Objective To detect TCRγ gene rearrangements in skin lesions and peripheral blood of patients with parapsoriasis, and to study their clinical significance. Methods Totally, 20 patients with parapsoriasis were included in this study. BIOMED?2 multiplex PCR was performed to detect TCRγgene rearrangements in lesional skin (n=20)and peripheral blood(n=11)samples from the patients with parapsoriasis. Statistical analysis was performed to assess the relationship of TCRγ gene rearrangements with clinical types of parapsoriasis as well as general information and histopathological manifestations(including non?specific manifestations and atypical manifestations)of patients. Results TCRγ gene rearrangements were positive in lesional skin from 7 of the 20 patients, in peripheral blood from 3 of 11 patients, and in both lesional skin and peripheral blood from 2 patients. Positive TCRγ gene rearrangements in skin lesions were significantly correlated with mycosis fungoides(MF)?related atypical histopatho?logical manifestations(P<0.05), but those in neither skin lesions nor peripheral blood were correlated with gender and age of patients or clinical course and types of parapsoriasis(all P>0.05). During an average follow?up time of 44.85 ± 18.48 months, 1 case progressed into MF, and 2 were cured. Conclusions Positive TCRγgene rearrangements in skin lesions of patients with parapsoriasis may be correlated with MF?related atypical manifestations. The presence of TCRγgene rearrangements and atypical histopathological manifestations may suggest the possibility of progression from parapsoriasis into MF.

Objective\ By inhibiting approach of apoptosis of T cells to observe if it could reduce the attacks of tumor cells upon lymphocytes and its possible role in tumor treatment. Methods\ The expressions of Fas/FasL genes in ovarian carcinoma cells were detected by using flow cytometry and RT\|PCR method.The construction of pcDNA3\|antisense Fas,the constructed vector was transfected into Jurkat cells with lipofectin,the change in expression of Fas gene was determined by flow cytometry.By means of Annexin\|V and MTT the effect of apoptosis in transduced Jurkat cells was investigated,and also using MTT cytotoxic test to investigate how 3AO cells kill Jurkat cells. Results\ Fas/FasL were expressed in 6 ovarian carcinomal cells.The expression level of Fas protein in Jurkat cells transduced with the constructed vector was decreased.Apoptosis was reduced in antisense Fas\|transfected Jurkat cells after anti\|Fas treatment. Conclusion\ FasL expression in ovarian carcinoma may be one of the reasons for ovarian carcinoma to escape immunosurveillance and attacking lymphocytes.Blocking Fas expression in lymphocytes can partially inhibit Jurkat cells apoptosis induced by anti\|Fas and reducing the attacks of tumor cells upon lymphocyte.\;

0.05). Conclusions The high expression of GATA3, a Th2 inducer, may correlate with the disease acti vity of SLE, while T-bet, a Th1 inducer, may not correlate with the disease acti vity.

Objective To analyze the correlation of interleukin(IL)-12B gene single nucleotide polymorphism(SNP)rs6887695 with clinical phenotypes(including age at onset,family history,clinical types,gender)of psoriasis vulgaris in Chinese Han population.Methods This study recruited 575 patients with psoriasis vulgaris and 1403 healthy controls.DNA samples were obtained from these subjects.PCR with Taqman fluorescent probe(ABI 7900 system)was performed to analyze the genotype of SNP rs6887695 in IL-12B gene.Statistical analysis was carried out by using the software SPSS 14.0,and Chi-square test was conducted to compare the frequency of the SNP rs6887695 genotypes and alleles between the patients and controls as well as between patients with different clinical phenotypes of psoriasis.Results The frequency of GG,GC and CC genotype of the SNP rs6887695 was 42.61％,45.39％ and 12.0％ respectively in the patients,compared to 34.42％,47.83％ and 17.75％ in the healthy controls(x2 =16.31,P ＜ 0.01);the frequency of G and C allele of the SNP rs6887695 was 65.30％ and 34.70％ respectively in the patients,compared to 58.34％ and 41.66％ respectively in the healthy controls(x2 =16.54,P＜0.01).Significant differences were observed in the distribution of genotypes and alleles of the SNP rs6887695 between patients with chronic plaque psoriasis(n =543)and those with acute guttate psoriasis(n =32,x2 =18.11,12.19,both P ＜ 0.01).Increased frequency of G allele and GG genotype of the SNP rs6887695 were noted in the patients with psoriasis vulgaris compared with the healthy controls,and in the patients with plaque psoriasis compared with those with guttate psoriasis.However,there was no statistical difference in the distribution of SNP rs6887695 genotypes or alleles between 540 patients with adult onset psoriasis and 35 patients with child onset psoriasis,between 102 patients with family history and 440 patients without family history,or between 341 male patients and 234 female patients(all P ＞ 0.05).Conclusions The IL-12B SNP rs6887695 may be associated with the susceptibility to psoriasis vulgaris in Chinese Han population,especially with the susceptibility to plaque psoriasis,but seems unassociated with the age at onset,family history or gender of patients.

