This paper reported a new type of amylase (neoamylase) secreted by a Bacillus strain ZX99. The enzyme was a kind of ectoenzyme that could catalyze starch into isomalto-oligosaccharide effectively, but could not act on pullulan as substrate. The strain Bacillus ZX99 was mutated by ultraviolet ray and a mutant strain BS3.232 was screened. The activity of the neoamylase produced from BS3.232 increased by 60% over that from ZX99 under the same conditions. The results of thin-layer chromatography of products from starch and pullulan catalyzed by the enzyme demonstrated that the enzyme was different from neopullulanase and can be used to produce isomaltooligosaccharide from starch, including isomaltose, panose, isomaltotriose, isomaltotetose.

Background Studies showed that macophenolic acid (MPA)down-regulates and inhibits the expression and secretion of tissue growth factor and inflammatory factor,and further impacts the proliferation and inflammation process.Pterygium is an inflammatory and proliferative lesion.Whether MPA has an inhibitory effect on pterygium is unclear.Objective This study was to investigate the antifibrotic effects of macophenolic acid on pterygium fibroblasts(PFBs) in vitro and discuss its mechanism.Methods Pterygium tissue was obtained from pterygium patient during the surgery.PFBs were cultured using explants and identified with vimentin immunohistochemisty.0,0.125,0.250,0.500,1.000 μmol/L MPA were added into the culture medium,respectively,and the cells were cultured in the medium without MPA as the control group.MTT colorimetry was used to find the optimization effective concentration of MPA and evaluate their inhibitory effect on PFBs,and BrdU fluorescence staining was used to assess the growth statue of PFBs.Expressions of nuclear factor-κB(NF-κB),p65 and inhibitor of NF-κB-α(IκB-α) in the cells were detected by Western blot.Results The cells was spindle in shape 3 days after cultured and showed the vortex and radial arrangement with the positive response to vimentin.With the increase of MPA,the proliferative value of PFBs (A560)showed gradually decline,with a significant difference among the five groups (F =42.874,P＜0.01).In addition,the proliferative value of PFBs (A560) significantly lowed as the prolong of MPA active time(F=26.038,P＜0.01).BrdU fluorescence staining showed a significant decrease of DNA synthesis of PFBs with the elevation of MPA dose among the five groups(F=175.279,P＜0.05),and the A560of PFBs DNA synthesis in different concentrations of MPA groups was lower than that of the control group (all at P＜0.05).No apoptotic and necrotic cell was found after MPA action by DAPI staining.The expression level of p65 in the PFBs was 0.886±0.072 and 1.542±0.124 in the MPA group and the control group,indicating a declined value in the MPA group(P＜0.05).However,the expression value of IκB-α in the cytoplasm PFBs was significantly higher in the MPA group compared with the control group(2.141 ±0.305 vs.1.559±0.267) (P＜0.05).Conclusions MPA has an inhibitory effect on the growth of PFBs,which probably is related to the arresting of NF-κB pathway.

Animals ; Carps ; genetics ; Cloning, Molecular ; Fish Proteins ; genetics ; Hydroxylation ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; Lakes

Bronchiolitis ; diagnosis ; microbiology ; pathology ; Female ; Humans ; Infant ; Leukotriene D4 ; analysis ; Male ; Nasopharynx ; chemistry ; microbiology ; pathology

OBJECTIVETo evaluate the impact of platelet membrane glycoprotein (GP)-specific autoantibodies on high-dose dexamethasone therapy in patients with idiopathic thrombocytopenic purpura (ITP).

METHODSModified direct monoclonal antibody immobilization of platelet antigen assay (MAIPA) was used to detect platelet GPIIb/IIIa and/or GPI b specific autoantibodies. All patients received oral dexamethasone 40 mg/d for 4 days.

