Search

A new sensing platform based on red emitting carbon dots has been developed, which has promising potential for use in targeting nucleolus and lysosomes, as well as pH sensing in vivo and zebrafish bioimaging.

Nitrogen-doped orange emitting carbon dots have been developed with excellent potential for applications in β-carotene sensing, lysosomal monitoring and zebrafish bioimaging.

Hydrogel has been widely used in modern biotherapeutics due to its excellent biocompatibility, degradability, and high drug loading capacity. Among them, the construction of a phototherapy system including photosensitizer and hydrogel has aroused great interest in tumor therapy. Unfortunately, complex modifications are necessary to integrate different photosensitizers into the hydrogel. In this work, an injectable hydrogel was proposed by the Schiff base reaction between HA-CHO and carbon dots (CDs), which can realize PTT and PTT simultaneously. Notably, the CDs with rich -NH

MicroRNAs (miRNAs) are a family of endogenous noncoding RNAs with the functions of gene regulation, which serve as promising markers for a range of diseases such as diabetic foot ulcers, cancers, etc. In this work, we engineered a roll-to-roll DNA nanomachine for highly sensitive electrochemical detection of miRNA. A dumbbell-structured DNA probe could be transitioned to be wheel-structured conformation upon target recognition, which rolls around track strands on the surface of gold nanoparticles (AuNPs) in the presence of nicking endonuclease. The resulting single strands on AuNPs are activated for the second round of rolling at the DNA-modified electrode interface, leading to the variation of electrochemical responses. The roll-to-roll amplification behavior allows a wide detection range with a limit of detection as low as 10 aM. The practicability is also demonstrated by the application in human serum samples with satisfactory results. It is expected that the proposed electrochemical method offers a new paradigm to develop miRNA assays based on DNA nanotechnology.

The increasingly serious problem of heavy metal ion pollution has caused great harm to human health and the environment. It is urgent to develop a simple and feasible method to remove heavy metal ions in wastewater. In this paper, we prepared blue-green fluorescent carbon nanoparticles (chi-CNPs) with chitosan as the raw material, which realized the efficient removal of heavy metal ions. In addition, the removal effect of heavy metal ions can be evaluated by the fluorescence changes of chi-CNPs. Finally, we applied chi-CNPs to the treatment of industrial wastewater. The value of the total dissolved solids (TDS) was used to evaluate the treatment effect. The removal rate of heavy metal ions by chi-CNPs was 54.6%, which was much higher than that of other carbon dots or semiconductors. Because of its simplicity, safety and cleanliness, it is suitable for large-scale production and is expected to become a general method to solve the pollution of heavy metal ions in wastewater.

Ferric(III) ions (Fe3+) are one of the most abundant metal ions in environmental and biological systems. The determination of Fe3+ has attracted great attention for healthcare concerns. In this work, we have developed a novel fluorescence method for the sensing and intracellular imaging of Fe3+ based on the prepared red-emissive carbon nanodots. The nanoprobes are synthesized via a microwave method using ammonium fluoride and o-phenylenediamine as carbon precursors, which exhibit excellent optical properties and low toxicity. More importantly, the carbon nanodots show high selectivity towards Fe3+ against other interfering ions. The sensitivity is also high with the limit of detection as low as 0.05 μM. Meanwhile, the carbon nanodots have been successfully used for fluorescence imaging of cells and could be quenched by intracellular Fe3+. These results suggest that the red-emissive carbon nanodots have diverse potential utilities in biomedical fields.

