15-hydroxyeicosatetraenoic acid (15-HETE) plays an important role in hypoxia-induced pulmonary vasoconstriction. Release of nitric oxide (NO) is apparently decreased and activity of endothelial nitric oxide synthase (eNOS) is impaired in chronic hypoxia. However, little is known whether 15-HETE contributes to eNOS/NO pathway in the constriction induced by 15-HETE. We examined the response of rat pulmonary artery (PA) rings to 15-HETE, the production of NO, total eNOS expression and the phosphorylation of eNOS in bovine pulmonary artery endothelial cells (BPAECs) stimulated by 15-HETE. Rat PA rings were divided into three groups: endothelium intact group, endothelium denuded group, and nitro-L-arginine methyl ester (L-NAME, 0.1 mmol/L, an inhibitor of eNOS) group. Constrictions to 15-HETE were significantly enhanced in endothelium denuded group and L-NAME group (both P< 0.05 vs endothelium intact group, n= 9); BPAECs were incubated in different conditions to test nitrite production by Greiss method. Nitrite production was significantly reduced by 1 mumol/L 15-HETE (P<0.05), and increased by the lipoxygenase inhibitors, 10 mumol/L cinnamyl 3,4- dihydroxy-[alpha] -cyanocinnamate (CDC, P< 0.05) and 0.1 mmol/L nordihydroguiairetic acid (NDGA, P< 0.01 ); Western blot analysis of extracts from BPAECs incubated with 15-HETE in different time was carried out to test total eNOS expression, and the expression was changed unobviously. Immunoprecipitation (IP) and Western blot analysis of cell extracts from BPAECs treated with 2 mumol/L 15-HETE in different length of time were accomplished, using phospo-eNOS-threonine 495 (Thr495, an inhibitory site) antibody for IP, and eNOS or 15-lipoxygenase (15-LO) antibodies for Western blot. 15-HETE depressed eNOS activity by increasing the levels of phospho-eNOS-Thr 495. The data suggest that eNOS/NO pathway is involved in PA constrictions induced by 15-HETE and that 15-HETE depresses eNOS activity by phosphorylation in Thr495 site. The protein interaction between phospho-eNOS (Thr495) and 15-LO is discovered for the first time.
Animals ; Cattle ; Down-Regulation ; drug effects ; Endothelium, Vascular ; cytology ; drug effects ; enzymology ; Hydroxyeicosatetraenoic Acids ; pharmacology ; In Vitro Techniques ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Pulmonary Artery ; cytology ; enzymology ; physiology ; Rats ; Rats, Wistar

OBJECTIVETo investigate the in vitro effect of iron overload on the generation of reactive oxygen species (ROS) and of bone marrow (BM) cell function.

METHODSBM mononuclear cells (BMMNCs) were cultured with ferric citrate (FAC) at different concentrations and for different time to create iron overload and confirmed by the detection of cellular labile iron pool (LIP). The changes of ROS, apoptosis, hematopoietic colony formation (CFU-E, BFU-E, CFU-GM and CFU-mix) and the percentage of the CD34 + cells percentage were analyzed. The differences of these index were tested after the iron overload treated with deferasirox (DFO) or antioxidants (N-acetyl-L-cysteine, NAC).

RESULTS1) When BMMNCs were cultured with FAC, the LIP was found to increase in a time and concentration dependent manner. The intracellular LIP reached maximum at 400 micromol/L of FAC for 24 hours. 2) The ROS of total cells, leukocytes and erythrocytes increased to 1.77, 1.75 and 2.12 fold respectively compared with that of normal control when cells were cultured at 400 micromol/L of FAC for 24 hours . DFO and NAC could reduce the ROS efficiently (P<0.05). 3) The apoptotic rates of the FAC treated cells [(24.80 +/- 2.99)%] increased significantly compared with that of normal control [(8.90 +/- 0.96)%]. The capacity of hematopoietic colony formation in FAC treated cells decreased markedly compared with that of normal control (P<0.05). The percentage of CD34+ cells of FAC treated cells [(0.39 +/- 0.07)%] also decreased significantly compared with that of normal control [(0.91 +/- 0.12)%]. And these changes could be recovered by addition of NAC or DFO.

CONCLUSIONIron overload can affect the hematopoiesis by inducing the generation of ROS and this damage could be corrected by removing the excess iron and ROS of the BM cells. These findings might improve the treatment of dyshematopoiesis in patients with iron overload.

