1.Serum adiponectin and resistin levels in patients with polycystic ovarian syndrome and their clinical implications.
Yuxia, WANG ; Xingmei, XIE ; Weijie, ZHU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(5):638-42
In this study, serum adiponectin and resistin levels were determined in 46 patients with polycystic ovarian syndrome (PCOS), and their correlation with serum sexual hormones and insulin resistance (IR) were examined. The subjects included 26 obese patients with body mass index (BMI)>25 and 20 non-obese patients with BMI[Symbol: see text]25, with 25 obese and 25 non-obese healthy volunteers without PCOS serving as controls. Serum adiponectin and resistin levels in all subjects were measured, and endocrinal and metabolic indices were also analysed. Our results showed that the serum adiponectin levels in both obese and non-obese PCOS groups were significantly lower than their controls, while the serum resistin levels in obese and non-obese PCOS group were significantly higher than in their controls (P<0.001). The serum adiponectin level was significantly lower and serum resistin level significantly higher in the non-obese PCOS group as compared with the obese control group (P<0.05). Serum adiponectin level was negatively correlated with FIN, HOMA-IR, LH and LH/FSH (P<0.05), but serum resistin level was positively correlated with FIN, HOMA-IR, LH and LH/FSH (P<0.05). We are led to conclude that PCOS patients have obvious IR, low serum adiponectin and high serum resistin, and adiponectin and resistin might play important roles in the pathogenesis of IR in PCOS patients.
2.Analysis of the function of the UBE gene based on bioinformatics
Yang YAO ; Jie SU ; Di WEI ; Xingmei ZHU ; Biping CHENG
Journal of Medical Postgraduates 2014;(11):1156-1159
[Abstract ] Objective Bioinformatics provides a lot of valuable information for online prediction of new genes.In this study, we predicted the biological function of ubiquitin-conjugating enzyme 2S ( UBE2S) based on bioinformatics. Methods The UBE2S gene was screened and cloned from the cDNA library of human HepG2 cells.The relationship of the structure and function of UBE2S was explored based on the full-length cDNA library.MEGA5.05, CLUSTALW2 and SWISS-MODEL were used to study the phylogeny, conservation, and 3D structure of UBE2S. Results The UBE2S gene encoded a polypeptide of 241 residues with a predicted molec-ular weight of 23 770 and an isoelectric point of 8.81.The UBE2S protein contained no transmembrane locus and the probabilities of their functions of growth factors, cation channel and structural protein were 8.904, 0.313, and 0.291.The analysis of BLASTp showed that the isolated UBE2S had a 90-97%identity with the other species. Conclusion Analysis of the structure and function of the UBE2S protein can not only provide more information about its gene family but also pave the way for further experimental studies on the molecular mechanism of the consequent hepatocellular carcinoma.
3.Denervation effects on bone regeneration and expression of Runx2 during distraction osteogenesis
Ke ZHENG ; Donghui SONG ; Xingmei FENG ; Songsong ZHU ; Jing HU ; Bin YE
Chinese Journal of Tissue Engineering Research 2015;(37):5988-5992
BACKGROUND:During the healing of fractures, removal of sciatic nerve can result in insufficient mechanical rigidity of newborn woven bone. However, there are less reports concerning the denervation effects during distraction osteogenesis. OBJECTIVE:To observe the effect of removal of the sciatic nerve on bone regeneration and the expression of Runt-related transcription factor 2 (Runx2) protein during distraction osteogenesis in a rabbit model. METHODS:Twenty-four adult male New Zealand rabbits were selected and underwent left tibial osteodistraction to construct animal models of distraction osteogenesis. Before distraction, the animals were randomly divided into group R (resecting the left sciatic nerve) and group I (intact left sciatic nerve). Six weeks after completion of distraction, the animals were kil ed and the lengthened tibias were harvested for radiography, three-dimensional CT reconstruction, histological evaluation, connectivity density (Conn.D) evaluation. RESULTS AND CONCLUSION:New regenerated bone was present and Runx2 protein was expressed in the distraction gaps of al animals at the end of the study, as revealed by radiography, three-dimensional CT reconstruction, and histological observation. However, less new bone formation and a lower degree of mineralization and expression of Runx2 protein were observed in group R compared with group I. The results suggest that the denervation appears to have an inhibitory effect on bone formation and the expression of Runx2 protein during distraction osteogenesis.
