Dysregulated Dermal Mesenchymal Stem Cell Proliferation and Differentiation Interfered by Glucose Metabolism in Psoriasis

Background and Objectives@#Psoriasis is a chronic inflammatory skin disease, which the mechanisms behind its initiation and development are related to many factors. DMSCs (dermal mesenchymal stem cells) represent an important member of the skin microenvironment and play an important role in the surrounding environment and in neighbouring cells, but they are also affected by the microenvironment. We studied the glucose metabolism of DMSCs in psoriasis patients and a control group to reveal the relationship among glucose metabolism, cell proliferation activity，and VEC (vascular endothelial cell) differentiation in vitro, we demonstrated the biological activity and molecular mechanisms of DMSCs in psoriasis. @*Methods@#and Results: We found that the OCR of DMSCs in psoriatic lesions was higher than that in the control group, and mRNA of GLUT1 and HK2 were up-regulated compared with the control group. The proliferative activity of DMSCs in psoriasis was reduced at an early stage, and mRNA involved in proliferation, JUNB and FOS were expressed at lower levels than those in the control group. The number of blood vessels in psoriatic lesions was significantly higher than that in the control group (p＜0.05), which the mRNA of VEC differentiation, CXCL12, CXCR7, HEYL and RGS5 tended to be increased in psoriatic lesions compared to the control group, in addition to Notch3. @*Conclusions@#We speculated that DMSCs affected local psoriatic blood vessels through glucose metabolism, and the differentiation of VECs, which resulted in the pathophysiological process of psoriasis.

Arabidopsis thaliana as Bioindicator of Fungal VOCs in Indoor Air

In this paper, we demonstrate the ability of Arabidopsis thaliana to detect different mixtures of volatile organic compounds (VOCs) emitted by the common indoor fungus, Aspergillus versicolor, and demonstrate the potential usage of the plant as a bioindicator to monitor fungal VOCs in indoor air. We evaluated the volatile production of Aspergillus versicolor strains SRRC 108 (NRRL 3449) and SRRC 2559 (ATCC 32662) grown on nutrient rich fungal medium, and grown under conditions to mimic the substrate encountered in the built environment where fungi would typically grow indoors (moist wallboard and ceiling tiles). Using headspace solid phase microextraction/gas chromatography-mass spectrometry, we analyzed VOC profiles of the two strains. The most abundant compound produced by both strains on all three media was 1-octen-3-ol. Strain SRRC 2559 made several terpenes not detected from strain SRRC 108. Using a split-plate bioassay, we grew Arabidopsis thaliana in a shared atmosphere with VOCs from the two strains of Aspergillus versicolor grown on yeast extract sucrose medium. The VOCs emitted by SRRC 2559 had an adverse impact on seed germination and plant growth. Chemical standards of individual VOCs from the Aspergillus versicolor mixture (2-methyl-1-butanol, 3-methyl-1-butanol, 1-octen-3-ol, limonene, and β-farnesene), and β-caryophyllene were tested one by one in seed germination and vegetative plant growth assays. The most inhibitory compound to both seed germination and plant growth was 1-octen-3-ol. Our data suggest that Arabidopsis is a useful model for monitoring indoor air quality as it is sensitive to naturally emitted fungal volatile mixtures as well as to chemical standards of individual compounds, and it exhibits relatively quick concentration- and duration-dependent responses.

Health Effects of Small Volatile Compounds from East Asian Medicinal Mushrooms

Medicinal fungi, taken whole or as various forms of extracts, have been used to alleviate, cure or prevent human ailments since pre-historic times. In particular, Asian cultures have incorporated a variety of mushrooms into their medical practices. Chemically pure, bioactive metabolites from fungi have been a mainstay of modern pharmacological research and in addition to antibiotics, include anticancer agents, immunosuppressants, enzyme inhibitors, antagonist and agonists of hormones, and a variety of psychotropic substances. However, to date not many studies have focused on the possible health benefits of odorant volatile organic compounds (i.e., gas phase compounds). An analysis of these compounds for their health related effects will expand the range of compounds available for the treatment of chronic and acute diseases. This review highlights phenolic acids and monoterpenes from Asian medicinal mushrooms (AMMs), which not only produce pleasant odors but also have antioxidant and antibacterial effects. Odorant bioactive volatile phase compounds from medicinal mushrooms remain an essentially untapped source for future medicines, and AMMs remain a promising resource for future pharmacological research.

