pH-stimulus response dye-cucurbituril sensor for amino acids in aqueous solution.

The host-guest inclusion complexes that comprise an inverted cucurbit[7]uril (iQ[7]) and a quinoline derivative, 4-(4-dimethylaminostyryl) quinoline (DSQ) at different pHs were exploited as multiple supramolecular sensors to sense l-α-amino acids. DSQ complexation inside iQ[7] at different pHs leads to increased fluorescence and formation of different-colored iQ[7]-DSQ complexes. The enhanced fluorescence of DSQ after iQ[7] encapsulation may be attributed to limited dimethylamine rotation and the formation of a twisted internal charge transfer (TICT) state. The DSQ@iQ[7] sensors have different affinities for l-α-amino acids at different pHs. Therefore, we propose a pH-stimulus response supramolecular sensor for the discrimination of structurally similar l-α-amino acids in aqueous solution.

Elevated Th22 cells correlated with Th17 cells in patients with rheumatoid arthritis.

BACKGROUND: T-helper (Th) 22 and Th17 cells are implicated in the pathogenesis of autoimmune diseases. The roles of Th22 cells in the pathophysiology of rheumatoid arthritis (RA) remain unsettled. MATERIALS AND METHODS: CD4(+)IFNγ(-)IL17(-)IL-22(+) T cells (Th22 cells), CD4(+)IFNγ(-)IL-22(-)IL17(+) T cells (pure Th17 cells), CD4(+)IL17(+) T cells (Th17 cells), and CD4(+)IFNγ(+) T cells (Th1 cells) in RA, osteoarthritis patients, and healthy controls were examined by flow cytometry. Plasma IL-22 and IL-17 levels were examined by enzyme-linked immunosorbent assay. RESULTS: Th22 cells, pure Th17 cells, Th17 cells, and interleukin-22 were significantly elevated in RA patients compared with osteoarthritis and healthy controls, but there were no significant differences regarding Th1 cells and interleukin-17. Th22 cells showed a positive correlation with interleukin-22 as well as pure Th17 cells or Th17 cells in RA patients. Additionally, the percentages of Th22 cells, pure Th17 cells as well as Th17 cells correlated positively with both C-reactive protein levels and 28-joints disease activity score. CONCLUSION: Together, our results indicated a possible role of Th22 pure Th17 cells and Th17 cells in RA, and blockade of the interleukin-22 may be a reasonable therapeutic strategy for RA.

OPN promotes survival of activated T cells by up-regulating CD44 in patients with oral lichen planus.

Oral lichen planus (OLP) is a chronic inflammatory disorder of oral mucosa, which represents cell-mediated autoimmune diseases. Pathological study demonstrated that abundant T lymphocytes infiltrated the oral mucosa, in which the activated T cells that trigger apoptosis of oral epithelial cells is an important mechanism for OLP. However, to date the molecular mechanisms underlying the T lymphocytes infiltration and accumulation in OLP remain unclear. In this paper, we found that the levels of plasma OPN were elevated and were associated with the up-regulated expressions of CD44 in OLP patients. In vitro, the addition of exogenous OPN can suppress the apoptosis of activated CD8(+) T cells via CD44, and this T cell resistance to apoptosis may be attributed to the reduction of endogenous mature granzyme B. Our results suggested that the abnormally elevated levels of OPN may contribute to the abnormal infiltration and accumulation of the activated T cells by up-regulating CD44 in OLP.

Expression of BAFF in the trophoblast and decidua of normal early pregnant women and patients with recurrent spontaneous miscarriage.

BACKGROUND: BAFF, the B cell activation factor, is a member of the tumor necrosis factor (TNF) ligand family that binds to BCMA, TACI, and BAFF-R. Previous studies have shown that members of the TNF family are detected in human placental trophoblast cells, but the expression patterns of BAFF involved in human decidua and the differential expression of BAFF between normal pregnancy and miscarriage are still incompletely documented or unknown. This study was designed to investigate the expression of BAFF and BAFF-R in the trophoblast and decidua of normal early pregnant women and recurrent spontaneous abortion (RSA) patients. METHODS: Forty-five patients with RSA and 45 normal pregnant women were included in this study. By reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and immunohistochemical experiments, we explored the expression of BAFF and BAFF-R in the maternal-fetal interface of normal early pregnant women and RSA patients. RESULTS: Analysis by RT-PCR and Western blotting revealed that BAFF was detected in both trophoblast and decidua of all the samples, and the expression level was higher in the tissues of normal early pregnant women (P<0.05) than that of recurrent spontaneous abortion patients under the same gestational weeks. Messages for BAFF-R were absent. Immunohistochemical experiments showed that expression of BAFF was cell-specific which was localized to villous cytotrophoblast and syncytiotrophoblast cells in trophoblast and to stromal cells in decidua. Whereas BAFF was prominent on the trophoblast and decidua of normal early pregnant women, it was decreased in the tissues of RSA patients. CONCLUSIONS: BAFF might steer maternal leukocytes away from a harmful immune response and toward a favorable one and play a potentially vital role for successful pregnancy.