RESULTSThe response rate of high-dose dexamethasone in GPIIb/IIIa and/or GPIb specific autoantibody-negative patients was significantly different from that of antibody-positive patients (P<0.05). The response rate of GPIIb/IIIa specific autoantibody-positive patients was lower than that of antibody-negative patients (P<0.05). GPIb specific autoantibody had no significant impact on the efficacy of high-dose dexamethasone (P>0.05).

CONCLUSIONPlatelet membrane GPIIb/IIIa-specific autoantibody can be a potential negative indicator for ITP patients'response to high-dose oral dexamethasone.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Autoantibodies ; blood ; Dexamethasone ; administration & dosage ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; immunology ; Platelet Glycoprotein GPIb-IX Complex ; immunology ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; immunology ; Treatment Outcome ; Young Adult

OBJECTIVETo analyze the characteristics of atrial tachycardia originating from the atrioventricular cingulum.

METHODThe electrophysiological mechanism, ablation site graph and nerve distribution of 16 cases of atrial tachycardia originating from the atrioventricular cingulum or adjacent atrial muscle which were proved by electrophysiological monitoring and radiofrequency ablation.

RESULTSAtrial tachycardia from peri-cingulum represented 23.2% of atrial tachycardia treated by radiofrequency ablation during the same period. The ratio of left to right atrioventricular cingulum was 3:16. There was no difference of the surface ECG characteristics and electrophysiological mechanism between the atrial tachycardia originating from atrioventricular cingulum and that from other positions. Both A and V components were recorded at all the successful ablation sites. The ratio of amplitude of A to V was between 2:3 and 6:1. Atrial potential in the target site was 20-46 (38.6 +/- 6.7) ms earlier than P'wave in surface ECG. The success rate of ablation was 87.5% and the recurrent rate 7.1%.

CONCLUSIONSPeri-cingulum atrial tachycardia accounts for a certain proportion in all atrial tachycardia. The exciting sites originating from right cingulum are more common than those from left cingulum. Its electrophysiological mechanism has no difference from other positions. Cingulum mapping and ablation have important practical meanings.

Adolescent ; Adult ; Aged ; Catheter Ablation ; Child ; Electrophysiologic Techniques, Cardiac ; Female ; Humans ; Male ; Middle Aged ; Tachycardia, Ectopic Atrial ; physiopathology ; surgery ; Young Adult

OBJECTIVETo detect the variation of gene expression profile of G1/S transition and elucidate the role of related genes regulating cell cycle from G1 to S phase in gastric cancer.

METHODSNocodazole-thymidine and double thymidine methods were used to synchronize gastric cells at G2/M and G1/S point, cDNA microarray chips was applied to examine the gene expression profile at G1 early and late phase, S early and late phase during the cell cycle, hierarchy analysis was conducted by a professional software package.

RESULTSA total of 2001 genes were detected available, 959 genes appeared to be upregulated or downregulated, including 147 genes upregulated at G1 late phase. These 147 genes are involved in DNA metabolism, transcription and translation,posttranslational modification, ubiquitination, signal transduction etc, which all affected cell cycle from different aspects.

CONCLUSIONComplex multiple gene processes, such as DNA metabolism, transcription and translation, posttranslational modification, ubiquitination, signal transduction etc,are implicated in and also essential for G1/S transition during gastric cancer cell cycle, part of these genes are significantly associate with overproliferation in gastric cancer.

Cell Division ; G1 Phase ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Humans ; S Phase ; genetics ; Signal Transduction ; Stomach Neoplasms ; genetics ; physiopathology ; Transcription, Genetic ; Tumor Cells, Cultured

OBJECTIVETo investigate clinical features of disk displacement during the course of condylar fracture and to explore the techniques of disk reposition and suturation.

METHODS32 patients (10 females and 22 males) who had disk displacements with condylar fractures were followed up. Reduction and reposition of the dislocated disks simultaneously with fixation of fractures were performed. 7 patients underwent intermaxillary fixation with elastic bands for 1 to 2 weeks.

