Objective To screening up-regulated protein kinases and their inhibitors in order to provide new targets for molecular thera-py of nasopharyngeal cancer.Methods GEO database and SAM software were employed to screen the up-regulated protein kinase gene in nasopharyngeal cancer.Based on DAVID database,the regulating functions of kinases were identified.The inhibitors of up-regulated kinase genes were identified by Selleckchem database.Literature mining was used to screen the potential anti-cancer drugs.Results Totally 2360 differentially expressed genes including 21 up-regulated protein kinases (CHEK1,CHEK2,PRKDC,AURKA,VRK2,STK17A,MELK,NU-AK1,TRPM7,MASTL,AXL,BUB1,BUB1B,CDK4,TTK,CDC7,CASK,AKT3,TBK1 and PBK)were identified in the whole genome profi-ling (Fold Change≥2,P <0.05).The results of function analysis showed the up-regulated genes were enriched in 10 function terms such as‘protein amino acid phosphorylation’‘phosphorylation’‘phosphate metabolic process’‘mitotic cell cycle’‘cell cycle phase’‘regulation of cell cycle’,and so on.The Selleckchem database analysis showed there were 9 up-regulated protein kinases equipped with 51 inhibitors which were proved already.The results of literature mining showed that 18 inhibitors of them had a few studies (less than 10 literatures)in cancer terms,and there was a potential to become new drugs to treat nasopharyngeal cancer.Conclusion A total of 21 up-regulated protein kinases were identified,and they might promote the nasopharyngeal carcinoma by regulating functions such as the cell-cycle control pathway.Their ki-nase inhibitors may have a potential role in anti-cancer treatment,which provided a new target point for molecular therapy of nasopharyngeal cancer.

Objective Based on protein-protein interaction network,gene modules were identified to provide new targets for molecular therapy of nasopharyngeal carcinoma.Methods GEO dataset (GSE12452)and SAMsoftware were employed to screen the differentially ex-pressed gene in nasopharyngeal carcinoma.Protein-protein interaction network was established by using String database.Based on the net-work,the gene modules were identified by using bioinformatics gene module analysis method.GO analysis was used to analyze the function of gene modules.Results In this study,2 634 differentially expressed genes were identified in nasopharyngeal carcinoma.There were 4 729 protein-protein interaction pairs among the differentially expressed genes according to the String database.We established the protein-protein interaction network based on these pairs.Seven gene modules were identified by bioinformatics methods.GO analysis results showed that the function of the gene modules including regulation of cell cycle,glycosylation,cell adhesion,oxidation reduction and so on.Conclusion There are 7 gene modules in protein-protein interaction network in nasopharyngeal carcinoma.These modules may play important roles in the progression and development of nasopharyngeal carcinoma.Our finding can provide a new sight for molecular diagnose and therapy of nasopharyngeal carcinoma.

To establish induction and liquid culture system for hairy roots of Danshen (Salvia miltiorrhiza), Agrobacterium rhizogenes A4, LBA9402, 15834 as test bacterium were used to infect aseptic leaves of Danshen. The hairy roots were induced and positive transgenic hairy roots were selected with PCR using rolB and rolC as the target gene. Then hairy roots of S. miltiorrhiza were harvested and salvianolic acids were extracted with 70% methanol containing 1% formic acid. The content of salvianolic acid B (SalB) and rosmarinic acid (RA) were determined by HPLC. According to the above research results, the Danshen hairy roots induced by A. rhizogenes LBA9402 were inoculated into the following group of culture media: MSOH, MS, B5, and 6,7-V liquid media. Then the same methods of extraction and determination for the content of Danshen hairy roots were adopted. Last, the hairy roots of S. miltiorrhiza induced by A. rhizogenes LBA9402 were inoculated into the MSOH liquid media with different pH values. The content of salvianolic acid were extracted with 70% methanol containing 1% formic acid and determined by HPLC. As a result, three kinds of A. rhizogenes A4, LBA9402, 15834 could induce hairy roots and Ri plasmids were integrated into the genome of S. miltiorrhiza by PCR. Danshen hairy roots induced by A. rhizogenes LBA9402 and A4 produced much more salvianolic acid, which were (3.27 ± 0.37)% [including (1.04 ±0.36)% of RA and (2.22 ± 0.29)% of SalB] and (3.17 ± 0.20)% [including (0.92 ± 0.31)% of RA and (2.25 ± 0.26)% of SalB], respectively. Hairy roots induced by A. rhizogenes LBA9402 when they were cultured in MSOH liquid media produced much more salvianolic acid, which was (4.56 ± 0.36)%, including (1.12 ± 0.26)% of RA and (3.44 ± 0.23)% of SalB. Hairy roots induced by A. rhizogenes LBA9402 produced the most salvianolic acid when they were cultured in MSOH liquid media with the pH value 4.81, which was 4.85%, including 1.16% of RA and 3.69% of SalB. So Danshen hairy roots induced by A. rhizogenes LBA9402 and A4 produced much more salvianolic acid when they were cultured in MSOH liquid media with the pH value 4.81. The research had established the foundation on genetic engineering to improve the quality of S. miltiorrhiza.
Agrobacterium ; physiology ; Benzofurans ; analysis ; metabolism ; Cell Culture Techniques ; instrumentation ; methods ; Cinnamates ; analysis ; metabolism ; Culture Media ; chemistry ; metabolism ; Depsides ; analysis ; metabolism ; Drugs, Chinese Herbal ; analysis ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Salvia miltiorrhiza ; chemistry ; growth & development ; metabolism ; microbiology