Photodynamic therapy, as an effective treatment for superficial tumors, has attracted more and more attention. The development of safe, biocompatible and in vivo photosensitive materials is helpful to promote photodynamic therapy. Here we report green fluorescent carbon quantum dots prepared from a natural vitamin, riboflavin (VB2), as a photosensitizer. The VB2-based carbon dots have excellent water solubility and biocompatibility, and their singlet oxygen generation ability is much stronger than that of riboflavin itself. Through endocytosis, the carbon dots can easily enter the cells and show bright green fluorescence. In vivo experiments show that after photodynamic therapy the carbon dots can significantly inhibit the growth of tumors, and will not have toxic and side effects on other organs.

miRNAs are small non-coding RNAs for gene regulation, which serve as promising biomarkers for the diagnosis of certain diseases. In this contribution, we have proposed a convenient electrochemical biosensing strategy based on the interaction between DNA modified gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs). In principle, citrate capped AuNPs and AgNPs can be co-decorated on the electrode successively. However, with the modification of DNA on AuNPs surface, a strong negative layer is formed. AuNPs@DNA modified electrode could then inhibit subsequent adsorption of AgNPs due to the electrostatic repulsion and steric hindrance effect. As a result, electrochemical response from AgNPs is significantly decreased. On the other hand, in the presence of target miRNA, DNA on AuNPs hybridizes with miRNA and can thus be cyclically digested by duplex-specific nuclease (DSN). Without the shield of DNA, AgNPs can be relaunched at the AuNPs modified electrode. By analyzing the silver stripping peak, highly sensitive detection of miRNA can be achieved. This biosensor exhibits the limit of detection as low as 0.62 fM and a broad linear range from 1 fM to 1 pM. It may hold great potential utility for miRNA assay in the applications of biomedical researches and early clinical diagnosis.

Carbon dots without heteroatoms-doping and surface modifications were designed to be a novel chemosensing strategy on the quantitative detection of uric acid (UA) with the aid of uricase-induced enzymatic reaction and Fenton reaction. In this work, ascorbic acid (AA)-derived carbon dots (A-CDs) were prepared in the mixture of ethanol and water via one-step hydrothermal synthesis at a relatively low temperature (120 °C) for 10 h. The resultant A-CDs were proved to be excitation-independent. When excited at the wavelength of 420 nm, the nanodots displayed green fluorescence (535 nm) which was then linearly quenched as UA concentration increased in the range of 0-56 μM, according to which the detection limit was calculated to be 0.49 μM. With regards to the excellent sensitivity and selectivity to UA, real sample assay was performed on the A-CDs detection system, which provided relatively reliable recoveries of UA contained in human serum/urine. Besides, in view of the high quantum yield, the A-CDs were applied to live-cell imaging assay and were considered to become an alternative tracer tool in biomedical imaging.

Abnormal miRNA expressions are closely related to the occurrence and development of cancers. It is of great significance to monitor miRNA expression levels for early diagnosis and therapy of the diseases. This study presents two independent colorimetric strategies for simultaneously monitoring multiple miRNAs based on cross-linking or non-cross-linking aggregations of gold nanoparticles (AuNPs). By introducing a Y shaped DNA structure and two types of DNA modified AuNPs, a triple-input DNA AND logic gate is facilely developed with the cross-linking aggregation of AuNPs as the signal output. To improve the sensitivity and shorten reaction time, the logic gate is modified by further employing a three DNA strands formed duplex and hybridization chain reaction. Non-cross-linking aggregation of AuNPs is used to evaluate the concentration of initial miRNA inputs. This strategy does not require DNA modification of AuNPs and ultrahigh sensitivity is achieved with the amplification of hybridization chain reaction. The present work may provide powerful tools for multiple miRNAs diagnostics and inspire further development of DNA based logic gates.

