Preparation of a miR-155-activating nucleic acid nanoflower to study the molecular mechanism of miR-155 in inflammation.

At present, the molecular mechanisms underlying inflammation remain unclear. In recent years, research on inflammation has focused on stimulating cell inflammation by using exogenous pro-inflammatory substances such as lipopolysaccharide (LPS) or inflammatory factors. To investigate the molecular mechanism of inflammation from a new perspective, we designed a nucleic acid nanoflowers (NFs) complex to directly activate inflammatory genes to study the inflammatory response without the need for external microbial factors to trigger an inflammatory response. An RNAa-type target gene-activated NFs was designed. Human umbilical vein endothelial cells (HUVECs) were transfected with NFs carrying small activating RNA (saRNAs) to directly co-activate microRNA (miR)-155 and SHIP1 genes. After RNA activation (RNAa)-type NFs were transferred into HUVECs, the expression of miR-155 and pro-inflammatory and cancer-related factors increased, anti-inflammatory factors were reduced, cell proliferation increased, and cell migration was promoted. IL-1ß protein levels were decreased and SHIP1 expression was downregulated. When miR-155 and its target SHIP1 were both activated, the expression of both was unaltered, maintaining cell homeostasis. This points towards miR-155 overexpression can trigger inflammation, and that miR-155 and its target genes act as a molecular switch role in the development of inflammation.

Environmental innovation ambidexterity and customer relationship performance: evidence from the manufacturing industry in China.

Cleaner production and pollution prevention are not only essential for enterprises to improve resource utilization efficiency and competitiveness, but also realistic requirements for enterprises to comply with green manufacturing policies and regulations. Many previous studies have investigated the impacts of environmental innovation on products and environmental performance, but few of them have explored the impact of environmental innovation on customer relationships. The main purpose of this study is to investigate the basic strategies to enhance customer relationship in the context of green development. Building upon the resource-based view, the dynamic capability perspective, and the upper echelons theory, this study develops a conceptual model focusing on the effects of environmental innovation ambidexterity (EIA) on customer relationship performance (CRP), the mediating effects of green product innovation (GPI), and the moderating role of top management's environmental awareness. The results of a survey of 285 high-tech manufacturing enterprises in China show that EIA has a positive effect on CRP. The results of the study also indicate that GPI mediates the relationship between EIA and CRP, and top management's environmental awareness moderates the impact of EIA on CRP. The results offer novel insights for suppliers to improve CRP, and provide theoretical guidance for high-tech manufacturing enterprises to effectively implement pollution prevention and GPI.

[Response surface methodology for optimizing the technology of induction and isolation of dendritic cells by multiple cytokines].

Objective To promote the induction and separation efficiency of bone marrow-derived dendritic cells (BMDCs) in vitro through optimizing the inducing and isolating process by multiple cytokines. Methods The factors to be optimized in single factor tests included recombinant mouse granulocyte macrophage-colony stimulating factor (rmGM-CSF), recombinant mouse interleukine 4 (rmIL-4), lipopolysaccharide (LPS), recombinant mouse tumor necrosis factor α (rmTNF-α) and inducing time. The numbers of immature dendritic cells (imDCs) and mature dendritic cells (mDCs) were investigated as the indicators. Box-Behnken experimental design-response surface methodology was used to analyze and verify the data. Morphological changes were observed using the inverted microscopy and the transmission electron microscopy. Surface molecules including CD11c and CD86 were detected using the flow cytometry. Results The optimum inducing conditions for imDCs were obtained as follows: rmGM-CSF was 46 ng/mL, rmIL-4 was 24 ng/mL, inducing time was 6 days, and the number of imDCs was (4.58±0.28)×106 cells, and the relative deviation was 4.00%. The optimum inducing conditions for mDCs were as follows: LPS was 1.4 µg/mL, rmTNF-α was 30 ng/mL, inducing time was 1 day, and the number of mDCs was (4.21±0.15)×106 cells, and the relative deviation was 3.80%. Sufficient typical imDCs and mDCs were obtained within 5-7 days of induction in vitro. Also, flow cytometry showed that the amplified imDCs had a high expression of CD11c (68.62%±2.3%) and a low expression of CD86 (37.95%±1.8%), and the mDCs had high expressions of both CD86 (90.34%±1.4%) and CD11c (82.05%±1.6%). Conclusion The combination of single factor tests and Box-Behnken design -response surface methodology could optimize the inducing and isolating method for DCs in vitro by multiple cytokines rapidly and efficiently, which provided basic experiment materials for further studies.

