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We developed a novel enzyme-free amplified SERS immunoassay by combining silver nanoparticle (AgNP)-linked immunoreaction and SERS transduction for the detection of disease biomarkers. As a proof of concept, our method was successfully illustrated with the disease biomarker α-fetoprotein with the detection limit of 3.3 × 10-13 g mL-1 and a double-blind experiment consisting of tens of serum samples was performed to confirm its reliability.

Point-of-care test (POCT), which allows for rapid and sensitive screening of drugs abuse, is essential and can significantly reduce the clinical, economic and social impact of the opioid crisis worldwide. However, the traditional gold nanoparticle-based lateral flow immunoassay strip is not sensitive enough for detection of trace drugs in hair sample. Herein, we aimed to develop a more specific system using a composite polymer-based nanobead that is deeply dyed with phthalocyanine or similar oil soluble dyes, and termed as deeply dyed nanobead (DDNB). The prepared composite nanobeads displayed a clear core-shell structure and the core/shell ratios were readily controlled by polymer/dye feeding ratios. The absorbance stable nanobeads capped with carboxyl groups were covalently conjugated with antibodies, and were employed for preparation of lateral flow immunoassay strips for sensitive detection of drugs in hair with naked eye. The developed platform allows the detection of drugs such as morphine and methamphetamine in hair samples within 13 min (including hair sample processing ∼5 min). The cut-off value of DDNB strip for methamphetamine detection with naked eye is down to concentration of 8.0 ng/mL, which is about 3.1 times more sensitive than the traditional gold nanoparticles based lateral flow immunoassay. Moreover, the colorful DDNB system has the potential for multiplexing detection of analytes at point-of-care settings and with low cost.

Purpose: As part of the effort to establish a general profile for solid tumors, the aim of this study was to develop a real-time polymerase chain reaction (RT-PCR)-based assay to assess colorectal ...

Background Highly efficient capture and detection of circulating tumor cells (CTCs) remain elusive mainly because of their extremely low concentration in patients’ peripheral blood. Methods We present an approach for the simultaneous capturing, isolation, and detection of CTCs using an immuno-fluorescent magnetic nanobead system (iFMNS) coated with a monoclonal anti-EpCAM antibody. Results The developed antibody nanobead system allows magnetic isolation and fluorescent-based quantification of CTCs. The expression of EpCAM on the surface of captured CTCs could be directly visualized without additional immune-fluorescent labeling. Our approach is shown to result in a 70–95% capture efficiency of CTCs, and 95% of the captured cells remain viable. Using our approach, the isolated cells could be directly used for culture, reverse transcription-polymerase chain reaction (RT-PCR), and immunocytochemistry (ICC) identification. We applied iFMNS for testing CTCs in peripheral blood samples from a lung cancer patient. Conclusions It is suggested that our iFMNS approach would be a promising tool for CTCs enrichment and detection in one step. Graphic abstract

Highly efficient capture and detection of circulating tumor cells (CTCs) remain elusive mainly because of their extremely low concentration in the peripheral blood of patients. Herein, we present an approach for the simultaneous capturing, isolation, and detection of CTCs using an immuno-fluorescent magnetic nanobead system (iFMNS) coated with a monoclonal anti-EpCAM antibody. The developed antibody nanobead system allows magnetic isolation and fluorescent-based quantification of CTCs. The expression of EpCAM on the surface of captured CTCs could be directly visualized without additional immune-fluorescent labeling. Our approach is shown to result in a 70 - 95% capture efficiency of CTCs, and 95% of the captured cells remain viable. Using our approach, the isolated cells could be directly used for culture, reverse transcription-polymerase chain reaction (RT-PCR), and immunocytochemistry (ICC) identification. We applied iFMNS for testing CTCs in peripheral blood samples from a lung cancer patient, which suggested that this approach would be a promising tool for CTCs enrichment and detection in one step.

Additional file 1. Characterization of nanoparticles: transmission electron microscopy analysis of FMN; Measurement of hydrodynamic diameter of FMN; Magnetization measurement of FMN; Determination concentration of FMN. Statistical analysis method. Figure S1. Surface xy and Z-stack confocal laser scanning microscopic image of tumor cells captured by iFMNS. Figure S2. Microscope images of control and captured cells for growth of 12 h, 24 h and 36 h. Figure S3. Capture efficiency comparison of FMN and FMN stored at 4 °C for 1 year. Figure S4. Comparison of fluorescent (FL) intensity of quantum dots nanobeads (QDNBs) and FMNs. Table S1. Comparison of iFMNS capturing efficiency at different tumor cell concentration and in various spiking in medium.