Blood clotting time monitoring is of great importance in predicting the risk of haemorrhage and cardiovascular diseases. In this study, a new P-Lamb wave sensor was developed to measure activated partial thromboplastic time (aPTT) with enhanced sensitivity. The sensor utilizes a piezoelectric and hydrophobic material, Parylene-C, as a waveguide for the P-Lamb wave sensor in order to monitor the aPTT in whole blood samples. With a smooth surface, the P-Lamb wave sensor reduced energy loss and prevented leaky modes, with a decrease of 33% ± 1.25% in the surface roughness and an increase of 2.75% ± 0.15% in the signal. The experimental results demonstrated that, compared to a quartz crystal microbalance (QCM), a P-Lamb sensor with high sensitivity (the frequency shift of a P-Lamb sensor is approximately 200 times greater than that of a QCM) improved stability and enhanced repeatability. Parylene-C worked as a phonon coupling layer, which improved the stability and sensitivity of the Lamb sensor. The relative standards deviation (RSD) of the aPTT measurement was 2.11%, which was 1.22 times lower than reported values. The P-Lamb wave sensor was also tested for its sensitivity to different concentrations of heparin and was found to exhibit an increase in coagulation time with an increasing heparin concentration. Compared to the SYSMEX CS 5100 haematology analyser, the clinical coefficient index (R2) was 0.99467. Considering its compact size, low cost, and mass production of the chip unit, the developed P-Lamb wave sensor is a promising device for point-of-care diagnosis of haemostasis and personal health monitoring.

Transmembrane serine proteases have been implicated in the development and progression of solid and hematological cancers. Human airway trypsin‐like protease 4 (HAT‐L4) is a transmembrane serine protease expressed in epithelial cells and exocrine glands. In the skin, HAT‐L4 is important for normal epidermal barrier function. Here, we report an unexpected finding of ectopic HAT‐L4 expression in neutrophils and monocytes from acute myeloid leukemia (AML) patients. Such expression was not detected in bone marrow cells from normal individuals or patients with chronic myeloid leukemia, acute lymphocytic leukemia and chronic lymphocytic leukemia. In AML patients who underwent chemotherapy, persistent HAT‐L4 expression in bone marrow cells was associated with minimal residual disease and poor prognostic outcomes. In culture, silencing HAT‐L4 expression in AML–derived THP‐1 cells by short hairpin RNAs inhibited matrix metalloproteinase‐2 activation and Matrigel invasion. In mouse xenograft models, inhibition of HAT‐L4 expression reduced the proliferation and growth of THP‐1 cell–derived tumors. Our results indicate that ectopic HAT‐L4 expression is a pathological mechanism in AML and that HAT‐L4 may be used as a cell surface marker for AML blast detection and targeting.

Pre-eclampsia (PE) is a chronic inflammatory disease in pregnancy, which is associated with enhanced blood coagulation and high thrombotic risk. To date, the mechanisms underlying such an association are not fully understood. Previous studies reported high levels of plasma deoxyribonucleic acid (DNA) in PE women, but the cellular source of the circulation DNA remains unknown. In this study, we tested the hypothesis that activated neutrophils undergoing cell death, also called NETosis, may be responsible for the elevated plasma DNA levels in PE women. We analysed plasma samples from non-pregnant, normal pregnant and PE women and found high levels of double-stranded DNA, myeloperoxidase (an abundant neutrophil granular enzyme) and histones (the major nucleosome proteins) in PE-derived samples, indicating increased NETosis in the maternal circulation. The high plasma DNA levels positively correlated with enhanced blood coagulation in PE women. When isolated neutrophils from normal individuals were incubated with PE-derived plasma, an elevated NETosis-stimulating activity was detected. Further experiments showed that endothelial micro-particles, but not soluble proteins, in the plasma were primarily responsible for the NETosis-stimulating activity in PE women. These results indicate that circulating micro-particles from damaged maternal endothelium are a potent stimulator for neutrophil activation and NETosis in PE women. Given the pro-coagulant and pro-thrombotic nature of granular and nuclear contents from neutrophils, enhanced systemic NETosis may represent an important mechanism underlying the hyper-coagulability and increased thrombotic risk in PE.

