The mechanism of S100A7 inducing the migration and invasion in cervical cancer cells / 中华肿瘤杂志

Objective: To investigate the mechanism of S100A7 inducing the migration and invasion in cervical cancers. Methods: Tissue samples of 5 cases of cervical squamous cell carcinoma and 3 cases of adenocarcinoma were collected from May 2007 to December 2007 in the Department of Gynecology of the Affiliated Hospital of Qingdao University. Immunohistochemistry was performed to evaluate the expression of S100A7 in cervical carcinoma tissues. S100A7-overexpressing HeLa and C33A cells were established with lentiviral systems as the experimental group. Immunofluorescence assay was performed to observe the cell morphology. Transwell assay was taken to detect the effect of S100A7-overexpression on the migration and invasion of cervical cancer cells. Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to examine the mRNA expressions of E-cadherin, N-cadherin, vimentin and fibronectin. The expression of extracellular S100A7 in conditioned medium of cervical cancer cell was detected by western blot. Conditioned medium was added into Transwell lower compartment to detect cell motility. Exosomes were isolated and extracted from the culture supernatant of cervical cancer cell, the expressions of S100A7, CD81 and TSG101 were detected by western blot. Transwell assay was taken to detect the effect of exosomes on the migration and invasion of cervical cancer cells. Results: S100A7 expression was positively expressed in cervical squamous carcinoma and negative expression in adenocarcinoma. Stable S100A7-overexpressing HeLa and C33A cells were successfully constructed. C33A cells in the experimental group were spindle shaped while those in the control group tended to be polygonal epithelioid cells. The number of S100A7-overexpressed HeLa cells passing through the Transwell membrane assay was increased significantly in migration and invasion assay (152.00±39.22 vs 105.13±15.75, P<0.05; 115.38±34.57 vs 79.50±13.68, P<0.05). RT-qPCR indicated that the mRNA expressions of E-cadherin in S100A7-overexpressed HeLa and C33A cells decreased (P<0.05) while the mRNA expressions of N-cadherin and fibronectin in HeLa cells and fibronectin in C33A cells increased (P<0.05). Western blot showed that extracellular S100A7 was detected in culture supernatant of cervical cancer cells. HeLa cells of the experimental group passing through transwell membrane in migration and invasion assays were increased significantly (192.60±24.41 vs 98.80±47.24, P<0.05; 105.40±27.38 vs 84.50±13.51, P<0.05) when the conditional medium was added into the lower compartment of Transwell. Exosomes from C33A cell culture supernatant were extracted successfully, and S100A7 expression was positive. The number of transmembrane C33A cells incubated with exosomes extracted from cells of the experimental group was increased significantly (251.00±49.82 vs 143.00±30.85, P<0.05; 524.60±52.74 vs 389.00±63.23, P<0.05). Conclusion: S100A7 may promote the migration and invasion of cervical cancer cells by epithelial-mesenchymal transition and exosome secretion.

Effect of Sijunzi Tang on Behavior and Energy Metabolism of Hippocampal Neurons in Rats with Alzheimer's Disease / 中国实验方剂学杂志

Objective: To observe the changes of behavior and mitochondria in Alzheimer's disease(AD) rats, and to explore the mechanism of Sijunzi Tang in the treatment of AD rats' behavior and the changes of mitochondrial energy metabolism in hippocampal neurons. Method: The 60 male SPF grade rats were randomly divided into normal group, model group, low, medium and high dose group of Sijunzi Tang(3.24, 6.48,12.56 g·kg-1), and dihydroergot group (0.27 mg·kg-1), 10 rats in each group. The normal group received no intervention and had a normal diet. The rest of the rats were injected with D-galactose to the abdomen at a dose of 200 mg·kg-1 once a day for 6 weeks. Morris water maze experiment was used to detect the cognitive function of rats. Tm-Vision behavioral experiment system was used to observe the behavioral changes of rats, and the hippocampal neuronal line structure was observed by transmission electron microscope. Mitochondrial complex colorimetry was adopted to detect rats CⅠ, CⅡ, CⅢ, CⅣ activity.Western blot was used to detect the expression of mitochondrial adenosine monophosphate-activated protein kinase(AMPK) protein in the hippocampus. Result: Compared with normal group, model group rats latent escape period time increasing, through fewer, movement distance, movement time increasing, center residence time increased (PPPPPPPPPPPPConclusion: The abnormal behavior of AD rats may be related to the decrease of central hippocampal energy metabolism and mitochondrial function. Sijunzi Tang has therapeutic effect.

