Three-dimensional structure model of azoreductase AZR of Rhodobacter sphaeroides was con- structed using homology modeling method. It is a flavodoxin adopting ?/? structure. Structure alignment of two different types of flavin-dependent azoreductases revealed that they possessed high similarity. Based on sequence and structure analysis, site-directed mutagenesis of K109H and K109A were performed. The opti- mal pH values are pH 6 and pH 9 for K109H and K109A mutant protein, respectively. The optimal tempera- ture (30℃) is not affected by mutagenesis. Positively charged residues at position 109 is necessary for the binding of methyl red, while K109H is not a conserved mutagenesis for the binding of NADPH. K109 may only be involved in the binding of the 2’-phosphate group of NADPH and have no effect on the binding of NADH.

For the efficient interception performance of the membrane, some biological aspect in membrane bioreactor are different from those in the activated sludge process. The physiological and biochemical characteristics of microbial community, activated sludge, and microbial products in the membrane bioreactor are illustrated in this paper.

OBJECTIVETo investigate the in vitro effect of erythropoietin (EPO) on hepcidin of monocytes and its molecular mechanisms.

METHODSHepcidin and signaling molecules including C/EBPalpha, Smad1/5/8, p-Smad1/5/8 and p-STAT3 were detected by real time PCR and Western blot. THP-1 monocytes were stimulated by interleukin-6 (IL-6) or lipopolysaccharide (LPS). EPO receptor (EPOR) antibody was added to observe its antagonistic effect on EPO and impact on the signaling proteins.

RESULTSEPO suppressed mRNA expression of THP-1 hepcidin of monocytes induced by 20 ng/ml IL-6 or 1 microg/ml LPS in both dose and time dependent manner. The most decrease of hepcidin expression was observed at 2 IU/ml EPO for 6 hours. EPO also down-regulated hepcidin protein induced by 20 ng/ml IL-6. At 2 IU/ml EPO for 6 hours hepcidin protein was down-regulated, as was C/EBPalpha, p-Smad1/5/8 and p-STAT3. Antibody to EPOR antagonized the down-regulation of EPO on hepcidin and signaling proteins.

CONCLUSIONSMonocytes hepcidin can be reduced by EPO when stimulated by IL-6 or LPS. The mechanism of which may be at least in part, via suppression of C/EBPalpha, p-Smad1/5/8 and p-STAT3 signaling.

Antimicrobial Cationic Peptides ; metabolism ; Cells, Cultured ; Erythropoietin ; pharmacology ; Hepcidins ; Humans ; Interleukin-6 ; pharmacology ; Lipopolysaccharides ; pharmacology ; Monocytes ; drug effects ; metabolism ; Signal Transduction

This study was purposed to investigate the effect of multiple myeloma patients' sera on hepcidin mRNA expression of Hep-3b hepatoma cell line and effect of human interleukin-6 (IL-6) antibody or recombinant human erythropoietin (rhEPO) on hepcidin mRNA expression. The clinical information and serum of multiple myeloma patients were collected. Their sera of a final concentration of 10% were added into Hep-3b cell medium. The mRNA from Hep-3b cells was extracted, and hepcidin mRNA expression was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). A final concentration of 10 ng/ml human IL-6 antibody and 2 U/ml rhEPO were added into the medium respectively. The results showed that the sera of untreated multiple myeloma patients elevated hepcidin mRNA expression of Hep-3b cells, compared with healthy controls and iron deficiency anemia patients. This effect was fully neutralized by human IL-6 antibody or rhEPO. The hemoglobin (Hb) level was stable during the follow up of regularly treated multiple myeloma patients and the effect of MM patient serum on Hep-3b cell hepcidin mRNA expression was reduced. It is concluded that the hepcidin mRNA expression of Hep-3b cell can be increased by untreated multiple myeloma patient serum. This promotive effect can be antagonised by IL-6, which suggests that IL-6 may be possible to elevate expression level of hepcidin in Hep-3b cells and results in anemia of chronic disease (ACD). The above mentioned promotive effects also can be suppressed by rhEPO, which indicates that the rhEPO may possess curative effect for ACD disease. During short-term follow-up of treated patients with multiple myeloma the Hb level is stable, the influence of patients serum on hepcidin mRNA of Hep-3b cells decreases, which shows the stabilization of disease and amelioration of ACD patient status.
Adult ; Aged ; Antibodies, Monoclonal ; pharmacology ; Antimicrobial Cationic Peptides ; genetics ; Cell Line, Tumor ; Erythropoietin ; blood ; pharmacology ; Female ; Hepcidins ; Humans ; Interleukin-6 ; immunology ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; metabolism ; RNA, Messenger ; genetics

OBJECTIVETo explore the feasible age limits in Chinese elderly patients with non-Hodgkin's lymphoma (NHL).

