Purpose To study the effect of overexpressing either wild type or a familial Alzheimer disease mutant presenilin 1 (mPS1) on tau phosphorylation in neuroblastoma NG-108 cells. Methods Three different plasmids transfected NG-108 cells respectively. Immunostaining and confocal microscopic technique were used to study the distribution of presenilin 1 and phosphorylated tau. Immunoblot test was applied to investigate the change of tau phosphorylation. Results Immunostaining showed that in brain of sporadic Alzheimer disease, PS1 mainly distributed in neuron and partially colocalized with the phosphorylated tau. Immunoblot tests showed that the cells transected either wild type PS1 or mPS1 contained more phorphorylated tau than the control cells. However, MTT test showed no significant difference between mock transfected cells and the wPS1 or mPS1 transfected cells. In addition, after transfection of the constructed PS1-EGFP vector, overexpressed EGFP-PS1 was located at cell surface membrane and subcellular organelles at earlier time at 12 hr, then EGFP-PS1 diffused in cytosol. Immunocytochemical observations demonstrated that some of the PS1-EGFP transfected cells contained more phosphorylated tau protein, which formed aggresome with PS-1-EGFP. When treated with phosphotase inhibitor okadaic acid, in the PS1-EGFP transfected cells accumulated more phosphorylated tau than the un-transfected cells. Conclusions Wild type PS1 is possibly involved in tauopathy in sporadic Alzheimer's disease.

Objective To evaluate the surgical procedures and clinical results to treat the bleph arochalasis.Methods A series of 43 blepharochalasis in 23 patients were surgically treated.A double eyelid fold incision was made.Surgeon clippd redundant skin of upper eyelid with forceps,so that would be excised was marked out.Then redundant skin and muscles as well as hernia septal fat were removed.The lacrimal glands were found and repositioned into lacrimal glands fossa behind superolateral orbital rim.We choose different methods to perform plastic operation in 23 cases of blepharochala sis according to different topographic anatomic characteristics.Results Postoperatively,not only the patients recovered in a followed-up period of 1-3 years,but all lacrimal glands were normally rep soitioned.Conclusions These methods are effective in patients with blepharochalasis and meet the cosmetic demands.Good clinical results in both function and appearance are achieved.

Objective:To construct eukaryotic expression plasmid ecording a novel surface protein Fba of GAS, and to explore its impact on host immune responses.Methods:Fba gene was amplified by PCR using strain SSI-9 (GAS M1 serotype isolates) as the template, then cloned into pcDNA3.1 for constructing eukaryotic expression plasmid pcDNA3.1/fba and sequenced. Female CD1 mice were randomly individed into 6 groups, and immunized respectively with Fba protein, M protein, pcDNA3.1/fba + Fba protein, pcDNA3.1/fba, pcDNA3.1 and PBS as control. Blood was obtained from the mice and specific antibody of IgG was detected by ELISA. Spleen cells were assessed with lymphocyte proliferation assays.CD4+T cell and CD8+T cell were detected by flow cytometry (FCM). Assay results were analyzed with SPSS10.0.Results:The IgG against Fba protein kept hightest levels in group immunized with Fba protein. The levels of lymphocyte proliferation, CD4+, CD8+T cell were significantly high in the group pcDNA3.1/fba. Conclusion:(1) Just as M protein, the antibody to Fba protein could be efficiently induced by immunization with Fba protein, which showed that Fba protein was hopefully to be a candidate protein for vaccine against GAS.(2) Eukaryotic expression plasmid pcDNA3.1/fba was successfully constructed and this recombinant plasmid could efficiently induce antibody and CD4+ T cell for againsting GAS.