Preferential expressions of peripheral blood T cell receptor beta chain variable region subfamilies in patients with psoriasis vulgaris / 中华皮肤科杂志

Objective To assess the preferential expressions of peripheral blood T cell receptor beta chain variable region (TRBV) subfamilies in patients with psoriasis vulgaris(PV), and to estimate their role in the pathogenesis of psoriasis. Methods Thirty-three upstream primers were designed to target the human functional TRBV genes, downstream primers to target the common T cell receptor beta constant (TRBC) gene,with T cell receptor alpha constant (TRAC) gene as the internal reference. Total RNA was extracted from the peripheral blood T cells of 10 health human controls and 10 patients with PV, and transcribed into cDNA.Then, TRBV genes were amplified by real-time fluorescence quantitative PCR (RFQ-PCR) and the fluorescence intensity of each samples was detected. The expression levels of TRBV genes in the control group were used to calculate the cut-off values (mean expression levels of TRBV subfamilies in the 10 normal controls + 3 standard deviations). When the expression level of a TRBV subfamily from patients with PV was equal to or higher than the cut-off value, it was considered as the preferentially expressed TRBV subfamily. Results The threshold cycle (Ct) value varied from 21 to 24 for TRAC gene. The difference in the Ct value between TRBV subfamily genes and TRAC gene in patients with PV was 2.98 for TRBV2 gene, 3.24 for TRBV5-7 gene, 2.52 for TRBV6-6/6-9 gene, 2.04 for TRBV 12 gene, 3.56 for TRBV 24 gene, and 4.12 for TRBV 29 gene, and the expression levels of these subfamily genes were significantly higher than those in the normal controls (all P ＜ 0.05). According to the above standard, TRBV6-6/6-9, TRBV12 and TRBV29 were considered to be preferentially expressed subfamilies. Conclusions There is a preferential expression of TRBV gene subfamilies in peripheral blood of patients with psoriasis vulgaris, which may play a vital role in the abnormal T cell-mediated immune responses in psoriasis.

Expressions of Notch receptors and target genes in bone marrow CD34+ cells from patients with psoriasis / 中华皮肤科杂志

Objective To study the expressions of Notch receptors (Notch1 and 2) and target gene Hes-1 in bone marrow CD34+ cells from patients with psoriasis. Methods CD34+ cells were isolated from the bone marrow of 12 patients with psoriasis and 19 normal human controls. Western blot was conducted to detect the expressions of Notchl, Notch2 and Hes-1 proteins in CD34+ cells, with the house-keeping gene β-actin as an internal reference. Results In contrast with normal controls, a significant increment was observed in the expressions of Notch1 and Hes-1 proteins in psoriatic patients (0.9488 ± 0.3221 vs. 0.6693 ± 0.1465, 0.8579 ±0.3729 vs. 0.5728 ± 0.1787, both P ＜ 0.05). No significant difference was observed in the expression of Notch2 between psoriatic patients and normal controls (0.7568 ± 0.3461 vs. 0.8312 ± 0.2887, P ＞ 0.05).Conclusion The overexpression of Notch1 and Hes-1 may be associated with the low proliferation activity of psoriatic hematopoietic cells.

Levels of stem cell factor and granulocyte colony-stimulating factor secreted by bone marrow stromal cells in psoriasis patients / 中国组织工程研究

BACKGROUND:It is discovered in our preliminary study that the activity of psoriatic bone marrow hematopoietic cells is abnormal.OBJECTIVE:To reveal level of stem cell factor(SCF) and granulocyte colony-stimulating factor(G-CSF) secreted by bone marrow stromal cells in psoriasis patients.DESIGN,TIME AND SETTING:This case-control observation experiment was performed at the Laboratory of Department of Dermatology,Taiyuan Central Hospital from October 2007 to August 2008.PARTICIPANTS:Twenty-four common-psoriasis outpatients were selected at the Department of Dermatology,Taiyuan Central Hospital,including 16 males and 8 females,aged 16-59 years old.Twenty controls were selected from Department of Hematology,Taiyuan Central Hospital,whose age and gender were matched with psoriasis patients.METHODS:Bone marrow mononuclear cells were isolated from psoriasis patients and controls using the density gradient centrifugation.Bone marrow stromal cells were cultured by the adherent method.When the cells were cultured for three passages and 72 hours,bone marrow stromal cells and supernatant liquid were collected.MAIN OUTCOME MEASURES:The phenotypes of cells were identified by flow cytometry and the level of SCF and G-CSF were detected by enzyme linked immunosorbent assay kit(ELISA).RESULTS:Flow cytometry assay showed that over 90% bone marrow stromal cells were positive for CD29,but negative for CD34,CD45 and HLA-DR.That is,the purity of bone marrow stromal cells was over 90%.The levels of SCF and G-CSF in psoriasis patients were significantly higher than that in controls(P

