In recent years the interaction between host and gut microbiota has attracted increasing attention. However, intestinal flora dysbiosis may lead to many diseases, and there is increasing evidence that the intestinal microbiota in patients with chronic kidney disease (CKD) is associated with the pathophysiological status of the host. "Gut-kidney axis" provides a better explanation of the two-way communication between intestinal flora and CKD. Impaired kidney function leads to dysbiosis of intestinal flora and an altered intestinal flora can damage the intestinal mucosal barrier and facilitate the entry into the bloodstream of harmful bacteria, which can induce chronic inflammation and thus accelerate renal injury. In addition, the accumulation of nephrotoxic metabolites from an altered intestinal flora can aggravate CKD in the "gut-kidney axis". Among them, p-cresol sulfate, indoxyl sulfate and trimethylamine oxide are the most widely studied metabolites of nephrotoxicity, and their renal toxicity has been widely confirmed in basic research and clinical studies. Current studies show that the intestinal microbiota-metabolite network is closely related to the occurrence and development of chronic kidney disease. Thus, intervention in the intestinal microbiota may provide a new approach to the prevention and treatment of chronic kidney disease.

Benzene is classified as Group 1 carcinogen by IARC. It has been found that benzene induces hematotoxicity even in low dose exposure. The identification of key events during benzene induced hematotoxicty leads to adjustment of occupational exposure limits of benzene. In this review, we focus on the exposure, metabolism, target organs, key epigenetic changes, toxicty effects and end points of low-dose chronic benzene exposure induced hematotoxicity and finally discuss the perspectives on the future study of this area.
Benzene ; toxicity ; Carcinogens ; toxicity ; Epigenesis, Genetic ; Humans ; Occupational Exposure

Adenocarcinoma/surgery* ; Esophageal Neoplasms/surgery* ; Esophagogastric Junction/surgery* ; Gastrectomy ; Humans ; Margins of Excision ; Neoadjuvant Therapy ; Retrospective Studies ; Stomach Neoplasms/surgery*

OBJECTIVETo ascertain the frequency of prothrombin (FII) gene 3'-untranslated region (3'-UT) G20210A variant and to explore whether this mutation is related to arterial thrombosis in Chinese Zhuang population.

METHODSSeventy-six patients with cerebral thrombosis, 23 patients with myocardial infarction and 106 healthy Chinese Zhuang persons were studied. The G20210A mutant allele of the prothrombin gene in all blood specimens was investigated by DNA extraction, polymerase chain reaction amplification, Hind III digestion and polyacrylamide gel electrophoresis.

RESULTSThe patients and normal control subjects were all homozygous for the normal G20210G allele, and there was no FII G20210A variant.

CONCLUSIONFactor II gene 3'-UT G20210A mutant allele is absent in the 99 Chinese Zhuang ethnic patients with ischemic stroke and myocardial infarction and is absent in 106 normal healthy Zhuang people. FII G20210A mutation may not be a major risk factor for thrombogenesis in ethnic Chinese.

3' Untranslated Regions ; genetics ; Adult ; Aged ; Aged, 80 and over ; Base Sequence ; China ; DNA Mutational Analysis ; Ethnic Groups ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Humans ; Intracranial Thrombosis ; genetics ; Male ; Middle Aged ; Mutation ; Myocardial Infarction ; genetics ; Polymerase Chain Reaction ; Prothrombin ; genetics

BACKGROUNDA nonsteroidal anti-inflammatory drug, indomethacin, has been shown to have anti-leukemic activity and induce leukemic cell apoptosis. This study was to elucidate the mechanism of indomethacin-induced K562 cell apoptosis.

METHODSK562 cells were grown in RPMI 1640 medium and treated with different doses of indomethacin (0 micromol/L, 100 micromol/L, 200 micromol/L, 400 micromol/L, 800 micromol/L) for 72 hours. The cells were harvested, and cell viability or apoptosis was analyzed using MTT assay and AO/EB stain, combining laser scanning confocal microscopy (LSCM) technique separately. For the localization and distribution of intracellular caspase-3 or caspase-8 protein, immunofluorescence assay was carried out. To reveal the activation of caspase-3 or caspase-8 in indomethacin-treated cells, Western blot detection was used. The change in intracellular free calcium was determined by Fluo-3/Am probe labeling combined with LSCM.

