To determine the expression profile and potential roles of CD24 in oral squamous cell carcinoma and explore the values of CD24 function as a potential target of clinical therapy.Semi-quantitative immunohistochemistry was used to construct the expression profile of CD24 in 78 human oral tissues and 59 Hamster buccal pouch tissues. Real-time RT-PCR and Western blot were used to analyze the CD24 expression levels in oral DOK4 cells, oral cancer CAL-27 and WSU-HN6 cells. Then these two cancer cell lines were selected to evaluate the effect of all-trans retinoic acid (ATRA) and CD24 antibody on CD24 expression, and the proliferation and tumorsphere formation capacity of these two cell lines.CD24 expression was found significantly elevated in both human and animal tissues compared with normal and benign tissues (P<0.05), as well as in oral cancer CAL-27 and WSU-HN6 cells compared with DOK cells (P<0.05). CAL-27 and WSU-HN6 cells possess increased proliferative and specific tumorsphere formation capability compared with DOK cells (P<0.05). Both ATRA and CD24 antibody were able to effectively inhibit the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells (P<0.05). Among them ATRA at least involved partially in the proliferation by down-regulating the CD24 expression (P<0.05), while CD24 antibody blocking had no effect on the CD24 expression.CD24 was upregulated in oral cancer and functioned as a potential factor that promoted the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells. Both ATRA and CD24 antibody might effectively inhibit the proliferation and tumorsphere formation of CAL-27 and WSU-HN6 cells and function as a potential therapy target.
The first human infection with novel H7N9 influenza virus was reported in March 2013 in China. To date, the H7N9 influenza viruses possess a potential pandemic threat to public health worldwide and have caused severe infection and high mortality in humans in China. The latent period of human infection of H7N9 influenza virus is generally within 7 days. Patients usually show flu like symptoms, such as runny nose, sore throat, fever, cough, headache and muscle pain. Severe patients have rapid development, characterized by severe pneumonia and dyspnea. Antibodies play a major role in protective immunity against the influenza virus infection, and passive immunization with monoclonal antibodies (MAbs) specific to viral proteins might be a potential option for humans against lethal infection of the influenza viruses. It is known that hemagglutinin (HA), is the major viral antigenic protein on the influenza virus particle and the primary target of neutralizing antibodies against influenza virus. In this study, in order to obtain the HA MAbs of influenza virus H7N9 (A/Shanghai/2/2013), study their biology characteristics, and evaluate the prophylactic and therapeutic efficacy of the HA MAbs in mouse model. The BALB/c mice were immunized with inactivated H7N9 vaccine by subcutaneous administration for three times, and boosted with inactivated H7N9 vaccine by the tail vein three days before fusion. Finally, 28 MAbs were obtained and identified by ELISA and hemagglutination inhibition test (HI). Eight IgG subtype MAbs (1A8, 2A12, 2H3, 4A11, 4C8, 4G5, 6A3, 7G2) were selected to identify their biology characteristics. The ELISA titers of ascites of the eight MAbs were 102–106. The eight Mabs exhibited high affinity which can reach nmol/L level and meet the requirement of affinity for monoclonal antibody drug. The immunofluorescence assay indicated that the eight MAbs had excellent specificity for HA protein expressed in 293T cell and were able to recognize the H7N9 influenza virus particles. In addition, all the eight MAbs could inhibit HA activity and the MAbs 2H3, 4A11, 4G5 and 6A3 can also present neutralization activity against the H7N9 live virus. The highest neutralizing titer was up to 1:14125. At the same time, we evaluated the prophylactic and therapeutic efficacy of these MAbs against the homologous H7N9 strain. The activity against H7N9 virus was evaluated by clinical syndromes, survival rates and lung viral titers. The results showed that in the prophylactic and therapeutic experiments, MAb 6A3 can inhibit effectively H7N9 influenza virus infection, provided 100% protection and remarkably reduced the lung viral titers in mouse model. The HA monoclonal antibodies against influenza virus H7N9 were successfully obtained. The HA MAbs not only may be as a prophylactic and therapeutic measure to prevent H7N9 influenza virus infection, but also laid the foundation for us to research the H7N9 HA antigen epitope and develop diagnostic reagents.
