Armillariella tabescens EJLY2098 was induced to produce ?-mannanase with konjac fine flour (Amorphopallus rivieri) as single carbon source. This induced enzyme was then purified using DEAE ion exchange chromatography and named atMAN47. Zymologic analysis showed that the molecular weight of this ?-mannanase was approximately 47 kD. The enzyme was stable when pH ranged from 5.0 to 6.5 and could be activated by Na+ and Ba2+. With an optimal temperature of 50?C. Action mode analysis of TLC revealed that the enzyme belonged to the endo-?-mannanase family. Being a meta-acid endo-?-mannanase, it was suitable to be applied to feed industry with a promising future as an enzyme preparation.

During oral glucose tolerance test(OGTT),endothelium-dependent vasodilation(EDD)at different time points in impaired glucose tolerance(IGT)group was lower than that in normal control group.EDD at 60 and 120 min in IGT + vitamin C group was higher than that in IGT group(all P＜0.05).There was a negative relationship between blood glucose level and EDD during OGTT in IGT patients.

Objective To evaluate the effects of various polysorbates(PS)on the stability of different types of monoclonal antibody(mAb)drugs.Methods Three types of monoclonal antibodies mAbA(IgG1 proantibody drug),mAbB(IgG1 mAb)and mAbC(IgG1 mAb with Fc N297A mutation)were used as model proteins,and different kinds or contents of PS were added into the mAb formulations respectively to investigate the influencing factors.The effects of PS on the stability of mAb drugs were evaluated comprehensively by detecting the changes of quality attributes,such as protein aggregates and insoluble particles.Results PS20 and PS80 showed no significant difference in inhibiting the formation of aggregates and charge variants in the three mAbs(P>0.05),while the addition of PS80 in mAbB and PS20 in mAbC significantly inhibited the increase of insoluble particles respectively(P<0.05);The content of PS20 showed a significant effect on the detection indexes of charge variants and insoluble particles in mAbC(P<0.05).Conclusion Different types of mAbs have different sensitivities to various kinds and contents of PS.Therefore,when designing the formulation of mAbs,it is necessary to select appropriate kinds and contents of PS to further improve the stability of mAb drugs.

OBJECTIVETo assess if the modulating effect of platelet-derived growth factor (PDGF)-BB on p21(WAF1) was mediated by upregulating transforming growth factor (TGF)-beta(1) expression in vascular smooth muscle cells (VSMC).

METHODSTGF-beta(1) mRNA and protein expressions were measured by reverse transcription-PCR and ELISA, the protein expressions of p21(WAF1) and the downstream TGF-beta signalling including TGF-beta type I receptor (ALK-5 in VSMC), Smurf2, pSmad2/3, Smad4, Smad7 were detected by Western blot.

RESULTSPDGF-BB significantly upregulated the expressions of TGF-beta(1) at mRNA (0.79-fold) and protein (1.98-fold) levels in VSMC, significantly inhibited the expression of p21(WAF1) (-67 +/- 12)%, and enhanced the expressions of ALK-5, pSmad2/3, Smad4, Smurf2 protein by 1.21-fold, 0.95-fold, 0.69-fold and 2.55-fold respectively, inhibited Smad7 expression (-65 +/- 9)%, these alterations were partially restored by anti-TGF-beta(1) neutralizing antibody.

CONCLUSIONSThese findings suggested that PDGF-BB inhibited p21(WAF1) expression in VSMC partially through upregulating TGF-beta(1) expression via PDGF-BB and TGF-beta signalling pathways.

Animals ; Cell Division ; Cells ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Male ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; Platelet-Derived Growth Factor ; pharmacology ; Proto-Oncogene Proteins c-sis ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Transforming Growth Factor beta1 ; metabolism

Objective:To observe the effect of acupuncture in regulating ubiquitin-proteasome pathway (UPP),and discuss the action of acupuncture in intervening heroin-induced brain damage.Methods:Thirty male Sprague-Dawley (SD) rats were divided into a control group,a model group and an acupuncture group by using the random number table.Rats in the model and acupuncture groups received intramuscular heroin injection for successive 8 d at a progressively increased dose.Afterwards,the injection was suspended for 5 d for withdrawal.The heroin relapse rat model was established by repeating the drug addiction and withdrawal process for 3 times.The control group followed the step of the model establishment,but was given intramuscular injection of normal saline at the stage of addiction and no intervention at the stage of withdrawal;the model group was given intramuscular heroin injection at a progressively increased dose at the addiction stage and no intervention at the withdrawal stage;the acupuncture group was dealt in the same way as the model group at the addiction stage,but received acupuncture at Baihui (GV 20) and Dazhui (GV 14) at the withdrawal stage,with the needles retained for 30 min each time,1 session a day,for successive 5 d.On the 39th day,brain tissues were extracted from the hippocampus and ventral tegmental area (VTA) of the three groups of rats.The apoptosis of brain nerve cells was detected by using terminal deoxynucleotidyl transferase-mediated nick and labeling (TUNEL).The mRNA and protein expressions of ubiquitin (Ub),ubiquitin protein ligase (E3) and 26S were examined by immunohistochemistry and quantitative real-time polymerase chain reaction (RT-qPCR).Results:Compared with the model group,rat's hippocampus and VTA in the acupuncture group showed significantly fewer cells positively stained by TUNEL staining (P＜0.01),and its mRNA and protein expressions of Ub,E3,26S were significantly lower (P＜0.01).Conclusion:Reducing nerve cell apoptosis and regulating the mRNA and protein expressions of Ub,E3 and 26S in rat's hippocampus and VTA are possibly one of the action mechanisms of acupuncture in intervening heroin-induced brain damage.

