Objective To explore the health-relate quality of life(QOL) status of adult persistent allergic rhinitis (PAR);the change of QOL of pro-post specific immunotherapy (SIT) and pharmacotherapy .Methods Skin prick tests(SPT) were performed on PAR patients .According to the results ,80 adult cases that were allergic to dermatophagoides were enrolled in ENT outpatient clinic of affiliated hospital of Nantong University from April to August 2011 .The patients were randomly allocated to receive either specific immunotherapy(n=40) or pharmacotherapy (n=40) ,all of them were given RQLQ before and after half-year treatment ;40 cases without any allergic diseases were chosen from ENT in-patient department ,and were given RQLQ .The scores of previous treatment of the PAR group were compared with health control group ,then compared with the scores of post-treatment ,and also compared the scores of post-treatment between the immunotherapy group and pharmacotherapy group .Results The scores of the PAR patients were higher than that of health control patients in all dimensions of RQLQ (P< 0 .05) ,and the most troublesome problems were nasal symptoms .The scores of the patients who received SIT were evidently lower than that of pro-treatment in all dimensions of RQLQ(P<0 .05) ,the scores of the patients who received medical treatment were also lower than before (P<0 .05) , and the scores of the SIT group were lower than the pharmacotherapy group (P<0 .05) .Conclusion The QOL of adult patients with PAR was improved after SIT or drug treatment ,and QOL improvement is more obvious treat by SIT .

The immune effect of CD4~+CD28~-T cells on Graves'ophthalmopathy(GO)was investigated.The expressions of interferon-γ(IFN-γ),interleukin-2(IL-2),and IL-4 in CD4~+ CD28~-T ceils were assayed by flow cytometry in GO patients,Graves'disease(GD)patients without ophthalmopathy,and healthy control subjects.The results showed that the percentage of CD4~+CD28~-T cells significantly increased in GO patients(P<0.05),with increased IFN-γ expression(P<0.05)and decreased IL-2 expression(P<0.05).These changes were closely correlated with clinical activity score(P<0.05).There were no significant differences in IL-4 expression among three groups.The resuh suggests that CIM~+ CD28~- T cells which hishly secrete IFN-γare related to the pathological lesion of GO.

Objective To study the effect of soluble, refolded, recombinant extracellular domain of the human Fc gamma receptor Ⅱ a (huFcγRⅡa) on the binding of human IgG to cells. Methods Extra-cellular domain of the huFcγRⅡ a gene was amplified from recombinant plasmid pe3huR Ⅱ by PCR and then cloned into pET-28a vector. The recombinant plasmid pETshuR Ⅱ was transformed into E. coli BL21 (DE3) after identified by PCR and doubly digested. The inclusion bodies of fusion protein were extracted and purified by washing, dissolved in 6 mol/L guanidine buffer, and refolded by rapid dilution technique. The refolding protein activity was tested by ELISA and flow cytometry. Results Extraceilular domain of the huFcγRⅡa gene was successfully cloned into pET-28a. The results of SDS-PAGE showed that the molecular mass (Mr) of the expressed protein was 24.8 × 103, and the expression rate was 30%. The purity of recom-binant shuR Ⅱ was up to 90% after washing. ELISA showed that the recombinant shuR Ⅱ was able to bind human IgG in a dose dependent manner, shuRⅡ could competitively inhibit the binding of human IgG to huFcγRⅡa expressed on the surface of COS-7 cells by flow cytometry. Conclusion The results demon-strate that it is possible to obtain large quantities of recombinant shuR Ⅱ which has comparable binding prep-erties to those of the whole membrane bound huFcγR Ⅱ a.

Objective To extract the loci of murine MHC gene and construct plasmids.Methods The RNA of mice was extracted and reversely transcribed into cDNA.By using nested PCR,the products were connected with T vector,cloned,and sequenced.Subsequently,the genes were digested by endonucleases,connected with expression vector,and sequenced again to choose the correct clones.Results After the nested PCR,the products were approved by sequencing.After being connected with the vectors,they were approved again by sequencing and the correct clones were chosen.Conclusion All of the loci of the MHC gene can be obtained by nested PCR.The plasmids from the correct clone can be used in the further experiments of transferring the gene to mitigate the transplantation rejection.

