Chronic psychological stress aggravates intestinal barrier damage and promotes enteritis development through inhibiting Wnt/β-catenin pathway / 中华微生物学和免疫学杂志

Objective:To investigate the mechanism by which chronic psychological stress aggravates intestinal barrier damage and promotes the development of enteritis through inhibiting Wnt/β-catenin pathway, so as to provide a new therapeutic strategy for the clinical diagnosis and treatment of inflammatory bowel disease (IBD).Methods:A comorbidity model of chronic psychological stress and enteritis was established using C57BL/6J mice. HE staining was used to analyze the effects of chronic psychological stress on the intestinal pathological damage in mice with enteritis. ELISA was used to detect the expression of proinflammatory cytokines. The ultrastructural changes of colonic cells and the state of intestinal mucus layer were observed under transmission electron microscope and scanning electron microscope. The secretion of mucoprotein 2 (MUC2) and the expression of cell proliferation marker Ki67 were detected by immunofluo rescence staining. The numbers of goblet cells were detected by Alcian blue-periodic acid-Schiff (AB-PAS) staining. Western blot was performed to analyze the expression of tight junction protein between intestinal epithelial cells, β-catenin which was a key protein of Wnt/β-catenin pathway maintaining crypt proliferation, and downstream protein c-myc.Results:The sugar water consumption ratio decreased, but tail suspension immobility time, the swimming immobility time and the expression of corticotropin releasing hormone (CRH) in hypothalamus increased (all P<0.05) in the stress group as compared with those in the control group. Chronic psychological stress promoted weight loss and colonic shortening in mice with enteritis, exacerbated pathological damage and enhanced the release of pro-inflammatory factors. Moreover, increased disappearance of intestinal epithelial microvilli and severe cellular ultrastructural damage were also observed in the stress+ dextran sulfate sodium salt (DSS) group. There was no pathological damage in the control and stress groups. Chronic psychological stress aggravated intestinal barrier injury and inhibited intestinal barrier repair by inhibiting Wnt/β-catenin pathway. Conclusions:In the mouse model of DSS-induced enteritis, chronic psychological stress preconditioning inhibited the Wnt/β-catenin pathway, weakened the repair ability of intestinal epithelium, aggravated the loss of mucus layer of intestinal barrier and the damage of tight junction structure, and promoted the development of enteritis. In the absence of enteritis, chronic psychological stress had no significant effects on the Wnt/β-catenin pathway and the intestinal barrier.

Chemokine receptor CX3CR1 promotes local remodeling of monocyte-derived Langerhans cell subsets to maintain chronic skin inflammation / 中华微生物学和免疫学杂志

Objective:To investigate the role of chemokine receptor CX3CR1 in chronic skin inflammation and its regulatory mechanism.Methods:Wild type (WT) C57BL/6 mice and Cx3 cr1 GFP/GFP mice were induced by DNFB to establish acute and chronic allergic contact dermatitis (ACD) model. Ear inflammation and swelling were observed with hematoxylin-eosin (HE) staining. Flow cytometry (FCM) was used to detect the changes in classical Langerhans cell (LC) and monocyte-derived LC (Mo-LC), as well as the expression of major histocompatibility complex Ⅱ (MHCⅡ), inducible nitric oxide synthase (iNOS) and TNF-α. Changes in epidermal LC in UV irradiation-induced dermatitis models were also analyzed. In human chronic skin inflammation, CX3CL1 expression was detected using immunohistochemistry, RT-PCR and Western blot and CD1a, CD14 and CD207 expression was observed with immunofluorescence staining. Results:In the chronic ACD model, Cx3 cr1 GFP/GFP mice showed significantly alleviated ear inflammatory and swelling as compared with WT mice, but no significant difference was found in the acute ACD model. The percentages of Mo-LC were decreased in the chronic ACD model and after three weeks of UV irradiation. Moreover, MHCⅡ, TNF-α and iNOS expressed by Mo-LC were significantly upregulated as compared with those by classical LC. CX3CL1 expression was significantly upregulated and the numbers of CD14 + monocytes and CD1a + langerin - Mo-LC were dramatically increased in human chronic skin inflammation. Conclusions:CX3CR1 might maintain inflammatory response by regulating local remodeling of Mo-LC in chronic skin inflammation.