OBJECTIVETo evaluate the effect of the aluminum hydroxide (Al-OH) adjuvant on the 2009 pandemic influenza A/H1N1 (pH1N1) vaccine.

METHODSIn a multicenter, double-blind, randomized, placebo-controlled trial, participants received two doses of split-virion formulation containing 15 μg hemagglutinin antigen, with or without aluminum hydroxide (Al-OH). We classified the participants into six age categories (>61 years, 41-60 years, 19-40 years, 13-18 years, 8-12 years, and 3-7 years) and obtained four blood samples from each participant on days 0, 21, 35, and 42 following the first dose of immunization. We assessed vaccine immunogenicity by measuring the geometric mean titer (GMT) of hemagglutination inhibiting antibody. We used a two-level model to evaluate the fixed effect of aluminum Al-OH and other factors, accounting for repeated measures.

RESULTSThe predictions of repeated measurement on GMTs of formulations with or without Al-OH, were 80.35 and 112.72, respectively. Al-OH significantly reduced immunogenicity after controlling for time post immunization, age-group and gender.

CONCLUSIONThe Al-OH adjuvant does not increase but actually reduces the immunogenicity of the split-virion pH1N1 vaccine.

Adjuvants, Pharmaceutic ; chemistry ; Adolescent ; Adult ; Aluminum Hydroxide ; chemistry ; Antibodies, Viral ; blood ; Child ; Child, Preschool ; China ; Data Interpretation, Statistical ; Double-Blind Method ; Female ; Hemagglutination Inhibition Tests ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza Vaccines ; adverse effects ; chemistry ; immunology ; Influenza, Human ; epidemiology ; immunology ; prevention & control ; virology ; Male ; Middle Aged ; Models, Statistical ; Pandemics ; Young Adult

OBJECTIVECongenital arteriovenous malformations(AVM) are considered to be the most difficult and challenging problems in the treatment of hemangiomas and vascular lesions. This study focused on the natural history, the clinical classification, the choice and effectiveness of various treatments.

METHODSThis retrospective review included 83 patients with extracranial arteriovenous malformations, who were referred to our department over the past 6 years. The anatomic patterns, clinical staging, respective treatments, influential factors of endovascular treatment, causes of recurrence were analyzed.

RESULTSAccording to clinical manifestations, arteriovenous malformations were categorized as three clinical stages: the quiescence, the expansion and the decompensation stages. Most AVMS in the quiescence stage only require endovascular treatment while those in the decompensation stage require surgical resection. Angiography was performed not only for diagnosis of AVM but also as an initial therapeutic step in the form of embolization, which might be the only means to some AVM without surgical possibility or necessity.

CONCLUSIONThe new concept of staging and management is expected to be helpful for diagnosis and treatments of AVM.

Acrylates ; therapeutic use ; Angiography ; Arteriovenous Malformations ; classification ; surgery ; therapy ; Embolization, Therapeutic ; Endoscopes ; Female ; Follow-Up Studies ; Humans ; Male ; Polyvinyls ; therapeutic use ; Retrospective Studies ; Treatment Outcome

BACKGROUNDPrevious studies showed that anti MHC-II monoclone antibody (MAb) only had partial inhibiting effect of alloreactive mixed lymphocyte reaction (MLR) in vitro and it was unsteady and non-persistent. The aim of this research was to determine whether radioactive isotope (188)Re marked MHC-II antibody could benefit the allograft acceptance in transplantation as compared to normal MHC-II antibody.

METHODS188Re was incorporated to 2E9/13F (ab')(2) which is against swine MHC class II antigen (MAb-(188)Re). Porcine peripheral blood mononuclear (PBMC) cells were examined for proliferation and cytokine mRNA expression after stimulation with MHC-II MAb or MAb-(188)Re.

RESULTSThe proliferative response of recipient PBMCs in mixed lymphocyte reaction (MLR) to donor alloantigen showed that the stimulation index of MAb-(188)Re group was significantly lower than the MHC-II MAb group and control (P < 0.05). mRNA expression of interleukin 2, interferon Υ and tumor necrosis factor α (type 1 cytokines) was lower in MAb-(188)Re group than the MHC-II MAb group, while interleukin 10 (type 2 cytokines) was higher in MAb-(188)Re group in the first 24 hours.

CONCLUSIONMAb-(188)Re could help the graft acceptance by inhibiting T cell proliferation, lowering the expression of type 1 cytokines and elevating the type 2 cytokines produced by PBMC.