RESULTSIndomethacin could lead to K562 cell apoptosis and inhibit cell viability in a concentration-dependent manner. An increased expression of intracellular caspase-3 or caspase-8 was observed at higher doses of indomethacin (400 - 800 micromol/L). Western blot results showed upregulation and activation in both caspase-3 and caspase-8 protein. Under indomethacin intervention, the levels of intracellular free calcium showed a significant increase. Blocking the activity of cyclooxygenase did not abolish the effects of indomethacin on K562 cell apoptosis.

CONCLUSIONSActivation and upregulation of caspase-3 or caspase-8 protein were responsible for Indomethacin-induced K562 cell apoptosis. Variation of intracellular free calcium might switch on the apoptotic pathway and the proapoptotic effect of indomethacin might be cyclooxygenase-independent.

Apoptosis ; drug effects ; Calcium ; metabolism ; Caspase 3 ; Caspase 8 ; Caspases ; genetics ; metabolism ; Cyclooxygenase Inhibitors ; pharmacology ; Enzyme Activation ; Gene Expression Regulation, Enzymologic ; drug effects ; Humans ; Indomethacin ; pharmacology ; K562 Cells

This study was aimed to observe whether expressions of JAK2, STAT1, STAT5 proteins in indomethacin-treated CML cells involved in the proliferation inhibition of CML cells, and elucidate the pharmacological mechanism of indomethacin anti-leukemia. MTT assay and trypan blue dye exclusion test were used to detect the inhibitory effect of indomethacin on CML cells proliferation. JAK2, STAT1, STAT5 proteins were analyzed by Western blot; the subcellular distribution of STAT1, STAT5 were detected with indirect immunofluorescence technique. The results showed that indomethacin at >or= 400 micromol/L significantly inhibited the proliferation of CML cells and down-regulated the expression of STAT1, STAT5 protein, no JAK2 change was observed. STAT1 and STAT5 were located in cytoplasm. It is concluded that indomethacin inhibits the proliferation of CML cells and the mechanism may be related to down-regulated expression of STAT, or blockage of cells growth signals.
Antineoplastic Agents ; pharmacology ; Blotting, Western ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Fluorescent Antibody Technique ; Humans ; Indomethacin ; pharmacology ; Janus Kinase 2 ; metabolism ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism ; pathology ; STAT1 Transcription Factor ; metabolism ; STAT5 Transcription Factor ; metabolism ; Signal Transduction ; drug effects

OBJECTIVETo investigate the effects of indomethacin on HL-60 cell proliferation and apoptosis, and elucidate partly the molecular mechanism about the anti-leukemia effect of indomethacin by studying beta-catenin signal transduction pathway.

METHODSHL-60 cells were treated with indomethacin at different concentrations (0, 25, 50, 100, 200, 400 micro mol/L). Cells viability and proliferation were determined by Trypan blue staining and cell counting respectively. DNA ladder pattern and cell morphology were used to identify cell apoptosis. The expression and cleavage of caspase-3, beta-catenin, c-myc were detected by Western blot techniques.

RESULTSIndomethacin could significantly inhibit HL-60 cells proliferation and induce cells apoptosis with a time and dose dependent manner. An up-regulating expression and cleavage of caspase-3 were observed. Indomethacin could inhibit beta-catenin expression and induce its degradation, c-myc protein exhibited a down-regulation with a concentration dependent manner.

CONCLUSIONIndomethacin could inhibit HL-60 cell proliferation and induce cells apoptosis. Anti-leukemia effect of indomethacin was associated with the inhibition of "beta-catenin --> c-myc" signal pathway.

Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Caspase 3 ; Caspases ; metabolism ; Cytoskeletal Proteins ; metabolism ; Dose-Response Relationship, Drug ; HL-60 Cells ; Humans ; Indomethacin ; pharmacology ; Proto-Oncogene Proteins c-myb ; metabolism ; Signal Transduction ; drug effects ; Time Factors ; Trans-Activators ; metabolism ; beta Catenin

OBJECTIVETo study the plasma HCII activity and antigen level variations and their relationship with arterial and deep venous thrombotic diseases.

METHODSSeventy-five patients with brain infarction (BI), 50 myocardial infarction (MI), 36 deep venous thromboembolic disease (DVT) and 50 healthy controls were entered in this study. Plasma HCII activity was measured with chromogenic substrate method and the HCII antigen level by Western blotting assay. Plasma antithrombin (AT) activity was detected for the HCII deficiency individuals with DVT using chromogenic substrate method.

