To study the chemical constituents from the the deep-sea fungus Alternaria sp. F49. Seven compounds were isolated from the EtOAc extract by using silica gel, Sephadex LH-20, ODS and HPLC methods. Based on the spectroscopic analysis, their structures were identified as (8R)-5-O-methyl-orcinotriol (1), orcinotriol (2), α-acetylorcinol (3), 3'-hydroxyalternariol 5-O-methyl ether (4), altenusiol (5), altenusin (6), and 5'-methoxy-6-methyl-biphenyl-3,4,3'-triol (7). (8R)-5-O-Methyl-orcinotriol (1) is a new phenolic compound which has never been reported in the literature. Compounds 4-7 showed strong DPPH free radical scavenging activity; whereas compounds 1-7 showed strong ABTS free radical scavenging activity.

BACKGROUNDMast cells are implicated in the development of irritable bowel syndrome (IBS), which is associated with the activation of the "neural-immune" system. The aim of this study was to investigate the role of mast cells in the remodeling of cholinergic and peptidergic neurotransmitters induced by acute cold restriction stress (ACRS) post infection (PI) using mast cell deficient rats (Ws/Ws) and their wild-type controls (+/+).

METHODSTransient intestinal infection was initiated by giving 1500 Trichinella spiralis (T.S.) larvae by gavage. ACRS was induced for 2 hours at day 100 PI. Samples of terminal ilea were prepared for H&E staining, mast cell counting and activation and assessment of IL-1beta and IL-10.

RESULTSWhen infected, both strains of rats experienced an acute infectious stage followed by a recovery. Histological scores were significantly higher in infected rats compared with those of the non-infected controls at day 10 PI (10 day-PI vs. control: +/+: 2.75+/-0.17 vs. 0.42+/-0.09; Ws/Ws: 2.67+/-0.67 vs. 0.50+/-0.34; P<0.01). In +/+ rats, post-infection ACRS induced the formation of low-grade inflammation, represented by the imbalance of IL-1beta and IL-10 (IL-1beta: PI+ACRS vs. control: (1812.24+/-561.61) vs. (1275.97+/-410.21) pg/g, P<0.05; IL-10: PI+ACRS vs. control: (251.9+/-39.8) vs. (255.3+/-24.7) pg/g, P>0.05), accompanied by hyperplasia and activation of mast cells (PI+ACRS vs. control: 58.8+/-19.2 vs. 28.0+/-7.6; P<0.01). The balance between acetylcholine (ACh) and substance P (SP) was also disturbed (ACh: PI+ACRS vs. control: (743.94+/-238.72) vs. (1065.68+/-256.46) pg/g, P<0.05; SP: PI+ACRS vs. control: (892.60+/-231.12) vs. (696.61+/-148.61) pg/g, P<0.05). Nevertheless, similar changes of IL-1beta/IL-10 and ACh/SP were not detected in Ws/Ws rats.

CONCLUSIONThe imbalance of ACh/SP, together with the activation of mucosal immunity induced by post-infection ACRS were lacking in mast cell deficient rats, which supports the premise that mast cells play an important role in cholinergic and peptidergic remodeling in the ileum of rats.

Acetylcholine ; metabolism ; Animals ; Enzyme-Linked Immunosorbent Assay ; Ileum ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Intestines ; immunology ; metabolism ; parasitology ; Male ; Mast Cells ; cytology ; metabolism ; physiology ; ultrastructure ; Microscopy, Electron, Transmission ; Neurotransmitter Agents ; metabolism ; Radioimmunoassay ; Rats ; Substance P ; metabolism ; Trichinella spiralis ; physiology ; Trichinellosis ; immunology

This experimental study was designed to explore the possible mechanisms of Cariporide, a kind of Na+/H+ exchanger inhibitor, for protecting the lung from warm ischemia/reperfusion injury (WI/RI) of isolated rat lung model. Thirty isolated rat lungs were established on the Langendorff apparatus and randomly divided to three groups (n = 10, each): control group (C group), ischemia/reperfusion group (IR group) and Cariporide group (CP group). Mean pulmonary artery pressure (MPAP) and peak airway pressure (pAwP) were monitored continuously. At the end of reperfusion, right bronchoalveolar lavage was performed, bronchoalveolar lavage fluid (BALF) recovery rate (BALFRR) was recorded, and protein content in BALF was measured. Lung water content (LWC), malondialdehyde (MDA) and superoxide dismutase (SOD)of left lung tissue were measured; histomorphology evaluation was performed under light microscope and transmission electron microscope. In comparison with the data from IR group, BALF protein concentration, LWC, MDA content and MPAP content of reperfusion were significantly decreased, but SOD activity was increased in CP group. Histomorphologic feature also showed that pathological change significantly reduced in CP group. In this rat WI/RI model, the mechanism by which the selective Na+/H+ exchanger inhibitor (Cariporide) attenuates lipid peroxidation induced by WI/IR may be: preventing Ca2+ overload via inhibiting the transport of Na+/H2 exchanger-1 (NHE1) in the context of the coupled exchanger, thereby reducing the activation of xanthine oxidase pathway and oxygen free radical liberation which is dependent on certain intracellular Ca2+ concentration, and lastly promoting the endogenous antioxidative mechanism.
Animals ; Antioxidants ; pharmacology ; Guanidines ; pharmacology ; In Vitro Techniques ; Ischemic Preconditioning ; Lung ; blood supply ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; prevention & control ; Sodium-Hydrogen Exchangers ; antagonists & inhibitors ; Sulfones ; pharmacology ; Superoxide Dismutase ; metabolism ; Warm Ischemia