Animals ; Antibodies, Monoclonal ; chemistry ; pharmacology ; Cell Proliferation ; drug effects ; Interleukin-10 ; genetics ; Interleukin-2 ; genetics ; Isoantigens ; immunology ; Leukocytes, Mononuclear ; drug effects ; radiation effects ; Lymphocyte Culture Test, Mixed ; Mitomycin ; pharmacology ; Radioisotopes ; Reverse Transcriptase Polymerase Chain Reaction ; Rhenium ; Swine ; Tumor Necrosis Factor-alpha ; genetics

Methylation, a methyl group-consuming reaction, plays a key role in the degradation (i.e., inactivation) of monoamine neurotransmitters, including catecholamines, serotonin and histamine. Without labile methyl groups, the methylation-mediated degradation cannot take place. Although high niacin (nicotinic acid and nicotinamide) intake, which is very common nowadays, is known to deplete the body's methyl-group pool, its effect on monoamine-neurotransmitter degradation is not well understood. The aim of this article was to investigate the effect of excess nicotinamide on the levels of plasma serotonin and histamine in healthy subjects. Urine and venous blood samples were collected from nine healthy male volunteers before and after oral loading with 100 mg nicotinamide. Plasma N(1)-methylnicotinamide, urinary N(1)-methyl-2-pyridone-5-carboxamide (2-Py), and plasma betaine levels were measured by using high-performance liquid chromatography (HPLC). Plasma concentrations of choline, serotonin and histamine were measured using commercial kits. The results showed that the plasma N(1)-methylnicotinamide level and the urinary excretion of 2-Py significantly increased after oral loading with 100 mg nicotinamide, which was accompanied with a decrease in the methyl-group donor betaine. Compared with those before nicotinamide load, five-hour postload plasma serotonin and histamine levels significantly increased. These results suggest that excess nicotinamide can disturb monoamine-neurotransmitter metabolism. These findings may be of significance in understanding the etiology of monoamine-related mental diseases, such as schizophrenia and autism (a neurodevelopmental disorder).
Betaine ; blood ; Choline ; blood ; Chromatography, High Pressure Liquid ; Histamine ; blood ; Humans ; Male ; Niacinamide ; administration & dosage ; analogs & derivatives ; blood ; Pyridones ; urine ; Serotonin ; blood

OBJECTIVETo analyze the clinical data and result of voiding cystourethrography (VCUG) in high-risk children with vesicoureteral reflux (VUR) for better awareness of VUR, and to assess the usefulness of non-radioactive voiding ultrasonography (VUS) in the diagnosis of VUR.

METHODNinety-three high-risk children with VUR who were hospitalized from July 2007 to April 2010 were studied. The study included 58 cases of urinary tract infection (UTI) and 35 cases of fetal or postnatal hydronephrosis detected on a B ultrasound scan. The results of urinalysis, urine culture, renal function, B ultrasound and VCUG were evaluated. Part of patients underwent VUS followed by VCUG immediately.

RESULT(1) Sixty-two boys and 31 girls (aged 1 month to 11.5 years, mean age 2 years) were included. VUR was detected in 26 patients (28%) by VCUG. In terms of kidney-ureter units, VUR was detected in 36 of 186 kidney-ureter units, including 6 grade I, 3 grade II, 6 grade III, 15 grade IV and 6 grade V. (2) VUR was detected in 20 of 58 UTI patients (34.5%) by VCUG. The proportion of VUR in recurrent UTI group was 61.1%, much higher than that in first UTI group (22.5%). Thirteen of 20 VUR (65%) occurred in UTI patients under 1 year of age (M/F 10/3), with more bilateral VUR and severe grades of VUR than the older group. VUR was detected in 6 of 35 fetal or postnatal hydronephrosis patients (17.1%) by VCUG. (3) Twenty-two patients underwent both VUS and VCUG. VUR was detected in 4 patients and 6 kidney-ureter units by VCUG, while in 6 patients and 9 kidney-ureter units by VUS. Taking VCUG as the reference standard, VUS had a sensitivity of 100%, specificity of 92.1%, positive predictive value of 66.7%, and negative predictive value of 100%. There was a concordance rate of 93.2% between VUS and VCUG.

CONCLUSIONIt is important to early screen VUR in UTI, fetal or postnatal hydronephrosis patients. There are more VUR, especially more bilateral VUR and severe grades of VUR, occurred in UTI patients under 1 year of age compared to older children. The incidence of VUR in recurrent UTI group was much higher than that in first UTI group. VUS is an accurate, reliable and radiation-free technique for the detection of VUR. It could be used to screen high-risk children for VUR and do the evaluation in the follow-up of VUR.

Child ; Child, Preschool ; Diagnostic Imaging ; Female ; Humans ; Hydronephrosis ; diagnosis ; diagnostic imaging ; Infant ; Male ; Ultrasonography ; Urinary Tract Infections ; diagnosis ; diagnostic imaging ; Urography ; Vesico-Ureteral Reflux ; diagnosis ; diagnostic imaging