Role of liver dendritic cell subsets in liver homeostasis and injury repair / 中华微生物学和免疫学杂志

Dendritic cells (DC) represent a heterogeneous cell population including many subsets. DC subpopulations with different characteristics and functions have been identified. The liver, as an immunological organ, is important in induction of immune tolerance. The unique anatomical architecture and immune properties of the liver have given DC the ability to maintain liver homeostasis and respond quickly to liver tissue damage. This article reviewed the role of hepatic DC subsets in the maintenance of tissue homeostasis and repair of damaged tissue in the liver.

Mechanism and research progress on radiotherapy combined with immunotherapy / 中华放射肿瘤学杂志

In recent years, the incidence rate of malignant tumors has been ever increased. As the persistent advancement of various therapeutic techniques, the therapeutic plans of cancer have been improved. Radiotherapy takes effect mainly by killing the topical tumor cells by radiation. During radiotherapy, the anti-tumor immune response can be induced or enhanced. Appropriate radiotherapy dose and segmentation model combined with certain immunotherapy plays a more and more significant role in the treatment of tumors. In this article, the underlying mechanisms of radiation-enhanced anti-tumor immune response and the current status and research prospects of radiotherapy combined with immunotherapy were reviewed.

Progress in regulation of intestinal immune homeostasis / 中华微生物学和免疫学杂志

A healthy gut consists of commensal flora,epithelial layer and gut-associated lymphoid tissue (GALT). GALT is hyporesponsive to commensal flora and dietary antigens,but can recognize and at-tack pathogens. Accumulating evidence suggests that dendritic cells (DCs) play a crucial role in managing this paradoxical situation and maintaining the complex homeostasis in gut. Influenced by intestinal epithelial cells (IECs) and commensal flora,intestinal DCs possess unique properties that enable them to regulate T-helper 2 (Th2) cells,regulatory T cells (Tregs) and immunoglobulin A (IgA)-producing cells in a steady state. During infection,intestinal DCs are involved in the induction of effector lymphocytes, although they are also responsible for initiating pathogenic responses in inflammatory bowel diseases (IBDs). Therefore, intestinal DCs are associated with not only the maintenance of immune tolerance to commensal flora,but also the induction of protective immune responses against pathogens. This review outlines the roles of commensal flora, epithelial layer, and GALT in mucosal homeostasis and inflammation and summarizes recent progress in DCs-mediated intestinal immune homeostasis.

Properties of Hydrogel and Its Applications in Biomedicine / 生物医学工程学杂志

Hydrogel is a creative polymeric biomaterial which can resemble extracellular matrix (ECM) in vitro. Hydrogel is also a material with intrinsic bioinert, but it can offer mechanical support and developmental guide for cell growth and new tissue organization by designing physicochemical and biological properties of hydrogels precisely. This review mainly introduces design of hydrogels, properties and applications in tissue engineering and regenerative medicine, drug delivery, stem cell culture and cell therapy.

Effect of 5-FU on expression of Bmi-1, Sca-1 and Oct-4 in human breast cancer cell line MDA-MB-468 / 临床与实验病理学杂志

