Huadong Medical Institute of Biotechnology

Nanjing, China

Huadong Medical Institute of Biotechnology

Nanjing, China
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Cai B.,Anhui Medical University | Wang M.,Anhui Medical University | Wang M.,Nanjing Jingdu Hospital | Zhu X.,Huadong Medical Institute of Biotechnology | And 7 more authors.
International Journal of Molecular Sciences | Year: 2015

Lipopolysaccharides (LPS) can induce acute inflammation, sepsis, or chronic inflammatory disorders through the Toll receptor 4 (TLR4) signaling pathway. The TLR4/MD2 (myeloid differentiation protein 2) complex plays a major role in the immune response to LPS. However, there is not a good method to suppress the immune response induced by LPS via this complex in macrophages. In this article, we aimed to evaluate the effects of humanized anti-TLR4 monoclonal antibodies on LPS-induced responses in mouse macrophages. The peritoneal macrophages of mice were incubated with anti-TLR4 monoclonal antibodies and stimulated with LPS. The expression levels of cytokines were analyzed by quantitative polymerase chain reaction and enzyme-linkedimmunosorbent assays. Additionally, activation of various signaling pathways was evaluated by Western blotting. The results showed that the humanized anti-TLR4 monoclonal antibody blocked the inflammatory cytokines expression at both the mRNA and protein level. We also found that the Fab fragment significantly inhibited the nuclear factor kappaB signaling pathway by reducing the phosphorylation of the inhibitor of kappaBalpha and decreasing the translocation of p65, resulting in the suppression of p38, extracellular signal-regulated kinase 1/2, c-Jun N-terminal kinase 1/2, and IFN-β regulatory factor 3 phosphorylation. Therefore, our study showed that this humanized anti-TLR4 monoclonal antibody could effectively protect against LPS-induced responses by blocking the TLR4 signaling pathway in mouse peritoneal macrophages. © 2015 by the authors; licensee MDPI, Basel, Switzerland.


PubMed | Anhui Medical University, Nanjing Jingdu Hospital, Huadong Medical Institute of Biotechnology and Key Laboratory of Antibody Technique of the Ministry of Health
Type: Journal Article | Journal: International journal of molecular sciences | Year: 2015

Lipopolysaccharides (LPS) can induce acute inflammation, sepsis, or chronic inflammatory disorders through the Toll receptor 4 (TLR4) signaling pathway. The TLR4/MD2 (myeloid differentiation protein 2) complex plays a major role in the immune response to LPS. However, there is not a good method to suppress the immune response induced by LPS via this complex in macrophages. In this article, we aimed to evaluate the effects of humanized anti-TLR4 monoclonal antibodies on LPS-induced responses in mouse macrophages. The peritoneal macrophages of mice were incubated with anti-TLR4 monoclonal antibodies and stimulated with LPS. The expression levels of cytokines were analyzed by quantitative polymerase chain reaction and enzyme-linked immunosorbent assays. Additionally, activation of various signaling pathways was evaluated by Western blotting. The results showed that the humanized anti-TLR4 monoclonal antibody blocked the inflammatory cytokines expression at both the mRNA and protein level. We also found that the Fab fragment significantly inhibited the nuclear factor kappaB signaling pathway by reducing the phosphorylation of the inhibitor of kappaBalpha and decreasing the translocation of p65, resulting in the suppression of p38, extracellular signal-regulated kinase 1/2, c-Jun N-terminal kinase 1/2, and IFN- regulatory factor 3 phosphorylation. Therefore, our study showed that this humanized anti-TLR4 monoclonal antibody could effectively protect against LPS-induced responses by blocking the TLR4 signaling pathway in mouse peritoneal macrophages.


