This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.

This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via differentially expressed proteins identified. Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells. The mRNA and protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and Western blotting. The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation were identified with MALDI-TOF/TOF. The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus. The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.

Before the technique of advanced high-throughput sequencing comes up , less is known about the human gut microbiota .It has been understood that trillions of microbes , in which 99% are bacteria , inhabit the human gut, forming a complicated ecological community .The gut microbiota has a great impact on human physiology and suscepti -bility to disease through its integrative metabolic activities and interactions with the host .In physiology , gut microbiota con-tributes to the host acquisition of nutrition and energy from diets , promoting development and maturation of gastrointestinal tract and immune system , and protecting host from invasion of enteropathogens .In pathology , dysbiosis underlying altered gut microbiota is associated with the susceptibilities to various diseases , including inflammatory bowel disease , type 1 dia-betes, asthma, obesity, metabolic syndrome , autism and cancer .Understanding of the factors that underlie alterations in the composition and function of gut microbiota will be helpful in the development of drugs and the design of therapies that target it.This goal is formidable .It is because that the compositions of gut microbiota are immensely diverse , varying be-tween individuals in a population and fluctuating over time in an individual , especially during early development and disea-ses.Viewing the gut microbiota with an ecological perspective will provide new insights into how to improve our health by targeting this microbial community in clinical treatments .

Given its population of CCR5-expressing, immunologically activated CD4 +T cells, the gastrointestinal (GI) mucosa is uniquely susceptible to human immunodeficiency virus (HIV)-1 infection. Recent studies have shown that, as in macaques infected with simian immunodeficiency virus (SIV), intestinal CD4 +T cells are selectively and rapidly depleted in the intestine of HIV-infected patients. Depletion of intestinal CD4 +T cells occurred at all stages of infection regardless of highly active antiretroviral therapy (HAART). Here we discuss the important implications of the recent findings for our understanding of HIV pathogenesis, treatment, and vaccine design. The major significance is that it supports a simple hypothesis to explain the pathogenesis of HIV infection, that most HIV replication occurs in the intestine and that disease progression may correlate with turnover of specific cell subsets in mucosal tissues.

AIM: To explore the correlation between development of CD4~+CD25~+ regulatory T cells (CD4~+CD25~+ Tr) and thymus CD4~-CD25~+ cells. METHODS: The ratios of CD4~+CD25~+ regulatory T cells to CD4~+ T cells in thymus, spleen, lymph node and peripheral blood of mice from birth to mature and also the ratios of CD4~-CD25~+cells to CD4~-T cells in thymus were measured by flow cytometry. Purified CD4~+CD25~+ T cells and CD4~+CD25~- T cells were labeled with CFDA-SE, and then stimulated with various kinds of stimulators. RESULTS: The percentages of CD4~+CD25~+ Tr in mouse spleen, lymph nodes and peripheral blood increased gradually, but not in thymus, from day one to week 10 of the age with rapid rising from day one to week 1. The percentages of CD4~-CD25~+ cells in mouse thymus were quite high on day one after birth, and decreased rapidly from day one to week 1. Both CD4~+CD25~+ Tr and CD4~+CD25~- T cells showed no proliferation in response to ConA, while CD4~+CD25~+ Tr showed a transient enlargement of cell size. Both CD4~+CD25~+ Tr and CD4~+CD25~- T cells underwent proliferation in response to PDB plus ionomycin. CD4~+CD25~- T cells, but not CD4~+CD25~+ Tr, showed a proliferative response to the stimulation of coated anti-CD3 plus soluble anti-CD28 antibody, however, CD4~+CD25~+ Tr showed significant proliferation and CD4~+CD25~- T cells showed a stronger response in addition of high dose of IL-2. CONCLUSION: The thymus CD4~-CD25~+ cells are probably the precursor of CD4~+CD25~+ Tr during cell development.