The pathogenic effects of crops from Keshan disease area were studied by rat-feeding. As double controls, another two groups were fed with crops from non-endemic area and synthetic diet. Those from disease area caused necrosis of heart muscle vascular changes of myocardial mitochondria and fatty degeneration of liver. Compared with the controls, the free radical and lipid peroxide content in heart and liver were significantly higher, while the ATP and AN contents in these organs were apparently lower. The multielement contents and its constituent patterns in the feeds, whole blood, heart, liver, kidney and brown adipose tissue of rats in experimetal group were apparently different from those in the control groups. The statistic analysis of multiple factors showed that eight elements Se, Mo, Cu, Mn, Cr, Fe, Zn and Pr might be the perilous factors, that is, under the condition of low selenium, the constituent pattern of selenium and the eight elements might be the complicated factors of endemic crops.

Objective To express the novel fibronectin-binding protein Fba of group A streptococcus(GAS)and analyze its immunogenicity,so that to evaluate the immune responses to GAS infection.Methods fba gene was amplified by polymerase chain reaction(PCR)and confirmed by sequencing.Then it was cloned into pGEX4T-2 vector and Fba protein was expressed in E.coli BL21.The protein expression was identified by enzyme-linked immunosorbent assay(ELISA)and Westernblot.The sera from mice infected with GAS and anti-streptolysin-O positive patients were detected using microtiter plates coated with purified Fba protein as antigen.Afterward Balb/C mice were immunized with this purified protein and the sera were collected after the third immunization for the detection of IgG titer.Results It was confirmed by ELISA and Western blot that the recombinant Fba protein had a specific affinity with anti-Fba sera of rabbit.The anti-serum IgG titer of mice imrnunized with Fba protein was up to 1:4800.Conclusions GAS infection or Fba protein immunization are able to induce high serum titer of anti-Fba which could react specifically with the recombinant Fba protein.It indicates that Fba protein has good immunogenicity and antigenicity.So Fba protein could be a GAS candidate vaccine and an important tool to detect anti-GAS titer in GAS infected patients.

Objective To analyze adherence to short message service (SMS) based obesity intervention in overweight and obesity adults in Beijing.Methods Sixty-three participants received a 24-week (3 stages) SMS obesity intervention,and were then required to answer questions about their performance of 3 individualized weight loss goals daily via SMS.Three group sessions and monthly coaching call were also conducted.Adherence was graded according to the SMS reply rate and goal score.Logistic regression analysis was computed to analyze the influence factors of adherence.Results Among 55 intervention completes,the rate of loss to follow-up was 12.7％.Adherences of 55 participants were ranked as 3 levels:high level (SMS reply rate 5-7 d/wk and score ＞ 9) 56.4％,middle level 36.3％ and low level 7.3％.Association between body mass index (BMI) change and adherence was statistically significant (r =-0.241,P =0.026),similar to relationship between waist circumference (WC) change and adherence (r =-0.303,P =0.005).At 24-week,BMI and WC of the high level group were reduced by 2.26％ and 3.80％,respectively,and the changes were statistically different among 3 groups (BMI:F =3.659,P =0.033 ; WC:F =4.699,P =0.013).Logistic regression analysis showed that advanced age (odds ratio (OR) =1.108,95％ CI:0.997-1.231),obesity (OR =12.974,95％ CI:1.245-135.195),more suitable goals(OR =1.451,95％ CI:0.974-2.162),weight lost early (OR =10.982,95％ CI:1.608-75.007),usually replied SMS in the morning (OR =6.725,95％ CI:1.098-41.201)were favorable factors of adherence.High expectation to weight loss (OR =0.055,95％ CI:0.005-0.626)was the negative factor of adherence.Conclusions SMS is a promising tool to promote self-monitoring adherence for Chinese overweight and obesity adults,and good adherence indicates more weight loss.Many factors could influence SMS adherence,and more strong evidence on adherence to SMS-based obesity intervention are desired.