METHODSThe clinical data of 507 patients with NHL who were admitted to Peking Union Medical College Hospital (PUMCH) from January 1990 to December 2007 were retrospectively analyzed. They were further followed up by reviewing medical records or by phone. The deadline of follow-up was October 2008.

RESULTSThe 5-year/8-year overall survival (OS) rates were 64.6%/45.7%, 53.0%/ 44.1%, 32.8%/17.5%, 40.0%/22.8%, and 19.8%/0, respectively, in patients aged < 60 years, 60-64 years, 65-69 years, 70-74 years, and > or = 75 years. The OS rate was significantly different between patients aged > or = 75 years and other age groups, and between patients aged 65-70 years and patients younger than 60 years (P < 0.05). Only age, serum albumin, and hemoglobin affected the survival status in elderly NHL patients.

CONCLUSIONSixty-five years can be regarded as the age limit in Chinese NHL patients.

Age Factors ; Aged ; Aged, 80 and over ; China ; epidemiology ; Female ; Follow-Up Studies ; Humans ; Lymphoma, Non-Hodgkin ; mortality ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Survival Rate

Disorders of iron utilization caused by abnormal elevation of hepcidin levels are the main mechanism of anemia of chronic disease. Hepcidin is mainly produced by the liver. Recently it has been found that monocytes are another source of hepcidin. The increased hepcidin in serum and urine of multiple myeloma patients may be one cause of anemia of chronic disease (ACD). However it is unclear whether the peripheral blood monocyte hepcidin is involved in the pathogenesis of anemia of chronic disease. This study was purposed to investigate the role of monocyte hepcidin in multiple myeloma patients with anemia of chronic disease. The clinical data and peripheral venous blood of multiple myeloma patients were collected.Serum concentration of IL-6 and TNF-α was detected by ELISA. Peripheral blood monocytes were isolated by CD14(+) magnetic beads. Hepcidin, IL-6 and TNF-α mRNA of monocytes were detected by real time quantitative PCR. The results showed that the expression level of monocyte hepcidin mRNA in myeloma patients was higher than that in normal controls. In untreated patients, the expression level of monocyte hepcidin mRNA was negatively correlated with hemoglobin, and positively correlated with serum ferritin and IL-6 levels, but unrelated with TNF-α levels.It is concluded that the increased monocyte hepcidin levels in multiple myeloma patients may play an etiologic role in ACD.
Adult ; Aged ; Anemia ; etiology ; Case-Control Studies ; Chronic Disease ; Female ; Ferritins ; blood ; Hepcidins ; blood ; Humans ; Interleukin-6 ; blood ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Monocytes ; metabolism ; Multiple Myeloma ; blood ; complications ; Tumor Necrosis Factor-alpha ; blood