A breakthrough has recently been made in the studies on pathogenesis of HIV disease,which result in some new theories.New strategies for HIV drug and vaccine development are emerging in the impact of these new understandings.The intestinal infection hypothesis proposes that HIV disease can be regarded as some kind of infectious disease of gut immune system as the major HIV infection is located in intestine.The acute catastrophe hypothesis suggests that the subsequent pathological changes are the fallout from a mucosal catastrophe of acute intestinal HIV infection,in which the majority of CD4+ T cells are deleted due to infection.The general immune overactivation hypothesis proposes that the general immune overactivation is harmful to patient as it increases the cell susceptibility to HIV infection and supportiveness of HIV replication.The host proteins related to HIV replication can be new targets for HIV drug discovery.The mucosal vaccine strategies,which attempt to induce HIV specific neutralizing antibodies on mucosal surface may be the first choice in development of prophylactic HIV vaccines.Induction of tolerance may be one of the strategies for HIV therapeutic vaccines because HIV disease can be regarded as autoimmune disease caused by HIV infection.

AIM: The aim of the present study is to explore the effects and mechanisms of HIV-1 glycoprotein gp120 on calcium-influx and ERK-phosphorylation of human microglia.METHODS: The level of intracellular calcium of human microglia grown on coverslip,which was loaded by calcium-probe,Fluo-4,and then treated in various experimental processing,was detected by confocal microscopy with time resolution mode.The binding of gp120 to human microglia was determined with confocal microscopy or flow cytometry after treatment with gp120 and stained with anti-gp120-FITC antibody.Phosphorylation of ERK within human microglia with or without gp120 stimulation was analyzed with confocal microscopy following the direct immuno-staining with anti-phosphorylated ERK antibody.RESULTS: The results of confocal microscopy showed that calcium-influx response was triggered by HIV-1 glycoprotein gp120 in human microglia.The results from analysis by confocal microscopy and flow cytometry showed that gp120 was able to bind to human microglia.ERK phosphorylation was enhanced in human microglia stimulated with gp120.CONCLUSION: HIV-1 glycoprotein gp120 induces calcium-influx in human microglia and enhances ERK phosphorylation in human microglia,indicating that gp120 is an activator of human microglia.So gp120 may be involved in the pathogenesis of HIV-associated dementia.

Objective:To study the effect of isoflavone and genistein on activation of T lymphocytes in order to develope new immuno intervention reagent.Methods:Fluorescence conjugated monoclonal antibodies and flow cytometer were used to detect the expression rate of CD69 by activated T cells in vitro in response to Phytohemagglutinin(PHA) and Phorbol 12,13 dibutyrate(PDB),with some samples pre incubated with 10,50 or 100 ?mol/L of genistein,after 2 h and 6 h of incubation in whole blood culture system.Results:After 2 h of culture,the inhibitory effect in PHA group was stronger than PDB group(P

The study investigated the effects of red clover extract (RCE) on mouse T macrophages and lymphocytes in vitro. The cell toxic effect of RCE was estimated by MTT assay. Multiple-fluorescence staining plus flow cytometry were used to detect the effect of RCE on CD69/CD25/CD71 expression of mouse T lymphocytes stimulated by Con A; CFDA-SE staining plus flow cytometry were used to analyze the effect of RCE on proliferation of T lymphocytes activated by Con A; The effect of RCE on nitric oxide(NO) secretion of mouse macrophages stimulated by lipopelysaccharide (LPS) for 24 h was assayed by Griess reagent system. We found that RCE had potent anti-inflammatory effects on mice. RCE had littl ecell toxic effect on mouse lymphocytes and macrophages. RCE strongly inhibited the excessive production of inflammatory mediators ( NO, CD69, CD25, CD71 ), in a dose-dependent manner, like cyclosporine A injection. RCE could inhibit proliferation of CD3<'+> T lymphocytes. These data suggested that RCE migh texhibit anti-inflammatory effect by inhibiting the activation and proliferation of mouse lymphocytes and the NO secretion of mouse macrophages.

Aim To investigate effects of genistein (5, 7, 4′-trihydroxyisoflavone) on rat costochondral growth plate chondrocyte (RGC) collagen and Sox9 expression. Methods Primary cultured RGC, effects of genistein on collagen synthesis, col2a1 mRNA transcription and Sox9 protein expression of 1st passage RGC were detected by isotope incorporation, RT-PCR and western blotting. Results Genistein inhibited collagen synthesis of 1st passage RGC (P