Influence of histamine on interleukin-2 production and proliferation of T lymphocytes in vitro / 中国免疫学杂志

Objective:To investigate the influence of histamine on intedeukin(IL-2) production and proliferation of CD4+ T lymphocytes and CD8+ T lymphocytes.Methods:Density gradient centrifugalization and absorption technique were respecitively applied to detach peripheral blood mononuclear cells(PBMC) and peripheral blood lymphocytes(PBLC). CD4+ and CD8+ T lymphocytes divided with anti-CD4+ and anti-CD8+ antibody were cultured in vitro. IL-2 levels in supernatant were quantified by enzyme-linked immunosobent assay (ELBA), and proliferation index were measured by MTT methods. Results:IL-2 levels in supernatant and proliferation rate of CD4+ and CD8+ T lymphocytes treated with histamine were significantly lower than those of their corresponding un-stimulated controls(P

Clinical observe of the middle ear malformation / 临床耳鼻咽喉头颈外科杂志

Objective: To analyze the diagnosis and therapeutic effect of the middle ear abnormalities.Method: 65 ears from 1967 to 1996 were reviewed. The degrees of the ossicular abnormalities were analyzed,operative effect were valued. Result:In the 65 ears,incus abnormalities was 57 ears (87.7％),stapes was 55 ears(84.6％), mixed malformation was dominant. 47 ears were operated with ossicular chain reconstruction. 44 ears(93.6％) had hearing improvement, 32ears (68.1％) had obviously improvement (＞20 dB). They werefollowed up over one year, 44 ears had steadily heraring and had not any complications. Conclusion:If diagnosisand interference were all proper, operation would obtain good result.

Human bone marrow CD34~+ cells differentiate to T cells in thymic and bone marrow stromal microenvironment in vitro / 中国免疫学杂志

Objective:To study methods for inducing CD34+ cells of human bone marrow to differentiate into T cells in vitro to provide theory and method basics for the investigation of activity of T cells derived from psoriatic bone marrow CD34+ cells and establish a technological platform to investigate T lymphopoiesis activity of hematopoietic cells.Methods:Bone marrow CD34+ hematopoietic progenitor cells were isolated by immunomagnetic cell selection and induced differentiate into T cells in the bone in the marrow and thymic stromal microenvironment.Immunfluorescence dying method and flow cytometry analysis were performed to detect CD1-CD3+ cells,CD3+CD4+CD8-cells and CD3+CD4-CD8+ cells dynamically.Results:In the first week,the non-adhension cells were composed mostly of immature CD1+CD3-cells and CD1+CD3+ cells and small proportion of mature CD1-CD3+ cells.In the following analysis,the proportion of immature cells rapidly decreased and CD1-CD3+ cells increased.After 1 week culture,CD4+CD8+ double positive T cells and a small population CD4+CD8-and CD4-CD8+ could be detected among the CD3+ cells.In the following culture,the proportion of CD4+CD8+ double positive T cells decreased significantly and single positive T cells increased gradually.However,small proportion of mature T cells could be detected in the early stage and cann't be found after 4 weeks in the culture system without thymic stromal cells.Conclusion:Mature single positive T cells can develop from CD34+ hematopoietic progenitor cells in the bone marrow and thymic stromal microenvironment and the thymic stromal cells are vital for T lymphopoiesis.

The study of the colony formation of HPP-CFC from bone marrow-derived hematopoietic cells of psoriatic patients and the methylation of p21 gene promotor in HPP-CFC / 中国免疫学杂志

Objective:To investigate the colony formation of high-proliferative potential colony-forming cells(HPP-CFC) from bone marrow-derived hematopoietic cells of psoriatic patients and p21 gene promotor methylation in HPP-CFC,and probe into the relationship between the colony formation and the methylation status of p21 gene promoter.[WT5"HZ] Methods:[WT5"BZ]Bone marrow-derived mononuclear cells were separated by density gradient centrifugation.The cells were cultured in methycellulose semi-solid culture medium with SCF,GM-CSF,IL-3 and IL-6 for 14 days, and then high-proliferative potential colony-forming cells(HPP-CFC) were counted.The HPP-CFC were collected and their genomic DNA was isolated . DNA was subjected to bisulfite treatment,and the modified DNA was studied by using the methylation-specific polymerase chain reaction (MSP).[WT5"HZ]Results:[WT5"BZ]In methycellulose semi-solid culture system, the number of HPP-CFC in bone marrow of psoriatic patients was significantly less than that of normal control. The positive frequency of methylation of p21 gene promoter in HPP-CFC of normal contrasts was higher than that of psoriatic patients. [WT5"HZ]Conclusion:[WT5"BZ]The activity of methylation status of p21 gene promoter of bone marrow derived hematopoietic cells of psoriatic patients is abnomal. The lower positive frequency of methyllation of p21 gene promotor in HPP-CFC perhaps play a role in lower colony-forming capability of HPP-CFC of psoriatic patients.