Spore surface display is one of attractive microorganism surface display systems. With the advantage of resistance attribute and specific assembly pattern, the technology of spore surface display now is attracting more and more attention. According to the current reports and main achievements of spore surface display, the structure and assembly of spores, the principle for construction and some existing spore surface display systems were elaborated in this paper. Now with the unique property of spores, the technique is not only widely used in production of vaccines but also has great applied potential in the field of biocatalysis and cell-factory.
Bacillus subtilis ; genetics ; metabolism ; Biocatalysis ; Biotechnology ; methods ; Gene Expression Regulation, Bacterial ; Genetic Engineering ; methods ; Recombinant Proteins ; genetics ; metabolism ; Spores, Bacterial ; genetics ; metabolism ; Tetanus Toxoid ; genetics ; immunology

Objective:To identify monoclonal antibody McAb2 recognizing epitope of Fba of GAS.Methods:The overlapped peptides were synthesized and their abilities to bind McAb2 were detected by dot-ELISA.The predominance amino acids specific for McAb2 were screened using phage 7 peptide library.Results:The result by dot-ELISA analysis demonstrated that the synthetized peptide,amino-acid residues 100-112~(th),could bind McAb2 with high affinity.The predominance amino acids specific for McAb2 were ITPDL,which was located in 100-110~(th)aa of Fba by panning with phage 7 peptide library.Conclusion:The domain and the predominance amino acids of Fba recognized by McAb2 is determined.The results would contribute to the research of the role of Fba on the pathogenic mechanism of GAS,the identification of function of McAb2,and the development of epitope-peptide vaccine.

Objective The truncated fibronectin-binding proteins A (Fba protein) were cloned and expressed, then animals were immunized with Fba protein and subsequently challenged with group A streptococcus (GAS) to further investigate protective antibodies induced by each domain and determine the most immunogenic domain. Methods Fba proteins, which were divided into four overlaps based on the structural domains, were truncated and expressed. The fba truncated genes were amplified by polymerase chain reaction (PCR) with SSI-9 of GAS as template, and cloned into prokaryotic expression plasmid pGEX-2T and expressed in E. coli BL-21. The products were confirmed by Western blot and purified by affinity chromatography column. Female BALB/c mice were immunized with the four truncated proteins respectively, with phosphate buffered solution (PBS) as control. The serum IgG of mice was detected by enzyme-linked immunosorbent assay (ELISA). After the third immunization, mice were challenged with fatal dose of GAS (M+ Fba+ ) to evaluate the protective rates in each group. The data were compared by analysis of variance and Fisher's exact test.Results Prokaryotic expression plasmids of pGEX-2T/FbaAl, pGEX-2T/FbaA2, pGEX-2T/FbaA3 and pGEX-2T/FbaA4 were successfully constructed and the four truncated proteins were expressed and purified successfully. Serum levels of IgG in each experimental group gradually increased with immunization with Fba protein more times. After the third immunization, the IgG titer against FbaA2 1290.2, P<0. 01). After GAS challenge, four out of eight mice were protected in FbaA2 protein group, while two out of eight mice in FbaAl protein, FbaA3 protein and FbaA4 protein groups,respectively (P<0. 05). Conclusions Four truncated Fba proteins are constructed and expressed successfully. Truncated FbaA2 protein could be able to induce strongest protective immune response.

Objective:To observe cellular,humoral and mucosal immune responses induced by DNA-or protein-based of FimH of UPEC type 1.Methods:After mice were immunized respectively with recombinant plasmid pcDNA3.1/fimH or pcDNA3.1/fimC,and the combinant FimH and FimC protein,the anti-FimH protein IgG of sera and SIgA in bladders were detected by ELISA.The lymphocyte phenotypes of CD3,CD4 and CD8 were analyzed by FCM.Results:The titers of IgG in sera and SIgA in the bladders were all low in the group immunized by recombinant FimH plamid,but the percentage of CD4+T cells in spleen was high,which revealed that recombinant FimH plamid was able to trigger better cellular immune response.The titers of IgG were very high in the group immunized by FimH protein,which suggested that the FimH protein was able to trigger better humoral immune response,but SIgA in the bladders was not detectable.Conclusion:The DNA for FimH can induce humoral,mucosal and cellular immune response.FimH protein can only induce humoral immune response.FimC protein is able to enhance the immunogenicity of FimH protein.