Objective To identify small non-coding RNAs encoded by plasmid pPCP1 and investigate their roles in biofilm formation, stress tolerance and/or virulence in Yersinia pestis.Methods Seven plasmid pPCP1-encoded sRNAs were identified by RNA-seq results in Y.pestis in our previous studies.Northern blot was used to validate the presence of the seven sRNAs.The sRNA-deletion mutants were constructed via λ-Red homologous recombination system.The biofilm formation, high salt tolerance and virulence of the phenotypes were compared between Y.pestis WT strain and sRNA mutants.Results and Conclusion The expression of seven pPCP1-encoded sRNAs was validated and the transcript length detected by Northern blotting corresponded to the length observed by RNA-seq.On this basis, five sRNA-deletion mutants were obtained.The capacity of biofilm formation was weakened upon deletion of sR3446.The tolerance of sR3446, sR3457, sR4338 and sR4340 mutants was found weakened in vitro compared to that of wild-type strain,but the tolerance of sR6143 was found increased.Slight virulence attenuation was found in two sRNA mutants ( sR4338 and sR4340 ) .The results suggest that pPCP1-deriving sRNA might be implicated in stress response, biofilm and virulence in Y.pestis.

Objective To study the changes in some important surface markers of Langerhans cell (LC) in the skin of SLE patients. Methods The normal-appearing skin and lesional skin of 9 SLE patients were studied. ABC immunohistochemistry and the monoclonal antibodies against HLA-DR, CD54, CD68, CD1a and CD4 were used. Results ①In SLE skin lesions decreased LC density was shown,and there were also changes in the morphology and surface markers of LC. ②HLA-DR expression on keratinocytes was shown in most lesional specimens and a few on non-lesional specimens. ③Neither CD4 nor CD54 expression was shown on both lesional epidermis and non-lesional epidermis, CD4+cells were only observed in the dermal infiltrates. ④Two types of CD68+dendritic cells were seen in lesional and non-lesional epidermis, and more CD68+dendritic cells were seen in the infiltrates of lesional skin. ⑤Fibrillar CD68+materials around the basal KC were observed, and some of such fibrillar materials were connected with epidermal dendritic cells while others were not. Conclusions There are some differences in LC surface marker expression on lesional skin and non-lesional skin of SLE patients, which need to be further studied.

Objective:To study the effect of astragalus polysacchrides on anticardiolipin, antiphosphatidyl choline, antiphosphatidyl serine, antiphospha-tidyl inositol, antiphosphatidic acid and antiphosphatidyl ethanolamine antibodies (aCL, aPC, aPS, aPI, aPA, aPE) in SLE mice. Methods:19 NZB?NZW F1 female mice were divided randomly into 3 groups: group PG2I (25 mg/kg/d), group PG2II (50 mg/kg/d), group NS (0.2 ml/d). Select BXSB and C57BL/6 female mice as control. The level of antiphospholipid antibodies in these mice were examined by ELISA. Results:Compared with group NS, the A value in group PG2II were decreased significantly, however, A value in group PG2I were elevated slightly. There were no difference among group PG2II, BXSB and C57BL/6. The level of antiphospholipid antibodies in group NS and PG2I were increased significantly compared with that of BXSB and C57BL/6 mice. Conclusion:The generation of antiphospholipid antibodies can be improved by low dose of PG2 and inhibited by high dose of PG2.

Objective:To observe the distribution,morphology and density of monocytes/macrophages and dendritic cells in the normal human dermis.Methods:Normal skin from 6 locations such as the face,trunk,proximal limbs,distal limbs,and palms and soles of 8 subjects were collected for the study.The horizontal and longitudinal sections of the skin were stained with an ABC immunoperoxidase procedure with anti-CD1a and anti-CD68 monoclonal antibodies.Results:In the superficial dermis CD68 positive monocytes/macrophages form a dense network with a density in a 6-micron section ranging from 361/mm~2 to 562/mm~2.These network of CD68 positive cells continued on to surround the blood vessels and skin appendages.Lower densities of CD68 positive dendritic cells were found in the deep(reticular) dermis,dispersed between collagen bundles.The CD68 positive cells were detected within the superficial dermis with variable densities: distal limbs 562/mm~2,trunk 517/mm~2,face 509/mm~2,palms 507/mm~2,proximal limbs,472/mm~2,and soles 361/mm~2.Conclusion:There exists in the superficial dermis a relatively dense network of CD68 positive monocytes/macrophages.Such a distribution might indicate the clear polarity of the dermal monocytes/macrophages,with their direction of defense towards to the dermal-epidermal junction.

Objective To establish a reconstructed human epidermis model in vitro, and to study the process of re-epithelialization. Methods A dermal substrate devoid of epidermis was prepared; a 2 mm skin biopsy explant was transplanted onto the dermal substrates. Visualization of epidermal cell migration was carried out by fluorescence imaging. The proliferation and differentiation of the new epithelial cells were observed using histopathological and immunohistochemical staining (Ki67). EGF was added to the culture medium of the experimental samples but not to that of the controls. Results After 3 days of culture, re-epithelialization was observed on the surface of the dermal substrate. A complete structure resembling regular epidermis was noted in 10 days. As compared to control samples, EGF-treated samples had larger area of re-epithelialization (t= 3.02, P

Objective To compare the intensity and isotype of anticardiolipin(ACL)antibodies in the sera from patients with syphilis and SLE.Methods IgG and IgM type of ACL antibodies were examined by ELISA in99syphilis patients and75SLE patients.Results Although similar positive percentage of IgG ACL antibodies was shown in syphilis and SLE patients,stronger absorbance(A value)was seen in syphilis patients(P

