0.05)between the images from the two groups.Conclusion When scanning the heart with volume CT(VCT),the application of ECG modulated mA can effectively reduce the exposure dosage without sacrificing the image quality.

A thiosulfate-oxidizing bacterium TX was isolated from polysulfide wastewater treatment system.It was preliminarily identified as Halothiobacillus sp.according to its morphological,physiological proper-ties and similarity analysis of 16S rRNA gene sequence(GenBank accession No.EU871645).This bacte-rium was an obligate chemolithoautotrophic sulfur-oxidizing bacterium since it could utilize thiosulfate,elemental sulfur,tetrathionate,sulfide or sulfite as sole energy source but did not show growth on the me-dium with glucose,sucrose,fructose,lactose,mannose or yeast extract as sole carbon source.The optimal growth condition for thiosulfate oxidation was 30?C~35?C and pH 3.0~5.0 respectively.Significant reduc-tion in medium pH from 4.0 to 1.2 was observed due to oxidation of thiosulfate to sulfate during the batch culture incubation period.During the course of thiosulfate oxidation accumulation of tetrathionate was ob-served either in synthetic medium or in the mixed liquor from aeration tank of the polysulfide wastewater treatment system.This indicates that the "S4 intermediate(S4I)" pathway was responsible for thiosulfate oxidation in this bacterium.

Background Conjunctivochalasis is an age-related inflammatory ocular surface disease manifesting redundant,loose conjunctiva folds.Studies showed that conjunctivochalasis often accompanies with inflammatory response,indicating that a variety of inflammatory cytokines are involved in pathogenesis of conjunctivochalasis.Objective This study attempted to investigate the expressions of pentraxin-3 (PTX-3) and tumor necrosis factor-α stimulated gene-6 (TSG-6) proteins in fibroblasts of conjunctivochalasis.Methods The protocol was approved by Ethic Committee of Putuo Hospital affiliated Shanghai University of Traditional Chinese Medicine.Thirteen eyes of 13 patients with conjunctivochalasis,16 eyes of 16 patients with age-related cataract and 16 eyes of 15 patients with pterygium were included in Putuo Hospital Affiliated to Shanghai University of Traditional Medicine from January,2011 to June,2012.The samples of conjunctival tissue were collected in the patients during surgery under the informed consent,and fibroblasts were cultured and passaged by explant culture method.Content of PTX-3 and TSG-6 proteins in cell supernatant was detected by ELISA.Results Translucent poor,dark black area was around the cell overflow.Cultured cells of third generation showed long shuttle type,uniform size,radiated out in all directions,and oval nuclei was sited in the cytoplasm,surrounded by varying the length of cross-linking of cell processes.The concentrations of PTX-3 protein in cell supernatant were (2 182.33 ± 738.68) pg/ml in the conjunctivochalasis group,which were higher than (738.32±335.00) pg/ml in the age-related cataract group and (981.37±416.04)pg/ml in the pterygium group,showing a significant difference among the three groups (F =6.474,P=0.032).The contents of TSG-6 proteins in cell supernatant were (59.10±6.52) pg/ml,(53.99± 11.16) pg/ml and (35.01±5.33)pg/ml in the conjunctivochalasis group,pterygium group and age-related cataract group,with a significant difference among the three groups (F =7.421,P =0.024),and contents of TSG-6 proteins in the agerelated cataract group was lowest,however,no significant difference was found in the TSG-6 contents between the conjunctivochalasis group and the pterygium group (P＞0.05).Conclusions Inflammatory reaction participates in the pathogenesis and development of conjunctivochalasis,up-regulation of PTX-3 and TSG-6 expression in fibroblasts might partakes in the pathogenesis of conjunctivochalasis.