Development of simple, robust, and reliable detection strategy of disease biomarkers holds tremendous promise for early clinical diagnosis and prognosis of diseases. In this work, through combining a silver nanoparticle (AgNP) linked immunoassay and aggregation induced emission (AIE)-based fluorogenic Ag+ probe, we developed a silver-amplified fluorescence immunoassay for the detection of disease biomarkers. This method overcame the intrinsic limitations of enzymes as the dissolution of AgNPs generated numerous Ag+, which could switch on the fluorogenic Ag+ probe driven by tetrazolate-Ag+ complexation. As a proof of concept, our method could be used for determining α-fetoprotein (AFP) with a linear relationship in concentrations ranging from 0.1 ng mL−1 to 5 μg mL−1 and a low limit of detection of 42 pg mL−1. Our method was successfully confirmed for the detection of AFP in real serum samples from hepatocellular carcinoma (HCC) patients, demonstrating the great potential for clinical diagnosis.

Current strategies for the detection of disease biomarkers often require enzymatic assays that may have limited sensitivity due to inferior stability and vulnerable catalytic activity of the enzyme. A new enzyme-free amplification method for identifying suitable biomarkers is necessary to lower the limit of detection and improve many critical diagnosis applications. Here, we presented an enzyme-free amplified plasmonic immunoassay that enhanced the detection sensitivity of disease biomarkers by combining a novel plasmon-induced silver photoreduction system with a silver nanoparticle (AgNP)-linked immunoassay. The key step to achieving ultrasensitivity was to use Ag

A highly sensitive quantum dot (QD)-based western blot assay with extended dynamic range was developed. Bimodal size distribution QD (BQ) immunoprobes composed of small size single QD (7.3 nm) and big size QD nanobead (QB) (82.9 nm) were employed for fluorescent western blot immunoassay on a membrane. Small size QD immunoprobes contributed to wider dynamic range of assay, while big size QB immunoprobes provided higher detection sensitivity. This BQ-based western blot assay can achieve a wide dynamic range (from 7.8 to 4000 ng IgG) and is nearly as sensitive as commercial available ultrasensitive chemiluminescent methods, just using a simple gel imager with UV light (365 nm) excitation and red light filter (610 nm). The fluorescent signals of BQ western blot were stable for 10 min, while chemiluminescent signals faded after 1 min. Moreover, this BQ immunoprobe was utilized for the detection of housekeeping protein and specific target proteins in complex cell lysate samples. The limit of detection of housekeeping protein is 0.25 μg of cell lysate, and the signal intensities were proportional to loading protein amount in a wide range from 0.61 to 80 μg. We believe that this new strategy of bimodal size distribution nanoparticles can also be expanded for other functional nanoparticle-based biological assays to improve the sensitivity and extend the dynamic range. Graphical abstract.

The incidence of aggressive neuroendocrine prostate cancers (NEPC) related to androgen-deprivation therapy (ADT) is rising. NEPC is still poorly understood, such as its neuroendocrine differentiation (NED) and angiogenic phenotypes. Here we reveal that NED and angiogenesis are molecularly connected through EZH2 (enhancer of zeste homolog 2). NED and angiogenesis are both regulated by ADT-activated CREB (cAMP response element-binding protein) that in turn enhances EZH2 activity. We also uncover anti-angiogenic factor TSP1 (thrombospondin-1, THBS1) as a direct target of EZH2 epigenetic repression. TSP1 is downregulated in advanced prostate cancer patient samples and negatively correlates with NE markers and EZH2. Furthermore, castration activates the CREB/EZH2 axis, concordantly affecting TSP1, angiogenesis and NE phenotypes in tumor xenografts. Notably, repressing CREB inhibits the CREB/EZH2 axis, tumor growth, NED, and angiogenesis in vivo. Taken together, we elucidate a new critical pathway, consisting of CREB/EZH2/TSP1, underlying ADT-enhanced NED and angiogenesis during prostate cancer progression., Neuroendocrine prostate cancers are very difficult to treat and often arise after androgen-deprivation therapy. Here the authors show that ADT activates CREB that acts by increasing EZH2 activity to promote neuroendocrine differentiation in prostate cancer, providing alternative avenues for therapy.