[Effect of hypoxia inducible factor-1 alpha on vascular endothelial growth factor expression in human tongue squamous carcinoma cells (Tca8113) under hypoxia].

OBJECTIVE: To investigate the effect of hypoxia inducible factor-1 alpha (HIF-1 alpha) on vascular endothelial growth factor (VEGF) expression in Tca8113 cells under hypoxia. METHODS: The expression of the mRNA of HIF-1 alpha and VEGF in Tca8113 cells was examined by RT-PCR technique at different culture times (1/2 h, 1 h, 3 h, 6 h, 12 h, 24 h) under normoxic and hypoxic conditions. RESULTS: The expression of HIF-1 alpha under hypoxia showed the trend of increasing first and then decreasing, and was higher than that of the control (normoxic group) at 6h and 12 h (P < 0.05). The expression of VEGF under hypoxia was higher than that of the control group at 1/2 h, 1 h, 3 h, 12 h, 24 h (P < 0.05). The expression of hypoxia-induced VEGF mRNA increased with the increased expression of HIF-1 alpha mRNA in the cell lines tested at the initial stage of hypoxia. But no statistical significant association was observed between HIF-1 alpha and VEGF expression within 24 h under hypoxia (rs = 0.5750, P > .005). CONCLUSIONS: The increased expression of VEGF in Tca8113 cells might be mediated by multiple factors, including HIF-1 alpha.

Membrane type 1-matrix metalloproteinase is involved in the migration of human monocyte-derived dendritic cells.

Dendritic cells (DC) are highly mobile APC. The trafficking of both immature and mature DC is crucial for their functions, which depends mainly on chemotactic attraction and matrix metalloproteinases (MMP) activity. MMP that are in a transmembrane form belong to membrane type (MT)-MMP, among which MT1-MMP has been shown to possess strong proteolytic activity that is capable of degrading extracellular matrix molecules. Although it is well established that MMP are zinc-dependent endopeptidases that collectively degrade most components of the extracellular matrix, relatively little is known about MT-MMP-mediated matrix degradation during DC migration. In this study, we showed that MT1-MMP was expressed in human monocyte-derived immature and mature DC by semi-quantitative reverse transcription PCR and western blotting analyses. Moreover, immunofluorescence microscopic studies showed that MT1-MMP was expressed on the membrane surface of DC. Blocking of MT1-MMP activity greatly reduced the invasion capacity of immature DC in Matrigel, whereas mature DC mobility was not affected. Taken together, our results show a novel functional link between MT1-MMP and DC motility and suggest that MT1-MMP may play an important role in modulating the migration of immature DC.

Hypoxia inhibits the migratory capacity of human monocyte-derived dendritic cells.

Hypoxia, a prominent characteristic of inflammatory tissue lesions and solid tumour microenvironments, is a crucial stimulus capable of modulating the expression of specific genes involved in leucocyte recruitment. Although studies have shown that hypoxia can affect leucocyte migration by influencing the expression of migration-related genes, such as matrix metalloproteinases (MMP) and their endogenous tissue inhibitors of matrix metalloproteinases (TIMP), it remains unclear whether hypoxia can affect the migration of dendritic cells (DC). In this study, we showed that human monocyte-derived DC under hypoxic conditions in a transwell system have significantly reduced migratory capacity compared to normoxic controls. A moderate phenotypic change of hypoxic DC was observed. In hypoxic DC, we detected a twofold increase in TIMP-1 transcript levels, and downregulated expression of MMP-9 and membrane type 1-MMP genes by threefold and 17-fold, respectively. Our results suggest that hypoxia may inhibit DC migratory activity by regulating the balance between MMP and TIMP gene expression.

[Hepatocellular carcinoma associated antigen HCA520, a novel Ca2+-binding protein].

AIM: To analyze the binding activity of the hepatocellular carcinoma associated antigen HCA520 to Ca2+. METHODS: The HCA520 gene was gained by PCR. Then, it was cloned into the prokaryotic expression vector pGEX-4T-3. The GST-HCA520 fusion gene was induced to express in E. coli and the expressed product was purified via GST-agarose affinity resin. The fusion protein was confirmed by Western blot analysis. The Ca2+ binding capability of the fusion protein was analyzed by dot blot. RESULTS: The GST-HCA520 fusion gene was constructed and the protein was successfully expressed and purified, which was identified by DNA sequencing and Western blot respectively. The fusion protein could bind to Ca2+ depend on dosages. CONCLUSION: HCA520, a novel hepatocellular carcinoma associated antigen, is a novel Ca2+ binding protein.