Objective: : EID3 (EP300-interacting inhibitor of differentiation) was identified as a novel member of EID family and plays a pivotal role in colorectal cancer development. However, its role in glioma remained elusive. In current study, we identified EID3 as a novel oncogenic molecule in human glioma and is critical for glioma cell survival, proliferation and invasion. Methods: : A total of five patients with glioma were recruited in present study and fresh glioma samples were removed from patients. Four weeks old male non-obese diabetic severe combined immune deficiency (NOD/SCID) mice were used as transplant recipient models. The subcutaneous tumor size was calculated and recorded every week with vernier caliper. EID3 and AMP-activated protein kinase α1 (AMPKα1) expression levels were confirmed by real-time polymerase chain reaction and Western blot assays. Colony formation assays were performed to evaluate cell proliferation. Methyl thiazolyl tetrazolium (MTT) assays were performed for cell viability assessment. Trypan blue staining approach was applied for cell death assessment. Cell Apoptosis DNA ELISA Detection Kit was used for apoptosis assessment. Results: : EID3 was preferentially expressed in glioma tissues/cells, while undetectable in astrocytes, neuronal cells, or normal brain tissues. EID3 knocking down significantly hindered glioma cell proliferation and invasion, as well as induced reduction of cell viability, apoptosis and cell death. EID3 knocking down also greatly inhibited tumor growth in SCID mice. Knocking down of AMPKα1 could effectively rescue glioma cells from apoptosis and cell death caused by EID3 absence, indicating that AMPKα1 acted as a key downstream regulator of EID3 and mediated suppression effects caused by EID3 knocking down inhibition. These findings were confirmed in glioma cells generated patient-derived xenograft models. AMPKα1 protein levels were affected by MG132 treatment in glioma, which suggested EID3 might down regulate AMPKα1 through protein degradation. Conclusion : Collectively, our study demonstrated that EID3 promoted glioma cell proliferation and survival by inhibiting AMPKα1 expression. Targeting EID3 might represent a promising strategy for treating glioma.

Prostate cancer (PCa) patients who progress to metastatic castration-resistant PCa (mCRPC) mostly have poor outcomes due to the lack of effective therapies. Our recent study established the orphan nuclear receptor ROR

Objective:To obtain a high specificity and high affinity anti-human PD-L1 monoclonal antibody which can be used for clinical diagnosis and block PD-L1 and PD-1 binding.Methods:BALB/c mice were immunized with recombinant PD-L1 protein.The positive cell clones stably secreting anti-human PD-L1 monoclonal antibody were obtained by classical hybridoma cell fusion technique.The specificity,affinity,subtype and other characteristics of the antibody were identified by ELISA.Immunofluorescence and indirect immunofluorescence were used to detect the tumor cells.Antibody blocking activity was confirmed by tumor killing test.Results:Two cell strains stably secreting monoclonal antibodies against human PD-LI were screened out.Abl and Ab2 had high titer and affinity.The antibody titers were 1:2.56×106 and 1:3×105,and the affinity was 1.5×109 L/mol and 2.5×10s L/mol respectively.There was no cross reaction between these two antibodies and PD-L2.Immunoblotting,indirect immunofluorescence confirmed that the antibody can be used to the diagnosis.Experiment showed that PD-L1 antibodies can increases tumor-killing activity of CIK cells.Conclusion:Two hybridoma cell lines capable of stably secreting highly specific and high affinity anti-human PD-L1 monoclonal antibody are obtained.They can specifically bind to PD-L1 molecules on tumor cells and can be used to the diagnosis of tumor phenotype and prognosis.Antibody blocking function can be applied to combined CIK cell immunotherapy.

Objective:To identify enolase,47 kD allergen,from Litopenaeus vannamei by Mass spectrometry. Methods: The proteins were extracted from Litopenaeus vannamei tissue with acetone precipitation method. The protein components were analyzed by SDS-PAGE and Western blot. By using Matrix-Assisted Laser Desorption/Ionization time of flight mass spectrometry ( MALDI-TOF/TOF-MS) ,the 47 kD allergen from Litopenaeus vannamei was identified as enolase. Results:By SDS-PAGE,we proved that the native protein components from Litopenaeus vannamei were completely. According to the Western blot result more than 14 components could react with the positive serum. MALDI-TOF/TOF analysis results showed that the suspected proteins were enolase. A sensitization frequency was 55%. Conclusion:Enolase was identified as a new allergen of Litopenaeus vannamei.