Detection of biomarkers is extremely important for the early diagnosis of diseases. Here, we developed an easy and highly sensitive fluorescence detection system for the determination of biomarkers by combining the rapid separation of magnetic beads and silver nanoparticles labeled antibodies. An ultrasensitive silver ions fluorescence probe 3', 6'-bis (diethylamino)-2-(2-iodoethyl) spiro[isoindoline-1, 9'-xanthen]-3-one (Ag+-FP) was applied to immunoassay. A significant signal amplification was achieved as the AgNPs can be dissolved by H2O2 and generate numerous Ag+, which would turn "on" the fluorescence of Ag+-FP. Using α-fetoprotein (AFP) and C-reactive protein (CRP) as target analytes, good linear responses were obtained from 0.1 to 10 ng mL-1 and the limits of detection (LOD) were as low as 70 pg·mL-1 and 30 pg·mL-1, respectively. In addition, the developed system was further evaluated for the detection of real samples including 30 positive serum specimens obtained from hepatocarcinoma patients and 20 negative serum samples, and performs as well as the commercial electrochemiluminescence immunoassay (ECLI) method with less cost and more convenience. Thus, the designed detection system can be used as a promising platform for the detection of a variety of biomarkers and served as a powerful tool in clinical diagnosis.

A fluorometric and magnetic resonance (MR) dual-modal detection scheme is presented for determination of ascorbic acid (AA). It is based on the use of a blended Au/MnO2@BSA mixture that was prepared via a biomimetic strategy, using bovine serum albumin (BSA) as the template at physiological temperature. The MnO2@BSA fraction (one part of the composite) is not susceptible to MR but can be degraded to MR-active compounds upon a redox reaction with even ultralow concentrations of AA. In parallel, the blended Au/MnO2@BSA recovers its fluorescence because MnO2@BSA acts as a quencher of the fluorescence of circumjacent Au@BSA (the other part of the composite). Fluorescence typically is measured at excitation/emission wavelengths of 470/625 nm. Leveraging on this redox reaction between MnO2 and AA, a dual-mode detection scheme for AA was developed. Both the fluorescence and the MR signal increase with the concentration of AA. The lowest limit for the detection of AA is 0.6 μM in the fluorometric mode and 0.4 μM in the MR mode. Analysis of AA-spiked serum samples showed that the recoveries obtained by either the fluorometric and MR mode can reach 94%. This is the first report of the use of blended nanoparticles with their inherent cross-validation regularity.

Determination of disease biomarkers in clinical samples is of crucial significance for disease monitoring and public health. The dominating format is enzyme-linked immunosorbent assay (ELISA), which subtly exploits both the antigen-antibody reaction and biocatalytic property of enzymes. Although enzymes play an important role in this platform, they generally suffer from inferior stability and less tolerant of temperature, pH condition compared with general chemical product. Here, we demonstrate a metal-linked immunosorbent assay (MeLISA) based on a robust signal amplification mechanism that faithfully replaces the essential element of the enzyme. As an enzyme-free alternative to ELISA, this methodology works by the detection of α-fetoprotein (AFP), prostatic specific antigen (PSA) and C-reactive protein (CRP) at concentrations of 0.1 ng mL(-1), 0.1 ng mL(-1) and 1 ng mL(-1) respectively. It exhibits approximately two magnitudes higher sensitivity and is 4 times faster for chromogenic reaction than ELISA. The detection of AFP and PSA was further confirmed by over a hundred serum samples from hepatocellular carcinoma (HCC) and prostate cancer patients respectively.

Antibodies with high targeting capability after being labelled by quantum dots (QDs) have been of great importance for in vitro diagnostic, cellular imaging and in vivo theranostic applications. However, such work concentrating on the conjugation of QDs and antibodies is still insufficient. It is encouraged to develop reliable bioconjugation strategies to improve detection sensitivity of QD–antibody bioconjugates. In this study, two bioconjugation approaches were used for antibody and QDs covalent coupling. Their detection sensitivity was parallelly determined and compared. It was experimentally found that succinimidyl valerate-PEG-maleimide (SMPEG) as the cross linker for oriented bioconjugation of antibodies to QDs with a flexible spacer arm can significantly improve the detection sensitivity of the resulting QD–antibody bioconjugates, compared with those prepared with traditional carbodiimide chemistry using EDC·HCl as the cross linker. Gel electrophoresis suggests antibodies can be successfully conjugated with QDs by both the EDC method and the SMPEG method, but immunofiltration assay shows QD–antibody bioconjugates obtained by the SMPEG strategy have lower detection limits and better linear scope patterns for in vitro molecular diagnosis. In vitro cancer cell imaging further shows that QD–antibody bioconjugates prepared by the SMPEG strategy have superior cell targeting capability without observable nonspecific binding, while the EDC-mediated QD–antibody bioconjugates have low signal to noise ratios with obvious nonspecific cellular binding. Moreover, the number of antibodies conjugated on the surface of QDs was determined by the chemiluminescent dot blot assay, showing 2.13 ± 0.5 and 0.76 ± 0.2 of antibodies per QD by EDC-mediated and SMPEG-mediated coupling, respectively. It further verifies that the SMPEG-mediated coupling strategy can significantly maintain the biological activity of the linked antibodies, even though it has low labelling efficiency. It could be therefore claimed that oriented bioconjugation via the SMPEG coupling strategy is favored by immunoassays, with improved detection sensitivity.

