OBJECTIVE To construct siRNA eukaryotic expression vector targeting protein kinase CK2?and to investigate its inhibitory effect on invasion of the HEp-2 cell line in human laryngeal carcinoma. METHODS siRNA expression vector psiRNA-hH1neo-CK2 targeting protein kinase CK2?was constructed by gene recombination,and then was transfected into the HEp-2 cells by lipofectamine methods. Protein kinase CK2?mRNA and protein of the transfected cells were detected by reverse transcription polymerase chain reaction(RT-PCR) and Western blot,respectively. The invasion of the transfected cells was measured by Boyden chamber.SP method was used to examine the expressions of MMP2 and TIMP2 protein of the transfected HEp-2 cells. RESULTS Protein kinase CK2?siRNA expression vector was successfully constructed by gene recombination. Compared with non-specific interfering groups and blank groups, protein kinase CK2?mRNA and protein were significantly decreased respectively in the psiRNA-hHIneo-CK2 groups(P

This study was aimed to identify Chrysanthemi Flosbefore and after sulfur fumigation based on its different odour by the electronic nose technology.It was expected to explore a new method for the Chrysanthemi Flos identification according to the odour.The electronic nose technology was used in the detection of peak response values of Chrysanthemi Flos on sensor array.The principal component analysis (PCA) and 10 machine learning (ML) ways were used in the analysis of response values and establishment of optimized identification models.The results showed that there was a significant difference in the odour between sulfur fumigated Chrysanthemi Flos and non-sulfur fumigated ones.The identification models were successful with high correct judge rate by PCA and 6 ML ways including BF Tree,J48 and Random Tree.It was concluded that the electronic nose technology can be used for the accurate identification of sulfur fumigated Chrysanthemi Flos and non-sulfur fumigated ones.The electronic nose technology combined with multiple ML methods can be introduced in the quality evaluation of Chrysanthemi Flos.It provided more ideas for the application of electronic nose in data mining for traditional Chinese medicine (TCM) studies.

Objective Fructus Amomi(Sharen) is derived from the dry ripe fruit of Amomum villosum Lour., A.villosum Lour. var. xanthioides T.L. Wu et Senjen and A.longiligulate T.L.Wu, which is widely utilized for its clinic effects on digestive system. However, Fructus Amomi from different species and habitats, possessing different quality, is difficult to identify. In this study, we aim to develop a simple, rapid and reliable method for authentication of Fructus Amomi. Methods Twenty-five batches of samples of Fructus Amomi were collected and electronic nose was introduced into analyzing their odor with multiple mathematical statistics methods. Na?ve bayes network (NBN), radical basis function (RBF) and random forest (RF) were applied to establish different classifiers while BestFirst+CfsSubsetEval (BC) was used to screen the attributes for searching sensor array with higher contributions. Results Firstly, after attribute-screening via BC, the established discriminative models via NBN, RBF and RF could successfully identify genuine and non-genuine samples, presenting correct judging ratios of 78% and 84% through ten-fold cross validation and external test set validation, respectively. Besides, quantity predictive models were constructed as well. In case of content of bornyl acetate, one of the effective components in Fructus Amomi, values were higher than 3.5 mg/g and lower than 1.8 mg/g with sensor response of 0.04 and 0.03, respectively. Conclusion In this paper, quality discriminative model and quantity predictive model of Fructus Amomi were established via electronic nose and multiple mathematical statistics methods. It indicates that electronic nose could be a promising method for quality evaluation of Chinese material medica.

In order to study the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in human laryngeal squamous cell carcinoma (LSCC) and its significance, the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues (both P < 0.01), and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ill + IV tissues of LSCC as compared with the stage I + II tissues of LSCC (P < 0.01). There was a high positive correlation between VEGF and COX-2 expression in LSCC (r = 0.756, P < 0.01). These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC.
Carcinoma, Squamous Cell/*metabolism ; Cyclooxygenase 2/*biosynthesis ; Cyclooxygenase 2/genetics ; Laryngeal Neoplasms/*metabolism ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics ; Tumor Markers, Biological ; Vascular Endothelial Growth Factor A/*biosynthesis ; Vascular Endothelial Growth Factor A/genetics

In order to study the effect of 5, 6-Dichloro-1-beta-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose-and time-dependent manner. After being treated with 0, 10, 20, 40, 80 microm mol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68+/-0.19)%, (1.95+/-0.12)%, (8.51+/-0.26)%, (11.26+/-0.17)% and (14.99+/-0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 microm mol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.

