3.MicroRNA in oral squamous cell carcinoma.
Chinese Journal of Stomatology 2013;48(6):376-380
Apoptosis
;
Carcinoma, Squamous Cell
;
diagnosis
;
genetics
;
metabolism
;
therapy
;
Cell Proliferation
;
Chromosome Aberrations
;
Early Diagnosis
;
Epigenesis, Genetic
;
Humans
;
MicroRNAs
;
genetics
;
metabolism
;
Mouth Neoplasms
;
diagnosis
;
genetics
;
metabolism
;
therapy
;
Neoplasm Invasiveness
;
Neoplasm Metastasis
;
Polymorphism, Genetic
4.Effects of RNA Interference Combined with Ultrasonic Irradiation and SonoVue Microbubbles on Expression of STAT3 Gene in Keratinocytes of Psoriatic Lesions
RAN LI-WEI ; WANG HAO ; LAN DONG ; JIA HONG-XIA ; YU SI-SI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(2):279-285
The most effective sequence of small interfering RNA (siRNA) silencing STAT3 of psoriatic keratinocytes (KCs) was screened out,and the effects of the most effective siRNA combined with ultrasonic irradiation and SonoVue microbubbles on the expression of STAT3 of KCs and the dose-and time-response were investigated.Three chemically-synthetic siRNAs targeting STAT3 carried by Lipofectamine 3000 were transfected into KCs,and the effects on STAT3 expression were detected,then the most effective siRNA was selected for the subsequent experiments.The negative controls of siRNA (siRNA-NC) labeled with Cy3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles were transfected into KCs,then the optimal parameters of ultrasonic irradiation were determined.The most effective siRNA carried by Lipofectamine 3000 combined with ultrasonic irradiation at the optimal parameters and SonoVue microbubbles was transfected into KCs,and the dose-and time-response of RNA interference was determined.The effect of RNA interference by the most effective siRNA at the optimal time and dose carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (LUS group) was compared with that only carried by Lipofectamine 3000 (L group).The results showed that siRNA-3 achieved the highest silencing efficacy.0.5 W/cm2 and 30 s were selected as the parameters of ultrasonic irradiation.The siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue mierobubbles could effectively knock down the STAT3 expression at mRNA and protein levels in dose-and time-dependent manners determined at 100 nmol/L with maximum downregulation on mRNA at 48 h,and on protein at 72 h after transfection.The LUS group achieved the highest silencing efficacy.It was concluded that siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles could effectively knock down the STAT3 expression in psoriatic KCs,and the optimized transfection condition and the sequence of siRNA-3 could serve for further research on gene therapy of psoriasis.
6.Determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials by LC-MS/MS.
Hai-hong SI ; Yan-jing LI ; Jia XUE ; Wen-zhe HUANG ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2015;40(14):2832-2836
To develop a LC-MS/MS method for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials, the column was Agilent ZORBAX Eclipse plus C18 (3.0 mm x 50 mm, 1.8 µm), and the mobile phase consisted of methanol-water (containing 0.2% formic acid) (95:5) at a flow rate of 0.5 mL · min(-1). The multiple reaction ion monitoring (MRM) with an ESI interface in the negative ion mode was selected. The results showed that the linear ranges of five kinds of ginkgolic acids were in the range of 0.2-36.0 µg · L(-1) (r ≥ 0.999 5). The lowest limit of quantification (LOQ) of ginkgo acid C13: 0, C15:1, C17:2, C15:0 and C17:1 were 0.18, 0.18, 0.21, 0.10 and 0.20 µg · L(-1), respectively. The average recovery was between 73.28% and 87.56%, and the average content of total ginkgolic acids in three batches of samples was in the range of 0.023-0.028 µg · g(-1), which was much lower than 2 µg · g(-1) prescribed in drug registration standards. This method is simple and rapid with high sensitivity, which can be used for the determination of five kinds of trace ginkgolic acids in diterpene ginkgolides meglumine injection materials.
Chromatography, Liquid
;
methods
;
Ginkgolides
;
analysis
;
Injections
;
Limit of Detection
;
Salicylates
;
analysis
;
Tandem Mass Spectrometry
;
methods
7.Influences of chronic hypoxia on the gene expression of Kv1.3,Kv2.1 and Kv3.1 induced by acute hypoxia in PASMC of rats
Zhigang HONG ; Si JIN ; Wei KONG ; Dixun WANG ; Qiling CHEN ; Bingyong SUN
Chinese Journal of Pathophysiology 2000;0(10):-
AIM and METHODS: Total RNA was extracted from 6th rat subcultured pulmonary artery smooth muscle cells(PASMC) exposed to continual chronic hypoxia or normoxia and the effects of chronic hypoxia on the changes of Kv1.3,Kv2.1,Kv3.1 mRNA in cultured PASMC induced by acute hypoxia were studied by semiquantitative RT-PCR in vitro. RESULTS:①Kv1.3,Kv2.1,Kv3.1 genes were found to be expressed in PASMC of rats exposed either to hypoxia or normxia.②The expression of Kv2.1 and Kv3.1 in 6th subcultured of PASMC in normaxia group could be upregulated by exposure to acute hypoxia,the levels of Kv2.1 and Kv3.1 mRNA were significantly increased from 0.646?0.092, 0.782?0.104 to 1.059?0.134, 0.985?0.116,respectively (P
8.A review on the application of animal models in preclinical research of the treatment of liver diseases with mesenchymal stem cells
Xufeng FU ; Bingbing CHEN ; Yaping YAN ; Hong WANG ; Junfeng WANG ; Bingrong ZHENG ; Wei SI
Acta Laboratorium Animalis Scientia Sinica 2017;25(1):102-106
Chronic liver diseases can further develop to liver fibrosis and cirrhosis. Currently, there is no effective treatment except liver orthotopic transplantation at this point. The extreme shortage of liver organ source forced people to find alternative treatment strategies. Mesenchymal stem cells ( MSCs) have the abilities of immunomodulatory, hepatocyte differentiation, promotion of liver cells regeneration in situ and inhibiting the activation of hepatic stellate cells. Therefore, MSCs transplantation provides a very broad prospect for cell therapy. It is important to provide preclinical evaluation of the efficacy and safety before the application of cell therapy in clinical trials. The progress of various animal models of human liver diseasees and significance of using MSCs to treat liver diseases in preclincal studies based on these animal models were reviewed in this paper.