The presence of excessive ROS can cause much harm to the human body and can even cause diseases. Therefore, it is important to detect and remove ROS, but there is no ideal method available for this at present. In this research, using procyanidins, a type of plant extract with strong reducibility, as raw materials, fluorescent carbon dots (CDs) were prepared by a hydrothermal method. The proanthocyanidin-based carbon dots (PCDs) emit a light-green colored light under UV irradiation. The PCDs retain the strong reducibility of procyanidins and are highly water-soluble compared with procyanidins. The PCDs, in addition to having good biocompatibility, also have the superior properties of radical scavenging activity and cell imaging. In in vitro experiments, 1,1-diphenyl-2-picrylhydrazyl (DPPH; 100 μM) was reduced by 30% when PCDs were added up to a concentration of 87.5 μg mL-1. At the same time, the fluorescence quenching correlates with the concentration of hypochlorite and hydrogen peroxide and has a good linearity in the range of 250-2250 nM and 60-180 μM with a detection limit of 3.676 nM and 0.602 μM, respectively. Based on the previously described advantages, PCDs have potential as a biomedicine.

Immunomagnetic beads are important tools for the isolation and detection of circulating tumor cells (CTCs). However, the current immunomagnetic bead technique provides poor CTC separation purity due to nonspecific binding of background cells. Furthermore, immunomagnetic beads have not been appropriately functionalized for enabling CTC analysis and quantification. In this work, bimetallic magnetic gold nanoparticles were prepared and coated with leukocyte membranes to form leukocyte membrane-camouflaged nanoparticles. After conjugation with the antibody of epithelial cell adhesion molecule (EpCAM), the biomimetic immunomagnetic gold nanoparticles (CM-Fe3O4@Au-Ab) showed a high specific recognition ability on mock (EpCAM-positive) CTCs and a reduced interaction with leukocytes. We subsequently optimized the conditions for CTC separation, including the concentration of nanoparticles and the incubation time. Under the optimized conditions, CM-Fe3O4@Au-Ab exhibited high CTC capture efficiency with negligible background cell binding in mock clinical blood samples. More importantly, gold probes were tagged on the surface of these separated CTCs. When coupled with ICP-MS analysis, the number of CTCs and gold signals exhibited a good linear relationship, and a low limit of detection was obtained, enabling us to estimate the number of CTCs in blood samples. Hence, we expected that CM-Fe3O4@Au-Ab could provide an opportunity to surmount the limitations of current CTC detection.

Membrane-bound proteases are essential for epidermal integrity. Human airway trypsin-like protease 4 (HAT-L4) is a type II transmembrane serine protease. Currently, its biochemical property, cellular distribution and physiological function remain unknown. Here we examined HAT-L4 expression and function in vitro and in vivo. In Western analysis, HAT-L4 expressed in transfected CHO cells appeared as a 48-kDa protein. Flow cytometry confirmed HAT-L4 expression on the cell surface with the expected membrane topology. RT-PCR and immunostaining experiments indicated that HAT-L4 was expressed in epithelial cells and exocrine glands in tissues including skin, esophagus, trachea, tongue, eye, bladder, testis and uterus. In the skin, HAT-L4 expression was abundant in keratinocytes and sebaceous glands. We generated HAT-L4 knockout mice by disrupting the Tmprss11f gene encoding HAT-L4. HAT-L4 knockout mice were viable and fertile. No defects were found in HAT-L4 knockout mice in hair growth, wound healing, water repulsion and body temperature regulation. Compared with wild-type controls, HAT-L4-deficient newborn mice had greater body fluid loss and higher mortality in a trans-epidermal body fluid loss test. In metabolic studies, HAT-L4-deficient adult mice drank water more frequently than wild-type controls did. These results indicate that HAT-L4 is important in epidermal barrier function to prevent body fluid loss.

Sensitive and specific detection of protein is of great significance for early diagnosis and prognosis of many diseases. However, great challenges remain unsolved including relative low sensitivity, high cost, long testing time, complicated instrument and laborious operation. To improve the performance of protein detection methods, development of fine reaction interface for recognition and signal amplification is of great importance. In this work, we construct a novel mode of DNA assembly at electrode interface based on a tripodal surface anchor and an electrochemical aptasensor for protein assay is developed. The orientation of the immobilized DNA is optimized, which promises the efficiency of protein recognition. In addition, hybridization chain reaction is employed for further signal amplification. Therefore, this detection method shows high sensitivity with excellent specificity. The strategy can be universally applicable by simply modifying the sequences of used DNA probes.