The 5-year survival of colorectal cancer (CRC) has had no obvious improvement during the past decades, although a significant progress in treatments has been established. The molecular mechanisms that drive the progression of CRC are still unclear. This study aims to determine the biological activities of linc01615 and its regulatory microRNAs in CRC cells.The expression of linc016150 and miR-491-5p was measured in six CRC cell lines and one normal colon mucosal epithelial cell line. Their effects on cell proliferation, apoptosis, invasion, and migration were tested in Caco-2 cells.Linc01615 mRNA expression was upregulated in six colorectal cancer cell lines compared to the normal colon mucosal epithelial cell line, which was negatively regulated by miR-491-5p in Caco-2 cells. Silencing of linc01615 gene expression significantly decreased cell proliferation, increased apoptosis, and inhibited invasion and migration in Caco-2 cells. Overexpression of linc01615 exhibited an opposite effect on silencing of linc01615 expression. Transfection with miR-491-5p mimics downregulated linc01615 expression and inhibited the biological activities of linc01615. In contrast, the inhibitor of miR-491-5p up-regulated linc01615 expression and subsequently enhanced the biological activities of linco1615. Also, overexpression of linc01615 can block the effects of miR-491-5p mimics in Caco-2 cells.Linc01615 functions as an oncogene while miR-491-5p functions as a tumor suppressor in colorectal cancer cells through negatively regulating each other; both are involved in cell proliferation, apoptosis, invasion, and migration in colorectal cancer cells.
The wbsJ gene from Escherichia coli O128:B12 encodes an alpha1,2-fucosyltransferase responsible for adding a fucose onto the galactose residue of the O-antigen repeating unit via an alpha1,2 linkage. The wbsJ gene was overexpressed in E. coli BL21 (DE3) as a fusion protein with glutathione S-transferase (GST) at its N-terminus. GST-WbsJ fusion protein was purified to homogeneity via GST affinity chromatography followed by size exclusion chromatography. The enzyme showed broad acceptor specificity with Galbeta1,3GalNAc (T antigen), Galbeta1,4Man and Galbeta1,4Glc (lactose) being better acceptors than Galbeta-O-Me and galactose. Galbeta1,4Fru (lactulose), a natural sugar, was furthermore found to be the best acceptor for GST-WbsJ with a reaction rate four times faster than that of lactose. Kinetic studies showed that GST-WbsJ has a higher affinity for lactose than lactulose with apparent Km values of 7.81 mM and 13.26 mM, respectively. However, the kcat/appKm value of lactose (6.36 M(-1) x min(-1)) is two times lower than that of lactulose (13.39 M(-1) x min(-1)). In addition, the alpha1,2-fucosyltransferase activity of GST-WbsJ was found to be independent of divalent metal ions such as Mn2+ or Mg2+. This activity was competitively inhibited by GDP with a Ki value of 1.41 mM. Site-directed mutagenesis and a GDP-bead binding assay were also performed to investigate the functions of the highly conserved motif H152xR154R155xD157. In contrast to alpha1,6-fucosyltransferases, none of the mutants of WbsJ within this motif exhibited a complete loss of enzyme activity. However, residues R154 and D157 were found to play critical roles in donor binding and enzyme activity. The results suggest that the common motif shared by both alpha1,2-fucosyltransferases and alpha1,6-fucosyltransferases have similar functions. Enzymatic synthesis of fucosylated sugars in milligram scale was successfully performed using Galbeta-O-Me and Galbeta1,4Glcbeta-N3 as acceptors.
Deficiency of muscle basement membrane (MBM) component laminin-α2 leads to muscular dystrophy congenital type 1A (MDC1A), a currently untreatable myopathy. Laminin--α2 has two main binding partners within the MBM, dystroglycan and integrin. Integrins coordinate both cell adhesion and signalling; however, there is little mechanistic insight into integrin's function at the MBM. In order to study integrin's role in basement membrane development and how this relates to the MBM's capacity to handle force, an itgβ1.b−/− zebrafish line was created. Histological examination revealed increased extracellular matrix (ECM) deposition at the MBM in the itgβ1.b−/− fish when compared with controls. Surprisingly, both laminin and collagen proteins were found to be increased in expression at the MBM of the itgβ1.b−/− larvae when compared with controls. This increase in ECM components resulted in a decrease in myotomal elasticity as determined by novel passive force analyses. To determine if it was possible to control ECM deposition at the MBM by manipulating integrin activity, RGD peptide, a potent inhibitor of integrin-β1, was injected into a zebrafish model of MDC1A. As postulated an increase in laminin and collagen was observed in the lama2−/− mutant MBM. Importantly, there was also an improvement in fibre stability at the MBM, judged by a reduction in fibre pathology. These results therefore show that blocking ITGβ1 signalling increases ECM deposition at the MBM, a process that could be potentially exploited for treatment of MDC1A.