Prediction of ETA oligopeptides antagonists from Glycine max based on in silico proteolysis / 中国中药杂志

Oligopeptides are one of the the key pharmaceutical effective constituents of traditional Chinese medicine(TCM). Systematic study on composition and efficacy of TCM oligopeptides is essential for the analysis of material basis and mechanism of TCM. In this study, the potential anti-hypertensive oligopeptides from Glycine max and their endothelin receptor A (ETA) antagonistic activity were discovered and predicted based on in silico technologies.Main protein sequences of G. max were collected and oligopeptides were obtained using in silico gastrointestinal tract proteolysis. Then, the pharmacophore of ETA antagonistic peptides was constructed and included one hydrophobic feature, one ionizable negative feature, one ring aromatic feature and five excluded volumes. Meanwhile, three-dimensional structure of ETA was developed by homology modeling methods for further docking studies. According to docking analysis and consensus score, the key amino acid of GLN165 was identified for ETA antagonistic activity. And 27 oligopeptides from G. max were predicted as the potential ETA antagonists by pharmacophore and docking studies.In silico proteolysis could be used to analyze the protein sequences from TCM. According to combination of in silico proteolysis and molecular simulation, the biological activities of oligopeptides could be predicted rapidly based on the known TCM protein sequence. It might provide the methodology basis for rapidly and efficiently implementing the mechanism analysis of TCM oligopeptides.

Protein composition analysis and cytotoxicity of gulbulin hydrolysates of Brucea javanica seeds / 中国中药杂志

To analyze the protein composition of Brucea javanica seeds and evaluate the cytotoxicity of its gulbulin hydrolysates. Four protein fractions of albumin, gulbulin, prolamin and glutelin were sequentially extracted and then quantified by Kjeldahl method. Different kinds of proteases were applied to hydrolyze B.javanica gulbulin, and MTT assay was used to evaluate the cytotoxicity of low molecular weight hydrolysates (≤3 kDa) on human breast cancer MCF-7 cell. The results showed that： the total protein content of B.javanica seeds was 17.47%, albumin, gulbulin, coxin, glutelin and residue protein accounted for 15.01%, 8.11%, 2.47%, 44.92% and 23.62% of the total protein content, respectively. The hydrolysates (≤3 kDa) of B.javanica globulin produced by pepsin showed significant growth inhibitory activity on MCF-7 cells, and the IC₅₀ value was（6.52±0.01） mg•L⁻¹ after 72 h of incubation. Protein was abundant in B.javanica seeds, and its peptides demonstrated specific cytotoxicity on MCF-7 cell line in vitro, suggesting antitumor active ingredient can be further generated from B.javanica seeds.

Analysis of the expression of matrix metaUoproteinase and tissue inhibitor of metalloproteinase in gestational trophoblastic diseases / 中华全科医师杂志

Objective To explore the relationship between the expression of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) in gestational trophoblastie diseases and its invasiveness.Methods The expression and distribution of MMP-7,MMP-9,TIMP-1 and TIMP-2 were determined by immunohistochemistry PV-6000 in 49 cases of hydatidiform mole (eight of them with malignant transformation),six cases of invasive hydatidiform mole,two cases of choriocarcinoma and 18 cases with normal villi in early pregnancy.Results The expression of MMP-7,MMP-9,TIMP-2 and MMP- 9/TIMP-1 was significantly higher in choriocareinoma,invasive mole and malignant transformed mole than that in normal villi in early pregnancy and non-malignant transformed mole (F=10.950,7.760,7.304, 3.442;P