Objective To detect the expression of CD70 in peripheral T lymphocytes of patients with systemic lupus erythematosus (SLE) and the effect of azacitidine, an inhibitor of DNA methylation, on it. Methods Blood samples were obtained from 10 patients with active SLE (SLEDAI score ≥5), 10 patients with nonactive SLE (SLEDAI score < 5) and 10 normal human controls. Peripheral T lymphocytes were isolated and cultured for 72 hours. A part of the T lymphocytes from normal controls, which were cultured in the presence of azacitidine at 1 mol/L, served as the methylation-inhibited group. Semiquantitative reverse transcription PCR and flow cytometry were applied to detect the mRNA expression of CD70 and frequency of CD70+CD4+ cells in the cultured lymphocytes, respectively. Results The frequency of CD70+CD4+ lymphocytes was 14.55%±5.49% in normal control group, 85.25%±14.08% in active SLE group, 77.65% ±18.77% in nonactive SLE group, and 81.54%±8.71% in methylation-inhibited group. Compared with the normal control group, a significant increase was observed in both the frequency of CD70+CD4+ lymphocytes (all P < 0.01) and the expression of CD70 expression (all P < 0.05) in other three groups. There was a positive correlation between the frequency of peripheral CD70+CD4+ lymphocytes and disease activity of SLE in patients (r = 0.72, P < 0.05). Conclusions The elevated expression of CD70 appears to play a significant role in the immunologic disarrangement in SLE, and may act as a indicator of disease activity of this disease.

Objective To explore the characteristics of DNA antigen deposited in the skin lesions of patients with lupus erythematosus (LE). Methods Thirty-two LE skin specimens were collected. The deposited immune complexes were obtained by cryoprecipitation methods. Then the samples were digested by protease. Finally DNA was extracted with phenol and chloroform. The size of DNA fragments was detected on agarose gel electrophoresis. Ten kinds of probes were used to analyze the origin of these DNA molecules by dot hybridization. Results DNA fragments were successfully isolated from 27 skin specimens with four kinds of different sizes including band-I (20 000 bp), band-Ⅱ (1 300 bp), band-Ⅲ (800-900 bp) and band-Ⅳ (100-120 bp). In 15 specimens most of DNA was identified as band-I. In 2 specimens band-Ⅰ, -Ⅱ and -Ⅲ were all noticed, while all four bands were detected in 10 specimens. There was a positive correlation between small-sized fragments (100-200 bp) and disease activity (P

In order to study the affection of neutralizing antibody on the therapeutic effect of IFN? -2b,NBA was used to detect the neutralizing antibody to against IFN in the sera from 27 patients with chronic hepatitis B.The results indicated that neutralizing antibody against IFN was present in 15 of 27 patients who had been treated with IFN?-2b,and the overall frequency of neutralizing antibody was 55.6%(15/27).At the end of IFN therapy,HBV DNA was undetectable in 4 of the 15 patients(4/15, 26.7%)with neutralizing antibody.By contrast.HBV DNA became undetectable in 10 of the 12 patients (10/12,83.3 % ) without neutralizing antibody (P

Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.

Objective To observe the effects of hepatitis B virus(HBV)X gene muhifunctional protein(HBx)on the biological characteristics of QSG7701 and the transformational effects on QSG7701 cell.Methods QSG7701 ceils were stably transformed by recombinant plasmid pCMV/X and eukaryotic expressed plasmid pRc/CMV2 by liposome-based assay,respectively.Non-transfeeted QSG7701 cells were employed as controls.The expressions of HBx,c-Myc and Bel-2 proteins in QSG7701 cells were detected by Western blot.MTT colorimetric analysis,flow cytometry and soft agar clone-forming assay were performed tO detect the biolo~dcal activity of cells.Results HBx Drotein was highly expressed in pCMV X/QSG7701 cells.The expression level of c-Myc protein in the pCMV X/QSG7701 cells was much higher than those in the other tWO groups of cells.The expression of Bcl2 protein was detected in the three groups of cells,but the expression levels were similar.Percentage of S stage cells in pCMV X/QSG7701 ceils was significantly higher than those in pRcCMV2/QSG7701 and non-transfected QSG7701 cells E(28.80±2.32)%,(15.5±2.64)%and(21.5±3.66)%,LSD 0.05=3.95%,LSD 0.01=5.47%,P<0.01].While percentage of G1 stage cells in pCMV X/QSG7701 cells was significantly lower than those in pRcCMV2/QSG7701 and non-transfected QSG7701 cells[(62.30±3.85)%,(78.70±4.12)%and(78.10q±4.45)%,LSD 0.05=5.63%,LSD 0.01-7.79 %,P<0.01].The apoptosis rate of pCMV X/QSG7701 cells was much higher than those in pRcCMV2/QSG770t and non-transfected QSG7701 cells[(14.90±1.01)%,(8.91±0.48)%and (4.03±0.47)%,LSD 0.05=O.94%,LSD 0.01=1.31%,P<0.01].The population doubling time of pCMV X/QSG7701 cells was shorter than those in pRcCMV2/QSG and non-trarmfected cells(14 h,29 h,38 h,respectively).The cloning ratio in soft agar of pCMV X/QSG7701 cells WaS significantly higher than those of pRcCMV2/QSG and non-transfected QSG7701 cells[(19.83±1.96)%,(1.76±0.03)%and (1.33±0.18)%,LSD 0.05=1.53%,LSD 0.01=2.11%,P<0.01].Conclusion HBx may transform human non-tumor hepatoeyte QSG7701,which makes the cell growth malignizeA.