Investigation of psoriatic T lymphocytes action on Ki67,c-Myc and Bcl-xL protein expression of keratinocytes in vitro / 中国免疫学杂志

Objective:To reveal the action of psoriatic peripheral blood T lymphocytes on keratinocytes proliferation and the significance in the pathogenesis of psoriasis.Methods:Keratinocytes were cocultivated with psoriatic peripheral blood T lymphocytes in comparison with those cocultivated with normal T lymphocytes. Immunohistochemical technique was performed to detect the expression of Ki67, c-Myc and Bcl-xL proteins.Results:There were significant overexpression of Ki67, c-Myc and Bcl-xL proteins in keratinocytes cocultivated with psoriatic T lymphocytes compared with the uncocultivated group and normal T lymphocytes group. Expression of Ki67, c-Myc and Bcl-xL proteins in keratinocytes cocultivated with normal T lymphocytes were not significantly different from the uncocultivated group.Conclusion:Psoriatic T lymphocytes, which have specific activity, can induce keratinocytes abnormal pattern of proliferation. One of the important mechanisms might be its action on the expression of c-Myc and Bcl-xL proteins.

The Correlation between Normal or Abnormal Vestibule Function and Prognosis of the Sudden Deafness / 听力学及言语疾病杂志

Objective To study the correlation between prognosis and vertigo and status of the vestibule functions with the patients of the sudden deafness (SD).Methods For 161 patients of the SD, the parameters of the hearing before treatment, vertigo and vestibule functions, were analyzed to establish a statistic model by means of Logistic regression with single factor analysis of variance, Logistic regression analysis and Chi square test.Results The single factor analysis of variance and Chi square revealed statistical differences between prognosis of the SD and hearing before treatment(P0.05).Conclusion If the patients with SD had more severe hearing loss, the prognosis would be less successful. The normal or abnormal caloric tests had influence for the prognosis of the SD, but vertigo showed influence on prognosis only when caloric test was abnormal. The degree of hearing loss was not related to that of vertigo or vestibule functions while the sudden deafness accompanied with vertigo was not due to the damage of vestibular functions.

Clinical Analysis of the Characters of Posturography in Patients with Sudden Deafness / 听力学及言语疾病杂志

Objective To investigate the clinical characters of posturographic test in patients with sudden hearing loss.Methods The postural map and correlative parameters value were examined by dynamic posturography in 100 cases of normal subjects and 120 cases of sudden hearing loss.According to state of the vestibule function,120 patients were divided into normal(67 cases) and abnormal(53cases) groups.The parameters of the length,velocity and Romberg of postural sway were compared between two groups.Results The postural map was most prominent centre patten(51%) in normal subjects,the patient's map was most prominent diffuse patten(60.83%).Length,velocity and Romberg of postural sway of the patients were significantly increased in comparison with normal subjects(P0.05).Conclusion The results of posturographic test indicate that the vestibular system in patients with sudden deafness is damaged.

The activity of T cells differentiated from the bone marrow CD34~+ cells of psoriatic patients with family histories / 中华皮肤科杂志

Objective To study the in vitro activity of T cells differentiated from bone marrow CD34+ cells of psoriatic patients with family histories. Methods Bone marrow CD34+ cells were isolated from psoriatic patients with family history and normal persons by immunomagnetic cell selection and induced to differentiate to T cells in thymic stromal microenvironment in vitro. CD3+ T cells were obtained by CD3+ cell selection system and the proportions of CD4+CD8- and CD4-CD8+ cells were measured by flow cytometry. The proliferation activity of T cells was measured by MTT colorimetry in both spontaneous proliferating group and superantigen (SAg)-stimulated group. And the levels of IL-4, IL-8 and IFN-? were quantified by enzyme-linked immunosorbent assay in culture supernatant. Results Mature CD4+CD8- cells and CD4-CD8+ cells were detected from CD3+ cells, which were derived from bone marrow CD34+ cells. When the psoriatic patients and normal controls were compared, there was no significant difference in the proportion of CD4+CD8- cells or that of CD4-CD8+ cells in CD3+ cells. The proliferation activity of T cells was significantly higher in the psoriatic patients than that in the normal controls. There was no statistical difference of IL-4?IL-8 or IFN-? levels in the supernatant of T cells between the spontaneous group and the normal control. However, the IFN-? and IL-8 levels in the supernatant of T cells were higher in the psoriatic patients than those in the normal controls after the SAg stimulation. Conclusion The abnormal activity of peripheral blood T cells from psoriatic patients with family history may be related to the bone marrow hematopoietic cells.