A fluorometric and magnetic resonance (MR) dual-modal detection scheme is presented for determination of ascorbic acid (AA). It is based on the use of a blended Au/MnO

Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (EGFR-TKIs) have been used to treat non-small cell lung carcinoma (NSCLC) patients that have EGFR-activating mutations. EGFR-TKI monotherapy in most NSCLC patients with EGFR mutations who initially respond to EGFR-TKIs results in the development of acquired resistance. We investigated the role of fibroblasts in stromal cell-mediated resistance to gefitinib-induced apoptosis in EGFR-mutant NSCLC cells. While gefitinib induced apoptosis in EGFR-mutant NSCLC cells, apoptosis induction was diminished under stromal co-culture conditions. Protection appeared to be mediated in part by Aurora-A kinase (AURKA) upregulation. The protective effect of stromal cells was significantly reduced by pre-exposure to AURKA-shRNA. We suggest that combinations of AURKA antagonists and EGFR inhibitors may be effective in clinical trials targeting mutant EGFR NSCLCs.

Simple and sensitive detection of infectious disease at an affordable cost is urgently needed in developing nations. In this regard, the dot blot immunoassay has been used as a common protein detection method for detection of disease markers. However, the traditional signal reporting systems, such as those using enzymes or gold nanoparticles lack sensitivity and thus restrict the application of these methods for disease detection. In this study, we report a simple and sensitive detection method for the detection of infectious disease markers that couples the dot-blot immunoassay with quantum dots nanobeads (QDNBs) as a reporter. First, the QDNBs were prepared by an oil-in-water emulsion-evaporation technique. Because of the encapsulation of several QDs in one particle, the fluorescent signal of reporter can be amplified with QDNBs in a one-step test and be read using a UV lamp obviating the need for complicated instruments. Detection of disease-associated markers in complex mixture is possible, which demonstrates the potential of developing QDNBs into a sensitive diagnostic kit.

Although B cells play important roles in the humoral immune response and the regulation of adaptive immunity, B cell subpopulations with unique phenotypes, particularly those with non-classical immune functions, should be further investigated. By challenging mice with Listeria monocytogenes, Escherichia coli, vesicular stomatitis virus and Toll-like receptor ligands, we identified an inducible CD11ahiFcγRIIIhi B cell subpopulation that is significantly expanded and produces high levels of IFN-γ during the early stage of the immune response. This subpopulation of B cells can promote macrophage activation via generating IFN-γ, thereby facilitating the innate immune response against intracellular bacterial infection. As this new subpopulation is of B cell origin and exhibits the phenotypic characteristics of B cells, we designated these cells as IFN-γ-producing innate B cells. Dendritic cells were essential for the inducible generation of these innate B cells from the follicular B cells via CD40L-CD40 ligation. Increased Bruton's tyrosine kinase activation was found to be responsible for the increased activation of non-canonical NF-κB pathway in these innate B cells after CD40 ligation, with the consequent induction of additional IFN-γ production. The identification of this new population of innate B cells may contribute to a better understanding of B cell functions in anti-infection immune responses and immune regulation.