Determination of blood clotting time is essential in monitoring therapeutic anticoagulants. In this work, Parylene-C passivated on quartz crystal microbalance (P-QCM) was developed for the activated partial thromboplastin time (aPTT) measurement. Compared with typical QCM, P-QCM possessed a hydrophobic surface and sensitive frequency response to viscoelastic variations on electrode surface. Fibrin could be adsorbed effectively, due to the hydrophobicity of the P-QCM surface. Comparing with typical QCM, the peak-to-peak value (PPV) of P-QCM was increased by 1.94% ± 0.63%, which indicated enhancement of signal-to-noise ratio. For P-QCM, the coefficient of variation (CV) of frequency decrease and aPTT were 2.58% and 1.24% separately, which demonstrated improvement of stability and reproducibility. Moreover, compared with SYSMEX CS 2000i haematology analyzer, clinical coefficient index (R2) was 0.983. In conclusion, P-QCM exhibited potential for improving stability, reproducibility and linearity of piezoelectric sensors, and might be more promising for point of care testing (POCT) applications.

The immune costimulatory molecule B7-H3 has been shown to be involved in the regulation of murine bone formation. However, the role of B7-H3 in bone metabolic diseases remains unknown. In our study, matrix metalloproteinase 2 (MMP-2) and soluble B7-H3 (sB7-H3) were found to be correlatively up-regulated in the sera of osteoporosis patients. Furthermore, our results showed that MG63 cells treated with MMP-2 inhibitors produced lower amounts of sB7-H3 while cells with recombinant MMP-2 had an increased membrane B7-H3 (mB7-H3) shedding. Therefore, elevated MMP-2 levels resulted in an elevation of serum sB7-H3 and reduction of osteoblastic mB7-H3. B7-H3 knockdown in MG63 cells significantly decreased osteoblastic markers and substantially decreased the number of mineralized nodules after 21days. Thus, B7-H3-deficient MG63 cells exhibited impaired bone formation. These results suggest that mB7-H3 is required for the later phases of osteoblast differentiation and that MMP-2/B7-H3 plays a negative regulatory role in osteoporosis.

Reactive oxygen species (ROS) in the body play an important role in various processes. It is well known that harmful high levels of ROS can cause many problems in living organisms in a variety of ways. One effective way to remove intracellular ROS is to use reducing materials that can enter the cell. Herein, we developed a strong reducing carbon nano-dot from a natural product, lutein, as an initial raw material. This is a hydrothermal synthesis method with the advantages of simplicity, high yield, mild reaction conditions, and environmental friendliness. The prepared carbon dots exhibit bright blue fluorescence, and have good water solubility and biocompatibility. In particular, the carbon dots can easily enter the cell and effectively remove ROS. Therefore, the carbon dots are thought to protect cells from oxidative damage by high levels of ROS.