Bone Marrow Cells ; physiology ; Cells, Cultured ; Culture Media ; chemistry ; Erythrocytes ; Ferric Compounds ; pharmacology ; Hematopoiesis ; Humans ; Iron Overload ; Reactive Oxygen Species ; metabolism

This study was aimed to detect the expression of leukemia stem/progenitor cell (LSPC) related genes (ABCB1, BMI-1, HOXB4) in the patients with acute leukemia, and to explore its clinical significance in acute leukemia. Bone marrow samples were collected from de novo acute leukemia patients (41 cases), patients with complete remission (CR, 16 cases) and the patients with non-malignant hematologic diseases (10 cases) respectively. And the expressions of ABCB1, BMI-1, HOXB4 genes were detected by comparative real-time quantitative PCR (RQ-PCR) with SYBR Green assay. The results showed that the expressions of ABCB1, BMI-1, HOXB4 were not detected in the patients with non-malignant hematologic diseases, but were higher (relative expressive level: 4.26 ± 2.26, 3.72 ± 1.91, 3.74 ± 2.38) in de novo acute leukemia patients and lower (relative expressive level: 2.14 ± 1.47, 2.07 ± 0.99, 1.47 ± 0.89) in the acute leukemia patients with CR (p < 0.05). The expressions of LSPC related genes were lower (relative expressive level: 1.77 ± 1.29, 2.09 ± 1.26, 1.78 ± 1.49) in the patients acquired CR/partial remission (PR) than those in the patients not acquired CR/PR (relative expressive level: 7.23 ± 1.78, 3.96 ± 0.92, 4.48 ± 2.57) (p < 0.01). Univariate analysis revealed that there were more cases with the expression of LSPC immunophenotype (CD34(+)CD38(-)CD96(+) and CD34(+)CD38(-)CD123(+)) and more hyperleukocytosis cases in patients with any higher expression of LSPC related gene (p < 0.05). Analysis of multiple parameters discovered larger significance (p < 0.01). It is concluded that there is a good relationship between LSPC related genes (ABCB1, BMI-1, HOXB4) and LSPC immunophenotype. The expression of LSPC-related genes is higher in de novo acute leukemia patients, and lower in patients acquired CR/PR. The patients with higher expressed LSPC-related genes display worse response to chemotherapy, lower CR/PR rate and higher leukocytosis, the analysis of multiple parameters may be a good method for assessing the therapeutic efficacy/prognosis of acute leukemia.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Adolescent ; Adult ; Aged ; Female ; Gene Expression ; Homeodomain Proteins ; genetics ; metabolism ; Humans ; Immunophenotyping ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Middle Aged ; Neoplastic Stem Cells ; metabolism ; Polycomb Repressive Complex 1 ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism ; Young Adult

OBJECTIVETo evaluate the expression of CD117 in human testicular germ cell tumors and its value in the differential diagnosis of seminoma and nonseminoma.

METHODSSeventy-four human testicular germ cell tumor specimens were studied by ABC kit immunohistochemical staining detection using CD117 monoclonal antibodies. The immunoreaction scores (IRS) of all the specimens were calculated and analysed for their clinical significance.

RESULTSAmong the 74 germ cell tumors, 31 out of 32 (9 6.9%) seminomas showed positive staining of the CD117 mostly on the cell membrane. Four of 31 (12.9%) nonseminomas displayed a weak positive staining of CD117 only in the cytoplasm of a few cells. In 10 of 11 mixed germ cell tumors, a relatively weak expression of CD117 was shown only in the seminoma component. The CD117 expression was diagnostically decreased from seminoma to mixed seminoma and to nonseminoma successively, with IRS of 6.82 +/- 2.76, 1.25 +/- 0.42 and 0.60 +/- 0.16, respectively. There was a significant difference in the CD117 expression between seminoma and nonseminoma (P < 0.05). CD 117 proteins were positively expressed in all the 20 specimens of the normal testis.

CONCLUSIONThe detection of CD117 by immunohistochemical staining using CD117 monoclonal antibodies is a newly developed useful method for differentiating seminoma and nonseminoma.