BACKGROUNDTransforming growth factor-beta1 (TGF-beta1) exerts strong fibrogenic potential in culture-activated HSCs. Smad4 is a key intracellular mediator for the transforming growth factor-beta (TGF-beta) superfamily of growth factors. The aim of this study was to assess the effects of the antisense Smad4 gene on Ito cell line, LI90.

METHODSThe recombinant retroviral vector pLXSN-Smad4 was constructed by cloning the rat antisense Smad4 cDNA into the retroviral vector pLXSN. Retroviruses with or without the antisense gene were obtained by transfecting pLXSN-Smad4 and pLXSN vectors into PA317 cells. Human hepatic stellate cells (HSCs) LI90 were infected with these retroviruses followed by selection with G418. The expression of Smad4 was detected by Northern and Western blots. Cell biological characteristics, including cell growth curve, 3H-TdR and 3H-proline uptake by HSCs and the production of extracellular matrix were assessed.

RESULTSmRNA and protein expressions of Smad4 in LI90 cells transfected with retrovirus containing the antisense Smad4 gene were much lower than those in LI90 cells transfected with empty vector or parental LI90 cells. Cells hypoexpressing the Smad4 gene exhibited a slower rate of growth, a lower uptake of 3H-TdR and 3H-proline (P < 0.01), and smaller production of th extracellular matrix, compared with parental LI90 cells and cells transfected with empty retrovirus.

CONCLUSIONSThe antisense Smad4 gene can suppress the expression of the Smad4 gene, reduce endogenous production of Smad4 mRNA and protein, block TGF-beta1 signaling pathway, inhibit activation of Ito cells, obstruct the growth of Ito cells, decrease the production of the extracellular matrix (ECM). Our results may provide a basis for the development of antifibrotic gene therapy.

Cell Line ; DNA, Antisense ; pharmacology ; DNA-Binding Proteins ; antagonists & inhibitors ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Liver Cirrhosis ; therapy ; Retroviridae ; genetics ; Smad4 Protein ; Trans-Activators ; antagonists & inhibitors ; genetics ; Transfection ; Transforming Growth Factor beta ; physiology ; Transforming Growth Factor beta1

OBJECTIVETo compare apoptosis gene expression profiling of U937 cells in suspension culture with that cultivated with mesenchymal stem cells (MSCs), and find out the relationship between drug resistance of leukemia cells and hemopoietic microenvironment.

METHODSU937 cells were cultivated in adhesion culture with MSCs and in suspension culture for 48 hours. Cell cycle was determined by flow cytometry and gene expression profiling by cDNA microarray.

RESULTSCompared with that in suspension, G(0)/G(1) fraction of U937 cells increased in adhesion culture (45.3 +/- 3.1)% vs (32.6 +/- 2.1)%, respectively (P < 0.05), whereas G(2)/M fraction and apoptosis rate were decreased. After 48 h twenty-eight differential expression genes were screened out in 487 apoptosis-related genes, among which 27 were up-regulated and were mainly apoptosis-suppressor genes, apoptosis-promoter genes, cell cycle positive control genes and cell cycle negative control genes. But Bcl-XL was up-regulated most obviously. The only one gene down-regulated was an apoptosis promoter gene.

CONCLUSIONAdhesion culture with MSCs can lead to growth suppression and decrease natural apoptosis of U937 cells. The mechanism was multiple gene effects, but Bcl-XL may be of the most importance.