[ ABSTRACT] AIM: To detect basic fibroblast growth factor ( bFGF ) expression in clinical common malignant tumor ( non-small-cell lung cancer,breast cancer, colon cancer and melanoma) , and to identify relationship between the expression and tumor clinicopathological characteristics.METHODS:Immunohistochemical SP method was used to detect the expression of bFGF at protein level in 208 cases of paraffin-embedded tissue of primary malignant tumor patients ( 68 cases of lung cancer, 80 cases of breast carcinoma, 41 cases of colon cancer and 19 cases of melanoma) .RESULTS:The bFGF protein expression levels were significantly higher in low differentiated non-small-cell lung cancer with lymph node metastasis, and were positively correlated with TNM.In addition, no significant influence of the bFGF protein expression on the patients with median survival period was observed.The protein expression of bFGF was higher in advanced breast cancer with lymph node metastasis and was commonly found in the middle/higher differentiated colon cancer with regional lymph node metastasis.Meanwhile, bFGF protein was highly expressed in advanced melanoma patients with lymph node metastasis.CONCLUSION:bFGF may participate in the process of occurrence and progression of malignant tumor.Ex-pression of bFGF protein may be an effective parameter for evaluating metastasis and prognosis of malignant tumor.

Objective:To identify the allergens which can react with Chinese allergic patients in shrimp and crab ,and analysis their reaction-rates.This results would provide foundation for further research on allergen-detection and desensitization therapy.Methods:Allergen components in Metapenaeus ensis ,Macrobrachium rosenbergii and Charybdis feriata by using 46 portions of shrimp(crab) allergic patients’ serum IgE in Western blot.Results:The reactions between shrimps and allergic patients ’ serum IgE were stronger than that between crab and serum.32-38 kD Tropomysin (TM),40 kD Arginine kinase (AK),60-80 kD Hemocyanin (Hc) and 21 kD arcoplasmic calcium-binding protein(SCP) were the major allergens in shrimps.TM,AK and Hc were common major allergens among shrimps and crab and TM shared the highest reaction-rate.Compare to the results of some from American researchers , AK,Hc and SCP have higher reaction-rate when react with Chinese patients serum ,and we also found a new allergen in shrimp.Con-clusion:For Chinese patients , shrimps have higher reaction-rate than crabs and the allergens among shrimps and crabs which are roughly same.There are some differences in allergens among different human races.A new allergen with 48 kD was found in this re-search.

Objective:To determine the expression of miR-17-92 cluster in osteosarcoma tissue samples and explore its associa-tion with clinical significance. Methods: Quantitative polymerase chain reactiom analysis was used to examine the expression of miR-17-92 cluster in osteosarcoma tissues. Normal bone tissues from 63 patients were matched, and the relationships between the ex-pression of miR-17-92 cluster and the clinicopathological features and prognosis of osteosarcoma were explored. Results:The relative expression of miR-17-92 cluster in osteosarcoma tissues was significantly higher than those in adjacent normal tissues (P<0.05). The high expression of miR-17-92 had a significant correlation with reduced survival (P=0.027). Conclusion:The expression of miR-17-92 cluster closely correlates with the occurrence and progress of osteosarcoma and may be used as an indicator for osteosarcoma prognosis.

Listeria monocytogenes is a pathogenic bacterium, therefore, it is essential for food safety monitoring to establish a rapid and specific detecting method. In this study, immunomagnetic beads and selective medium were combined to detect Listeria monocytogenes at different concentrations (10(1)-10(5) CFU/mL). Other three types of Listeria spp., Staphylococcus aureus and Vibrio parahaemolyticus were also detected to conduct the cross-reaction analysis. Meanwhile, contaminated milk samples were prepared to explore the limit of detection of immunomagnetic beads combining with selective medium. Results showed that Listeria monocytogenes with the concentration of 10(3) CFU/mL and above was successfully detected. Milk samples were detected within 6 hours, with a detection limit of 0.7 CFU/mL. The method developed is capable of detecting milk samples within 30 h, which is 38 h faster compared with national standard method with the same sensitivity.
Bacteriological Techniques ; methods ; Culture Media ; chemistry ; Immunomagnetic Separation ; methods ; Listeria monocytogenes ; growth & development ; isolation & purification ; Sensitivity and Specificity

Objective To obtain the specific human scFv basic fibroblast growth factor(bFGF)using phage antibody library technology. Methods The library was panned with human recombinant bFGF for 4 rounds. The antigen binding activities of random clones were tested by ELISA in order to select specific antibodies, which were then examined by DNA sequence analysis. Results The positive clone selected from the 104 random clones was able to bind bFGF specifically, while not able to bind other growth factors,such as aFGF, VEGF(vascular endothelial growth factor). By competition ELISA assay we found one clone 44 could inhibit bFGF binding to FGFR1. Conclusion Seven specific human phage antibody against bFGF was obtained by phage display technique, one clone could inhibit bFGF binding to its high affinity receptor FGFR1.

AIM: To investigate the effect of Se-containing spirulina phycocyanin (Se-SPC) on liver injury of mice induced by carbon tetrachloride (CCl 4). METHODS: The mouse model was conducted by intragastric feeding with 2% CCl 4 oil for three times, meanwhile Se-SPC, spirulina phycocyanin (SPC) and Na 2SeO 3 were injected (ip) to various groups for 7 days. Then selenium (Se), glutathione peroxidase (GPx), superoxide dismutase (SOD), alanine aminotransferase (ALT), malondiaoldehyde (MDA) and nitric oxide (NO) levels in blood and liver were measured. RESULTS: The level of Se, GPx and SOD activities were obviously higher ( P