Objective: To investigate the effect and mechanism of helix B surface peptide (HBSP) on myocardial ischemia reperfusion injury (MIRI) in experimental mice.
Methods: The MIRI model was established by ligation of anterior descending coronary artery of the mice for 45 min and followed by corresponding treatment at 5 min before reperfusion. A total of 64 male ICR mice were randomly assigned to 4 group:①Sham group,②MIRI group, the mice received normal saline at 5 min before reperfusion,③HBSP group, MIRI mice received HBSP at 5 min before reperfusion and④HBSP+PD98059 group, MIRI mice received PD98059 (a speciifc blocker of ERK1/2) at 20 min before reperfusion and followed by HBSP at 5 min before reperfusion.n=16 in each group, all animals were treated for 2 hours. The area of myocardial infarction (MI) was detected by TTC-EB double staining method, the myocardial apoptosis rate was examined by TUNEL method, the levels of protein expression of ERK1/2 and phosphorylation of ERK1/2 were measured by Western blot analysis.
Results: Compared with MIRI group, HBSP group presented decreased MI area, decreased myocardial apoptosis rate and increased phopsphorylation level of ERK1/2, allP<0.05. Compared with HBSP group, HBSP+PD98059 group showed decreased phopsphorylation level of ERK1/2, increased myocardial apoptosis rate and increased MI area, allP<0.05.
Conclusion: HBSP may reduce the MI area via inhibiting myocardial apoptosis and therefore, protecting the experimental mice from MIRI; the mechanism might be related to the activation of ERK1/2 pathway.

Sudden chemical poisoning accidents have the characteristics of suddenness, groups, complexity and difficulty in rescue. It usually brings serious harm and far-reaching social impact. Establishing a good emergency management and rescue system from the four stages of prevention, response, rescue and recovery can reduce the adverse effects of accidents, casualties, and social burden. The prevention of sudden chemical poisoning accidents focuses on finding the cause and putting forward effective preventive measures. Emergency response can be carried out from three aspects: response and acceptation, hazard assessment and operation of corresponding emergency plans. Emergency rescue of sudden chemical poisoning accidents should fully consider both rescue technology and emergency management. The former includes pre-hospital patient screening and pre-hospital detoxification, while the latter includes dealing with the uncertainty of rescue process, allocation of medical resources and health care for emergency rescuers. In the recovery stage of the accidents, attention should be paid to long-term post-disaster monitoring of the population and emergency rescuers. Using the treatment time and development sequence as the framework, we can more comprehensively describe the characteristics of sudden chemical poisoning accidents, which is helpful in finding the key points of prevention and treatment of such accidents, and provide a basis for exploring the emergency managing methods of such accidents.

5-Flucytosine (5-FC) could be changed to 5-fluorouracil (5-FU) by cytosine deaminase (CD), the latter is able to kill cancer cells. However, there is no efficient method to deliver the CD gene into the tumor cells, which hampers the application of the suicide gene system. In this experiment, for the first time, the NDV has been utilized as a vector to deliver the CD gene into the cancer cells, the virus can infect the cancer cells specifically, replicate and assemble, while the cytosine deaminase is expressed. Then the CD converts the prodrug 5-FC into 5-FU to achieve the purpose of inhibiting tumor. Firstly, the whole genome of E. coli JM109 was extracted, and the CD gene was obtained by cloning method. Then the CD and IRES-EGFP were ligated into the pEE12.4 expression vector to become a recombinant pEE12.4IE-CD eukaryotic expression plasmid. The human liver cancer cells were transfected with the plasmid. The cells were treated with different concentrations of 5-FC, MTT method was used to determine the killing effect of CD/5-FC system on the human liver cancer cells. The cell deaths were 18.07%, 42.98% and 62.20% respectively when the concentrations of prodrug were at 10, 20 and 30 mmol x L(-1). In 5-FC acute toxicity experiment, Kunming mice were injected with different concentrations of 5-FC at intervals of 1:0.5. The LD50 of 5-FC through iv injection was determined by improved Karber's method, the LD50 was 507 mg x kg(-1) and the 95% confidence limit was 374-695 mg x kg(-1). According to the maximum LD0 dose of the LD50, the maximum safe dose was 200 mg x kg(-1). Body weight and clinic symptoms of the experimental animals were observed. These results laid the foundation to verify the antitumor effect and safety of CD/5-FC system in animal models. The CD gene was ligated into the NDV (rClone30) carrier, then the tumor-bearing animal was established to perform the tumor inhibiting experiment. The result showed that the recombinant rClone30-CD/5-FC system has a high antitumor activity in vivo. To summarize, CD gene has been cloned and its bioactivity has been confirmed in the mammalian cells. It is the first time in this study to utilize the recombinant NDV to deliver the CD gene into the tumor cells; our result proves the rClone30-CD/5-FC system is a potential method for cancer therapy.
Animals ; Antimetabolites, Antineoplastic ; metabolism ; pharmacology ; Cell Death ; drug effects ; Chick Embryo ; Cytosine Deaminase ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Flucytosine ; metabolism ; pharmacology ; Fluorouracil ; metabolism ; pharmacology ; Genetic Vectors ; Hep G2 Cells ; Humans ; Lethal Dose 50 ; Liver Neoplasms, Experimental ; pathology ; Mice ; Newcastle disease virus ; genetics ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection ; Tumor Burden ; drug effects

BACKGROUNDVolatile anesthetics (VAs) may affect varied and complex physiology processes by manipulating Ca(2+)-calmodulin (CaM). However, the detailed mechanism about the action of VAs on CaM has not been elucidated. This study was undertaken to examine the effects of VAs on the conformational change, hydrophobic site, and downstream signaling pathway of CaM, to explore the possible mechanism of anesthetic action of VAs.