Objective— Thrombophilia is a major complication in preeclampsia, a disease associated with placental hypoxia and trophoblast inflammation. Preeclampsia women are known to have increased circulating microparticles that are procoagulant, but the underlying mechanisms remain unclear. In this study, we sought to understand the mechanism connecting placental hypoxia, circulating microparticles, and thrombophilia. Approach and Results— We analyzed protein markers on plasma microparticles from preeclampsia women and found that the increased circulating microparticles were mostly from endothelial cells. In proteomic studies, we identified HMGB1 (high-mobility group box 1), a proinflammatory protein, as a key factor from hypoxic trophoblasts in stimulating microparticle production in human umbilical vein endothelial cells. Immunodepletion or inhibition of HMGB1 in the conditioned medium from hypoxic human trophoblasts abolished the endothelial microparticle-stimulating activity. Conversely, recombinant HMGB1 stimulated microparticle production in cultured human umbilical vein endothelial cells. The microparticles from recombinant HMGB1–stimulated human umbilical vein endothelial cells promoted blood coagulation and neutrophil activation in vitro. Injection of recombinant HMGB1 in pregnant mice increased plasma endothelial microparticles and promoted blood coagulation. In preeclampsia women, elevated placental HMGB1 expression was detected and high levels of plasma HMGB1 correlated with increased plasma endothelial microparticles. Conclusions— Our results indicate that placental hypoxia-induced HMGB1 expression and release from trophoblasts are important mechanism underlying increased circulating endothelial microparticles and thrombophilia in preeclampsia.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder that affects a variety of organ systems. Anti-dsDNA Abs and complement factors have been used as indicators of lupus activity for more than 50 years. A novel indicator of activation in SLE is reported in this paper. Anti-collagen type II (CII) Ab was obviously elevated in patients with SLE compared to those patients with ankylosing spondylitis (AS) and healthy controls (HCs). Anti-CII-Ab-positive patients with SLE showed significantly higher levels of serum IgG and higher titers of ANA but lower levels of C3 and C4 than controls. A positive correlation was demonstrated between anti-CII Ab and serum IgG in SLE patients ( r = 0.50, p

Membrane-bound proteases are essential for epidermal integrity. Human airway trypsin-like protease 4 (HAT-L4) is a type II transmembrane serine protease. Currently, its biochemical property, cellular distribution and physiological function remain unknown. Here we examined HAT-L4 expression and function in vitro and in vivo. In Western analysis, HAT-L4 expressed in transfected CHO cells appeared as a 48-kDa protein. Flow cytometry confirmed HAT-L4 expression on the cell surface with the expected membrane topology. RT-PCR and immunostaining experiments indicated that HAT-L4 was expressed in epithelial cells and exocrine glands in tissues including skin, esophagus, trachea, tongue, eye, bladder, testis and uterus. In the skin, HAT-L4 expression was abundant in keratinocytes and sebaceous glands. We generated HAT-L4 knockout mice by disrupting the Tmprss11f gene encoding HAT-L4. HAT-L4 knockout mice were viable and fertile. No defects were found in HAT-L4 knockout mice in hair growth, wound healing, water repulsion and body temperature regulation. Compared with wild-type controls, HAT-L4-deficient newborn mice had greater body fluid loss and higher mortality in a trans-epidermal body fluid loss test. In metabolic studies, HAT-L4-deficient adult mice drank water more frequently than wild-type controls did. These results indicate that HAT-L4 is important in epidermal barrier function to prevent body fluid loss.

B7-H3, a novel member of the B7 superfamily, plays a critical role during T cell activation; its functions are still unclear. In this study we obtained a novel anti-mouse B7-H3 monoclonal antibody (MAb) and characterized its biological functions. Our results demonstrated that this MAb could be used for flow cytometry and Western blot and immunohistochemistry analyses, suggesting that the performance of this MAb is much better than a commercial MAb (M3.2D7). Furthermore, data showed different expression profiles of mouse B7-H3 on various immune cells. We further showed that mouse B7-H3 protein was not expressed on normal tissues except for bladder epithelial cells using this MAb. Interestingly, the MAb could stimulate the proliferation and cytokine secretion of T cells. Taken together, this MAb might be of great value for further investigation of B7-H3 molecule.

