Bacterial ghost is intact bacterial envelope which is lysised by the lysis geneE of PhiX174. It can be used as vaccine directly. Foreign antigen can be targeted into outer membrane, inner membrane or the periplasmic space of bacterial, as a result, a recombinant bacterial ghost is constructed. Bacterial ghost, as a novel drug delivery system, is becoming more and more concerned, which can deliver DNA and protein vaccine or other drugs in order to have a better immune responses and therapeutic effects.

The paper reported two methods. a genetic probe and the solid phase radio-immunological assay, for the detection of the thermolabile toxin produced by Escherichia coli. The former is the application of a 32P-IabelIed probe of B subunit DNA prepared from Hind Ⅲ restriction fragment of LT-encoding DNA. and the latter is by using CNBr-activated paper as carrier and 125I-SPA to substitute the secondary antibody. Applying the methods to 160 strains of E. coli isolated from patients suffering from infantile diarrhea, the results showed that 42 out of 160 strains belong to the strain which produces the thormolabile toxin. These two methods not only have the advantages of being sensitive and specific, but also are beneficial favourable to the epidemiological survey and clinical diagnosis, because the testing bacteria are decomposed on the nitrocellulose membrane, so that hundreds of samples can be examined in one test.

Objective:To investigate the relationship between ADAM 33 polymorphisms and susceptibility of asthma of Chinese in Qingdao.Methods: The polymorphism of ADAM33 gene, they were rs2280090, rs487377 and rs2787094, were detected with SnaPshot method.Results: There was no significant difference between asthma group and control group in genotypic frequencies.Conclusion:The study shows that the rs 2280090 , rs487377 and rs2787094 were not associated with susceptibility of asthma in Han nationality in Qingdao .

Objective:To detection antibody against heat-stable enterotoxin by fusion protein.Methods:Mutant heat-stable enterotoxin precursor gene was ligated in vector pGEX-4T-2 to inductively express as a fusion protein GST/proST_m with glutathione S-transferase(GST).To investigate the antigenic action,serum and fecal antibodies against heat-stable enterotoxin was detected with this fusion protein.Results:The fusion protein was a about 32 kD protein.All the samples contain the antibody against ST.Conclusion:Such strategy was a promising method to detect antibody against heat-stable enterotoxin.

The expression of a soluble protein on cell surface is often desirable for study of a functional protein, wide application of a protein or investigation of protein-protein interaction. The expression of a soluble protein on the surface of a cell is often achieved by genetically linking a protein to the extra-cellular fragment of a transmembrane partner. In this study, the myc epitope was linked with N terminal of transmembrane proteins either A2TM or deltaLNGFR amplified by overlapping PCR. The plasmids expressing fusion protein were transfected into 293FT cells and the expression of target proteins was evaluated by fluorescent microscope, flow cytometry and Western blotting. The results of flow cytometry revealed that both A2TM and deltaLNGFR were expressed on the cell surface, but A2TM could only be detected with high copy number. Western blotting showed that the expression level of deltaLNGFR was very high and protein was heavily glycosylated, by contrast the expression of A2TM was hardly detected. The results indicate that glycosylated deltaLNGFR is a good candidate partner for the expression of a soluble protein on the cell surface.
Apoptosis Regulatory Proteins ; biosynthesis ; genetics ; Cell Membrane ; metabolism ; Genes, MHC Class II ; genetics ; HLA-A2 Antigen ; biosynthesis ; genetics ; Humans ; Membrane Fusion Proteins ; genetics ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism

OBJECTIVETo assess the association of polymorphisms of IL-4 gene (rs2243250, rs2243283) and IL-4R gene (rs1805012, rs1801275, rs1805010) with susceptibility to asthma among ethnic Chinese in Qingdao.

METHODSFor 400 asthma patients and 200 healthy subjects, above polymorphisms were detected with SnaPshot method.