Background Cardiomyocytes apoptosis was related to oxidative stress which has been shown to play an important role in ventricular remodeling after myocardial infarction(MI).Probucol is a hypolipidemic and antioxidant agents.Several reports demonstrated its cardioprotective effect after MI.However,the exact effect of probucol on the modification of the apoptosis related gene Bcl-2 and Bax is not clear.Objective To investigate the effects of probucol on mRNA and protein expression of Bcl-2,Bax and oxidative stress in myocardial infarcted rats.Methods Forty-one SD rats that survived 24 h after ligating left anterior descending coronary artery were randomly to receive placebo-saline(5 mL/d,n=20)or probucol(probucol 60 mg/kg?d,n=21).Twelve rats underwent sham operation were served as control(n=12).Six weeks after treatment,hemodynamic pararmeters and left ventricular function were measured with catheterization.Cardiomyocytes apoptosis were determined by TUNEL method.Myocardium mRNA of Bcl-2 and Bax expression levels in the non-infarcted myocardium were as- sessed by RT-PCR.The myocardium protein expression of Bcl-2 and Bax in the non-infarcted myocardium were de- termined by Western blot.Colorimetry was used to determine oxidative metabolism index in myocardium.Results 1)Compared with the sham rats,all MI rats showed marked decrease in Bcl-2 mRNA and protein expression in myo- cardium with increase of Bax mRNA and protein expression and apoptosis index(P

Aged ; Case-Control Studies ; Cerebral Infarction ; blood ; genetics ; Female ; Fibrinogen ; genetics ; metabolism ; Haplotypes ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic

Objective To investigate the feasibility of high?resolution microendoscopy( HRME) im?aging for animal gastrointestinal mucosa. Methods Mucosal tissues were harvested from the stomach, small intestine, and large intestine of Japanese big?ear white rabbits. The effects of HRME imaging of different lo?cations such as the gastric antrum and fundus, small intestine and large intestine were observed, and those of different exposure time were compared.Accuracy of HRME imaging was compared with pathology. Results The specific tissues of the gastrointestinal mucosa could be clearly distinguished from the HRME images. In the superficial layer of the fundic mucosa, numerous closely arranged glands as well as oval or elongated branched openings of the gastric pits and linear peripheral cracks were visible;the nuclei were arranged reg?ularly. In the superficial layer of the antral mucosa, irregular or tubular openings of the gastric pits and cracked glandular cavities were visible, with the cells surrounding the gastric pits regularly arranged and the nuclei small and densely distributed. In the superficial layer of the small intestine mucosa, stereoscopic thick?finger?shaped villi cluster was visible. The intervillous spaces were crack?like, and the surface was cov?ered by regularly arranged reflective, absorptive cells. In the superficial layer of the large intestine mucosa, many regularly arranged daisy?like round crypts of uniform size, as well as reflective, goblet cells surrounding the crypt and the interval space between crypts were visible. When the exposure time increased, the nuclei became brighter. An excellent correlation was noted between the results of histologic examination and those obtained by using HRME. Conclusion HRME can produce accurate images of the animal gastro?intestinal mucosae and may be a novel technique for further studies of human gastrointestinal pathology.

Objective To detect the expressions of miR-155,T helper type 17 (Thl7) cells,and Th17 cellspecific transcription factor RORγt and effector cytokine interleukin (IL)-17 in peripheral blood and skin lesions from,and to evaluate their relationship in,patients with atopic dermatitis (AD).Methods Peripheral blood was obtained from 37 patients with AD and 33 age-and sex-matched healthy controls,and biopsy specimens from the lesional and perilesional skin of five patients with severe AD as well as from the normal skin of five healthy human controls.Real-time fluorescence-based reverse transcription (RT)-PCR was employed to measure the mRNA expression levels of miR-155,RORγt and IL-17 in peripheral blood mononuclear cells (PBMCs) and skin specimens,flow cytometry to detect the percentage of Th17 cells in PBMCs,enzyme-linked immunosorbent assay (ELISA) to determine the plasma concentration of IL-17.Statistical analysis was done using independent sample's t test,one-way analysis of variance followed by the least significant difference test,and linear correlation analysis.Results Compared with the healthy controls,the patients with AD showed a significant increase in Th17 cell percentage (1.78％ ± 0.52％ vs.0.47％ ± 0.15％,P＜ 0.01),mRNA expression levels of miR-155 (5.78 ± 1.78 vs.1.82 ± 0.46,P＜ 0.01),RORγt (6.08 ± 1.04 vs.1.64 ± 0.52,P＜ 0.01) and IL-17 (7.09 ± 1.75 vs.1.71 ± 0.46,P＜ 0.01),as well as in the plasma concentration of IL-17 ((2.51 ± 6.15) pg/ml vs.(11.80 ± 2.24) pg/ml,P＜ 0.01).There was a sequential decrease in the expression levels of miR-155,RORγt and IL-17 mRNA from lesional skin,perilesional skin to normal skin (F =41.803,17.040 and 37.064 respectively,all P ＜ 0.01).The miR-155 mRNA expression level in PBMCs was positively correlated with the SCORing Atopic Dermatitis (SCORAD) index,Th17 cell percentage,RORγt and IL-17 mRNA expression levels as well as IL-17 plasma concentration (r =0.405,0.426,0.402,0.410 and 0.408 respectively,all P ＜ 0.05).Similarly,the miR-155 expression level was positively correlated with RORγt and IL-17 mRNA expression levels in lesional and paralesional specimens (r =0.428 and 0.435 respectively,both P ＜ 0.05).Conclusion The up-regulated expression of miR-155,Th17 cells and their effector cytokine IL-17 may be associated with the development of AD.