Along with the increase in fungal infections, Candida albicans prevention and control become the focus of anti-fungal infection at present. This study aims to discuss the effect monomer andrographolide (AG) on C. albicans biofilm dispersion. In the experiment, micro-well plates and medical catheter pieces were used to establish the C. albicans biofilm model. It was discovered by XTT assay and flat band method that 1 000, 500, 250 mg x L(-1) AG could impact the activity of C. albicans biofilm dispersion cells. The morphological structures of residual biofilms on catheter pieces were observed with scanning electron microscopy, which showed that 1 000, 500, 250 mg x L(-1) AG could induce C. albicans biofilm dispersion in a dose-dependent manner, and the dispersed cells were dominated by the yeast phase. According to the real-time fluorescence quantification PCR (qRT-PCR) test, AG could up-regulate HSP90 expression and down-regulate UME6 and PES1 expressions. This study demonstrates that AG could induce C. albicans biofilm dispersion to some extent.
Anti-Inflammatory Agents ; pharmacology ; Biofilms ; drug effects ; Candida albicans ; genetics ; physiology ; ultrastructure ; Diterpenes ; pharmacology ; Dose-Response Relationship, Drug ; Fungal Proteins ; genetics ; Gene Expression Regulation, Fungal ; drug effects ; HSP90 Heat-Shock Proteins ; genetics ; Microscopy, Electron, Scanning ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

OBJECTIVETo investigate the effects of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) , alone and in combination with fluconazole (FLZ) on FLZ-resistant Candida albicans.

METHODThe minimum inhibitory concentrations (MIC) and sessile MIC80 (SMIC80) of EAHD and FLZ to FLZ-resistant C. albicans were determined by CLSI M27-A3 microdilution method, and the synergy of EAHD combined with FLZ were examined by the checkerboard microdilution assay. Agar plate-method was adopted to observe the rate of antifungal activity according to time-kill curve. HPLC and qRT-PCR were utilized to evaluate the changes of ergosterol content and expressions of related genes, respectively.

RESULTMICs of EAHD ranged from 156 to 1,250 mg · L(-1), those of FLZ from 256 to above 2,048 mg · L(-1) with FICI approximate 0.066 in combination; SMIC80 of EAHD were higher than 1,250 mg · L(-1), SMIC80 of FLZ were higher than 512 mg · L(-1) and up to above 2,048 mg · L(-1). Combination group also showed synergy effect except one group showing addition effect. The results of T-K experiment also confirmed obviously fungicidal effect when treated for 12 h. When compared with control groups, the ergosterol was reduced 85% and 50% in the treatments of combination and EAHD alone by HPLC, respective- ly. The expressions of ERG1, ERG2, ERG6, ERG7 and ERG11 were upregulated, and ACS1, ACS2, MET6 were downregulated when exposed to FLZ. The expressions of the above genes were downregulated by treatment of EAHD. The expressions of ERG2, ERG6, ERG11 were upregulated, while ERG1, ERG7, ACS1, ACS2, MET6 were downregulated in combination group.

CONCLUSIONThe combination of EAHD and FLZ exhibited synergy against FLZ-resistant C. albicans through decreasing the synthesis of ergosterol, and resulting in the breakage of cell membrane.

Antifungal Agents ; pharmacology ; Candida albicans ; drug effects ; growth & development ; metabolism ; Drug Resistance, Fungal ; drug effects ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; Ergosterol ; biosynthesis ; Fluconazole ; pharmacology ; Microbial Sensitivity Tests

OBJECTIVETo investigate the effect of andrographolide derivative Yanhuning (YHN) on Candida albicans biofilms in rats.

METHODThe rat C. albicans biofilms subcutaneous catheter model was established by intraperitoneally injecting YHN (40, 20, 10, 5, 2.5 mg x kg (-1)), with the FLC (80 mg x kg(-1)) positive group as the control group. After 7 d, CFU counting and XTT assay were used to evaluate the effect of YHN on C. albicans biofllms in vivo. Scanning electron microscopy (SEM) was applied to observe the morphological changes in rat biofilms intervened by YHN. The real-time fluorescence quantification PCR was adopted to detect expressions of C. albicans adhesion-related genes, such as ALS1, ALS3, HWP1, EAP1 and MP65.

RESULTThe YHN group showed much less CFUs on catheter pieces and lower XTT metabolic activity than the blank group, with dosage dependence. SEM also showed that YHN could obviously decrease C. albicans adhesion on subcutaneous catheters in rats. According to qRT-PCR's results, YHN can down-regulate expressions of ALS1, ALS3, HWP1, EAP1 and MP65.

CONCLUSIONYHN could inhibit C. albicans biofilms in rats.

