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Liu W.,Shanghai JiaoTong University | Ma Y.,Shanghai JiaoTong University | Huang L.,Zhangjiagang First Peoples Hospital | Peng J.,Shanghai JiaoTong University | And 4 more authors.
Molecular Biology Reports

The identification of specific biomarkers for colorectal cancer would provide the basis for early diagnosis, prognosis, therapy, as well as clues for understanding the molecular mechanisms governing cancer progression. This study was designed to use comparative proteomics technology to find the differentially expressed proteins between human colorectal carcinoma and the corresponding normal tumor-adjacent colorectal tissues. We have used the highly sensitive two-dimensional gel electrophoresis (2-DE) coupled with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) for the identification of proteins differentially expressed in tumoral and neighboring normal mucosa. We have detected differences in abundance of 42 proteins with statistical variance of the tumor versus normal spot volume ratio within the 95th confidence level (Student's t-test; P < 0.05). 10 out of 42 analyzed proteins were unambiguously identified by MS coupled with database interrogation as being differentially expressed in colorectal cancer. Of the 10 newly implicated proteins, HSP27 was chosen for detailed analysis. Preliminary studies demonstrated that the differentially expressed proteins found by 2-DE could be confirmed and validated by western blotting and immunohistochemistry analyses in those few cases. The results suggest that HSP27 might be a potential biomarker for early diagnosis, prognosis, monitoring in the therapy of colorectal carcinoma. © 2009 Springer Science+Business Media B.V. Source

Shen Y.,Nanjing Southeast University | Dong Y.-M.,University of Arizona | Lu Q.,Nanjing Southeast University | Xu J.,Zhangjiagang First Peoples Hospital | And 3 more authors.
Journal of Obstetrics and Gynaecology Research

Aim To explore the effect of phenolic environmental estrogens (EE) on women with uterine leiomyoma (UL). Methods Urine and blood plasma samples were collected from 300 patients diagnosed with UL at the Affiliated Zhongda Hospital of Southeast University between December 2013 and December 2014. Control urine and blood plasma samples were collected from 300 women who are either patients without UL or healthy volunteers presenting to the same hospital for physical examination during the same period. Bisphenol A (BPA), nonylphenol (NP) and octylphenol (OP) concentration in these samples was measured using solid phase extraction (SPE) coupled with liquid chromatography-tandem mass spectrometry. Results The OP concentration in urine and blood plasma was significantly higher in the UL group compared with the control group (r = 0.224, P = 0.001). Urine BPA concentration was not significantly different between the UL group and the control group (r = 0.009, P = 0.896). There was also no statistically significant difference in urine NP concentration between the two groups (r = 0.057, P = 0.419). On logistic regression, exposure concentration of urine BPA (OR, 1.129; 95%CI: 1.081-1.179) and NP (OR, 1.165; 95%CI: 1.025-1.324) was associated with UL genesis (P < 0.05). Nevertheless, there was no significant difference in blood plasma concentration of BPA, OP and NP between the two groups (P > 0.05). Conclusion Urine and blood plasma EE exposure levels in women, especially the urine level, was related to the incidence of UL. © 2016 Japan Society of Obstetrics and Gynecology. Source

Shao X.,Soochow University of China | Cao X.,Zhangjiagang First Peoples Hospital | Song G.,Soochow University of China | Zhao Y.,Soochow University of China | Shi B.,Soochow University of China
Journal of Diabetes Research

Aims. To study the proliferation of osteoblasts and genes expression under normal glucose, high glucose, and metformin (Met). Methods. MG63 osteoblast-like cells were cultured in osteogenic medium supplemented with normal glucose (glucose 5.5 mmol/L) or high glucose (glucose 16.7 mmol/L) and metformin + high glucose (Met 300 mol/L + glucose 16.7 mmol/L). Proliferation was detected with CCK-8 assay at days 1, 3, and 7. Real-time PCR and Western blot were performed to compare the expression of collagen I (Col I), osteocalcin (OCN), osteoprotegerin (OPG), receptor activator for NF-B ligand (RANKL), and metal matrix proteinases 1 and 2 (MMP1, MMP2). Alkaline phosphatase (ALP) activity was also detected at days 6, 12, and 18. Results. Exposure to high glucose inhibited the proliferation of osteoblasts (P < 0.05), with suppressed OCN and OPG. Meanwhile, Col I, RANKL, MMP1, and MMP2 were unaffected. Metformin attenuated the suppression on proliferation with increased expression of Col I, OCN, and OPG, meanwhile suppressing MMP1 and MMP2. High glucose lowered the intracellular ALP, while metformin raised it. Metformin attenuated the downregulation of ALP completely at day 6, partly at day 12, but not at day 18. Conclusions. Metformin attenuated the suppression effect of high glucose to the osteoblast proliferation and gene expression, more prominently in earlier stage. © 2014 Xinyu Shao et al. Source