Apoptosis ; genetics ; Cell Adhesion ; Cell Cycle ; genetics ; Cells, Cultured ; Coculture Techniques ; Flow Cytometry ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; pathology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Oligonucleotide Array Sequence Analysis ; U937 Cells

BACKGROUNDThe hematopoietic microenvironment (HM) plays a critical role in malignant cell growth, patient survival, and response to chemotherapy in hematologic malignancies. However, mechanisms associated with this environmental influence remain unclear. In this study, we investigated the role of bone marrow derived mesenchymal stem cells (MSCs) in U937 cell line, to find out the relations between leukemia drug resistance and the MSCs.

METHODSU937 cells were cultured in suspension or grew adherently with MSCs. The cell growth curve was drawn and the cell cycle was measured by flow cytometry. Apoptosis and sensitivity of U937 to daunoblastina (DNR) were quantified by DNA ladder detection and trypan blue exclusion assays, respectively. The gene expression profile chip technology was used to determine and analyze the changes in apoptosis-related gene expression after adherent culture and the expression of MDR1 mRNA was assessed by reverse transcriptional polymerase chain reaction (RT-PCR) at the same time.

RESULTSIn the adherent culture, the proliferation of the U937 cells was inhibited, the G0/G1 phase cells increased (F = 64.9726, P < 0.0001), G2/M phase cells were decreased (F = 98.1361, P < 0.0001) and the natural apoptosis rate was decreased (F = 24.0866, P < 0.0001) compared with those in the suspended culture. U937 cell viability was enhanced and cell apoptosis was blocked during DNR treatment in adherent culture with MSCs. Thirty-nine differently expressed genes were screened from the 487 apoptosis related genes in the adherent culture U937 cells. Among the 37 upregulated genes, Bcl-XL was upregulated most significantly. Two genes were downregulated. Adherent culture did not induce MDR1 mRNA expression in U937 cells.

CONCLUSIONSMSCs play a role in modulating the proliferation of U937 cells and response of U937 cells to DNR, and Bcl-XL apoptosis-inhibiting gene may be most important in determining the sensitivity of leukemic cells to treatment, which is not related to MDR1.

Apoptosis ; drug effects ; Bone Marrow Cells ; physiology ; Cell Proliferation ; Daunorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Genes, MDR ; Humans ; Immunophenotyping ; Mesenchymal Stromal Cells ; physiology ; U937 Cells ; drug effects

Objective To explore recurrent mechanism and combined treatment for the patients with recurrent intracranial aneurysms. Methods Clinical manifestations and image data of 18 patients with intracranial aneurysms (13 anterior and 5 posterior communicating artery aneurysms)who were treated with aneurysm clipping or endovascular interventional therapy from 1997 to 2004 were comprehensively analyZed to establish individual combined treatment strategies.In the 18 recurrent cases,15 underwent again endovascular embolotherapy,4 were implanted intracranial stent due to wide-necked aneurysm, and 3 received clipping operation. Results Intravascular embolotherapy and clipping operation were both successful in these patients. All cases achieved satisfactory recovery except that 1 case died and 2 suffered from hemiplegia. Conclusions The combined treatment strategies based on individual characteristics of current aneurysms,such as surgical clipping,endovascular therapy combined with stenting,can improve post-operative quality,of patients'life and decrease mortality.

OBJECTIVETo investigate the sensitivity to bleomycin (BLM) in peripheral blood lymphocytes (PBL) among coke-oven workers.

METHODSNinty-four coke-oven workers with exposure to a high level of polycyclic aromatic hydrocarbons and 64 non-coke-oven workers (control) were recruited into this study. PBL was challenged by 8 microg/ml BLM, a known carcinogen, to induce certain amount of DNA damage, the difference of olive tail moment (TM) measured by comet assay before and after BLM treatment reflected the sensitivity towards mutagens.

RESULTSThe distribution of age, sex, and prevalence of smoking and drinking were not significantly different between these two groups. The geometric mean of urinary 1-hydroxypyrene (1-OHP) was significantly higher in coke-oven workers than in controls (9.0 versus 1.5 microg/L, t = -9.317, P < 0.01). The coke-oven workers showed significantly higher sensitivity to BLM than controls (17.7 versus 14.9, t = -2.583, P = 0.01). A large inter-group difference in sensitivity to BLM was observed in both controls and coke-oven workers. Stratification analysis revealed the significant association between high 1-OHP level (> 9.0 microg/L) and increased sensitivity to BLM (F = 4.001, P = 0.05) among coke-oven workers. Smoking subjects showed a significant higher value of sensitivity than nonsmokers in controls but not in coke-oven workers. No significant difference was observed between age, drinking status, coking history or external exposure class and BLM sensitivity.

CONCLUSIONExposure to coke oven emission could increase the sensitivity to mutagens, which might be a reason of high incidence of lung cancer among coke-oven workers.

