Regulation of RUNX3 gene on RAW264.7 macrophage polarization / 西安交通大学学报(医学版)

Objective To investigate the role of RUNX3 in the regulation of macrophage polarization so as to provide a new therapeutic approach for immunity-related diseases.Methods ① After RAW264.7 cells were stimulated by IFN-γ,LPS and IL-4,respectively,the expressions of their surface markers(arginase-1 and iNOS) were detected by RT-PCR to observe whether RAW264.7 cells polarized to M1 or M2 after stimulation by IFN-γ, LPS and IL-4.The cells stimulated by IFN-γ and LPS were named group M1 and those stimulated by IL-4 were group M2;the control group was group M0.② The expression of RUNX3 was detected by immunofluorescence and RT-PCR methods in each cell group(M1,M2 and M0).③ RUNX3 over-expression vector was established.The RUNX3 gene in RAW264.7 cells was silenced.Cell lines with stable expression were screened with G 418 culture medium.The expressions of cell surface markers iNOS and CD86 were detected by RT-PCR;cell secretion(TNF-α) was detected using ELESA method.Results ① Stimulation of RAW264.7 cells with IFN-γ and LPS could induce RAW264.7 cells to polarize into M1 type macrophages.The mRNA expression of iNOS in M1 group was higher than that in group M0 detected by RT-PCR(P= 0.002),while using IL-4 to stimulate RAW264.7 cells could induce RAW264.7 cells to polarize into M2 macrophages.The results of RT-PCR detection showed that the expression of arginase-1 was higher in M2 group than in group M0(P=0.021).② The expression of RUNX3 mRNA in the M1 cells group was higher than that in the M0 cells group(P= 0.001),but the expression in the M2 cells group was decreased(P=0.041).③ After silencing of RUNX3,the expressions of RAW264.7 cell surface markers CD86 and iNOS(P=0.005)and the cells secretion of TNF-α(P＜0.001)were decreased compared with those in the control group.Conclusion RUNX3 transcriptional activation may promote the differentiation of macrophages into M1 type.

Intervention effect and mechanism of compound Ginkgo biloba preparations on nonalcoholic fatty liver / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the intervention effect and mechanism of compound Ginkgo biloba (CGB) preparations on nonalcoholic fatty liver disease (NAFLD).</p><p><b>METHOD</b>The C57BL/6 mouse NAFLD model was induced with high fat diets. Since the 2nd week after modeling, the mice were orally administered with 600 and 200 mg x kg(-1) x d(-1) CGB for eight weeks. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), cholesterol (CHOL) and LPS in serum, as well as pathological changes and expression of tumor necrosis factor-alpha (TNF-alpha) in hepatic tissues were observed. Changes in intestinal tight junction proteins ZO-1, Occludin, Claudin-1 in intestinal tissues were determined under microscopy.</p><p><b>RESULT</b>Compared with the normal group, the model group showed obvious fatty degeneration in rat livers, with notable increase in TNF-alpha expression (P < 0.01), significant increases in ALT, AST, TG, CHOL and LPS in serum (P < 0.01, P < 0.05), injury in intestinal tight junction proteins, and remarkable declines in ZO-1, Occludin and Claudin-1 (P < 0.01). Compared with the model group, CGB high and low dose groups showed obvious relieves in fatty degeneration in rat livers and injury in intestinal tight junction proteins, significant reductions in TNF-alpha expression (P < 0.01, P < 0.05) and AST, TG, CHOL and LPS in serum (P < 0.01, P < 0.05) and remarkable increases in ZO-1 and Occludin expressions (P < 0.05).</p><p><b>CONCLUSION</b>CGB can protect intestinal tight junction proteins, reduce intestinal leakage, relieve fatty degeneration and inflammations in livers and prevent NAFLD occurrence and development.</p>

Advance in studies on food allergy mechanism based on gut barrier / 中国中药杂志

Food allergies, as a type of adverse immune-mediated reactions to ingested food proteins, have become a serious public health issue that harms children and adults health, with increasing incidence year by year. However, without effective therapy for food allergies, doctors-have mostly advised to avoid allergens and provided symptomatic treatment. According to the findings of many studies, allergic diseases are correlated with intestinal barrier function injury, as evidenced by the significant increase in the intestinal permeability among patients with food allergies. In this paper, recent studies on correlations between food allergies and intestinal barrier functions, intestinal barrier function injury mechanisms of allergic foods and food allergy intervention strategies based on intestinal barrier functions were summarized to provide reference for laboratory researches and clinical treatment of food allergic diseases.