4.Serum Adiponectin and Resistin Levels in Patients with Polycystic Ovarian Syndrome and Their Clinical Implicatlons
WANG YUXIA ; XIE XINGMEI ; ZHU WEIJIE
Journal of Huazhong University of Science and Technology (Medical Sciences) 2010;30(5):638-642
In this study, serum adiponectin and resistin levels were determined in 46 patients with polycystic ovarian syndrome (PCOS), and their correlation with serum sexual hormones and insulin resistance (IR) were examined. The subjects included 26 obese patients with body mass index (BMI)>25 and 20 non-obese patients with BMI≤25, with 25 obese and 25 non-obese healthy volunteers without PCOS serving as controls. Serum adiponectin and resistin levels in all subjects were measured, and endocrinal and metabolic indices were also analysed. Our results showed that the serum adiponectin levels in both obese and non-obese PCOS groups were significantly lower than their controls, while the serum resistin levels in obese and non-obese PCOS group were significantly higher than in their controls (P<0.001). The serum adiponectin level was significantly lower and serum resistin level significantly higher in the non-obese PCOS group as compared with the obese control group (P<0.05). Serum adiponectin level was negatively correlated with FIN, HOMA-IR, LH and LH/FSH (P<0.05), but serum resistin level was positively correlated with FIN, HOMA-IR, LH and LH/FSH (P<0.05). We are led to conclude that PCOS patients have obvious IR, low serum adiponectin and high serum resistin, and adiponectin and resistin might play important roles in the pathogenesis of IR in PCOS patients.
5.A practical method of expression, purification and identification of beta-site app-cleaving enzyme
Rui ZHU ; Qiaoyan WU ; Xingmei ZHANG ; Xiaofang GAO ; Peiru WEI ; Xinyu ZHANG ; Fang WANG
Chinese Journal of Neuromedicine 2015;14(6):604-608
Objective To introduce a practical method that can be used to efficiently express,purify and identify Alzheimer's disease (AD) related beta-site app-cleaving enzyme 1 (BACE1) in common eukaryotic cells.Methods BACE1 cDNA was fished out from human brain cDNA library and ligated into the pEGFP-c3 expression vector,and then,the recombinant plasmid was transfected into the HEK293 cells.The BACE1 protein was purified with TALON Mental Affinity Resins column.The target protein was identified by Western blotting and fluorescence resonance energy transfer (FRET).BACE1 Activity Assay Kit was employed to test the activity of purified BACE1 in vitro.The recombinant BACE1/pEGFP-c3 plasmid and amyloid precusor protein (APP)/pDsRed-Monomer-N1 plasmid were co-transfected to the HEK293 cells and the cleavage activity of BACE1 in the cells was identified by Western blotting.Results The sequencing data of the obtained BACE1 gene were identical with those in GenBank.Activity test showed that the fluorescent values of blank controls,expressed BACE1 and standard BACE1 were 55.013±3.597,1836.629±154.195 (n=3) and 2639.548±207.1901 (n=3),respectively;as compared with the control group,significant differences were noted in both of the two groups (F=78.681,P=0.000);however,there is no significant difference between expressed BACE1 and standard BACE1 groups (P>0.05).Westem blotting showed the co-transfected BACE1 could cleave APP in HEK293 cells and the CTF-APP band was detectable.Conclusion A practical protocol is established for high expression,purification and identification of BACE1 in HEK293 cells,which is helpful to obtain BACE1,an important molecular target in AD research and treatment.
6.Bioinformatic analysis of direct protein targets of aspirin against human breast cancer proliferation.
Xingmei ZHU ; Jiani YANG ; Enhu ZHANG ; Wei QIAO ; Xuejun LI
Journal of Southern Medical University 2019;39(10):1141-1148
OBJECTIVE:
To explore the molecular mechanism underlying the inhibitory effects of aspirin against human breast cancer cell proliferation through bioinformatics analysis.
METHODS:
Drug Bank 5.1.3 was searched to identify direct protein targets (DPTs) of aspirin, and the protein-protein interaction (PPI) network of the DPTs was constructed online using STRING and the signaling pathways involved were identified. The genetic alterations of 6 DPTs associated with human breast cancer was analyzed and visualized by cBio Portal and OncoPrint, respectively. The transcriptomic data of breast cancer and normal tissues were downloaded from TCGA database, and the overexpressed genes were analyzed by DECenter. The intersection between the genes associated with the DPTs obtained by STRING analysis and the differentially over-expressed genes in TCGA was determined to confirm the candidate DPTs as a potential target of aspirin, and GO functional enrichment analysis was performed using Gene Ontology. The potential targets of aspirin against the proliferation of human breast cancer cells were verified by Western blotting.
RESULTS:
Eleven DPTs of aspirin were identified. KEGG pathway enrichment indicated that 6 genes (EDNRA, IKBKB, NFKB2, NFKBIA, PTGS2 and TP53) were associated with the occurrence and development of cancer. A total of 10 220 differentially expressed genes were identified from the TCGA database, and among them 4 genes (, , , ) were found to be the potential targets for aspirin. These genes were involved mostly in the regulation of cell cycle and cell division. Western blotting showed that aspirin could down-regulate the expression levels of several pivotal proteins that regulated cell cycle and cell division, including , , and .
CONCLUSIONS
, , and may be potential targets for aspirin to inhibit the proliferation of human breast cancer cells, by affecting the progress of cell cycle and cell division.