Erythropoietin (EPO) is the major means of treating anemia of chronic disease (ACD) through stimulating hematopoiesis, inhibiting hepcidin and decreasing proinflammatory factors. Recently, it has been found that monocytes are another source of hepcidin. EPO can reduce the hepcidin stimulated by IL-6 in monocytes, it is assumed that EPO can reduce hepcidin indirectly by reducing IL-6. However, the specific mechanism of EPO inhibiting the proinflammatory cytokines in monocytes is unclear now. This study was purposed to investigate the effect of EPO on monocyte proinflammatory factors and its molecular mechanism. IL-6 mRNA and TNF-α mRNA were detected by real time PCR, level of signaling molecule PARP-1 protein was detected by Western blot. THP-1 monocytes were stimulated by 1 µg/ml lipopolysaccharide (LPS) to observe the impact of EPO at different concentrations (0.5, 1, 2, 5, 10 U/ml) for different time (0, 3, 6, 12, 24 hours) on the expression of IL-6 mRNA, TNF-α mRNA and PARP-1 protein. 1 µg/ml or 5 µg/ml EPO receptor (EPOR) antibody and/or 3-aminobenzamide (3-AB, PARP-1 inhibitor) were added to observe the antagonistic effect on EPO and the impact on PARP-1. The results showed that LPS could stimulate the THP-1 cells. EPO could decrease the levels of IL-6 and TNF-α stimulated by LPS in a dose- and time-dependent manners. The most significant decrease in IL-6 mRNA expression was observed in 2 U/ml EPO for 6 hours. And down-regulation of TNF-α mRNA expression was pronounced at 10 U/ml EPO for 3 hours. IL-6 mRNA expression could be stimulated by LPS, PARP-1 protein was induced at the same time. EPO inhibited the expression of IL-6 mRNA, while PARP-1 protein also decreased. Down-regulation of IL-6 mRNA and PARP-1 protein level was pronounced at 2 U/ml EPO for 6 hours. 3AB is a direct inhibitor of PARP-1. Similar to 3AB, EPO receptor antibody could antagonize the decline of IL-6 induced by EPO. It is concluded that EPO can inhibit the expression of IL-6 and TNF-α in monocytes, and the inhibition of IL-6 expression may be associated with decrease of PARP level.
Anemia ; metabolism ; Cell Line ; Erythropoietin ; pharmacology ; Humans ; Interleukin-6 ; metabolism ; Monocytes ; drug effects ; metabolism ; Poly (ADP-Ribose) Polymerase-1 ; Poly(ADP-ribose) Polymerases ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVE: To analyze the marginal roughness and marginal fitness of chairside computer-aided design and computer-aided manufacturing (CAD/CAM) laminate veneers with different materials and thicknesses, and to provide a reference for the clinical application of laminate veneers. METHODS: The butt-to-butt type laminate veneers were prepared on resin typodonts, the preparations were scanned, and the laminate veneers were manufactured by chairside CAD/CAM equipment. The laminate veneers were divided into four groups (n=9) according to the materials (glass-matrix ceramics and resin-matrix ceramics) and thickness (0.3 mm and 0.5 mm) of the veneers, with a total of 36. The marginal topo-graphies of each laminate veneer were digitally recorded by stereomicroscope, and the marginal rough-nesses of the laminate veneers were determined by ImageJ software. The marginal fitness of the laminate veneers was measured by a fit checker and digital scanning and measuring method. At the same time, the mechanical properties of glass-matrix ceramic and resin-matrix ceramic bars (n=20) were tested by a universal testing device. RESULTS: The marginal roughness of 0.3 mm and 0.5 mm glass-matrix ceramic laminate veneers was (24.48±5.55) μm and (19.06±5.75) μm, respectively, with a statistically significant difference (P < 0.001). The marginal roughness of 0.3 mm and 0.5 mm resin-matrix ceramic laminate veneers was (6.13±1.27) μm and (6.84±2.19) μm, respectively, without a statistically significant difference (P>0.05). The marginal roughness of the glass-matrix ceramic laminate veneers was higher than that of the resin-matrix ceramic laminate veneers with a statistically significant difference (P < 0.001). The marginal fitness of 0.3 mm and 0.5 mm glass-matrix ceramic laminate veneers were (66.30±26.71) μm and (85.48±30.44) μm, respectively. The marginal fitness of 0.3 mm and 0.5 mm resin-matrix ceramic laminate veneers were (56.42±19.27) μm and (58.36±8.33) μm, respectively. There was no statistically significant difference among the 4 groups (P>0.05). For glass-matrix ceramics, the flexural strength was (327.40±54.25) MPa, the flexural modulus was (44.40±4.39) GPa, and the modulus of resilience was (1.24±0.37) MPa. For resin-matrix ceramics, the flexural strength was (173.71±16.61) MPa, the flexural modulus was (11.88±0.51) GPa, and the modulus of resilience was (1.29±0.27) MPa. The flexural strength and modulus of glass-matrix ceramics were significantly higher than those of resin-matrix ceramics (P < 0.001), but there was no statistically significant difference in the modulus of resilience between the two materials (P>0.05). CONCLUSION The marginal roughness of CAD/CAM glass-matrix ceramic laminate veneers is greater than that of resin-matrix ceramic laminate veneers, but there was no statistically significant difference in marginal fitness among them. Increasing the thickness can reduce the marginal roughness of glass-matrix ceramic laminate veneers, but has no effect on the marginal roughness of resin-matrix ceramic laminate veneers.
Ceramics ; Computer-Aided Design ; Dental Porcelain ; Dental Veneers ; Materials Testing ; Surface Properties

This study was aimed to explore the possible mechanisms of hepcidin increase in multiple myeloma patients. The clinical information and peripheral venous blood of eligible patients with previously untreated multiple myeloma were collected. Serum concentration of IL-6 was detected by ELISA. Peripheral blood monocytes were isolated by CD14⁺ magnetic beads. The expression of hepcidin, IL-6 and C/EBPα mRNA of monocytes were detected by real time quantitative PCR. The results indicated that the hemoglobin level was reduced in 17 multiple myeloma patients enrolled in study (97.8 ± 27.5 g/L), showing the characteristics of anemia of chronic disease. The hepcidin and C/EBPα expression of peripheral blood monocytes significantly increased (P < 0.01), serum IL-6 was also higher than that in normal controls (P < 0.01). Serum IL-6 positively correlated with monocyte hepcidin and C/EBPα expression (P < 0.05); monocyte C/EBPα expression positively correlated with monocyte hepcidin expression (P < 0.05). It is concluded that the elevated IL-6 may induce hepcidin expression through up-regulating C/EBPα in untreated myeloma patients.
Anemia ; CCAAT-Enhancer-Binding Protein-alpha ; metabolism ; Chronic Disease ; Hepcidins ; metabolism ; Humans ; Interleukin-6 ; Monocytes ; Multiple Myeloma ; metabolism ; RNA, Messenger ; Up-Regulation