Apoptosis of multiple myeloma cells induced by gossypol acetic acid in vitro and its mechanism / 中国实验血液学杂志

This study was aimed to investigate the apoptosis effect of gossypol acetic acid on classic human multiple myeloma RPMI8226 cell line in vitro and its mechanism. The inhibitory effect on proliferation of RPMI8226 cells was evaluated by means of MTT assay. Cytotoxic effect and apoptosis was identified and analyzed with the aid of transmission electron microscopy, mitochondria membrane potential (MMP) and DNA gel electrophoresis. Meanwhile, Western-blot assay was used to detect the changes of several key cell apoptosis regulatory proteins such as BAX, caspase-3 and caspase-8 in these cells before and after treatment. The results showed that low concentrations of gossypol acetic acid (> 16 micromol/L) could suppress the proliferation and induce the apoptosis in RPMI8226 cells effectively. At the same time, gossypol acetic acid could also down-regulate the mitochondrial membrane potential, up-regulate the expression of the apoptosis-related protein such as BAX and caspase-3. It is concluded that the gossypol acetic acid can selectively induce proliferation inhibition and apoptosis of multiple myeloma RPMI8226 cells with a smaller dose.

Effects of HCMV on phenotypes of parotid duct epithelial cells and its mechanisms / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate the effects of HCMV infection on phenotypes of parotid duct epithelial cells and relative mechanisms.</p><p><b>METHODS</b>The expressions of immediate early antigen of HCMV, pan cytokeratin and cathepsin D etc. were detected by immunohistochemical staining in tissues of parotid cytomegalic inclusion disease.</p><p><b>RESULTS</b>Cytokeratin which acts as an epithelial marker became negative while staining of Cathepsin D was intensified in parotid duct epithelial cells after infected by HCMV.</p><p><b>CONCLUSION</b>It demonstrated that cytokeratin was lost through over-expression of Cathepsin D in parotid duct epithelial cells infected by HCMV.</p>

Effect of bone marrow stromal cells on the apoptotic sensitivity of HL-60 and HL-60/VCR cells / 中国实验血液学杂志

This study was aimed to investigate the effects of human bone marrow fibroblastoid stromal cell line (HFCL) on chemosensitivity of acute myeloid leukemia sensitive HL-60 cell line and multidrug-resistant (MDR) HL-60/VCR cell line in vitro co-culture. Setting up co-culture system of HL-60 or HL-60/VCR cells in direct contact with HFCL cells, or with HFCL cells separated by transwell, and exposing HL-60 or HL-60/VCR cells to different concentrations of topotecon (TPT), morphologic evidence for apoptosis was determined by staining with Wright-Giemsa stain and acridine orange/ethidium bromide (AO/EB). Cell cycle, sub-G(1) and annexin V FITC staining were detected by flow cytometry. The expression of active caspase-3, Bcl-2 and Pgp was detected by Western blot. The results showed that HL-60 or HL-60/VCR cells treated by TPT revealed characteristic apoptotic morphological changes by Wright-Giemsa and AO/EB staining. The percentage of annexin V-positive cells and apoptotic cells decreased when they were cocultured with HFCL cells. The proportion of G(0)/G(1) HL-60 or HL-60/VCR cells treated by TPT increased and the sub-G(1) appeared significantly, but apoptotic and sub-G cells reduced after direct contact with HFCL cells. Meanwhile, although HL-60 or HL-60/VCR cells treated by TPT expressed activated caspase-3, and the expression of Bcl-2 decreased, the expression of activated caspase-3 decreased and Bcl-2 increased after direct contact with HFCL cells. In conclusion, HFCL stromal cells can prevent TPT-induced apoptosis in HL-60 and HL-60/VCR cells via modulation of Bcl-2 and active caspase-3.

Human cytomegalovirus inhibits proliferation of duct epithelial cells of human salivary gland / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of human cytomegalovirus (HCMV) on the proliferation of duct epithelial cells of human salivary gland (HSG).</p><p><b>METHODS</b>The expression of proliferating cell nuclear antigen (PCNA) and p53 were studied in 11 cases of parotid cytomegalic inclusive disease (PCID) using immunohistochemical staining method. The effects of human cytomegalovirus (HCMV) on the proliferation of HSG were investigated by MTT method in vitro. The expression of PCNA in HSG infected by HCMV was examined using immunocytochemical staining and Western blotting.</p><p><b>RESULTS</b>PCNA was expressed weakly in most of megalic inclusion cells which were positive for HCMV, while all the megalic inclusion cells were p53 negative in all 11 cases of PCID. HCMV inhibited proliferation of HSG in vitro in a time dependent and dose dependent manner. Down-regulation of PCNA was shown in infected cells.</p><p><b>CONCLUSION</b>HCMV inhibits proliferation of HSG and down-regulation of PCNA may be an expression of the inhibition.</p>

Expressions of vimentin and NF-?B in Hashimoto′s thyroiditis / 中华内分泌代谢杂志

The expressions of vimentin and NF-?B were detected in thyroid tissue by immuno- histoehemistry in 26 cases with Hashimoto′s thyroiditis and 9 normal subjects.The results showed that there was a significantly increased expression of vimentin in Hashimoto′s thyroiditis as compared with the normal controls.It indicated that epithelial-mesenchymal transformation existed in follicular epithelial ceils of Hashimoto′s thyroiditis. Furthermore,the expression of vimentin was closely related with NF-?B.