Background HCV transmission is closely associated with injection drug use (IDU), and co-circulation of multiple subtypes has been found among injection drug users (IDUs) in China. Objectives To investigate HCV subtype characterizations among IDUs and general population (GP) in Suzhou, a city at the important “Hu-ning” transportation line. Study design During January 2010 to May 2011, 123 HCV positive plasma from IDUs and 131 stored HCV positive sera from general individuals were collected in Suzhou. HCV C/E2 and NS5B fragments were amplified using a new multiple RT-nested PCR strategy and subsequent sequenced. Genotypes were characterized by phylogenetic analyses. Results Eight HCV subtypes (1a, 1b, 2a, 3a, 3b, 6a, 6n, and 6u) were detected among Suzhou IDUs, and six subtypes (1b, 2a, 3a, 3b, 6a and 6n) among GP. HCV subtype distribution is distinct between IDUs and GP. Interestingly, we detected discrepancy of genotyping results between C/E2 and NS5B regions in one general individual, indicating the presence of HCV intersubtype recombinant in China. The recombinant belongs to a 3a/1b recombinant. We also detected dual infections in one general individual and two IDUs. They include dual infections between 1b and 3a, 3a and 6a, and two distinct lineages of 3b. Conclusions Complex patterns of HCV epidemic among IDUs, as well as GP, in Suzhou, might imply a spread of HCV from IDUs to GP. The finding of one HCV 3a/1b intersubtype recombinant might represent the first report of HCV recombination in China.

Oxymatrine (OMT) is the quinolizidine alkaloid extracted from the Chinese herb Sophora flavescens Ait. that has many pharmacological effects and is used for the treatment of some inflammatory diseases. In this study, RAW264.7 cells and THP-1 differentiated macrophages were pretreated with various concentrations of OMT at 2 h prior to treatment with lipopolysaccharide (LPS) (1.0 μg/mL) for different durations. We detected the anti-inflammatory effect of OMT in LPS-stimulated macrophages and investigated the molecular mechanism. We showed that OMT pretreatment significantly inhibited the LPS-induced secretion of nitric oxide (NO), interleukin-1 beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) in supernatant, attenuated the mRNA levels of inducible nitric oxide synthase (iNOS), IL-1β, TNF-α, and Toll-like receptor 4 (TLR4), increased TLR4 and phosphorylation of inhibitor of kappa B-alpha (p-IBα) in cytosol, and decreased the nuclear level of nuclear factor-κB (NF-κB) p65 in macrophages. In conclusion, OMT exerts anti-inflammatory properties in LPS-stimulated macrophages by down-regulating the TLR4/NF-κB pathway.

This study aimed to investigate molecule B7-H3 expression profiles of patients with ankylosing spondylitis (AS) and the clinical significance of B7-H3 in the pathogenesis of AS. Serum B7-H3 levels were measured by ELISA in patients with AS and healthy controls. The expression of B7-H3 protein and mRNA on CD14+ monocytes of peripheral blood mononuclear cells (PBMCs) and serum levels of T cell-associated cytokines were also analyzed. The serum B7-H3 levels in AS patients were significantly lower than in healthy controls. The expression of B7-H3 protein and mRNA on CD14+ monocytes of PBMCs and serum levels of TNF-α, IL-6, and IL-17A in AS patients were significantly higher than in controls. The reduced serum B7-H3 level was highly negatively correlated with AS Disease Activity Score (ASDAS), TNF-α, and IL-17A. Upregulated B7-H3 protein may play a role in the pathogenesis of AS by binding its receptor on T cells.

CD276 (B7-H3) is a costimulatory molecule that plays a potent role in T cell responses, however, the role of B7-H3 in autoimmune diseases has not been elucidated. We analyzed B7-H3 expression in rheumatoid arthritis (RA) for the first time and found B7-H3 was significantly up-regulated on monocytes in RA patients, while the levels of soluble B7-H3 in serum were lower than in controls (P < 0.0001). These differences correlated with clinical and laboratory disease parameters and informatory factor TNF-α. Through in vitro experiments, we demonstrated that B7-H3 promoted TNF-α secretion. In addition, a new polymorphism variant, B7-H3-T-A-C-T, was identified and shown to be associated with the incidence of RA and the decreased release of sB7-H3. These results suggest that B7-H3 may be a promising biomarker associated with the pathogenesis of RA. Notably, the new B7-H3-T-A-C-T polymorphism variant is associated with RA risk and might be associated with the release of soluble B7-H3.

