A novel biosensor for aflatoxin B1 detecting has been reported. The biosensor electrode for AFB1 detecting was assembled by immobilized aflatoxin-oxidoreductase using open-ended multi-walled carbon nanotubes as matrix. Its linear range was between 0.16?M and 3.2?M. And if the specific anti-aflatoxin B1 antibody and aflatoxin oxidoreductase were both immobilized on the electrode with Multi-Walled carbon nanotubes, the detection limit of the modified electrode could be 16 nM with a 10 times improved sensitivity. The aflatoxin enzyme biosensor assembled this way strode one step forward its practical application.

Objective Intranuclear forkhead/winged helix transcription factor p3(Foxp3) plays a key role in T cell-mediated immunosuppression.The present study was performed to investigate the effects of high mobility group box-1 protein(HMGB1) on Foxp3 gene as well as protein expressions in splenic regulatory T cells(Tregs) and their potential regulating mechanisms in mice.Methods CD4+CD25+Tregs isolated from the spleens of male BABL/c mice by magnetic beads were seeded on 96-well(1?105 cells/well) cell culture plates coated with anti-CD3(1 ?g/ml) and soluble anti-CD28(1 ?g/ml),and the cells were stimulated with HMGB1 at various intervals or at different concentrations.After being stimulated,the Foxp3 mRNA/protein expressions in the Tregs were determined.The time-dependent and dose-dependent responses between HMGB1 and intranuclear Foxp3 expression were analyzed by flow cytometry,and the expressions of Foxp3 mRNA of Tregs were analyzed by quantitative PCR of SYBR GREEN.Results After stimulation with HMGB1,the intranuclear Foxp3 protein and mRNA expressions of splenic Tregs in mice were markedly down-regulated in 24 h to 72 h(P

Objective Thoracic cavity fistula following esophagus carcinoma resection is a serious complication with a high mortality.This study aims at a better therapy for thoracic cavity fistula following esophagus carcinoma resection by summarizing the ex-perience with the four-tube strategy ( jejunal fistula tube, stomach tube, chest drainage tube, and nasal fistula tube) in the treatment of the complication. Methods We retrospectively analyzed the clinical data about 62 cases of thoracic cavity fistula following esopha-gus carcinoma resection, 35 treated with the four-tube strategy ( treatment group) and the other 27 with the three-tube ( stomach tube, chest drainage tube, and nasal fistula tube) method ( control group) .We compared the hospital days, wound healing time, mortality, and incidence of anastomotic stenosis at 6 months after operation between the two groups of patients. Results Compared with the controls, the treatment group showed remarkable decreases in the hospital days (P<0.05), wound healing time (P<0.05), and mortality (P<0.05), but no statistically significant difference was observed in the incidence rate of anastomotic stenosis at 6 months after operation between the two groups of patients ( P >0.05 ) . Conclusion Compared with the three-tube method, the four-tube strategy has the advantages of shorter healing time and lower mortali-ty, and therefore is preferable for the treatment of thoracic cavity fis-tula following esophagus carcinoma resection.

Objective To observe the expression law of cytokine signaling suppressors (SOCSs) mRNA in burn rats with Staphylococcus aureus sepsis and investigate their potential role in the pathogenesis of postburn sepsis. Methods Wistar rats were inflicted with 20% TBSA Ⅲ? scald, followed by Staphylococcus aureus challenge. Then, the expressions of SOCS1, SOCS2 and SOCS3 mRNA and interferon-? (IFN-?) levels in the liver and lungs were determined. Results With Staphylococcus aureus challenge after burn, IFN-? levels in the liver and lungs were significantly elevated and reached peak at the 0.5th and 6th hours, respectively (P

Objective To study the potential role of tumor necrosis factor-α (TNF-α) induction in the development of mucosal barrier dysfunction in rats caused by acute intestinal ischemia-reperfusion injury, and to examine whether pretreatment with monoclonal antibody against TNF-α (TNF-α MoAb) would affect the release of D(-)-lactate after local gut ischemia followed by reperfusion. Methods Anesthetized Sprague-Dawley rats underwent superior mesenteric artery occlusion for 75 min followed by reperfusion for 6 hr. The rats were treated intravenously with either TNF-α MoAb (20 mg/kg) or albumin (20 mg/kg) 30 min prior to the onset of ischemia. Plasma D(-)-lactate levels were measured in both the portal and systemic blood by an enzymatic spectrophotometric assay. Intestinal TNF-αmRNA expression as well as protein levels were also measured at various intervals. In addition, a postmortem examination was performed together with a macropathological evaluation based on a four-grade scoring system.Results Intestinal ischemia resulted in a significant elevation in D(-)-lactate levels in the portal vein blood in both the control and treatment groups ( P ＜0.05). However, animals pretreated with TNF-α MoAb at 6 hr after reperfusion showed significant attenuation of an increase in both portal and systemic D(-)-lactate levels when compared with those only receiving albumin (P ＜ 0.05). In the control animals, a remarkable rise in intestinal TNF-α level was measured at 0.5 hr after clamp release ( P ＜ 0.01); however, prophylactic treatment with TNF-α MoAb completely annulled the increase of local TNF-α levels seen in the control animals. Similarly, after anti-TNF-α MoAb administration, intestinal TNF-α mRNA expression was markedly inhibited, which showed significant differences when compared with the control group at 0.5 hr, 2 hr and 6 hr after the release of occlusion ( P ＜ 0.05-0.01 ). In addition, the pathological examination showed marked intestinal lesions that formed during ischemia, which were much worse upon reperfusion,particularly at the 6 hr time point. These acute injuries were obviously attenuated in animals receiving TNF-α MoAb.Conclusions It appeared that acute intestinal ischemia was associated with failure of the mucosal barrier, resulting in increased plasma D(-)-lactate levels in both portal and systemic blood. These results suggest that TNF-α appears to be involved in the development of local damage associated with intestinal ischemic injury. Moreover, prophylactic treatment with TNF-α MoAb exerts preventive effects on ischemia/ reperfusion-induced circulating D (-)-lactate elevation and gut injury. ( J Geriatr Cardiol 2004;1(2):119-124. )

SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.