RESULTSFor rs1805012, the frequency of TC genotype in the asthma group was significantly lower than the control group (8.8% vs. 15.5%, χ (2)= 6.498, P= 0.039), and so were the frequencies of TC+ CC genotypes (9.0% vs. 15.5%, χ (2) = 5.522, P= 0.019) and the C allele (4.6% vs. 7.7%, χ (2) = 4.729, P= 0.039). No significant difference was detected between the two groups in the frequency of the remaining four polymorphisms or the haplotypes formed by rs2243250 and rs2243283 (All P> 0.05).

CONCLUSIONThis study has indicated that rs1805012 polymorphism of IL-4R gene is associated with asthma in ethnic Han Chinese from Qingdao region. TC+ CC genotypes have a protective role against asthma compared with TT genotype. However, polymorphisms of IL-4 gene are not associated with susceptibility to asthma.

Alleles ; Asian Continental Ancestry Group ; genetics ; Asthma ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Interleukin-4 ; genetics ; Male ; Middle Aged ; Receptors, Interleukin-4 ; genetics

Thrombin-like enzymes (TLEs) are studied widely because of their therapeutic potential in myocardial infarction and thrombotic diseases. We synthesized the DNA fragment encoding thrombin-like enzyme calobin from Agkistrodon caliginosus (Korean Viper) venom by fusion PCR and expressed it in Pichia pastoris. After induction by 0.5% methanol for 48 h, the expression level of recombinant calobin reached 3.5 g/L in medium. The recombinant calobin was purified by Q-Sepharose Fast Flow ion-exchange chromatography and Sephacryl-S-100 gel filtration chromatography. Purified sample had a molecular weight of 32 kD shown in SDS-PAGE. It hydrolyzed fibrinogen and formed a light white hydrolysis circle in fibrinogen plate. SDS-PAGE analysis showed that recombinant calobin cleaved Aalpha-chain of fibrinogen specifically, and produced an appropriately 40 kD new band. However, we failed to find its fibrin-clot formation activity.
Agkistrodon ; Animals ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Serine Endopeptidases ; biosynthesis ; genetics ; Thrombin ; biosynthesis ; genetics ; Viper Venoms ; enzymology

OBJECTIVE To assess the association of polymorphisms of TNF-alpha gene (rs1799724, rs1800630, rs1799964 and rs769178) and IL-13 gene (rs2158177 and rs1295687) with susceptibility to asthma among ethnic Chinese in Qingdao region. METHODS For 400 asthma patients and 200 healthy subjects, above polymorphisms were detected with a SNaPshot method. RESULTS For rs2158177, the frequency of genotype of GG in the asthma group was significantly lower than the control group (2.8% vs. 5%, OR = 0.31, 95%CI: 0.12-0.82, P = 0.021). No significant difference was detected in the genotypic frequencies for the remaining 5 polymorphisms between the two groups (All P > 0.05). CONCLUSION The study has indicated that rs2158177 polymorphism of the IL-13 gene is associated with asthma in ethnic Han Chinese from Qingdao. No association has been found between polymorphisms of TNF-alpha gene with susceptibility to asthma.
Adolescent ; Adult ; Aged ; Asthma ; genetics ; Female ; Humans ; Interleukin-13 ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Tumor Necrosis Factor-alpha ; genetics

Objective To compare the effects of 3D laparoscopic hepatectomy (3D-LH) and open hepatectomy(OH)in glissonian access for hepatocellular carcinoma(HCC)on postoperative immunosuppression by analyzing cellular immune function. Methods During January 2014 to December 2017,sixty patients randomly scheduled to undergo hepatectomy were divided into 3D-LH group and OH group,with 30 patients in each group. All patients' peripheral blood was sampled to measure cell-mediated-immunologic markers(CD4+,CD8+T cell) before operation and in the mornings of the 1st,7thday after surgery. The effects of different operative methods on cellular immune function in patients organism were investigated. Results The decreased level in 3D-LH group was less than that in OH group in postoperative 1stday. The improvement in the 3D-LH group was more faster than that in the OH group on postoperative 7thday. There was statistical significance between the two groups(P<0.05), respectively. Conclusion When compared to open hepatectomy ,3D laparoscopy liver resection in glissonian approach for HCC,less effects in cellular immune function.