Allergens ; immunology ; Child ; Cross Reactions ; Humans ; Hypersensitivity ; immunology ; Latex Hypersensitivity ; immunology

During L-lactic acid fermentation with Rhizopus oryzae,there existed a branch pathway by which pyruvate was transformed to eth- anol catalyzed by pyruvate decarboxylase(PDC)and alcohol dehydrogenase(ADH),thus decreasing the flux of pyruvate to lactic acid.In this study,the spores of Rhizopus oryzae AS3.3462 mutagenized with nitrosoguanidine(NTG),the appropriate dosage was 0.15 mg/mL and the lethal rate was 70%～80%.Two mutants,named mut-1 and mut-2,with decreased ADH activity were screened out by yeast peptone dextrose(YPD)agar medium containing allyl alcohol.These two mutants had decreased ADH activities of 41.63% and 50.29% compared with the parent strain.The fermentation behavior after 72h showed that the yields of ethanol produced by mut-1 and mut-2 were 4.87g/L and 6.56g/L respectively,while the wild type strain was 28.9g/L,and the lactate concentrations of mut-1 and mut-2 also increased from 40.31g/L to 54.45g/L and 44.07 g/L,respectively.It is also found that mut-1 and mut-2 had a high reducing sugar consumption rate and biomass accumulation than its present strain

Objective:To study the feasibility of electronic gingivoplasty on the subjects suffering from high blood sugar or blood coagulating dysfunction on rat model.Methods:Electronic gingivoplasty was performed in 10 SD rats with high blood sugar and 10 with blood coagulating dysfunction on the right side of mandible and scalpel gingivoplasty on the left.For each group of diseased rats 10 healthy rats were used as the controls. Gingival tissue slices of the rats with high blood sugar and the controls were obtained 1,24, 72 hours, one and two weeks after operation respectively, and processed for histological observation. The bleeding condition were evaluated with paper tips during the operations on the rats with blood coagulating dysfunction and the controls.Results:The process of histological reaction in the tissues of operation area after electronic gingivoplasty was basically similar to that after scalpel gingivoplasty. Electronic surgery caused more slight inflammatary reaction than scalpel surgery in high blood sugar group. The amount of bleeding during electronic surgery was significantly smaller than that during scalpel surgery in blood coagulating dysfunction group.Conclusion:Electronic surgery is feasible for gingivoplasty on the subjects with high blood sugar or blood coagulating dysfunction.

Objective To investigate the changes of circulaling progenitor cells and endothelial progenitor cells(EPCs)in non-septic and septic shock patients using flow cytometry.Method A total of 27 sepsis patients hospitalized in emergency department of Zhongshan hospital during August 2007 to February 2008 were enrolled in this study.The patients were dividedinto septic shock group(n=12)and non-septic shock group(n=15).Ten healthy individuals and ten non-sepsis ICU patients were collected as controls.Peripheral blood mononuclear cells(PBMCs) were isolated by Ficoll density gradient centrifugation,and EPCs labelled with antibodies against CDl33,CD34 and VEGFR-2 were identified and isolated by three-color fluorescence flow cytometry.Differences within the groups were analyzed using One way ANOVA.Results The percentages ofprogenitor cells and EPCs in the PBMC fraction in healthy controls were(0.25%4-0.14%),(0.09%-I-0.02%),respectively,and those in ICU controls were(O.38%.4-0.29%),(0.12%.4-0.02%).The percentages of progenitor cells and EPCs were significantly higher in栅sel如c shock patients(0.57%±0.12%),(0.22%.4-0.10%)than in heathy and non-sepsis ICU controls(P<0.05).However.the percentages of progenitor cells and EPC8 in septic shock pa.tienta(O.20%±0.12%,0.04%-t-O.01%)was obviousely lower than those in healthy,ICU controls and ilionseptic shock patients(P<0.05).Sptic shock survivors had significantly higher numbers of cEPCs than nonsur.vivors(P<0.05).Conclusions The level of progenitor cells and EPC8 in peipheral blood of sepsis patients might be the valuable markers to as.se88 the severity and outcome ofthese ptienS.

The potency of an improved recombinant multi-epitope vaccine against FMDV type Asia1 was evaluated in this study .A multi-epitope gene based on FMDV type Asia1 was designed and a recombinant expression plasmid (pRE-oIgG) was constructed .The proteins ,RE-oIgG and 3D were expressed in E .coli cells and purified with Ni-NTA agarose resin by affinity chromatography .The proteins ,RE-oIgG ,3D and RE-oIgG plus 3D ,were emulsified in an oil adjuvant ISA 206 .Twenty-five female guinea pigs were randomly divided into five groups and intramuscularly vaccinated for with RE-oIgG ,3D ,RE-oIgG plus 3D ,an inactivated FMDV vaccine (type Asia1) ,and PBS .All animals were vaccinated for two times .Anti-FMDV specific an-tibodies ,neutralization antibodies ,protection potency ,and lymphoproliferation assay were detected by ELISA ,virus neutrali-zation assay ,challenge test ,and flow cytometry ,respectively .Results showed that RE-oIgG plus 3D elicited significant high-level anti-FMDV specific antibodies compared to RE-oIgG alone (P<0 .05) .All the vaccinated animals induced higher level lymphoproliferation responses in vitro except PBS .Both 3D alone and PBS produced the negligible neutralizing antibodies and anti-FMDV specific antibodies .RE-oIgG plus FMDV 3D not only elicited high levels of anti-FMDV neutralizing antibodies ,but also induced significant lymphoproliferation responses .More importantly ,RE-oIgG plus 3D conferred complete protection to guinea pigs against challenge with 1 000 GPID50 .Interestingly ,two of five vaccinated animals with 3D alone were full protected against challenge ,and other three animals significantly showed a delay of 2-3 days in the onset of clinical signs .Therefore ,we considered that RE-oIgG plus 3D induces strong humoral and cellular immune responses ,which may be used for control and prevention of FMD in the future .

