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 .

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.

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.

Nude mice can be used as important animal models to study thyroid cancer because of their deficiency of immunitv,The model is helpful study the sensitivity of thyroid cancer to drugs.The tumor cells may come from primary tumor tissue,or from the cell lines existed.There are three methods to transplant the tumor,including orthotopic transplantation,heterotopic transplantation and tail vein injection.

Object To study the effects of different growth regulators and organic additives on rooting of regenerated shoots from Dendrobium huoshanense C Z Tang et S J Cheng protocorm like bodies Methods In order to induce root development, the regenerated shoots were cultured on MS medium containing 3% sucrose and 0 8% agar and supplemented with NAA, IBA, BA, KT, banana mud or amino acids at different concentrations During rooting induction, the development of roots was observed and the frequency of rooted shoots was scored Results The frequency and the process of root initiation were significantly influenced by different growth regulators or organic additives For initiation and development of roots, the suitable concentration of NAA, IBA, BA, KT, banana mud or amino acids was 0 2 , 0 1, 0 8, 0 8 mg/L, 20% and 0 1%, respectively Conclusion In all media tested, NAA, IBA or banana mud display greater effects on rooting of regenerated shoots of D huoshanense than others and KT enables shoots to keep green

Synaptic injury exists in the early period of vascular cognitive impairment (VCI), and it is closely correlated with the cognitive dysfunction, however, its specific mechanism remains unclear. To study the plasticity of synaptic morphological structure, the plasticity of the efficiency of synaptic transmission as well as the role and mechanism of synapic proteins in the onset of VCI will help to further clarify the pathogenesis of VCI, and thus more effectively combat VCI.

Objective:The protein of missing in metastasis (MIM), a novel discovered actin-binding scaffold protein, is involved in actin cytoskeleton rearrangements, signal transduction and transcriptional activation, and has close association with tumor growth, invasion, and metastasis. Recently, much focus has been placed on the role of MIM performed in tumor progression. In recent years, more and more attention is focused on its action mechanism in various kinds of tumors, which has a wide foreground of investigation. In this paper, we make a comprehensive review of the association of MIM with cancer development, in order to provide the theoretical basis and new strategies for application of MIM proteins in cancer diagnosis and treatment.

BACKGROUND:Because macrophages play an important role in the body’s inflammatory response, the detection of the impact of biological materials on the behavior of macrophages can assess the immunogenicity of materials. OBJECTIVE: To analyze the activation effect of decelularized extracelular matrix materials on macrophages. METHODS: The peritoneal macrophages of BALB/c mouse were obtained and cultured by dividing into five groups. Control group was simple cel culture group, experimental group 1 was acelular matrix membrane material directly contacting with macrophage for culture, experimental group 2 was fresh pericardial material directly contacting with the macrophage for culture, experimental group 3 was acelular matrix membrane material indirectly contacting with macrophages for culture, experimental group 4 was fresh pericardium material indirectly contacting with macrophages for culture. After 24 hours of culture, the secretion of nitric oxide and cytokines in cel culture supernatant was determined. After 48 hours of culture, the absorbance value was determined by MTT method and the toxicity grading was determined. RESULTS AND CONCLUSION: The toxicity grading in experimental groups 1-4 was respectively grades 2, 4, 0, 2. The nitric oxide level in experimental groups 1 and 2 was higher than that in the control group (P < 0.05), and the nitric oxide level in the experimental group 2 was higher than that in the experimental group 1 (P < 0.05). There were no significant differences in interleukin-2, interleukin-4,interferon γ, interleukin-17A and interleukin-10 levels between these five groups. The interleukin-6 level in the experimental group 2 was higher than that in the control group (P < 0.05); The expression levels of tumor necrosis factors in experimental group 1, 2 and 4 were higher than those in the control group (P < 0.05), and experimental group 2 higher than the experimental group 1 (P < 0.05), experimental group 1 higher than the experimental group 4 (P < 0.05). These results show that acelular matrix material can activate macrophages in direct contact.

AIM:To explore a method of isolation,purification,culture and identification of human microglia.METHODS:The brain tissue from abortive fetus was sterilely obtained,then chopped and triturated gently.The suspension was filtered and centrifuged to separate and isolate mixed glia cells.The cell density was adjusted to 2?1010 cells/L with DMEM/F12 complete medium and cultured in 75 cm2 flask at 37 ℃ in CO2 incubator(marked as first 0 d).After 7-10 days culture,floating cells including microglia and oligodendrocytes appeared in the flask.The first time purification was carried out to obtain highly purified microglia,for which the flasks were shaken gently.The floating cells were collected and transferred into a new flask(marked as second 0 d).4-5 days later,when microglia and oligodendrocytes grew in confluent state,the second purification was performed,for which the mixed cultured cells were digested with trypsin-EDTA and cell suspension was centrifuged,then the cell pellet was suspended by DMEM/F12 complete medium and cultured in flask(marked as 0 h).After 2 h,non-microglial cells including oligodendrocytes and cell debris were removed,and fresh medium was added into the adherent cells.In this way,the whole procedure of microglia purification was completed and the highly purified microglial cells were obtained.After purification,the expression rate of CD45,CD11b and CD68 on/in microglia were detected by laser-confocal microscopy and flow cytometry on the basis of immuno-fluorescence staining to identify the purity of microglial cells.Microglial phagocytotic function was evaluated by phagocytosis of fluorescent microspheres.The level of activation was identified by the phenomena of membrane ruffling in combination with fluorescent phalloidin staining.RESULTS:More than 98% of the microglias that we isolated and purified expressed CD45,CD11b and CD68.Almost all of the microglias could phagocytize fluorescent microbeads CONCLUSION:We succeed in isolating and purifying human microglias,which express CD45,CD11b and CD68.Almost all of the cells exhibit phagocytic function.So these cells will be useful for further functional and proteomic studies.