Armillariella tabescens EJLY2098 was capable of secreting p-mannanase by konjac inducement. A 34 orthogonal design was applied to determine the optimum medium of inducing mannanase by Armillariella tabescens EJLY2098. The results suggested that Armillariella tabescens EJLY2098 secreted the high-activity enzyme in the optimum medium, which was composed of 2% konjac, 1% peptone, 25% potato juice,0.3% KH2PO4,15% MgSO4?7H2O, 0.01% VitB1. Purified by DEAE-anion exchange chromatography, two eluting peaks (P1 and P2) with the p-mannanase activity were obtained, and one of them (named?-mannanase P2) was a single band by the SDS-PAGE, and the molecular weight of?-mannanase P2 was 78. 9kDa. The isoelectric point of?-mannanase P2 was estimated to be 4.0-4. 1. The optimum activity for the enzyme was found at 60℃and pH4. 0 - 6. 0, and the enzyme was stable between pH4. 5 - 6. 0. The activity of?-mannanase P2 were enhanced by Na+ and Ba2+ . This?-mannanase can be used in feed industy. a new fungi secreting?-mannanase was obtained, providing an important base for cloning mannanase gene and constructing recombin microbe expressing?-mannanase .

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.

Absract: Objective To report the result of annual monitoring and analysis of nasopharyngeal virus in children with respiratory tract infections in Nanxiang, Shanghai District. Methods Nasopharyngeal secretions were collected from 4389 children with acute respiratory tract infection in outpatient department from January 2007 to September 2013, 9 common respiratory viruses were analyzed by Multiplex RT-PCR, including inlfuenza virus (FLU), parainlfuenza virus (PIV), respiratory syncytical virus (RSV) , adenovirus (ADV), human bocavirus(HBOV), human coronavirus(Cov), enterovirus(EV), human metapneumovirus(HMPV), and rhinovirus(HRV). The same analysis was done in 123 asymptomatic children during the same period. Results The positive rate of detected respiratory viruses in children with respiratory tract infections in nasopharyngeal secretions were 34.8% (1526/4389), including FLU 10.3% (453/4389), RSV 7.3% (320/4389), PIV 6.2%(274/4389), ADV 3.3%(146/4389), HBOV 2.7%(118/4389), EV 2.5%(110/4389), Cov 2.4%(105/4389), HRV 1.6%(72/4389), HMPV 1.5%(67/4389);two and more combined respiratory viral infection were found in 273 cases (6.2%). The virus detection
rate between age groups was signiifcantly different (χ2=41.91, P<0.001). The school-age group had the lowest positive rate of 23.4%and the positive rates in other three groups were all higher than 35.0%. The infant group had the higher positive rate of RSV and HRV. FLU detection rate in school-age group was 13.6%. Respiratory viruses in children with asthmatic disease has high detection rate. RSV infection rate was the highest 14.8%(30/204) in the asthmatic disease group, followed by HBOV 13.8% (28/204). In nasopharyngeal secretions of 123 asymptomatic children, virus-positive detection rate of 6.5% (8/123), which showed signiifcant difference from that in respiratory virus infection group (χ2=42.60, P<0.001). Conclusions In seven consecutive years of testing, the inlfuenza virus and respiratory syncytial virus play an important role in children with respiratory tract infections in this region. The detection rate of virus showed difference between different age groups and a higher detection rate of RSV in infants with respiratory tract infections was observed. The overall detection rate of virus was decreased with the increase of age excluding the inlfuenza virus.

Objective to investigate the clinical effect of negative pressure suction by double caping pipe on cervical esophagus fistula after esophageal carcinoma surgery,and search for the effective treatment of cervical anastomotic fistula after esophageal carcinoma surgery.Methods The clinical data of 140 patients with cervical esophagus fistula after esophageal carcinoma surgery in Nanjing General Hospital of Nanjing Command from September 2004 to September 2015 were retrospective analyzed.Among them,85 cases were treated with low negative pressure suction by double caping pipe as experimental group,55 cases were treated with conventional drainage and dressing as the control group.The length of hospital stay,healing time,dressing frequency,neck bleeding risk,anastomotic stenosis and mortality rate between two groups were analyzed and compared.Results The length of hospital stay,the time of wound healing,the frequency of wound change dressing,the rate of neck bleeding in experimental group were (15.94± 1.57)d,(8.00± 1.55)d,(6.22± 1.52)times,1.18％ respectively,significantly lower than that of control group ((23.64 ± 2.36) d,(15.64 ± 2.08) d,(27.56±3.58) times,12.24％;P=0.000,0.000,0.000,0.029).While the rate of anastomotic stenosis after half a year in experimental group was 17.65％,lower than that of the control group (23.64％),the difference was no significant(P=0.387).Conclusion The treatment on cervical esophagus fistula with low negative pressure suction by double caping pipe has superiorities on hospitalization and healing time,dressing frequency,neck bleeding risk,mortality,and does not increase the incidence of anastomotic stenosis,and it can achieve a better therapeutic effect compare with the conventional drainage and dressing.