Animals ; Biofilms ; drug effects ; growth & development ; Candida albicans ; cytology ; drug effects ; physiology ; Catheters ; microbiology ; Cell Adhesion ; drug effects ; Diterpenes ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Rats

Background Conjunctivochalasis (CCh) is a common age-related ocular surface diseases.Researches showed that the inflammatory factors are upregulated in conjunctiva and tear of CCh patients,inferring inflammation participates in the pathogenesis of CCh,however,its mechanism is unclear now.Studies also showed that the expression of matrix metalloproteinases (MMPs) in CCh conjunctiva is elevated.However,the association between inflammatory factors and MMPs is unknown.Objective This research was to observe the effects of tumor necrosisfactor-α (TNF-α) and interleukin-1 β (IL-1 β) on the expression of MMP-1,MMP-3,tumor necrosis factor-stimulatedgene-6(TSG-6) and pentraxin-3 (PTX3) in cultured CCh fibroblasts.Methods Conjunctival fibroblasts wereisolated and cultured from CCh-derived and cataract-derived human conjunctiva explants.The cells were treated using20 ng/ml PBS,TNF-α or IL-1β for 4 hours,respectively.The expression levels of MMP-1,MMP-3,TSG-6 and PTX3genes and proteins were detected by quantitative real time PCR and Western blot assay,respectively.Results Thesecond-generation cells showed a long and fusiform shape with ovoid nucleus and radial agreement,and the cellprocessors connected each other.The differences of the relative expression levels of MMP-1,MMP-3,TSG-6 and PTX3mRNA in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-αand IL-1 β (MMP-1 mRNA:Fgroup =2.611,P =0.116;Fi =161.564,P =0.000;MMP-3 mRNA:Fgroup =5.201,P =0.029;Fintervene =211.021,P =0.000;TSG-6 mRNA:Fgroup =47.209,P =0.000;Fintervene =119.340,P =0.000;PTX3 mRNA:Fgroup =40.512,P =0.000;Fintervene =93.935,P =0.000).Compared with the cataract group with PBS treatment,the expression levels of MMP-1,MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with PBS treatment or the cataract group with TNF-α and IL-1 β treatment (all at P＜0.05).Compared with the CCh group with PBS treatment,the expression levels of MMP-1,MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with TNF-α and IL-1β treatment (all at P ＜ 0.05).The differences of the relative expression levels of MMP-1,MMP-3,TSG-6 and PTX3 proteins in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-α and IL-1 β (MMP-1:Fgroup =84.702,P =0.000;Fint =48.900,P =0.000;MMP-3:Fgroup =112.818,P =0.000;Fint =194.980,P =0.000;TSG-6:Fgroup =56.867,P =0.000;Fint =70.356,P =0.000;PTX3:Fgroup =1.488,P =0.231;Fint =89.872,P =0.000),and the expression changes of MMP-1,MMP-3,TSG-6 and PTX3 proteins were coincident with the genes among the groups with various treatments.Conclusions The expressions of MMP-1,MMP-3,TSG-6 and PTX3 in conjunctival fibroblasts are upregulated in CCh eyes.The interaction of TNF-α and IL-1 β with MMPs is probably involved with the pathogenesis and development MMPs probably.

Objective To explore clinical characteristics of diagnosis and treatment of solid-cystic papillary tumor(SCPT) of the pancreas in children.Methods There were retrospectively analyzed about the 7 patients treated in our hospital for SCPT,with the ave-rage age of 11.5 years.All patients complained abdominal pain following a trauma(71.4%) or overeating(28.6%).The main presentation was abdominal mass.Of the 7 patients,4 received distal pancreatectomy,2 pancreatico-duodenectomy,and 1 only biopsy.Results The nicks of all patients were primany hed.All patients were pathologically confirmed as SCPT after operation.All patients were followed up for 4 monthes to 4.5 years,the recent result was well.Conclusions SCPT is a low-grade malignant tumor,which is often asymptomatic,but the patients with symptoms generally suffer from an abdominal mass or abdominal pain.The prognosis is excellent after operation.

Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases ; metabolism ; Bacterial Proteins ; metabolism ; Cyclization ; Cysteine Endopeptidases ; metabolism ; Enzymes, Immobilized ; metabolism ; Kinetics ; Peptides ; metabolism ; Peptides, Cyclic ; biosynthesis ; Staphylococcus aureus ; enzymology