Objective To investigate the distribution of antiphospholipid antibodies(APA) in patients with systematic lupus erythematosus (SLE) and normal controls and its significance. Methods Six kinds of APA were detected by enzyme linked immunosorbent assay(ELISA). The examined sera were collected from 82 cases of normal controls and 32 cases of SLE. Results ①The positivity rate of IgG type aCL, aPI, aPA, aPE, aPS and aPC in the serum from normal controls was 6.10%, 6.10%, 7.32%, 7.32%, 3.66% or 3.66%, respectively. ②The positivity rate of IgM type aCL, aPA, aPC, aPS, aPE aPI in serum from normal controls was 2.44%, 3.66%, 3.66%, 6.10%, 6.10% and 4.88%, respectively. ③Compared with normal controls, the A values and positivity rate of IgG type of aCL, aPA, aPS, aPC and aPI in SLE patients were significantly higher. ④Compared with normal controls, the A values and positivity rate of IgM type of aCL, aPA, aPS, aPE and aPC in SLE patients were significantly higher. ⑤The total positivity rate of IgG type and IgM type APA was 68.75% or 71.88%, respectively. Conclusion ①There is lower titer of APA in normal controls, with some relationships between different types of APA and they appear at the same time.②the measurment of APA in sera of SLE might be benefit to the diagnosis of antiphospholipid syndrome. [

Objecfive To study the effects of some cytokines such as TNF-α,IL-6 and IFN-γ as well as lipopolysaccharide on CD68 expression in HaCaT cells.Methods Human HaCaT keratinocytes were randomly divided into natural proliferation group (without stimulation),IFN-γ-stimulated group,TNF-α-stimulated group,LPS-stimulated group and IL-6 stimulated group.The work concentration of TNF-α,IL-6,IFN-γ and LPS was 50 mg/L.HaCaT cells were collected after 24-hour treatment with the cytokines followed by the examination of CD68 expression with flow cytometry,immunohistochemistry and reverse transcription(RT)-PCR,respectively.Results Compared with untreated HaCaT cells,the count of CD68-positive cells was elevated in cells stimulated by TNF-α(t=3.60,P<0.01),IL-6(t=3.93,P<0.01),IFN-γ(t=2.38,P<0.05)and LPS(t=2.52,P<0.05),and the effect of TNF-α and IL-6 was stronger than that of IFN-γ and LPS.Among the four cytokines,only IL-6 enhanced the mean fluorescence intensity of CD68-positive cells (t=8.34,P<0.01).After 24-hour treatment with TNF-α,IFN-γ and IL-6,CD68 expression was observed in the cytoplasm and on the membrane of HaCaT cells and was stronger in cells treated with TNF-α and IL-6 than in those with the other cytokines.A significant increase was observed in the CD68 mRNA expression after 24-hour treatment with TNF-α (t=4.34,P<0.01),IL-6 (t=7.52,P<0.01)and IFN-γ (t=2.81,P<0.05);TNF-α and IL-6showed a stronger promotive effect than IFN-γ.Conclusion IL-6,TNF-α,IFN-γ and LPS can upregulate the CD68 expression in HaCaT cells.

Objective To investigate the role of immunophenotyping in distinguishing mycosis fun- goides (MF) from lichen planus and psoriasis.Methods The expression of CD1a,CD4,CD8,ICAM-1, LFA-1,HLA-DR,CD30 and CD7 was measured by ABC immunohistochemical technique in specimens ob- tained from lesional skin of 15 cases of MF,17 cases of lichen planus and 17 cases of psoriasis,and in the skin of 6 healthy controls.Results In the lesional epidermis of MF,the density of cells positive for CD1a, CD30 or ICAM-1,was significantly higher (mononuclear cells,P＜0.001;dendritic cells,P＜0.01) than that in the lesional epidermis of lichen planus,psoriasis and in the skin of healthy controls.The density of cells positive for CD4 or CD8 and of dendritic cells positive for HLA-DR was higher in lesional epidermis of MF than in that of lichen planus.The linear density of CD1a-positive cells (P＜0.01),the percentages of cells positive for ICAM-1 (P＜0.05) or LFA-1 (P＜0.05) were all higher in the lesional dermis of MF than in that of lichen planus.As far as the CD7-positive cell density was concerned,it was higher in the lesional dermis of lichen planus and psoriasis than in that of MF and skin of healthy controls (P＜0.01), while no difference was found between the epidermis of MF and that of lichen planus or psoriasis.Conclu- sion There are differences in the expression of CD1a,CD4,CD8,ICAM-1,LFA-1,HLA-DR,CD30 and CD7 in the lesional skin of MF,lichen planus and psoriasis,which may provide a clue to the pathogenesis of these diseases.

Objective To investigate the expression of survivin and bcl-2 in human squamous cell carcinoma (SCC) lesions and cell line SCL-1. Methods Tissue samples from 60 patients with SCC and 10 normal human controls were immunohistochemically stained to detect the expressions of survivin and bcl-2.Western blot was used to measure the expressions of bcl-2 and survivin proteins in HaCaT human keratinocytes and SCL-1 human squamous cell carcinoma cells. Results In normal control tissues, there was no expressions of survivin or bcl-2, while in SCC, the expression rates of bcl-2 and survivin were 70% and 60%, respectively,and there was no statistical correlation between the expressions of bcl-2 and survivin (P >0.05). Neither the expression of survivin nor that of bcl-2 was correlated to patients' age, gender or lesional site (all P >0.05). A statistical correlation was observed between the pathological stage in patients and expression of bcl-2 as well as between lymph node metastasis and expression of survivin (both P < 0.05). Western blot analysis revealed a significant increase in the expression of survivin and bcl-2 in SCL-1 cells compared with HaCaT cells. Con-clusion In SCC, survivin and bcl-2 seem to play their roles via different anti-apoptotic pathways.