The influence of culture supernatant of psoriatic PBMCs on the colony forming of bone marrow-derived hematopoietic stem cells and progenitor cells / 中华皮肤科杂志

Objective To study the influence of culture supernatant of psoriatic peripheral blood mononuclear cells (PBMCs) on the colony-forming of bone marrow-derived hematopoietic stem cells and progenitor cells. Methods Bone marrow-derived mononuclear cells were separated by density gradient centrifugation. Methylcellulose semi-solid culture medium was used to culture the bone marrow mononuclear cells in culture systems. The PBMC culture supernatant from psoriatic patients or normal controls were added to the culture, to observe their influence on the marrow-derived high proliferative potential colony forming cells (HPP-CFCs), erythroid and granulocyte-macrophage colony forming units (CFUs-E and CFUs-GM) of bone marrow hematopoietic stem/progenitor cells from healthy individuals. Results The values of HPP-CFCs, CFUs-E and CFUs-GM were significantly lower in the bone marrow cells stimulated by the supernatant of cultured psoriatic PBMCs than those stimulated by the supernatant of cultured normal PBMCs and those in the spontaneous proliferation group (P 0.05). Conclusions Psoriatic PBMCs have specific biological activity and can inhibit the colony forming of bone marrow hematopoietic cells from healthy individuals.

Effects of TNF-?,sTNFRⅠon IL-8Production and Proliferation of Psoriatic Keratinocytes / 中华皮肤科杂志

Objective To study the effects of TNF -?and sTNFRⅠon IL -8production and proliferatio n of psoriatic keratinocytes.Methods In vitro cultured keratinocytes were treated with TNF -?,sTNFRⅠand an-ti -TNFRⅠ.IL -8levels in supernatants were me asured by enzyme-linked immunosobent assay.Expression of IL -8mRNA in keratinocytes was observed by semiquantitive RT -PCR and Southern blotting.Proliferation index wa s measured by MTT colorimetry.Results IL -8levels,mRNA expression and pro liferation index were significantl y higher in psoriatic patients than th ose in control group(P

Investigation of severe combined immunodeficient mice transplanted with human hematopoietic cells intraperitoneally and intravenously / 中国免疫学杂志

Objective:To compare the seeding efficiency and graft versus host disease (GVHD) of severe combined immunodeficient (SCID) mice transplanted with human hematopoietic cells intraperitoneally (ip) and intravenously (iv) and to explore the method to set up psoriatic animal model by xenogeneic bone marrow transplantation.Methods:Normal human bone marrow mononuclear cells (BMMNC) were separated by density gradient centrifugation and BMMNC (4?10~7) were injected into lethally irradiated SCID mice by ip or iv injection. GVHD symptom and periphery blood white blood cell resuming dynamics in mice were observed after xenotransplantation and flow cytometry was performed to detect human source CD45~+ cells proportion in periphery blood and bone marrow of mice.Results:The mice transplanted by tail intravenous injection presented obvious GVHD symptoms promptly 2 weeks after transplanting and only one mouse survived in 12 weeks. Among the mice received tail intravenous injection and dealed with cydosporin(CsA) and methotrexate(MTX),some of the mice showed slight GVHD symptom and survival rate was 80%(8/10) in 12 weeks. Slight GVHD symptoms appeared after human bone marrow transplantation by intraperitoneal injection and then most of mice returned to the normal and the survival rate was 70%(7/10) in 12 weeks. The periphery blood white blood cells resuming dynamics, CD45~+ cell proportion of periphery blood and bone marrow after transplantation show no significant difference between the groups transplanted by intravenous and intraperitoneal injection.Conclusion:Human hematopoietic cells could home to bone marrow in SCID mice and result in hematopoietic reconstitution. The transplantation method by intraperitoneal injection, which showed efficient seeding capability, can be used to both bone-marrow transplantation and reducing GVHD.