RESULTSThere was no significant difference in the mean plasma HCII activity and antigen levels between BI group [(99.97 +/- 21.14)% and 0.96 +/- 0.24], MI group [(98.18 +/- 29.35)% and 0.95 +/- 0.20] and healthy controls [(96.80 +/- 20.11)% and 0.93 +/- 0.19]. The plasma HCII activity and antigen concentrations in patients with DVT [(89.57 +/- 17.12)% and 0.87 +/- 10.18] tended to be decreased as compared with healthy controls, but they were not significant. No significant difference was found for the prevalence of HCII deficiency between patient groups and control group. The HCII deficiency individuals with DVT had normal AT activity and fibrinogen concentration.

CONCLUSIONSPlasma HCII deficiency may not be the risk factor for arterial thrombosis in the Han population of Hunan Chinese. It is needed to further confirm if decreased plasma HCII is correlated with venous thrombosis.

Adult ; Aged ; Blotting, Western ; Cerebral Infarction ; blood ; etiology ; Female ; Heparin Cofactor II ; analysis ; deficiency ; immunology ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; blood ; etiology ; Venous Thrombosis ; blood ; etiology

OBJECTIVETo screen phosphopeptide specific for acute leukemia.

METHODSMononuclear cells from bone marrow were collected from 16 newly diagnosed acute lymphoblastic leukemia (ALL) and 20 acute myeloid leukemia (AML) patients. Peptides were extracted and purified, analyzed by immunoprecipitation and liquid chromatography coupled with tandem mass spectrometry (LC-MS).

RESULTS(1) Non-receptor tyrosine kinase family members Fyn, Yes, Src widely expressed in acute leukemia; (2) Some phosphopeptides, including non-receptor tyrosine kinase family members Abl/iso1 and Abl, non-receptor Ser/Thr protein kinase family members Bcr, JNK2, JNK2 iso2, Adaptor/scaffold members Cas-L, Cbl, CrkL CENTD1 (Centaurin delta1) ZO2, transcriptor GFR-1 and phosphatase SHIP-2 were detected in Ph positive ALL, but not in other kinds of ALL. (3) Hck, Lyn and Fgr selectively expressed in AML (except AML-M(3)).

CONCLUSIONSome phosphopeptides were specific for ALL and AML, and may be useful for diagnosis and therapy of acute leukemia.

Chromatography, High Pressure Liquid ; Humans ; Immunoprecipitation ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Neoplasm Proteins ; analysis ; Phosphopeptides ; analysis ; Phosphorylation ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; metabolism ; Proteomics ; Tandem Mass Spectrometry

OBJECTIVETo explore the relationship between microRNA and drug resistance in leukemia treatment by screening and identifying the microRNAs which differentially express in K562 cell line and its adriamycin resistant cells-K562/A02 cell line.

METHODSThe drug resistance potency of K562/A02 cells was evaluated by MTT assay. P-gp expression of K562 and K562/A02 cells were detected by flow cytometry (FCM). The differentially expressed microRNAs in K562 and K562/A02 cells were analyzed by microarray technique and Real Time RT-PCR.

RESULTSThe resistance to adriamycin (ADM) of K562/A02 cells was 180 fold greater than that of K562 cells. P-gp expression rate of K562 and K562/A02 cells was 0.2% and 86%, respectively. Twenty-two microRNAs expressed differentially in K562 and K562/A02 cells (P < 0.01). As compared to K562 cells, expressions of miR-221, miR-155 and miR-451 were up-regulated by more than two fold, while expression of miR-98, miR-181a, let-7f, let-7g, miR-424 and miR-563 down-regulated by more than two fold in K562/A02 cells. The results of real time RT-PCR were consistent with that of microarray. Of note, differential expressions of miR-451, miR-155, miR-221, let-7f and miR-424 were remarkable.

CONCLUSIONK562/A02 cells show a different microRNA expression profile as compared to its parental K562 cells, suggesting microRNAs including miR-221, miR-155, miR-451, let-7f and miR-424 may be involved in the mechanism of drug resistance in leukemia. These differentially expressed microRNAs provide potential novel targets for overcoming drug-resistance.

Doxorubicin ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; genetics ; Humans ; K562 Cells ; MicroRNAs ; genetics