Adolescent ; Adult ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Male ; Neoplasms, Germ Cell and Embryonal ; diagnosis ; metabolism ; Predictive Value of Tests ; Proto-Oncogene Proteins c-kit ; biosynthesis ; Seminoma ; diagnosis ; metabolism ; Testicular Neoplasms ; diagnosis ; metabolism

OBJECTIVETo study the efficacy and safety of the coronary interventional therapy to prevent electrical storm (ES) in patients with coronary artery diseases (CAD) but without myocardial infarction or ischemic cardiomyopathy.

METHODSCoronary angiography and stent implantation were performed in CAD patients with ES as major symptom, according to the standardized methods. Holter Electrocardiography was recorded regularly during follow-up.

RESULTSSix patients, five male and one female, with mean age of 49.5 +/- 9.1 year-old, were hospitalized. In 2 patients with repetitive syncope, multiple episodes of ventricular tachycardia and/or ventricular fibrillation (VF) were documented by Holter recording. One patient developed VF during exercise test. Three patients experienced chest pain and multiple episodes of cardiac arrest. Before procedure, averaged 16.5 +/- 5.3 episodes of syncope or VF were documented in 6 patients. Coronary angiography revealed severe one or multi-vessel diseases. Total 8 stents, including 3 drug-eluting stents, were implanted in 6 patients. Symptom, ST-T changes and ES disappeared after coronary stenting. During 4 month to 6.5 year follow-up (mean 47.7 +/- 30.7 months), ES was not documented, no appropriated shock occurred in patients with implantable defibrillator for 6.5 years.

CONCLUSIONIn CAD patients without myocardial infarction, coronary stenting can relieve the ischemic substrate of ES, hence prevents sudden death effectively.

Adult ; Angioplasty, Balloon, Coronary ; Coronary Angiography ; Coronary Artery Disease ; physiopathology ; therapy ; Electrocardiography ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Stents

This study was aimed to investigate the relationship between cytogenetic evolution and disease progression in patient with MDS-RAEB. By a long term (6 years) follow-up of a patient with MDS-RAEB, peripheral blood cell count, bone marrow cell morphology and conventional cytogenetics were monitored regularly. In addition, fluorescence in situ hybridization (FISH) was applied to confirm the aberrant karyotype. The results indicated that this patient was failed with conventional chemotherapy of AML, but had response to ATRA and 6-MP in the 72 months follow-up. At initial diagnosis, the cytogenetics analysis showed normal karyotype, whereas 46, XY, 2q+[1]/46, XY[19] was found at 48 months, 46, XY, dup(1q)[3]/46, XY[7] at 56 months, and dup (1) as well as der (11) with complex karyotype at 68 months, which was accompanied by progressive decrease of platelet count. It is concluded that karyotype evolution is perhaps associated with progression of MDS.
Adult ; Chromosome Aberrations ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 11 ; Follow-Up Studies ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics

OBJECTIVETo study the indoor environmental factors associated with the prevalence of asthma and related allergies among school children.

METHODSA cluster sampling method was used and the ISAAC questionnaire was conducted. A total of 4612 elementary students under Grade Five of 7 schools were enrolled in the survey for the impact of indoor environmental factors on the prevalence of asthma and related allergies in several urban and suburban schools of Beijing.

RESULTSA total of 4060 sample were finally analyzed including 1992 urban and 2068 suburban. The prevalence of wheeze, allergic rhinoconjunctivitis and atopic eczema in the past 12 months was 3.1% (61/1992), 5.3% (106/1992), 1.1% (22/1992) among urban children while 1.3% (27/2068), 3.1% (65/2068), 1.0% (22/2068) among suburban children respectively. The prevalence of wheeze and allergic rhinoconjunctivitis of the past 12 months in urban were both significantly higher than that in suburban (χ(2) = 14.77, 11.93, P < 0.01). The incidences of having asthma and eczema ever among urban children (5.3% (105/1992), 29.4% (586/1992)) were significantly (χ(2) = 39.03, 147.22, P < 0.01) higher than that among suburban (1.7% (35/2068), 13.8% (285/2068)). Although the distributions of indoor environmental factors were similar in both areas, passive smoking and interior decoration had different influence on the prevalence of asthma and related allergies among school children in the two areas. The significant impact of passive smoking on having asthma ever among suburban children was observed (OR = 2.70, 95%CI = 1.17 - 6.23) while no significant result in urban (OR = 1.06, 95%CI = 0.71 - 1.58); the percentage of interior decoration was 84.0% (1673/1992) among urban children and 80.0% (1655/2068) among suburban children, there was significant impact of interior decoration on the prevalence of having eczema ever among urban children (OR = 1.57, 95%CI = 1.17 - 2.10) but no significant results were found in suburban sample (OR = 1.06, 95%CI = 0.76 - 1.48).