Purpose To explore the relation between the expression of Bmi-1 and cancer stem cells and its relation to chemotherapy re-sistance in breast cancer. Methods MTT method was applied to detect the inhibition effect on proliferation of different concentrations (0. 01, 0. 1, 1, 10 μg/ml) of 5-Fluorouracil (5-FU) to MDA-MB-468 cells with 48 and 72 hours culture, a curve of proliferation in-hibition rate was drawn, and a suitable experiment concentration of 5-FU was chosen. MDA-MB-468 cells was serially passaged under continuous interference with the suitable concentration of 5-FU, 6 generations of cells were collected, cells with 5-FU interference were designed as experiment group and a corresponding control group with no 5-FU was set. RT-PCR and Western blot were employed to ex-amine mRNA and protein expression of Bmi-1 and Sca-1, Oct-4 in the 6 generations of cells of both control group and experiment group. Results Results of the MTT test showed that 5-FU could inhibit the proliferation of human breast cancer cell line MDA-MB-468, the 5-FU concentration of 0. 1 μg/ml was chosen as the suitable experiment concentration. RT-PCR tests showed that the differ-ences between the relative mRNA expression values of Bmi-1 and Sca-1, Oct-4 in the 6 generations of MDA-MB-468 cells were not sta-tistically significant in control group (all P>0. 05) and that the differences between the relative mRNA expression values of Bmi-1 and Sca-1, Oct-4 in the 6 generations of MDA-MB-468 cells were of statistical significance in experiment group cells (all P<0. 05). The mRNA expression of Bmi-1, Sca-1 and Oct-4 showed the following tendency in the 6 generations of passaged cells:decrease (1st gen-eration)—increase (2nd generation)—continuous increase (3rd generation)—decrease (4th generation)—increase (5th genera-tion)—decrease (6th generation). Western blot tests indicated that the differences between the relative protein expression values of Bmi-1 and Sca-1, Oct-4 in the 6 generations of MDA-MB-468 cells were not statistically significant in control group and that the differ-ences between the relative protein expression values of Bmi-1 and Sca-1, Oct-4 in the 6 generations of MDA-MB-468 cells were of sta-tistical significance in experiment group cells (all P<0. 05). The expression of Bmi-1 was positively correlated with stem cell associat-ed factors Sca-1, Oct-4 (r=1, all P<0. 01). Conclusions The expression of Bmi-1 gene is positively correlated with expression of stem cell associated factors Sca-1 and Oct-4 in breast cancer cell line MDA-MB-468 cell, and Bmi-1 may be a novel marker for cancer stem cells in breast cancer. Administration of 5-FU can affect the expression level of Bmi-1 and the ration of cancer stem cells in breast cancer cell line MDA-MB-468. Bmi-1 gene may be associated with drug resistance to chemotherapy and recurrence in breast cancer.

The construction and application of the “opening immunity” teaching model / 中华医学教育探索杂志

Teaching reform is the strong requirement of the times and social development.Immunology department inTaishan Medical College integrated teaching and a variety of extra-curricular teaching,broke the constraints of traditional teaching model in time and space,played the leading role of teachers and inspired students' independent learning,thus improving the teaching effectiveness in immunology.

Effects of pulmonary stromal cells supernatant on differentiation of mature dendritic cells / 中华老年医学杂志

Objective To observe the effects of lung stromal cells on differentiation of mature dendritic cells (mDCs) deriving from mouse bone marrow. Methods The mDCs were cultured by complete medium, half of which was from the lung stromal cell cultures. One week later, the mDCs were induced into regulatory dendritic cells (rDCs). The morphological characteristics of rDCs were observed by Giemasa-Wright stain. The content of cytokines (IL-10 and IL-12p70) in the co-culture supernatants was detected by enzyme-linked immunosorbent assay. The expression level of cellular phenotype was tested with fluorescence-activated cell sorting, compared with mDCs. The difference of rDCs and mDCs was observed. Results Compared with mDCs, rDCs secreted higher level of IL-10 (P＜0.01), and lower level of IL-12p70 (P＜0. 01). In cellular phenotype, the expression of CD11b on rDCs increased (P＜0.01), while the expression of CDllc, Ⅰa and CD86 declined (P＜0.01).Conclusions Lung stromal cells microenvironment may induce the mDCs differentiate into a kind of DC with different biological characteristics.

Study of the mechanism of dendritic cells induce FBL-3 cells to differentiate into the monocytes in vitro / 国际肿瘤学杂志

Objective To investigate mechanism involved in the differentiation of FBL-3 cells(eryth-roleukemia cells)induced by dendritic cells(DC). Methods To cultural the FBL-3 cells together with the DC supematant of different concentration for 72 hours,then we used the Wright's staining methed to record the ma-ture monocyte cell population, the transmission electron microscope to observe the ultramicrostructure, the flow eytoraetry to detect the expression rate of the surface molecular CD14. Results The Wright's staining methed, the transmission electron microscope and the flow cytometry all presented that after being induced by the super-natant of the DC ,the FBL-3 cells can partly differentiate into monocytes,inversion monocyte consistent with the personal characteristics. And the intensity of the DC supernatant was connected with interleukin-12. Conclusion The DC supernatant can induce the FBL-3 cells into monocytoid cell, the differentiated cells correspond to monocyte in macro-appearance,uhramicrostructure and phenotype. The competence of the DC supematant are partly concerned with interleukin-12.