Objective To investigate the distribution of ompA gnne in 15 Chinese reference standard strains belonging to 15 serogroups of Leptospira interrogate, and to express recombinant OmpA ( rOmpA ) and to identify immunogenicity and immunoprotection of rOmpA. Methods Genomic DNAs from different leptospiral strains were extracted by phenol-chloroform method. Entire ompA gene fragments from the strains were amplified by PCR and then sequenced after T-A cloning. A prokaryotic expression system of ompA gene from L. interrogans strain 56601 was constructed, and the expression and yield of rOmpA were determined by SOS-PAGE plus Bio-Rad Agarose Image Analyser. Rabbits were immunized with rOmpA for obtaining antiserum, and immunodiffusion test was used to measure the antiserum's titer. Western blot assay was performed to determine the immunoreaetivity of rOmpA with the antiserum against rOmpA and antiserum against whole cell of L. interrogans strain 56601, while mi-croscopic agglutination test (MAT) was applied to detect the cross agglutination to the 15 L. interrogans strains. A leptospire adhering cell model and a leptospire infecting guinea pigs model were used to determine the adhesion-bloc-king effect of rOmpA antiserum and immunoprotection of rOmpA. Results All the 15 L. interrogans strains, but not L. biflexa strain Patoe Ⅰ , had sequence conserved ompA genes. The yield of rOmpA was approximate 20% of the total bacterial proteins, rOmpA could induce rabbits to produce antibody and immunodiffusion titer of the anti-serum was 1:4. Both antisera against rOmpA and against whole cell of L. interrogans strain 56601 were able to pro-duce positive Western blot signs to rOmpA, and the former offered 1 : 20-1 : 320 MAT titers to the 15 L. interrogans strains. 1: 10-1:160 dilutions of rOmpA antiserum could efficiently block L. interrogans strain 56601 adhering to J774A. 1 cells, and 100 μg and 200 μg rOmpA displayed 50.0% and 75.0% immunoprotective rates in the infee-ted guinea pigs. Conclusion ompA gene only exists in genomes of different pathogenic L. interrogans serogroups. rOmpA has relatively stronger antigenicity, cross immunoreactivity and certain immunoprotection, implying that this recombinant protein may be used as a candidate antigen for developing universal genetic engineering vaccine of L. interrogans.

Objective:To find out markers of endometriosis. Methods: The two dimensional gel images of proteins extracted from eutopic endometrium from endometriosis patients and controls were analyzed by software Phoretix 2D,and the proteins expressed differently were identified primarily by query of data base.The proteins extracted from ectopic endometrium of ovarian endometriosis were transferred from two dimensional gel onto nitrocellulose membranes,followed by incubation with sera from women with and without endometriosis. Analyzed by MALDI-TOF-MS,the proteins hybridized differently were identified through their Peptide Mass Footprints. Results: Having compared the reproducible two dimensional gel images of proteins from eutopic endometrium of women with and without endometriosis,we obtained 11 proteins expressed differently.Through Western Blot technique,we found three proteins hybridized differently which were identified as vimentin,?-actin and ATP synthase ? subunit respectively. Conclusion: The protein expression spectra of eutopic endometrium from patients with endometriosis are significantly different from those of the controls,and the anti-endometrial autoantibodies against vimentin, ?-actin and ATP synthase ? subunit may be induced.