Objective To determine whether HBx gene can directly induce hepatocellular carcinoma in vivo, and to explore the mechanism of transplantation tumor in nude mice.Methods pCMVX/QSG7701 cell lines were vaccinated into subcutaneous tissue of nude mice. pRcCMV2/QSG7701 and QSG7701 cell lines were used as controls. The sections of transplantation tumor were observed microscopically by HE staining. RT-PCR was used to detect the expression of mutant p53 and c-Myc mRNA in transplant tumor and an other 3 cell lines. Results The transplant tumor occurred within the subcutaneous tissue of the nude mice inoculated with pCMVX/QSG7701 cell lines at 2nd week after the vaccination. No metastatic tumor was found in other organs. Transplant tumor was not formed in all the controls. HE staining confirmed that the transplant tumor was hepatocellular carcinoma. The mutant p53 mRNA and c-Myc mRNA expression level of transplant tumor and pCMVX/QSG7701 cells was significantly higher than that of pRcCMV2/QSG7701 and QSG7701 cells, respectively (P＜0.01).Conclusion HBx gene can up-regulate the expression of mutant p53 and c-Myc genes, and directly induce hepatocellular carcinoma in vivo.

Objectives To study the serum level and the clinical significance of anti-ribosomal protein P0 antibody in discoid lupus erythematosus(DLE) and systemic lupus erythematosus(SLE) patients. Methods Serum anti-RPLP0 IgG antibody of 18 DLE patients and 23 SLE patients were tested by Enzyme-Linked Immunosorbent Assay (ELISA). Direct immunofluoreseence (DIF) was used to examined the immunoreaetants from skin lesion. Serum antibody and complement C3 were detected by conventional methods. Results Anti-ribosomal P0 antibody was higher in SLE patients (1.23 ± 0.62. mean ± SD) than in patients with DLE (0.53 ± 0.18, P<0.001) and healthy controls (0.72 ± 0.16, P<0.001), but was no difference in the later two groups (P=0.5). Among SLE patients , anti-ribosomal P0 protein antibody were much higher in patients with arthritis , nephritis and specific skin lesion than in those without these disorders (P<0.05). Anti-ribosomal P0 antibody was not associated with SLEDAI and CLASI(P=0.012). Conclusions There is no difference of serum anti-ribosomal P0 antibodies between healthy controls and DLE patients. SLE patients have higher level of serum anti-ribosomal P0 antibody , specially in those with specific skin lesion.

Objective To investigate the expression and secretion of mice DNaseI gene plasmid transfected into bone marrow (BM-MSCs) mesenchymal stem cells. Methods The plasmids of mouse DNaseI gene had been transfected into the BM-MSCs of mice by liposomes. The expression of DNaseI gene in the BM-MSCs was detected by western blotting and the DNaseI activity was measured by DNA-methyl green substrate colorimetry. Results About 30% BM -MSCs were transfected with mice plasmid DNaseI gene, DNaseI was expressed in the transfected BM-MSCs and active DNaseI could be detected in the supernatant of cell culture. Conclusion The mice DNaseI gene plasmid can be transfected into mice BM -MSCs by liposomes and DNaseI gene can be expressed by the transfected BM-MSCs and active DNaseI can be secreted. This may provide potential target for the treatment of SLE.

Objective To investigate clinical features and antifungal therapeutic effect of chronic severe hepatitis (CSH) patients with invasive fungal infection (IFI), and to improve the diagnosis and treatment.Methods Clinical manifestation, blood routine, imageology and mycetology characteristic, antifungal treatment perscription and therapeutic effect of 79 CSH patients with IFI were retrospectively analyzed. Antifungal therapeutic effect was compared between fluconazole and voriconazole. Results Thirteen (16.5%) patients received glucocorticoid or other immunodepressants for a relatively long time, 40 (50.6%) patients had invasive operation, and 61 (77.2 %) patients were administered 1-6 kinds of broad-spectrum antibiotics. Seventy-three patients had fever. Leucocytes and neutrophilic granulocyte increased in 96.2% of the patients. Lung (31.6%), intestinal tract (26.2%) and oral cavity (14%) infections were common. Fungus was found in 70.9% of the patients. Candida albicans (40.9%) and aspergillus (21.1%) were often seen. Halo signs and crescent signs on lung CT were relatively specific in 40% of the patients with fungal pneumonia. Voriconazole was more effective than fluconazole(71.4% vs. 39.0%, P<0.05). Twelve patients with lung aspergillus infection were administered voriconazole, 8 (66.7%) patients of whom was effective, and the other 4 (33.3%) patients died. Conclusion There are high risk factors in major CSH patients with IFI. The most common clinical manifestations of CSH patients with IFI are fever, leukocytosis, lung and intestinal tract infection. Candida albicans and aspergillus infection are common. Voriconazole is more effective than fluconazole, and can increase the survival rate of CSH patients with IFI.

0.05).Conclusions The higher level of SEB-specific IgM and IgE in AD and eczema indi cates the colonization of Staphylococcus aureus,which participates in the exace rbation of allergic inflammation,is involved in the pathogenesis of AD and ecz ema.