Adult ; Benzo(a)pyrene ; toxicity ; Bleomycin ; toxicity ; Coke ; Comet Assay ; DNA Damage ; DNA Repair ; Female ; Humans ; Lymphocytes ; drug effects ; Male ; Middle Aged ; Mutagens ; toxicity ; Occupational Exposure

OBJECTIVETo investigate the association of XRCC1 polymorphisms and chromosomal damage levels in peripheral blood lymphocyte in coke-oven workers.

METHODSThe study included 141 coke-oven workers who exposed to a high level of polycyclic aromahaplotpetic hydrocarbon and 66 non-exposed controls. Urinary 1-hydroxypyrene and chromosome damage in peripheral lymphocyte were measured. Four -tagging single nucleotide polymorphisms in XRCC1 gene, including C26304T, G27466A, G28152A and G36189A, were detected and the XRCC1 haplotypes were estimated by using an extension of Clark algorithm. The associations between haplotype pairs and micronuclei data were assessed by analysis of covariance in the exposed and non-exposed groups.

RESULTSThe geometric means of urinary 1-hydroxypyrene levels in coke-oven workers and the controls were 12.0 and 0.7 micromol/mol Cr respectively (P < 0.01). The cytokinesis-block micronucleus cytokinesis-block micronucleus frequencies (number of micronucleus per 1 000 binucleated lymphocytes) was significantly higher in coke-oven workers (0.95 +/- 0.66)% than in the controls (0.40 +/- 0.36)%, P < 0.01. The haplotype CGGG was associated with the decreased frequencies of total micronucleus, and the haplotypes TGGG (P = 0.01) and CGAG (P < 0.05) were associated with the increased frequencies of total micronucleus in the multivariate analysis with adjustment for covariates among coke-oven workers.

CONCLUSIONSThe genetic polymorphisms of XRCC1 gene could influence the chromosome damage levels in coke-oven workers.

Adult ; China ; Chromosome Breakage ; Coke ; poisoning ; DNA-Binding Proteins ; genetics ; Female ; Genetic Predisposition to Disease ; Haplotypes ; Humans ; Lymphocytes ; metabolism ; Male ; Micronucleus Tests ; methods ; Middle Aged ; Occupational Diseases ; etiology ; genetics ; urine ; Occupational Exposure ; adverse effects ; analysis ; Polymorphism, Single Nucleotide ; Pyrenes ; analysis ; X-ray Repair Cross Complementing Protein 1

Objective To investigate the effect of different serum sodium levels on heart function and prognosis in very elderly patients with heart failure. Methods Totally 152 elderly patients(aged ≥ 85 years)with hyponatremia were enrolled.Based on the serum sodium level ,they were divided into mild(125 mmol/L ≤ Na+ < 135 mmol/L) ,moderate(125 mmol/L ≤ Na+ < 135 mmol/L) ,and severe hyponatremia(Na+ < 115 mmol/L)groups ,and 76 patients with normal blood sodium level(Na+ ≥ 135 mmol/L)were selected as control group. The general data ,levels of serum creatinine and N-terminal pro-brain natriuretic peptide (NTproBNP)and prognosis were compared among the above groups. Results Compared with the control group ,patients with severe hyponatremia were much older ,with more diuretics administration ,higher level of serum creatinine , lowerlevelofeGFRandleftventricularejectionfractions[(38.1±3.9)％ vs.(45.2±9.7)％ ].There were higher incidences of coronary atherosclerotic heart disease [72.4％ (n=21)vs. 56.6％ (n=43)] ,diabetes[41.4％ (n=12)vs. 23.7％ (n=18)] ,chronic renal insufficiency [22.6％ (n=8)vs. 11.8％ (n=9)] ,stroke[20.6％ (n=6)vs. 9.2％ (n=7)]in patients with severe hyponatremia compared with other groups. The severe hyponatremia group showed significantly higher levels of NTproBNP [(4823. 9 ± 588. 1 )ng/L v s. (1124. 4 ± 349. 1 )ng/L ,(1836. 2 ± 369. 3 )ng/L ,(2894. 1 ± 687. 3 )ng/L ]and higher rates of mortality [27.6％ (n=8) vs.5.3％ (n=4) ,7.6％ (n=6) ,13.6％ (n=6)]as compared to the control group ,mild and moderate hyponatremia groups. Furthermore ,length of stay in hospital were markedly longer in patients with severe hyponatremia than those with mild and moderate hyponatremia[(11.1 ± 7.1)d vs. (19.6 ± 5.7)d ,(16.1 ± 4.2)d] . Conclusions Hyponatremia is a common type of electrolyte disorder in very elderly patients with heart failure ,and severe hyponatremia may predictand increased risk of death.