Hepatic ischemia-reperfusion (I/R) injury contributes to hepatic dysfunction and failure after liver transplantation, major hepatic resection, trauma, and hypovolemic shock. Therefore, reducing I/R injury is an important goal to improve the outcome of these procedures. Recently, high-mobility group box 1 protein (HMGB1) has been identified as a pathogenic mediator in several inflammatory diseases, including hepatic I/R. PNU-282987, a selective α7 nicotinic acetylcholine receptor agonist, prevents nuclear factor κB (NF-κB) activation and thereby inhibits cytokine secretion through a specific cholinergic anti-inflammatory pathway. Our study was designed to evaluate whether PNU-282987 would inhibit HMGB1 expression and prevent I/R-induced liver damage. C57BL/6 mice were randomly divided into 3 groups as follows: sham group, vehicle plus I/R group, and PNU-282987 plus I/R group. Mice were subjected to 70% partial hepatic I/R for 60 min and pretreated with either vehicle or with PNU-282987, and blood and hepatic tissue samples were collected at 3, 6, and 12 h following reperfusion. The results showed that pretreatment with PNU-282987 decreased serum transaminase levels and ameliorated liver injury after hepatic I/R. Moreover, pretreatment with PNU-282987 suppressed NF-κB activation, cytokine production (tumor necrosis factor α, interleukin 1β), and HMGB1 expression in liver after hepatic I/R. These observations suggest that PNU-282987 protects the liver from I/R injury possibly by inhibiting HMGB1 expression, suppressing cytokine production, and preventing NF-κB activation in mice.

OBJECTIVES: Recent association studies by the Australo-Anglo-American Spondyloarthritis Consortium (TASC) in Caucasian European populations from Australia, North America and the UK have identified a number of genes as being associated with ankylosing spondylitis (AS). A candidate gene study in a Han Chinese population was performed based on these findings to identify associated genes in this population. METHODS: A case-control study was performed in a Han Chinese population of patients with AS (n = 775) and controls (n = 1587) from Shanghai and Nanjing. All patients met the modified New York criteria for AS. The cases and controls were genotyped for 115 single nucleotide polymorphisms (SNPs) tagging IL23R, ERAP1, STAT3, JAK2, TNFRSF1A and TRADD, as well as other confirmation SNPs from the TASC study, using the Sequenom iPlex and the ABI OpenArray platforms. Statistical analysis of genotyped SNPs was performed using the Cochran-Armitage test for trend and meta-analysis was performed using METAL. SNPs in AS-associated genes in this study were then imputed using MaCH, and association with AS tested by logistic regression. RESULTS: SNPs in TNFRSF1A (rs4149577, p = 8.2 × 10⁻⁴), STAT3 (rs2293152, p = 0.0015; rs1053005, p = 0.017) and ERAP1 (rs27038, p = 0.0091; rs27037, p = 0.0092) were significantly associated with AS in Han Chinese. Association was also observed between AS and the intergenic region 2p15 (rs10865331, p = 0.023). The lack of association between AS and IL23R in Han Chinese was confirmed (all SNPs p > 0.1). CONCLUSIONS: The study results demonstrate for the first time that genetic polymorphisms in STAT3, TNFRSF1A and 2p15 are associated with AS in Han Chinese, suggesting common pathogenic mechanisms for the disease in Chinese and Caucasian European populations. Furthermore, previous findings demonstrating that ERAP1, but not IL23R, is associated with AS in Chinese patients were confirmed.

Compact PEGylated polymer-caged quantum dots were prepared by covalently cross-linking the surface ligands using bifunctional poly(ethylene glycol). CdSe/ZnSe/ZnS quantum dots prepared in organic solvent via conventional synthesis method were firstly transferred to water phase by surface ligands exchange with glutathione molecules, and then cross-linked by bifunctional PEG through the carboxyl-amine coupling by carbodiimide method. Compared with small molecules capped QDs, polymer-caged QDs using PEG crosslinker have shown improved colloidal and pH stability, lower cell toxicity and non-specific binding, and still maintain compact hydrodynamic diameters (

Pengfei Zhang,1,* Yan Bao,1,* Mohamed Shehata Draz,2,3,* Huiqi Lu,1 Chang Liu,1 Huanxing Han11Center for Translational Medicine, Changzheng Hospital, Second Military Medical University, Shanghai, People’s Republic of China; 2Zhejiang-California International Nanosystems Institute, Zhejiang University, Hangzhou, Zhejiang, People’s Republic of China; 3Faculty of Science, Tanta University, Tanta, Egypt*These authors contributed equally tothis workAbstract: Convenient and rapid immunofiltration assays (IFAs) enable on-site “yes” or “no” determination of disease markers. However, traditional IFAs are commonly qualitative or semi-quantitative and are very limited for the efficient testing of samples in field diagnostics. Here, we overcome these limitations by developing a quantum dots (QDs)-based fluorescent IFA for the quantitative detection of C-reactive proteins (CRP). CRP, the well-known diagnostic marker for acute viral and bacterial infections, was used as a model analyte to demonstrate performance and sensitivity of our developed QDs-based IFA. QDs capped with both polyethylene glycol (PEG) and glutathione were used as fluorescent labels for our IFAs. The presence of the surface PEG layer, which reduced the non-specific protein interactions, in conjunction with the inherent optical properties of QDs, resulted in lower background signal, increased sensitivity, and ability to detect CRP down to 0.79 mg/L with only 5 µL serum sample. In addition, the developed assay is simple, fast and can quantitatively detect CRP with a detection limit up to 200 mg/L. Clinical test results of our QD-based IFA are well correlated with the traditional latex enhance immune-agglutination aggregation. The proposed QD-based fluorescent IFA is very promising, and potentially will be adopted for multiplexed immunoassay and in field point-of-care test.Keywords: C-reactive proteins, point-of-care test, Glutathione capped QDs, PEGylation