OBJECTIVE: To study the effect on the expression of protein kinase CK2alpha and growth of human laryngeal carcinoma xenograft in nude mice by applying small interfering RNA (siRNA) specific to protein kinase CK2alpha. METHOD: Human laryngeal carcinoma Hep-2 cells were implanted under the skin of nude mice. After the tumors grew to a definite size, the tumors were injected with siRNA expression plasmid specific to protein kinase CK2alpha. The weight and volume of subcutaneous tumors were measured. The expression level of protein kinase CK2alpha mRNA and protein of tumors were measured with reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemical technique, respectively. RESULT: Protein kinase CK2alpha mRNA and protein expressions were significantly decreased in tumors transfected with siRNA expression plasmid specific to protein kinase CK2alpha (P<0.05). The tumor grew slowly after transfected with siRNA expression plasmid specific to protein kinase CK2alpha (P<0.01). CONCLUSION The siRNA expression plasmid specific to protein kinase CK2alpha may suppress the growth and the protein kinase CK2alpha expression of subcutaneous tumors. RNA interfering technology may be a new strategy for the treatment laryngeal cancer.
Animals ; Casein Kinase II ; genetics ; Cell Line, Tumor ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Plasmids ; RNA, Small Interfering ; genetics ; Transfection ; Xenograft Model Antitumor Assays

OBJECTIVE: To purify P0 protein from guinea pig's inner ear by preparative SDS-PAGE and study the possible role it may play in the etiology of autoimmune inner ear disease. METHOD: A mixture of membraneous proteins of inner ear was separated by preparative SDS-PAGE. The corresponding band at 30kd was cut and electrically eluted. The protein collected was identified by analytical SDS-PAGE and Western blot assay. A group of 20 guinea pigs were immunized with P0 protein emulsified in complete Freund's adjuvant, another 10 guinea pigs were immunized with complete Freund 's adjuvant only as control. The guinea pigs' hearing thresholds, serum IgG level and morphological changes in the inner ear were investigated. The distribution of P0 protein in the cochlear was detected by immunohistochemical technique. RESULT: The purity of the protein was demonstrated by a single band at the 30 kD site in SDS-PAGE, which was identified as P0 protein by western blot analysis assay. About 17.5% P0-immunized guinea pigs showed increased hearing thresholds, elevated IgG level (F =6.48, P <0. 01), as well as a decreased number of spiral ganglion cells and inflammatory cell infiltration in the cochlear nerve region. The P0 protein is distributed in the cochlear nerve and spiral ganglion only. CONCLUSION P0 protein from guinea pig's inner ear can be successfully purified by preparative SDS-PAGE and an animal model of experimental autoimmune inner ear disease induced by P0 protein is successfully established.
Adenoviridae ; genetics ; Animals ; Cochlea ; metabolism ; Disease Models, Animal ; Electrophoresis, Polyacrylamide Gel ; Gene Expression ; Gene Transfer Techniques ; Genes, Homeobox ; Genes, Reporter ; Genetic Vectors ; Guinea Pigs ; Myelin P0 Protein ; isolation & purification

In order to study the expressions of vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2) in human laryngeal squamous cell carcinoma (LSCC) and its significance,the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues (both P＜0.01), and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ⅲ + Ⅳ tissues of LSCC as compared with the stage Ⅰ + Ⅱ tissues of LSCC (P ＜0. 01). There was a high positive correlation between VEGF and COX-2 expression in LSCC (r=0. 756,P＜0.01). These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC.

In order to study the effect of 5, 6-Dichloro-1-β-D-ribofuranosyl-benzimidazole (DRB) on the biological characteristics of human laryngeal carcinoma Hep-2 cell line in vitro, Hep-2 cells cultured in vitro were treated with different concentrations of DRB. Changes in cell proliferation, apoptotic rate and invasiveness were detected by MTT assay, flow cytometry (FCM) and matrigel in vitro invasion assay, respectively. It was found that DRB inhibited the proliferation of Hep-2 cells in a dose- and time-dependent manner. After being treated with 0, 10, 20, 40, 80 μmmol/L DRB for 24 h, the apoptotic rate in Hep-2 cells was (0.68±0.19) %, (1.95±0.12)%, (8.51 ±0.26)%, (11.26±0.17) % and (14.99±0.32)%, respectively. The matrigel in vitro invasion assay revealed that DRB began to inhibit the invasion of Hep-2 cells at the concentration of 5 μmmol/L, and with the increase of DRB concentration, the inhibitory effect was enhanced. It was suggested that DRB could influence the essential biological characteristics of Hep-2 cells, inhibit Hep-2 cells proliferation, reduce invasive ability and induce apoptosis of Hep-2 cells.

Objective: To comprehensively evaluate the quality of Radix et Rhizoma Rhei based on gray correlationalanalysis and functional components, and to explore the difference of Radix et Rhizoma Rhei in different genuineproducing areas. Methods: HPLC was utilized to analyze 14 main compositions contained in the samples, includingemodin, rhein, chrysophanol, aloe-emodin, physcion, rheinoside, physcion glucoside, chrysophan, aloe-emodinglucoside, emodin methyl glycoside, sennoside, sennoside B, catechin and gallic acid. Then python 2.7 software wasemployed for gray correlation analysis of functional components closely related to the traditional efficacy of Radix et Rhizoma Rhei. Results: The qualities of Radix et Rhizoma Rhei grow in different areas were different. Tanggute Radix et Rhizoma Rhei grew in Tianzhu Gansu had the best effects of "expelling water retention and attacking the accumulation", and that grew in Yajiang Sichuan had the best effects of "clearing heat and removing toxin". Zhangye Radix et Rhizoma Rhei grew in Lixian Gansu had the best effect of"expelling stasis and unblocking the channels". Conclusion: Patternrecognition has broad prospects in the field of quality evaluation of traditional Chinese medicine. From the clinicalefficacy of traditional Chinese medicine, pattern recognition at the level of efficacy components can provide a new ideafor establishing a more complete and scientific quality evaluation system for traditional Chinese medicine.