RESULTSThe occlusions were satisfactory in all cases but one for the reason of ramus height loss. No TMJ symptom was found when examined 3 months post operation.

CONCLUSIONSAnterior disk displacements were most occurred with high condylar process fractures. Surgical reposition and suturation of disk play an important role for the later TMJ-function.

Adolescent ; Adult ; Female ; Fracture Fixation, Internal ; Humans ; Joint Dislocations ; etiology ; surgery ; Male ; Mandibular Condyle ; injuries ; Mandibular Fractures ; complications ; surgery ; Middle Aged ; Temporomandibular Joint Disc ; pathology

BACKGROUNDThe congenital Long QT syndrome (LQTS) is a hereditary cardiac channelopathy that is characterized by a prolonged QT interval, syncope, ventricular arrhythmias, and sudden death. The chromosome 7-linked type 2 congenital LQTS (LQT2) is caused by gene mutations in the human ether-a-go-go-related gene (HERG).

METHODSA Chinese family diagnosed with LQTS were screened for KCNQ1, HERG and SCN5A, using polymerase chain reaction (PCR), direct sequencing, and clong sequencing. We also investigated the mRNA expression of the HERG gene.

RESULTSWe identified a novel I414fs + 98X mutation in the HERG gene. The deletion mutation of 14-bp in the first transmembrane segment (S1) introduced premature termination codons (PTCs) at the end of exon 6. This mutation would result in a serious phenotype if the truncated proteins co-assembled with normal subunit to form the defective channels. But only the proband was symptomatic.

CONCLUSIONSWe found that the mRNA level of the HERG gene was significantly lower in I414fs + 98X carriers than in noncarriers. We found a novel I414fs + 98X mutation. The mRNA level supports that NMD mechanism might regulate the novel mutation.

Adult ; ERG1 Potassium Channel ; Ether-A-Go-Go Potassium Channels ; genetics ; Female ; Frameshift Mutation ; Humans ; Long QT Syndrome ; genetics ; RNA, Messenger ; analysis

BACKGROUNDRemodeling of the anterior cruciate ligament (ACL) graft usually takes longer than expected. Gene therapy offers a radical different approach to remodeling of the graft. In this study, the internal ribosome entry site (IRES) sequence was used to construct a new recombinant adenovirus which permits co-expression of transforming growth factor-beta1 (TGFbeta1) and vascular endothelial growth factor 165 (VEGF165) genes (named Ad-VEGF165-IRES-TGFbeta1). We investigated the effects of the new adenovirus on the migration of and matrix synthesis by ACL fibroblasts.

METHODSAdenoviral vector containing TGFbeta1 and VEGF165 genes was constructed. ACL fibroblasts were obtained from New Zealand white rabbits. After ACL fibroblasts were exposed to Ad-VEGF165-IRES-TGFbeta1, the expression of VEGF165 and TGFbeta1 proteins were assessed by enzyme-linked immunosorbent assay (ELISA) and Western blotting analysis. Bioassay of VEGF165 and TGFbeta1 proteins were assessed by Western blotting analysis. Proliferation and migration of ACL fibroblasts were assessed by in vitro wound closure assay. Gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers were assessed by real-time PCR.

RESULTSThe results showed the successful construction of a recombinant co-expression adenovirus vector containing TGFbeta1 and VEGF165 genes. Co-expression of TGFbeta1 and VEGF165 can induce relatively rapid and continuous proliferation of ACL fibroblasts and high gene expression of collagen type I, collagen type III, and fibronectin mRNA among matrix markers.

CONCLUSIONCo-expression of TGFbeta1 and VEGF165 genes has more powerful and efficient effects on the migration of and matrix synthesis by ACL fibroblasts.

Adenoviridae ; genetics ; Animals ; Anterior Cruciate Ligament ; cytology ; metabolism ; Cell Movement ; Cells, Cultured ; Collagen ; genetics ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Fibroblasts ; physiology ; Fibronectins ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; Wound Healing