Mao Y.,Jiangsu Province Official Hospital | Zhang D.-W.,Nanjing Medical University | Lin H.,Jiangsu Provincial Blood Center | Xiong L.,Nanjing Medical University | And 8 more authors.
Journal of Experimental and Clinical Cancer Research | Year: 2012

Background: Alpha B-crystallin (αB-crystallin) has been suggested to play an important role in the development of solid tumors. However, the association between αB-crystallin expression and clinicopathological characteristics of human laryngeal carcinoma is not well defined. This study aimed to examine the expression of αB-crystallin in human laryngeal squamous cell carcinoma (LSCC) and investigate the relationship between its expression and the prognosis of LSCC. Methods. Real-time polymerase chain reaction (six LSCC samples, six tumor-adjacent normal samples) and immunohistochemistry by tissue microarrays (109 LSCC samples and 28 tumor-adjacent normal samples) were performed to characterize expression of the αB-crystallin gene in LSCC. Kaplan-Meier survival and Cox regression analyses were carried out to evaluate the prognosis of LSCC. Results: Real-time polymerase chain reaction and immunohistochemistry analysis showed that the expression of αB-crystallin in LSCC was significantly higher than that in tumor-adjacent normal tissues. Moreover, the expression level of αB-crystallin protein in LSCC was significantly related to alcohol consumption (P = 0.022), tumor differentiation (P = 0.007), pTNM stage (P = 0.041) and 5 years' survival (P =0.030). COX multi-factor analysis showed that αB-crystallin (P = 0.013), as well as pTNM stage (P =0.027) and lymphatic metastasis (P = 0.015) were independent prognosis factors for LSCC. Conclusions: The data suggest that αB-crystallin expression is correlated with malignant phenotypes of LSCC and it may serve as a novel prognostic factor for LSCC. © 2012 Mao et al.


Lin H.,Jiangsu Provincial Blood Center | Mao Y.,Jiangsu Province Official Hospital | Zhang D.-W.,Nanjing Medical University | Li H.,Nanjing Medical University | And 4 more authors.
Anti-Cancer Agents in Medicinal Chemistry | Year: 2013

Melanoma-associated antigen (MAGE) is expressed on the surface of multiple tumor cell types and is a promising target of biotherapeutic drug delivery via the anti-MAGE-A1 antibody. In this study, a human single-chain variable fragment (scFv) antibody phage library was generated and applied to recombinant MAGE-A1-coated immunotubes by phage display technology. The soluble antiMAGE-A1 scFv was expressed and purified by immobilized metal-chelated affinity chromatography (IMAC). The anti-MAGE-A1 scFv could bind native MAGE-A1 confirmed by enzyme-linked immunosorbent assay (ELISA), dot blot, and immunoprecipitation (IP) analysis. The immunotoxin was expressed and purified by IMAC successfully. The results indicated that the human anti-MAGE-A1 immunotoxin could provide a valuable drug for clinic cancer therapy. © 2013 Bentham Science Publishers.


Mao Y.,Jiangsu Province Official Hospital | Mao Y.,Huadong Medical Institute of Biotechnology | Lu M.P.,Nanjing Medical University | Lin H.,Jiangsu Provincial Blood Center | And 8 more authors.
PLoS ONE | Year: 2013

Background: Epstein-Barr virus (EBV) infection has been associated with lymphoma development. EBV latent membrane protein 1 (LMP1) is essential for EBV-mediated transformation and progression of different human cells, including lymphocytes. This meta-analysis investigated LMP1 expression with prognosis of patients with lymphoma. Methods: The electronic databases of PubMed, Embase, and Chinese Biomedicine Databases were searched. There were 15 published studies available for a random effects model analysis. Quality assessment was performed using the Newcastle-Ottawa Quality Assessment Scale for cohort studies. A funnel plot was used to investigate publication bias, and sources of heterogeneity were identified by meta-regression analysis. The combined hazard ratios (HR) and their corresponding 95% confidence intervals of LMP1 expression were calculated by comparison to the overall survival. Results: Overall, there was no statistical significance found between LMP1 expression and survival of lymphoma patients (HR 1.25 [95% CI, 0.92-1.68]). In subgroup analyses, LMP1 expression was associated with survival in patients with non-Hodgkin lymphoma (NHL) (HR = 1.84, 95% CI: 1.02-3.34), but not with survival of patients with Hodgkin disease (HD) (HR = 1.03, 95% CI: 0.74-1.44). In addition, significant heterogeneity was present and the meta-regression revealed that the outcome of analysis was mainly influenced by the cutoff value. Conclusions: This meta-analysis demonstrated that LMP1 expression appears to be an unfavorable prognostic factor for overall survival of NHL patients. The data suggested that EBV infection and LMP1 expression may be an important factor for NHL development or progression. © 2013 Mao et al.