Immunohistochemical study for the expression of LHR and VEGFon the ovary of mice during peri-implantation / 解剖学报

Objective To explore the biological effects of the luteinizing hormone receptor (LHR) and vascular endothelial growth factor (VEGF) on the ovary of mice during peri-implantation. Methods The immunohistochemistry SABC method and image analysis were used to study the distribution and changes of the LHR and VEGF in Kunming mouse( n =28) ovary during estrous,pregnancy of day 1, day 4 and day 6 stage. Results The expression of LHR-immunoreactive substance and that of VEGF-immunoreactive substance had the same distribution and changes. Compared with other groups,the level of LHR-immunoreactive substance and that of VEGF-immunoreactive substance increased highly on the stroma cells around largergrowing follicles in estrous group ( P <0.05). Along with the pregnancy, the positive immunostaining for LHR and VEGF increased gradually on the granulosa lutein cells, and reached the highest level on day 6 of pregnancy. Positive immunostaining for LHR or VEGF on some endothelia and blood cells were observed in day 1 of pregnancy or estrous group respectively. Form day 1 of pregnancy, the theca cells had positive immunostaining for LHR. Conclusion The expression of LHR and VEGF is closely related with the process of follicle growing, ovulation and corpus luteum formation.

Screen and Identification of The Protein-protein Interactors in The Host Cell With The SARS Coronavirus Nucleocapsid Protein / 生物化学与生物物理进展

15 SARS-CoV N Protein Interacting Protein (NPIP) were selected from host cells using Yeast Two-hybrid system (Y2H). These are Angiogenin, acyglycerol kinase, cytochrome oxydase subunit I, CXC chemokine ligand 16, epidermal growth factor receptor pathway substrate 15, glutathione S-transferase kappa 1,integrin beta 1, jun oncogene, NIMA (never in mitosis gene a)-related kinase 10, protein tyrosine kinase 2 beta,homo sapiens SH3KBP1 binding protein 1 and ubiquitin specific peptidase 53. With the aid of immunological co-precipitation (CO-IP), it was confirmed that chemokine CXCL16 was the interactor with SARS-CoV N protein in host cells.

The exploration and practice of teaching reform in immunology education under the new medical model / 中华医学教育探索杂志

This paper illustrates the immunology teaching reform of higher medical education to meet demands of the 21st century. The teaching reform includes cultivating students’humanities quality,scientific consciousness,ability to research and the following-up of immunology progress. According to the teaching practice and the results from the questionnaire,the new teaching model is better than traditional teaching model and is approved by the students.

Culture of Dendritic Cells from Human Umbilical Cord Blood using Umbilical Cord Serum instead of Fetal Calf Serum / 中国输血杂志

Objective To investigate the effect of umbilical cord serum(UCS)during induction of dendritic cells derived from umbilical cord blood. Methods Umbilical cord blood was collected aseptically and interface cells were collected with density gradient centrifugation method,CD34 +cells were purified and collected with Mini-MACS technique. Pro-DC were obtained and were divided into 3 groups:①cultured in RPMI-1640 medium with 10%UCS and cytokine,②cultured in RPMI-1640 medium with 10%FCS and cytokine③cultured in RPMI-1640 medium with 10%FCS .Interleukin-4(IL-4),granulocyte-macrophage colony-stimulating factor(GM-CSF)and tumor necrosis factor(TNF-a)were used in the first 2 groups, but not in control group (group 3). Some cells were collected on the 8th d for phenotypes analysis. Results The expression of CD80 ,CD54 and HLA-DR on the surface of DC cultured in UCS medium had no significant difference compared with that of DC cultured in FCS medium. The expression of surface molecules in UCS-DC group and FCS-DC group were both enhanced compared with the control group. Conclusion Mature DC could be induced from pro-DC derived from umbilical cord blood when cultured with UCS instead of FCS. That provides a new approach for the clinical application of DC.