Association analysis of five SNP variants with gout in the Minnan population in China.

BACKGROUND/AIM: There is a very high prevalence of gout in the Minnan population in China. We aimed to explore the genetic characteristics and genetic mechanisms of gout in the Minnan population by studying the association of 5 single nucleotide polymorphisms (SNPs) with gout. MATERIALS AND METHODS: A total of 163 gout patients and 187 normal controls from Minnan were enrolled in this case-control study. SNPs (rs1165205, rs3733591, rs6855911, rs2231142, rs333049) were genotyped by allele-specific polymerase chain reaction (PCR) and analyzed with SPSS 16.0. RESULTS: Significant association with gout was found for rs2231142 (P < 0.001), consistent with our prior studies. An association between rs1333049 and gout was also found (P = 0.03) in the Minnan population. No association of SNPs rs6855911, rs3733591, and rs1165205 was found with gout in the Minnan population. CONCLUSION: Rs1333049 is associated with gout in the Minnan population, although rs2231142 shows an even stronger association with gout. The C allele of rs1333049 and the A allele of rs2231142 might be crucial risk factors for gout.

[Anti-inflammatory effect of recombinant human kallistatin in ulcerative colitis of mice].

This study was designed to evaluate the anti-inflammatory effect of recombinant human kallistatin (Kal) on ulcerative colitis (UC) in the mouse model. Acute colitis was induced by administration of 4% dextran sodium suffate (DSS) to KM mice for 7 days. The mice were then randomized into 5 groups: model control, Kal 0.2 mg·kg(-1)·d(-1), 1.0 mg·kg(-1)·d(-1) and 2.0 mg·kg-1·d(-1) group, salazosulfapyridine (SASP) group. Ten age-matched normal KM mouse were administered with saline in the normal control. The weight, colon length, inflammation factor (MPO/SOD/MDA) and TNF-α/IL-10 levels among the five groups of mice were determined. The results showed that histological index score and MPO/MDA/TNF-α levels of high-dose Kal treatment group and SASP group were significantly lower compared with the model group (P < 0.01), but the weight, colon length, IL-10 level and SOD activity were significant higher than the model group (P < 0.01), approaching the normal group. These parameters showed that Kal can significantly relieve the UC state in a dose-dependent manner. This study demonstrates that Kal significantly remits UC in mice, and participates in the regulation of inflammatory cytokines TNF-α/IL-10 levels and has some antioxidant activity.

Plasmid mediated kallistain gene expression via intramuscular electroporation delivery in vivo for treatment of NCI-H446 subcutaneous xenograft tumor.

Kallistatin (KAL) is a novel anti-tumor protein with anti-angiogenic activity. The aim of this study was to investigate whether intramuscular injection of KAL plasmid DNA by electroporation could inhibit NCI-H446 subcutaneous xenograft tumor growth in mice. The tumor model of BALB/c nude mice was induced by subcutaneous inoculation of 5×10(6) NCI-H446 cells into the mice right flank. The next day, naked plasmid pEGFP or pKAL was electrotransfered into the skeletal muscle of nude mice (n=6 for each group), with the optimized electroporation conditions. Tumor cells migration was assessed by E-cadherin staining; Proliferation was determined by anti-Ki-67 staining; and apoptosis was assayed via TUNEL, tumor microvessel density (MVD) was examined by anti-CD34 staining to evaluate the angiogenesis of tumor. Compared to the pEGFP treating group, tumor growth was inhibited by 85% (pEGFP group: 486 ± 187 mm(3), pKAL group: 71±33 mm(3)) at day 42, the MVD of tumor tissues was significantly decreased, and tumor cellular proliferation was also inhibited. The results indicate that this therapeutic strategy might serve as a promising approach for cancer clinical therapy.