Immunological T cells and associated cytokines have been shown to be involved in the pathogenesis of multiple myeloma (MM). However, the abnormal immune imbalance of T lymphocyte subsets on MM remains unknown. We investigate the proportions of T helper 1 (Th1)/Th2/Th17/T regulatory (Treg) cells in peripheral blood mononuclear cells (PBMCs) by flow cytometry (FCM), and serum levels of relevant cytokines in MM patients and controls were detected by enzyme-linked immunosorbent assay (ELISA). The messenger RNA (mRNA) expression of T-bet, STAT6, RORgammat, and Foxp3 was measured by real-time quantitative polymerase chain reaction (PCR). The CD4+ Th1 and CD4+ Th17 cells in patients with MM were significantly higher than those in health controls as well as the expression of T-bet and RORgammat mRNA. Furthermore, serum levels of interferon gamma (IFN-γ), IL-6, and IL-17A in MM group were greatly increased and significantly associated with each other. Significant differences on Th cells, cytokines, and transcription factors were observed on MM patients. The imbalance of T lymphocyte subsets was thought to contribute to the pathogenesis and underlying mechanisms of MM.

Staphylococcus aureus (S. aureus) is an important human pathogen, which commonly causes the acquired infectious diseases in the hospital and community. Effective and simple antibiotic treatment against S. aureus-related disease becomes increasingly difficult. Developing a safe and effective vaccine against S. aureus has become one of the world's hot spots once again. The key issue of developing the vaccine of S. aureus is how to find an ideal key pathogenic gene of S. aureus. It was previously suggested that EsxA might be a very important factor in S. aureus abscess formation in mice, but clinical experimental evidence was lacking. We therefore expressed EsxA protein through prokaryotic expression system and purified EsxA protein by Ni-affinity chromatography. ELISA was used to detect the anti-EsxA antibodies in sera of 78 patients with S. aureus infection and results showed that the anti-EsxA antibodies were positive in the sera of 19 patients. We further analyzed the EsxA positive antibodies related strains by antimicrobial susceptibility assay and found that all of the corresponding strains were multi-drug resistant. Among those multi-drug resistant strains, 73.7% were resistant to MRSA. The results indicated EsxA is very important in the pathogenesis of S. aureus. We suggested that the EsxA is very valuable as vaccine candidate target antigens for prevention and control of S. aureus infection.

B7-H3, a recently identified B7 family member, has different isoforms in human and mouse. Mouse B7-H3 gene has only one isoform (2IgB7-H3) with two Ig-like domains, whereas human B7-H3 has two isoforms (2IgB7-H3 and 4IgB7-H3). In this study a systematic genomic survey across various species from teleost fishes to mammals revealed that 4IgB7-H3 isoform also appeared in pigs, guinea pigs, cows, dogs, African elephants, pandas, megabats and higher primate animals, which resulted from tandem exon duplication. Further sequence analysis indicated that this duplication generated a new conserved region in the first IgC domain, which might disable 4IgB7-H3 from releasing soluble form, while 2IgB7-H3 presented both membrane and soluble forms. Through three-dimensional (3D) structure modeling and fusion-protein binding assays, we discovered that the duplicated isoform had a different structure and might bind to another potential receptor on activated T cells. In T cell proliferation assay, human 2IgB7-H3 (h2IgB7-H3) and mouse B7-H3 (mB7-H3) both increased T cell proliferation and IL-2, IFN-γ production, whereas human 4IgB7-H3 (h4IgB7-H3) reduced cytokine production and T cell proliferation compared to control. Furthermore, both h2IgB7-H3 and mB7-H3 upregulated the function of lipopolysacharide (LPS)-activated monocyte in vitro. Taken together, our data implied that during the evolution of vertebrates, B7-H3 exon duplication contributed to the generation of a new 4IgB7-H3 isoform in many mammalian species, which have carried out distinct functions in the immune responses.