Chen R.,Nanjing Medical University | Lu M.,Nanjing Medical University | Wang J.,Jiangsu Provincial Hospital | Zhang D.,Nanjing Medical University | And 9 more authors.
Virchows Archiv | Year: 2013

Human trophoblastic cell surface antigen 2 (Trop2) has been suggested to play an important role in the development of solid tumors. However, the expression of Trop2 in extranodal NK/T cell lymphoma, nasal type (ENKTL) and the relationship with the clinical characteristics of this disease remain poorly understood. In this study, one-step quantitative PCR reverse transcription-polymerase chain reaction and immunohistochemical staining with tissue sections were employed to evaluate the expression of Trop2 in ENKTL. Furthermore, the relationship between Trop2 expression and prognosis of ENKTL was investigated. Expression of Trop2 mRNA and protein was significantly higher in ENKTL tissue than in corresponding non-lymphomatous tissue (p = 0.04 and p < 0.001, respectively). Expression of Trop2 protein in ENKTL was associated with lymph node involvement and poor overall survival (p = 0.045 and p = 0.018, respectively). Kaplan-Meier analysis and the logrank test indicated that lymph node involvement (p = 0.0481), single therapy strategy (p = 0.0037), and high expression of Trop2 (p = 0.0042) are significantly correlated with poor prognosis of ENKTL patients. The data suggest that Trop2 expression reflects a more malignant phenotype and may serve as an unfavorable prognostic factor for ENKTL. © 2013 Springer-Verlag Berlin Heidelberg.


Xian H.,Nantong University | Zhang H.,Nanjing Medical University | Zhu H.,Nantong University | Wang X.,Nantong University | And 4 more authors.
Pathology | Year: 2014

A proliferation inducing ligand (APRIL) is a member of the tumour necrosis factor superfamily. High APRIL expression has been found to correlate with tumour development, suggesting that APRIL participates in oncogenesis. However, little is known about APRIL expression in gastrointestinal stromal tumours (GISTs) or the relationship between APRIL expression and the clinical characteristics of GIST. Therefore, we assessed the expression of APRIL immunohistochemically using a tissue microarray from 178 patients with GIST and evaluated the relationship between APRIL expression and patient prognosis. Strong APRIL expression was observed in 42.7% of GISTs, with APRIL expression significantly associated with tumour diameter, gross classification and tumour grade (p<0.05 each). Kaplan-Meier analysis suggested that low APRIL expression and tumour size <5cm were associated with longer overall survival. These findings indicate that APRIL expression is correlated with malignant GIST phenotypes and it may serve as an unfavourable prognostic marker in patients with GIST. © 2014 Royal College of Pathologists of Australasia.


Zhu H.,Nantong University | Lu J.,Nantong University | Wang X.,Nantong University | Zhang H.,Nanjing Medical University | And 3 more authors.
Medical Oncology | Year: 2013

Zonula occludens-1 (ZO-1) is a membranescaffolding protein that plays an important role in maintaining tight-junction integrity, which is disrupted in many invasive cancers and intestinal diseases. However, the expression of ZO-1 in gastrointestinal stromal tumor (GIST) and its relationship with clinical characteristics of this disease remain poorly understood. In this study, immunohistochemical analysis using tissue microarray was employed to evaluate the expression of ZO-1 in GIST and to investigate the relationship between its expression and GIST prognosis. High ZO-1 expression was displayed in 71.8 % of GIST patients, which was related to tumor diameter (p < 0.05). The Kaplan-Meier method and logrank test indicated that high ZO-1 expression, small tumor diameter, tumor position in the esophagus, and a borderline-to- intermediate tumor grade displayed significant correlations with longer survival of GIST patients. The data suggest that ZO-1 expression is correlated with malignant phenotypes of GIST and it may serve as a favorable prognostic factor for GIST. These results also support a role for ZO-1 as a tumor-suppressor gene in GIST. © Springer Science+Business Media New York 2013.

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