Objective To monitor the activities of orbicularis oculi muscle by analyzing the electroneurogram (ENG) of its innervating nerve. Methods The ENG was recorded in vivo by a cuff electrode implanted around a dog’s zygomatic branch of facial nerve. The differences between the ENG during relaxing period of the orbicularis oculi muscle and during the constricting period were analyzed to explore a way to monitor the muscle activities. Results The ENG signals indicating occurrences of eye blinking events were extracted in the study. Time-frequency analysis showed that there was no ENG difference when the orbicularis oculi muscle was in relaxing periods no mater whether it was in the nature eye-opening period or in the interval of eye blinking. The ENG of orbicularis oculi muscle in constricting period was different from the ENG in relaxing period at amplitude and frequency. Conclusion It is feasible to monitor the orbicularis oculi muscle activities by ENG analysis.

Objective To construct GST/HIF1α fusion protein expression vector and induce its expression in Escherichia coli (E.coli). Methods The coding sequence of hypoxia inducible factor-la (HIF-la) and its deletion fragments were amplified from the plasmid pcD-NA3.1-HIF-la by PCR and inserted into pGEX-4T-2 by BamHI and Not I. The positive recombinanls were identified by restriction endonu-clease digestion and DNA sequencing. Then they were transformed into E.coli BL21, induced by IPTG and identified by SDS-PAGE and Western blot. Results The prokaryotic expression plasmid pGEX-4T-2-HIF-lα and its deletion mutants were successfully constructed and confirmed by enzyme digestion and sequencing. The desired CST/HIF-1α fusion proteins were expressed and confirmed by Western blot. Conclusion The prokaryotic expression plasmid of HIF-la and its deletion mutants were successfully constructed and the expression of fu-sion proteins was confirmed. This study provides the basis for the further research on purifying HIF-la protein and the biological function of HIF-lα.

Objective To investigate the effects of interleukin -15 ( IL-15 ) on the cytotoxicity of Vδ2 γδ(Vδ2) T cells against K562 cells.Methods PBMCs were separated and cultured with zoledronate and interleukin-2 ( IL-2) to induce the proliferation of Vδ2 T cells.The obtained Vδ2 T cells were in vitro stimulated with IL-15.Flow cytometry analysis was performed to evaluate the changes of phenotypes of Vδ2 T cells and their cytotoxicity against K562 cells.Results Zoledronate effectively induced the massive prolifer -ation of V2δT cells.The Vδ2 T cells were highly activated and the expression of CD 107a and IFN-γby Vδ2 T cells were upregulated upon IL-15 stimulation.The cytotoxicity of Vδ2 T cells against K562 cells was greatly enhanced by the treatment with IL-15.Conclusion IL-15 could activate the Vδ2 T cells and en-hance their cytotoxicity against K562 cells.

Objective:To study the role of γδ T cells and Vδ1 and Vδ2 T subsets played in patients with chronic hepatitis C.Methods:The percentage of peripheral bloodγδT cells and Vδ1 and Vδ2 T subsets cells was assessed by flow cytometry ( FACS) . Results:There were no significant differences in the percentage of circulating γδ T cells and Vδ1 and Vδ2 T subsets between HCV-infected patients and healthy controls ( HCs).However,The number of peripheral blood Vδ2 T cells from HCV-infected patients was positively correlated with serum alanine aminotransferase ( ALT) levels,but showed no correlation with serum HCV RNA.Peripheral Vδ2 T cells from HCV-infected patients were in activated status.The expression of CD107a was enhanced in Vδ2 T cells from HCV-infected patients compared to HCs.Conclusion:Vδ2 T cells were involved in liver injury in chronic HCV-infected patients.

ObjectiveTo observe the effect of Integrated Chinese-western three-stage rehabilitation on activities of daily living (ADL) in stroke patients. Methods60 stroke patients were randomly divided into the trial group and control group with 30 cases in each group. Patients of control group were treated with routine neurological interventions and three-stage rehabilitation, while those of trial group were administered resuscitating acupuncture therapy, scalp and body acupuncture in addition. All the patients were assessed with modified Barthel index (MBI) at the beginning, 1 month, 3 months, 6 months respectively after stroke. ResultsThere was no significant difference in ADL between the trial and control groups at the beginning, and both improved at the following period (P<0.01). At each of the following period, either the ADL scores (P<0.05) or the increase of the ADL scores (P<0.05) of the trial group were significant higher than those of the control group. ConclusionIntegrated Chinese-western three-stage rehabilitation could improve the ADL further for the stroke patients.