METHODSReal-time second-harmonic generation (SHG) was performed to monitor the conformational change of CaM in the presence of VAs, each plus 100 µmol/L Ca(2+). A hydrophobic fluorescence indicator, 8-anilinonaphthalene-1-sulfonate (ANS), was utilized to define whether the VAs would interact with CaM at the hydrophobic site or not. High-performance liquid chromatography (HPLC) was carried out to analyze the activity of CaM-dependent phosphodiesterase (PDE1) in the presence of VAs. The VAs studied were ether, enflurane, isoflurane, and sevoflurane, with their aqueous concentrations 7.6, 9.5, 11.4 mmol/L; 0.42, 0.52, 0.62 mmol/L; 0.25, 0.31, 0.37 mmol/L and 0.47, 0.59, 0.71 mmol/L respectively, each were equivalent to their 0.8, 1.0 and 1.2 concentration for 50% of maximal effect (EC50) for general anesthesia.

RESULTSThe second-harmonic radiation of CaM in the presence of Ca(2+) was largely inhibited by the VAs. The fluorescence intensity of ANS, generated by binding of Ca(2+) to CaM, was reversed by the VAs. HPLC results also showed that AMP, the product of the hydrolysis of cAMP by CaM-dependent PDE1, was reduced by the VAs.

CONCLUSIONSOur findings demonstrate that the above VAs interact with the hydrophobic core of Ca(2+)-CaM and the interaction results in the inhibition of the conformational change and activity of CaM. This in vitro study may provide us insight into the possible mechanism of anesthetic action of VAs in vivo.

Adenosine Monophosphate ; analysis ; Anesthetics, Inhalation ; pharmacology ; Anilino Naphthalenesulfonates ; Calmodulin ; antagonists & inhibitors ; chemistry ; physiology ; Cyclic Nucleotide Phosphodiesterases, Type 1 ; analysis ; Fluorescence ; Humans ; Hydrophobic and Hydrophilic Interactions

OBJECTIVETo construct four different micro- and nano-phase titanium film models and investigate the characteristics of their surface micro-topography.

METHODSFour different titanium films were prepared on commercial titanium discs, by direct current magnetron sputtering, at ambient, 100, 250, 380 degrees C substrate temperature, respectively. Their surface topography and crystal sizes were investigated using atomic force microscope (AFM) and X-ray diffraction (XRD). The size of granule and surface roughness in different group was calculated and compared.

RESULTSAll samples were covered by a thin film consisting of dense round or ovaloid granules. The granules and crystals was growing as the substrate temperature increasing. The Ti substrate had greater effect on the surface topography of film compared with Si substrate. This kind of complex topography caused the surface roughness of Ti substrate group decreased as the granules growing.

CONCLUSIONIn our study, four different micro- and nano-phase titanium film models were constructed for our coming investigation of their topographical influence on biological reaction of proteins and cells. Basic data on surface features was obtained for next in vitro and in vivo experiment.

Surface Properties ; Titanium ; X-Ray Diffraction

OBJECTIVETo explore the differences of the silica-induced inhibition on cellular proliferation and hprt gene mutagenesis between lung fibroblasts and alveolar type II cells.

METHODSThe proliferation inhibitive cytotoxicity was detected by MTT (3-[4,5-Dimethylthiazolzyl]-2,5-Diphenyl Tetrazolium Bromide) colorimetric method. Mutation in the hprt gene was screened by culture in the presence of the toxic purine analog, 6-thioguanine (6-TG).

RESULTSUnder the same circumstances of silica exposure, alveolar type II cells was more sensitive than lung fibroblasts for proliferation inhibition. The median proliferation inhibition concentration (IC50) of silica on epithelial was 140 micrograms/cm2, whereas IC50 of silica on fibroblasts was 282 micrograms/cm2. At the same doses of silica, the hprt gene mutation frequency in type II cells (84.2 x 10(-6))-156.6 x 10(-6) was statistically higher than that in fibroblasts (67.6 x 10(-6)-114.3 x 10(-6), P < 0.05).

CONCLUSIONThere were significant differences of both silica-induced cell proliferation inhibition and hprt gene mutation between rat lung fibroblasts and type II epithelial cells. In vitro, cultured rat alveolar type II cells were more sensitive in cytotoxicity and hprt gene mutagenesis to silica dust than lung fibroblasts were.

Animals ; Cell Proliferation ; drug effects ; Epithelial Cells ; drug effects ; Fibroblasts ; drug effects ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Lung ; cytology ; drug effects ; metabolism ; Mutation ; Pulmonary Alveoli ; cytology ; drug effects ; Rats ; Silicon Dioxide ; toxicity