Song Y.,Zhangjiagang First Peoples Hospital | Zuo Y.,Zhangjiagang First Peoples Hospital
Oncology Letters

The present study aimed to observe the methylation status of the CpG islands at the human hedgehog interacting protein (HHIP) gene in gastric cancer tissues, peritumoral tissues and the AGS cell line, to analyze the association between the methylation status of the CpG islands and the tumorigenesis of gastric cancer. The HHIP mRNA expression in 60 human gastric carcinnoma tissues, peritumoral tissues and the gastric carcinoma AGS cell line were detected by reverse transcription polymerase chain reaction (RT-PCR). The HHIP methylation status of the promoter region in the gastric carcinnoma tissues and peritumoral tissues was detected by methylation-specific PCR (MSP). Prior to and following treatment with methyl transferase inhibitor 5-aza-2'-deoxycitydine (5-aza-dc), the HHIP mRNA expression level, the methylation status of the promoter region and the methylation site loci on the CpG islands in the AGS cells were detected by RT-PCR, MSP and bisulfite sequencing PCR (BSP), respectively. The correlation between the methylation status of the CpG islands at the HHIP promoter region and the HHIP mRNA expression level were analyzed. It was found that the expression level of the HHIP mRNA in the gastric carcinoma tissues was significantly lower than that in the adjacent tissues (0.82±0.38 vs. 1.60±0.26, respectively; P<0.001). No significant correlations were observed between the expression of HHIP mRNA and age, gender, tumor-node-metastasis stage, differentiation degree and presence of lymph node metastasis (P>0.05). The degree of methylation of the HHIP gene promotor in the peritumoral tissues (17.7±3.59%) was significantly lower than that in the gastric cancer tissues (62.9±6.14%) and in the AGS cells (99.7±0.67%) (P<0.05). Compared with prior to 5-aza-dc intervention, the HHIP mRNA expression level in the AGS cells was significantly increased subsequent to intervention (0.21±0.12 vs. 4.68±0.22; P<0.01), while the degree of methylation in the AGS cells was significantly decreased (90.2±0.67 vs. 10.1±0.21%; P<0.01), and the methylation sites in CpG islands were significantly reduced. The degree of HHIP methylation showed a negative correlation with the level of mRNA expression (r=-0.693; P<0.01). It can be hypothesized that a high degree of methylation of the HHIP gene promoter CpG islands in gastric cancer tissues and cells causes a decrease in HHIP mRNA expression, which may be involved in the carcinogenesis of gastric cancer. © 2014 Spandidos Publications 2014. All rights reserved. Source

Tu J.,Zhangjiagang First Peoples Hospital | Wu G.,Zhangjiagang First Peoples Hospital | Zuo Y.,Zhangjiagang First Peoples Hospital | Zhao L.,Jilin University | Wang S.,Zhangjiagang First Peoples Hospital
Archives of Pharmacal Research

Alloferons are a group of naturally occurring peptides primarily isolated from insects that are capable of stimulating mouse and human NK cell cytotoxicity toward cancer cells. In this study, we found that a modified antibacterial peptide had a broad range of action against both gram-positive and gram-negative bacteria. A time-course experiment showed that CFU counts rapidly decreased after ZL-2 treatment, with the bacteria nearly eliminated within 4 h. We also examined the synergy between the peptide and antibiotics. The peptide ZL-2 resulted in a significant synergistic improvement in the potencies of ampicillin, erythromycin and ceftazidime against methicillin-resistant bacteria. In addition, ZL-2 had no detectable cytotoxicity in mouse spleen cells or a mouse animal model. In the mouse model by i.p. inoculation with Escherichia coli, timely treatment of i.p. injection with ZL-2 resulted in 100-fold reduction in bacteria load in blood as well as 80 % protection from death in the inoculated animals. In conclusion, we successfully identified a modified peptide with maximal bactericidal activity. This study also provides a potential therapeutic for the treatment of E. coli septicemia by increasing the activity of antimicrobials. © 2015 The Pharmaceutical Society of Korea Source

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