CONCLUSIONThe prevalence of asthma and related allergies among school children is much higher in urban areas than that in suburban areas and the indoor environmental factors such as passive smoking and interior decoration may differently explain the prevalence of asthma and related allergies in the two areas.

Adolescent ; Air Pollution, Indoor ; analysis ; Asthma ; epidemiology ; etiology ; Child ; Child, Preschool ; China ; epidemiology ; Cities ; Environmental Exposure ; Female ; Humans ; Hypersensitivity ; epidemiology ; etiology ; Male ; Prevalence ; Risk Factors ; Students ; Suburban Population ; Surveys and Questionnaires ; Urban Population

OBJECTIVETo investigate the therapeutic effects and mechanisms of curcumin treatment on hepatic fibrosis.

METHODSA model of hepatic fibrosis was established using carbon tetrachloride intraperitoneal injections in rats. Curcumin was administered to one group of the model rats (curcumin group) and the other rats were used as controls (control group). Serum levels of ALT, AST, HA, LN, PCIII, and NO were measured, and Hyp, MDA, and SOD in liver tissues were measured. Liver tissue slides were stained with HE and Masson staining to study the pathological changes in the livers. Grades of hepatic fibrosis were evaluated according to a semiquantitative scoring system.

RESULTSIn the curcumin group, serum levels of ALT, AST, NO, HA, LN, PCIII, MDA, and Hyp, were (218.50+/-48.89) U/L, (376.60+/-79.13) U/L, (47.96+/-6.53) micromol/L, (289.96+/-60.43) mg/L, (107.35+/-27.24) mg/L, (148.95+/-28.63) microg/L, (236.10+/-30.54) nmol/g, (478.40+/-75.74) microg/g and all were lower than those of the control group (693.75+/-117.57) U/L, (892.50+/-105.69) U/L, (70.95+/-10.23) micromol/L, (468.22+/-93.45) mg/L, (346.44+/-75.08) mg/L, (279.82+/-54.00) microg/L, (402.25+/-39.16) nmol/g, and (752.50+/-77.62) microg/g. The differences were significant. In the curcumin group, the level of SOD (90.39+/-21.23) in the liver tissues was significantly higher than that of the control group (46.52+/-20.01). The hepatic fibrosis scores in the curcumin group were significantly lower than those of the control group. These effects were dose-dependent.

CONCLUSIONSCurcumin reduces rat hepatic fibrosis. Anti-peroxidation and regulation of collagen metabolism in liver tissues may be involved in the therapeutic effectiveness of curcumin on hepatic fibrosis.

Animals ; Curcumin ; therapeutic use ; Drugs, Chinese Herbal ; metabolism ; therapeutic use ; Lipid Peroxidation ; Liver Cirrhosis, Experimental ; drug therapy ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

To investigate the expression of phospholipase C-gamma1 (PLC-gamma1) in mouse embryonic tissues, serial tissue sections were prepared routinely for immunocytochemistry for PLC-gamma1. The results showed that PLC-gamma1 was expressed in the cartilage, skeletal muscles, myocardium, the collecting tubule of the kidney, connective tissues and the brain, suggesting the important role PLC-gamma1 and the related signal pathway may play in the development of mouse embryonic tissues.
Animals ; Brain ; embryology ; enzymology ; Cartilage ; embryology ; enzymology ; Embryo, Mammalian ; enzymology ; Female ; Fetal Heart ; enzymology ; Immunohistochemistry ; Kidney ; embryology ; enzymology ; Mice ; Muscle, Skeletal ; embryology ; enzymology ; Phospholipase C gamma ; biosynthesis ; Pregnancy

Shengmaiyin is widely used in the treatment of arrhythmia and has achieved a good effect. Due to the complexity of traditional Chinese medical formula, the pharmacological mechanism of Shengmaiyin in the treatment of arrhythmia is unclear. In this study, we used the internet-based Computation Platform (www.tcmip.cn) to explore the molecular mechanism. Shengmaiyin was found to treat the arrhythmia by modulating the pathway related to energy metabolism such as carbon metabolism, purine metabolism, carbohydrate metabolism, or by regulating the level of ATP. In this study, we find that the main active drug component in Shengmaiyin may be ginseng.