9.Expressions of matrix metalloproteinase-2 and extracellular matrix metalloproteinase inducer in retinoblastoma
Yu-Hong, CHENG ; Qiang, SHI ; Jia-Quan, SHEN ; Li-Lun, WANG ; Si-Wei, LIU
International Eye Science 2015;(7):1154-1157
AlM: To investigate expressions of matrix metalloproteinase-2 ( MMP-2 ) and extracellular matrix metalloproteinase inducer ( EMMPRlN) in retinoblastoma (Rb) and the relationships between MMP-2, EMMPRlN and tumor development.METHODS:lmmunohistochemical technique was used to detect expressions of MMP-2 and EMMPRlN in 39 cases of paraffin embedded Rb samples. Quantitative analysis of expressions of MMP-2 and EMMPRlN were assessed by measuring the mean gray scale of Rb tissue with LElCA lM50 Color Pathologic Analysis System. The differences of expressions of MMP-2 and EMMPRlN in each clinical and pathological stage were statistically analyzed, and the same step was also undertaken to study the relationship between Rb with MMP-2 positive expression and that with EMMPRlN positive expression.RESULTS:The positive expression rate of MMP-2 was 90% (Gray value: 109. 64 ± 14. 52; 35/39), and that of EMMPRlN was 85% (Gray value:108. 01±13. 60;33/39). The expressions of MMP - 2 and EMMPRlN were significantly higher in tumors of glaucomatous stage (Gray value:108. 21±11. 47 and 107. 56±14. 32) than those in intraocular stage ( Gray value: 121. 13 ± 11. 32 and 119. 34 ± 12. 66; P<0. 01 and P<0. 05). And the same conclusion can be concluded between those in extraocular stage (Gray value: 91. 03 ± 11. 71 and 92. 26 ± 12. 93) with those in glaucomatous stage (P<0. 01 and P<0. 05). The expressions of MMP-2 and EMMPRlN were significantly higher in tumors with optic nerve invasion (Gray value:103. 89±13. 39 and 105. 23±14. 00) than those without optic nerve invasion ( Gray value: 118. 39 ± 15. 11 and 117. 53±16. 13) (P<0. 01 and P<0. 05).CONCLUSlON:The positive expression levels of MMP-2 and EMMPRlN may correlate with tumor infiltration and metastasis.
10.18F-FDG PET/CT in diagnosis of malignant pericardial lesions
Dan, SHAO ; Shu-xia, WANG ; Chang-hong, LIANG ; Si-yun, WANG ; Wei-ping, XU
Chinese Journal of Nuclear Medicine 2010;30(5):333-335
Objective To assess the value of integrated 18 F-fluorodeoxyglucose (FDG) PET/CT in differentiation of malignant and benign pericardial effusion. Methods 18F-FDG PET/CT were performed in 23 patients with pericardial effusion. The detected soft tissue tumor or nodulous lession in pericardium or the thickened pericardium, with the maximum standardized uptake value( SUVmax ) ≥2.5, was defined as PET/CT-positive. The invaded lession in pericardium with SUVmax ≥2.5 was also as the positive. The difference of SUVmax of benign and malignant lesions was analyzed with two-independent-sample test of nonparametric tests. The final diagnosis was confirmed by biopsy or post-operative pathology. Results The diagnosis were confirmed with 14 malignant and 9 benign lesions. The median of SUVmax was 6.0 in malignancy group and 2.2 in benign group (z= -3. 279, P =0.001 ). According to the pathology results, there were one false negative case and two false positive cases with PET/CT imaging interpretation. The sensitivity, specificity,accuracy, positive predictive value ( PPV ) and negative predictive value ( NPV ) of 18 F-FDG PET/CT in diagnosis of benignity or malignance of pericardium effusion were 92.9% ( 13/14), 7/9, 87.0% (20/23),86.7% (13/15) and 7/8, respectively. Conclusion For the patients with pericardium effusion 18F-FDG PET/CT may be a helpful modality for malignancy differentiation