Bruton tyrosine kinase (Btk) is not only critical for B cell development and differentiation but is also involved in the regulation of Toll-like receptor-triggered innate response of macrophages. However, whether Btk is involved in the regulation of natural killer (NK) cell innate function remains unknown. Here, we show that Btk expression is up-regulated during maturation and activation of mouse NK cells. Murine Btk -/-NKcells have decreased innate immune responses to the TLR3 ligand, with reduced expressions of IFN-γ, perforin, and granzyme-B and decreased cytotoxic activity. Furthermore, Btk is found to promote TLR3-triggered NK cell activation mainly by activating the NF-κB pathway. Poly(I:C)-induced NK cell-mediated acute hepatitis was observed to be attenuated in Btk -/- mice or the mice with in vivo administration of the Btk inhibitor. Correspondingly, liver damage was aggravated in Btk -/- mice after the adoptive transfer of Btk +/+ NK cells, further indicating that Btk-mediated NK cell activation contributes to TLR3-triggered acute liver injury. Importantly, reduced TLR3-triggered activation of human NK cells was observed in Btk-deficient patients with X-linked agammaglobulinemia, as evidenced by the reduced IFN-γ, CD69, and CD107a expression and cytotoxic activity. These results indicate that Btk is required for activation of NK cells, thus providing insight into the physiological significance of Btk in the regulation of immune cell functions and innate inflammatory response. © 2012 by The American Society for Biochemistry and Molecular Biology, Inc., link_to_subscribed_fulltext

Vanadium, a potent toxic agent and carcinogen, is widely used in industry. Evidences show that exposure to vanadium is associated with an increased risk of lung cancer. But the mechanisms involved are far from fully understood. In this present study, we investigated that exposure of human bronchial epithelial cells (Beas-2B) to vanadium pentoxide resulted in an obvious induction of cyclooxygenase-2 (COX-2) expression and this induction was both dose- and time-dependent. Exposure of Beas-2B cells to vanadium pentoxide also led to significant activation of nuclear factor of activated T-cells (NFAT) on a time- and dose-dependent manner. Furthermore, we found that inhibition of NFAT by dominant negative mutant of NFAT (DN-NFAT) resulted in a dramatic inhibition of COX-2 expression induced by vanadium pentoxide, showing that NFAT activation was required for COX-2 induction by vanadium pentoxide in Beas-2B cells. Moreover, knockdown of COX-2 expression by COX-2-specific small interference RNA and blockage of NFAT pathway by DN-NFAT and NFAT3 small interference RNA showed an increased cell apoptosis in Beas-2B on vanadium exposure. Together, our results demonstrated that COX-2 expression could be induced by vanadium pentoxide in NFAT-dependent way and played an anti-apoptotic role in Beas-2B cells. From the results, we anticipate that the carcinogenesis of vanadium to human bronchial cells may result from anti-apoptosis mediated by the NFAT-dependent induction of COX-2, and we also assume that either pro-apoptotic or anti-apoptotic effect in certain type of cells after vanadium exposure may depend on the level of COX-2 induction.

To express and identify soluble liver antigen (SLA) and cytochrome P-450 (CYP 2D6).SLA cDNA and CYP 2D6 cDNA were obtained from human liver tissue poly (A)+RNA by RT-PCR. The cDNAs were inserted into fusion expression vector PQE-30 site of BamH I and Hind III. SLA and CYP 2D6 were identified by the SDS-PAGE and Western blot.SDS-PAGE analysis showed that there was a very strong stained band at about 47 kd and 50 kd, respectively. The products could specifically band to anti-SLA or anti-CYP 2D6 autoantibodies.The clone and expression of SLA and CYP 2D6 provide useful substances for the diagnosis and research of pathogenesis on autoimmune hepatitis.