Soils are a large source of atmospheric methane, which is a prevalent greenhouse gas. The vertical flux pattern of methane in soils is unclear. We investigated relationships between methane vertical fluxes in the soil, total organic carbon (TOC) and methanogens in a riparian buffer of the Living Water Garden, a sponge city park, from August 2018 to January 2020. Average surface methane fluxes were 81.86 mg m-2 h-1 and ranged from 20.42 mg m-2 h-1 to 190.75 mg m-2 h-1. Cumulative methane fluxes from studied area were 7.26 kg CO2eq m−2 year−1 and the global warming potential (GWP) was at a moderate level. Results of structural equation model (SEM) showed that vertical methane fluxes varied with soil depth and were mainly regulated by methanogenic communities (λ = 0.44) and methanogenic diversity (λ = -0.47). Remarkably, the contribution of methanogenic genes abundance (λ = -0.19) and soil total organic carbon (λ = -0.28) at each soil depth to vertical methane fluxes was low, implying that the effect of microbial source carbon on vertical methane fluxes should be considered. These results indicated that methanogenic communities composition and diversity are effective predictors of vertical methane fluxes and modulate the relationship between TOC and vertical methane fluxes. These findings provide direct monitoring evidence for controlling vertical methane fluxes by regulating TOC and methanogens activity in soil.
Psoriasis is one of the most common skin inflammatory diseases worldwide. The vitamin D3 analog calcipotriol has been used alone or in combination with corticosteroids in treating plaque psoriasis, but how it suppresses psoriatic inflammation has not been fully understood. Using an experimental mouse psoriasis model, we show that topical calcipotriol inhibited the pivotal IL-23/IL-17 axis and neutrophil infiltration in psoriatic skin, and interestingly, such effects were mediated through the vitamin D receptor (VDR) in keratinocytes (KCs). We further reveal that IL-36α and IL-36γ, which have recently emerged as key players in psoriasis pathogenesis, were effectively repressed by calcipotriol via direct VDR signaling in mouse KCs. Accordingly, calcipotriol treatment suppressed IL-36α/γ expression in lesional skin from patients with plaque psoriasis, which was accompanied by a reduced IL-23/IL-17 expression. In contrast, dexamethasone indirectly reduced IL-36α/γ expression in mouse psoriatic skin through immune cells. Furthermore, we demonstrate that calcipotriol and dexamethasone, in combination, synergistically suppressed the expression of IL-36α/γ, IL-23, and IL-17 in the established mouse psoriasis. Our findings indicate that the combination of calcipotriol and corticosteroid efficiently disrupts the IL-36 and IL-23/IL-17 positive feedback loop, thus revealing a mechanism underlying the superior efficacy of calcipotriol and corticosteroid combination therapy for psoriasis.
Vascular normalization is an emerging concept in cancer treatment, but its precise mechanisms are not completely understood. The polarization of tumor-associated macrophages (TAMs) is important in tumor angiogenesis and metastasis. However, little is known about the effect of anti-angiogenic agents on the polarization of tumor-associated macrophages. Therefore, we explore the changes of TAMs polarization in the development of tumor vascular normalization induced by endostatin.A murine xenograft model of lung cancer was treated with endostatin for 10 days. The morphology and function of tumor vasculature was examined using various techniques. Flow cytometry was carried out to assess the TAMs, and immunofluorescence was used to examine Tie-2-expressing monocytes (TEMs) in tumors. Levels of the histidine-rich glycoprotein (HRG) in tumors were measured by immunohistochemistry and Western blot.Tumor vessels became more normal and mature on day six in the endostatin-treated mice. During vascular normalization, the number of M2-like TAMs and TEMs in the tumors was significantly reduced, whereas the number of M1-like TAMs showed an increase on day six after endostatin treatment, although the latter was not statistically significant. The HRG in the tumors accumulated at an early stage after endostatin administration.The polarization of TAMs is associated with tumor vascular normalization induced by endostatin. These observations may be useful in the exploration of new strategies for anti-angiogenic treatment.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)