Objective:To investigate the expression of TLR4 mRNA?TLR2 mRNA in the peripheral blood mononuclear cells (PBMC) from the patients with Gram positive and Gram negative infections.Methods:TLR4 and TLR2 mRNA were detected in the patients with Gram negative infections ( n =20) and the patients with Gram positive infections ( n =15) and the normal persons as control ( n =24) with the method of Taqman Real-time PCR.Results:TLR4 mRNA expression in the patients with Gram negative infections were significantly higher than Gram positive infections patients and the normals ( P 0.05).Conclusion:The results showed that the increased expression levels of TLR4 mRNA in PBMC from the Gram negative infectious patients are associated with Gram negative infections and the detection of TLR4 mRNA expression may be a method of diagnosis for Gram negative infections in the early stage.

Objective To explore the repair possibilities of endothelial progenitor cells (EPCs)in peripheral blood in patients with different extents of obstructive sleep apnea (OSA) through measuring the levels of pro-angiogenic factors and different subgroups EPCs in peripheral blood in patients with OSA. Methods Ninety adult patients with OSA, 30 healthy controls with matched age and gender were enrolled for this study. The subjects performed Polysomnography, were divided in-to four group based on Apnea Hypopnea Index (AHI). The serum levels of HIF-1α, SDF-1αand VEGF were assessed by ELISA. Mononuclear cells were isolated from peripheral blood with density gradient centrifugation, and flow cytometry was used to detect levels of CD133+KDR+EPC, CD133+CD34+EPC, CD34+KDR+EPC and ALDHloCD34+KDR+EPC based on AL-DH activity, and CD133, CD34, PE-KDR related cell surface markers. Results The levels of CD133+KDR+EPC, CD133+CD34+EPC, CD34+KDR+EPC were higher in OSA groups than those of control group, both of which were higher in severe OSA group than those of in mild and moderate OSA groups. The levels of ALDHloCD34+KDR+EPC were higher in mild and moderate OSA groups than that of the control groups, and the levels of ALDHloCD34+KDR+EPC were significantly lower in se-vere OSA group than those of control, mild and moderate OSA groups. Serum levels of HIF-1α. VEGF were significantly high-er in OSA groups compared to those in control groups, both of which were higher in severe OSA group than those of mild and moderate OSA groups. Serum levels of SDF-1αwere significantly lower in severe OSA groups than those of mild, moderate OSA and control groups (P<0.05). Conclusion The mobilization and recruitment of different subtypes of EPCs are obvious-ly increased in patients with OSA, but ALDHloCD34+KDR+EPC with vascular repair capacity keeps to invariability, even de-creases in patients with severe OSA, which results in endothelial damage, and increases the risk of cardiovascular disease.

Objective To investigate the pathogenic significance of antigenic epitopes and their relevant antibodies in pemphigus vulgaris (PV) by neonatal mouse model. Methods The extracellular domain 1-2 (EC1-2) fusion protein was expressed and purified by glutathione affinity chromatography on the basis of construction of recombinant EC1-2 vector, and then the New Zealand white rabbits were immunized to obtain the specific antisera. The IgG fraction was transferred into the neonatal mice passively after it was purified from the antisera. After 15-18 hours of injection, the abdomen skin and the sera of the mice were examined by light microscopy, electron microscopy, direct immunofluorescence and indirect immunofluorescence. Results In the evaluation of the study group of mice, the intraepithelial vesicle formation was observed. Electron microscopy showed that intercellular spaces were widened, desmosome split and disappeared. In immunofluorescence, the fluorescence-labeled IgG deposied between the acantholytic cells. In the control group of mice there were no pathogenic changes observed, except very weak fluorescence between intercellular spaces. Conclusion The PV mouse model established shows that the EC1-2 epitope in PVA antigen and its relevant antibodies were pathogenic, and can be used as a tool in studying the pathogenesis of PV.

Because of the population aging,the increase of the stroke patients and the need for rehabilitation,the treatment only in the rehabilitation department of the hospital is far from the satisfaction of people's demands of the service of rehabilitation.It is important to extend the community-based rehabilitation.Compared with the rehabilitation in hospitals,it is more economy,efficiency and convenience for stroke patients in community-based rehabilitation services,and further improve the rehabilitation effect of stroke patients.