LPS group.The most typical pathological changes and the highest pathological scores were found in the OA+LPS group.Conclusions All the three different methods including OA+OA,LPS+LPS,and OA+LPS can be used to establish “two-hit” animal models of acute respiratory distress syndrome.The “two-hit” animal model of acute respiratory distress syndrome induced by OA+LPS is more closer to clinical ARDS and is useful for studies on the pathophysiology of ARDS,and is an ideal “two-hit” animal model of ARDS.

OBJECTIVETo investigate the effects of non-neuronal muscarinic receptors (NNMR) stimulation on atherosclerosis and endothelial cells activation.

METHODSAtherosclerosis model was established in ApoE-/- mice by a high fat diet for 7 weeks. During the experimental periods, animals were received a low (7 mg/kg/d) or a high (21 mg/kg/d) dose of arecoline by gavage. At the termination of the treatments, serum total cholesterol and NO levels were measured, and the aorta morphology was analyzed by hematoxylin and eosin staining. The gene expression of monocyte chemoattractant protein-1 (MCP-1) and adhesion molecules in the thoracic aortas was determined by RT-PCR, and the MCP-1 protein expression and NF-κB activity were detected by Western blot analysis. NO production, MCP-1 secretion in cultured rat aortic endothelial cells (RAECs), and monocyte-endothelium adhesion assay were also performed after arecoline treatments.

RESULTSArecoline efficiently decreased atherosclerotic plaque areas, increased serum nitric oxide (NO) content, suppressed the mRNA and protein expression of MCP-1, and modulated the IκB-α degradation and P65 phosphorylation in the aortae of ApoE-/- mice. Furthermore, arecoline promoted NO production and suppressed MCP-1 secretion in cultured RAECs after ox-LDL exposure, and either atropine or NG-nitro-L-arginine methylester could abrogate these effects. Arecoline also significantly inhibited the adherence of U937 monocytes to the ox-LDL injured human umbilical vein endothelial cells, which could be abolished by atropine.

CONCLUSIONOur results indicate that arecoline attenuates the progression of atherosclerosis and inhibits endothelial cells activation and adherence by stimulating endothelial NNMR. These effects, at least in part, are due to its modulation on NF-κB activity.

Animals ; Aorta ; cytology ; Apolipoproteins E ; Arecoline ; pharmacology ; Atherosclerosis ; physiopathology ; prevention & control ; Cell Adhesion Molecules ; metabolism ; Chemokine CCL2 ; metabolism ; Cholesterol ; blood ; Disease Progression ; Endothelial Cells ; cytology ; drug effects ; Endothelium, Vascular ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; I-kappa B Proteins ; metabolism ; Lipoproteins, LDL ; Mice ; Mice, Knockout ; Monocytes ; cytology ; NF-KappaB Inhibitor alpha ; Nitric Oxide ; blood ; Nitroarginine ; pharmacology ; Rats ; Receptors, Muscarinic ; physiology ; Transcription Factor RelA ; metabolism

OBJECTIVETo explore the role of toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in myocardial ischemia/reperfusion injury (MI/RI) by observing the dynamic expression changes at mRNA and protein levels early after myocardial ischemia/reperfusion (I/ R).

METHODSThe Wistar rats were randomly divided into Sham and I/R group (n = 42), and killed according to different reperfusion time (1, 2, 4, 6, 12, 24 h and 7 d). Structural and morphous changes of myocytes were observed under optical microscope. The mRNA and protein levels of TLR2 and TLR4 were detected using real-time PCR (RT-PCR). Monocyte chemokine protein-1 (MCP-1) and interleukine-6 (IL-6) mRNA levels were measured by reverse transcriptase-polymerase chain reaction (rt-PCR).

RESULTS(1) With the extension of reperfusion time, the myocardial infarct size increased smoothly, and reached the plateau at 4 h, then stayed in the platform. After reperfusion for 7 d, the ventricular had been remodeled. (2) At the beginning of reperfusion, myocardial structure showed no significant change in Sham group, but had different degrees of injury in I/R group. In rats of the group reperfused for 7 d the left ventricular remodeling could be visible. (3) Compared to sham group,TIR2, TLR4, MCP-1, IL-6 mRNA level were increased in myocardium in I/R group. TLR2 and TLR4 both peaked at 4 h of reperfusion, IL6 peaked at 6 h, followed by a gradually decrease. TLR4 and IL-6 mRNA levels rose again at 7 d. MCP-1 level in I/R group remained fairly with sham group at the beginning of reperfusion, and markedly elevated at 7 d.

CONCLUSIONExpression of TLRs mRNA in myocardium during early after myocardial ischemia/reperfusion increased rapidly and activated TLRs might play an important role in MI/RI through promoting the generation of inflammatory factors. At the late reperfusion, TLRs levels raise again and the expression of inflammatory factors increase once again, Those may probably affect the remodeling of ventricular, and injure myocardial structure and function.

Animals ; Chemokine CCL2 ; metabolism ; Disease Models, Animal ; Interleukin-6 ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; Rats ; Rats, Wistar ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism

OBJECTIVETo determine the advantage of U1 small nuclear RNA as a ribozyme vector (U1-Rz) to inhibit HCV replication in vivo.

METHODSThe 3rd stem-loop was substituted by HCV core specific ribozyme to construct an U1-Rz eucaryotic expression plasmid. Then it was co-transfected with pCMV/T7-NCRC Delta-luc into Huh7 cell line mediated by lipofectin. The cell lysis supernatant was subjected to Western blot and lumenometer to determine the luciferase levels.

RESULTSA U1 snRNA chimeric ribozyme was constructed successfully. Both Rz and U1-Rz inhibited luciferase expression in Huh7 by 48.64% and 87.46%, respectively.

CONCLUSIONRz has more efficacy in cells when using U1 snRNA delivery system. U1 can be an efficient vector for HCV specific ribozyme.

Base Sequence ; Genetic Therapy ; Genetic Vectors ; Hepacivirus ; physiology ; Humans ; Molecular Sequence Data ; RNA, Catalytic ; genetics ; RNA, Small Nuclear ; genetics ; Virus Replication ; genetics

OBJECTIVEHigh altitude pulmonary edema (HAPE), a life-threatening disease, has no biological markers used for the routine prevention, diagnosis and treatment. The aim of this study was to identify serum proteins differentially expressed in patients with HAPE for discovering essential biomarkers.

METHODSA complete serum proteomic analysis was performed on 10 HAPE patients and on 10 high altitude and 11 sea level healthy people as control using two-dimensional gel electrophoresis, followed by matrix-assisted laser desorption/ionization mass spectrometry and peptide mass fingerprinting. Finally, two most significantly changed proteins were validated by enzyme-linked immunosorbent assay (ELISA).

RESULTSEight protein spots stained with differential intensity, respresenting 5 distinct proteins were identified in patients compared with healthy controls through analysis of these composite gels. Among them, four proteins, namely alpha 1-antitrypsin(alpha1-AT), Haptoglobin(Hp), apolipoprotein A-1 (apoA-1) and Complement C3 increased remarkably, while one protein, apolipoprotein A-IV (apoA-IV) decreased significantly. The variation of alpha1-AT and Haptoglobin, as detected by ELISA, was consistent with the results from proteomic analysis.

CONCLUSIONSIt is well known that Hp, alpha1-AT and complement C3 are associated with inflammation and apoA-1 and apoA-IV play important roles in lipid absorption, transport and metabolism. Therefore, the significant expression changes of Hp, alpha1-AT and complement C3 and apoA-1 and apoA-IV between HAPE patients and their corresponding healthy controls highlight the role of inflammatory response system and lipid metabolism system in the pathophysiology of HAPE.

Altitude ; Biomarkers ; blood ; Blood Proteins ; metabolism ; Case-Control Studies ; Electrophoresis, Gel, Two-Dimensional ; Enzyme-Linked Immunosorbent Assay ; Humans ; Peptide Mapping ; Proteome ; Pulmonary Edema ; blood ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

To look for the toxicity fraction of Euphorbia pekinensis and discuss the vinegar processing mechanism. The level of intestinal edema, water content of intestine and stool, IC50 values of IEC-6 were applied to evaluate the toxicity of different fractions. RT-PCR was employed for detecting AQP1, AQP3 mRNA expression. The petroleum ether (PE) fraction and ethyl acetate (EtOAc) fraction could significant cause intestinal edema in mice, increase the water content of duodenum, colon and stool, inhibited the mRNA expression of AQP1 and increased the mRNA level of AQP3 in colon, and the petroleum ether (PE) fraction was more poisonous. After the petroleum ether (PE) fraction was processed with vinegar, the level of intestinal edema, water content of duodenum, colon, stool and inhibition ratio of cells line were reduced. And we compared the composition change after vinegar processing, finding that the conpekinensis.
Acetic Acid ; chemistry ; Animals ; Cell Line ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; toxicity ; Euphorbia ; chemistry ; toxicity ; Male ; Mice ; Mice, Inbred ICR ; Molecular Structure

BACKGROUNDProgrammed cell death 5 (PDCD5) is a novel apoptotic regulatory gene that promotes apoptosis in various tumor cells. Studies have shown that PDCD5 accelerates the apoptosis of synoviocytes in vitro, implying a potential role in rheumatoid arthritis (RA) pathogenesis. This study examined the expression of PDCD5 in serum and synovial fluid of RA patients, its effect on the expression of inflammatory cytokine, interleukin-17 (IL-17), and the assessment of disease activity in RA.

METHODSPDCD5 and IL-17 levels in serum and synovial fluid from 18 patients with RA and 22 patients with osteoarthritis (OA) were detected using enzyme-linked immunosorbent assay (ELISA). Concentrations of serum PDCD5 in 40 healthy people were also detected as controls. As disease activity indices, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), and X-ray grading scale were also evaluated.

RESULTSSerum and synovial fluid PDCD5 levels in RA patients were significantly higher than those in OA and healthy controls. Serum PDCD5 level was inversely correlated to CRP and ESR, and was significantly higher in the RF negative group than in the positive group. PDCD5 level was also negatively correlated with IL-17 levels both in serum and synovial fluid of RA patients. However, differences in synovial fluid PDCD5 level from RA patients at different Larsen stages were not detectable.

CONCLUSIONSPDCD5 affects RA pathogenesis. Insufficient apoptosis of fibroblast-like synoviocytes and inflammatory cells in RA could increase the expression of PDCD5 protein. As PDCD5 levels correlated negatively with disease activity indices and IL-17 level, PDCD5 could become a target in the diagnosis and treatment of RA.

Aged ; Apoptosis ; Apoptosis Regulatory Proteins ; analysis ; blood ; physiology ; Arthritis, Rheumatoid ; etiology ; Blood Sedimentation ; C-Reactive Protein ; analysis ; Female ; Humans ; Interleukin-17 ; analysis ; blood ; physiology ; Male ; Middle Aged ; Neoplasm Proteins ; analysis ; blood ; physiology ; Synovial Fluid ; chemistry