1.Effect of miRNA-218 on renal cell carcinoma in nude mice
Long HE ; Long LIU ; Hongwei YANG ; Yan ZHANG ; Lianhui FAN
Cancer Research and Clinic 2015;27(11):728-731
Objective To explore the effects of miRNA-218 on renal cell carcinoma of nude mice in vivo.Methods The pcDNA3.1-miR-218 and its control negative plasmids were stably transfected into renal cell carcinoma cell line A498 and 769-P.These cells were inoculated into nude mice in different groups to observe the changes of body and tumor and to detect the expression of miR-218 in the tissues of nude mice.Results In the A498 cells + pcDNA3.1-miR-218 transfected group,the weight loss of tumor bearing nude mice after 25 days was lower than that in the control group,and the tumor volume was smaller than that in the control group after 10 days (P < 0.05).In the 769-P cells + pcDNA3.1-miR-218 transfected group,the weight loss of tumor bearing nude mice was lower than that in the control group after 19 days,and the tumor volume was smaller than that in control group after 10 days (P < 0.05).The expression of miRNA-218 in bearing nude mice with A498 cells or 769-P cells transfected by miRNA-218 was significantly higher than that in the control group,and the difference was statistically significant (P < 0.05).Conclusion Up-regulation of miRNA-218 expression can inhibit the growth of renal cell carcinoma of nude mice in vivo.
2.Construction and application in renal cell carcinoma 769-P cells of miR-218 expression plasmids
Long HE ; Yan ZHANG ; Jun WANG ; Long LIU ; Lianhui FAN
Cancer Research and Clinic 2013;25(12):806-808,811
Objective To construct a stable expression plasmids miR-218,to lay the experimental foundation for the expression and mechanism of miR-218 in human renal cell carcinoma.Methods The primers were designed by Has-miR-218 and the miR-218 fragment were amplified by PCR,and then which was connected with the carrier pcDNA3.1 (+) to build a stable expression plasmids.The recombinant plasmids were trasfected into renal cell carcinoma cell line 769-P,and the expression of miR-218 in these cells were detected.Results The plasmids were constructat successfully,which was determind by enzyme digestion and sequencing results.The constructed expression plasmids pcDNA3.1-miR-218 transfection of human renal cell carcinoma cell line 769-P,miR-218 cxpression level of cells was significantly increased after trasfected by recombinant plasmids carrying miR-218 gene (relative expression quantity was 1.64).Conclusion The miR-218 expression plasmids pcDNA3.1-miR-218 were successfully constructed,the plasmids can be applied to the study of miR-218 function and mechanism in renal cell carcinoma.
4.Study of the syndromes of traditional Chinese medicine in terms of metabonomics technology: the current situation and considerations.
Jing ZHANG ; Long LIU ; Xin YAN ; Xiaoqiang YUE
Journal of Integrative Medicine 2012;10(10):1069-76
Syndromes constitute a core aspect in the study of Chinese medicine, and research on the concept of syndromes is important to the study of the process of modernization of traditional Chinese medicine. However, it is somewhat challenging to define a syndrome due to the complexity inherent in the subject, even with the assistance of the reductionism approach of modern medicine. Holistic and dynamic in nature and attaching much importance to functional changes, the newly emerging metabonomics is in many ways inline with the concepts of syndrome differentiation of pathological states in traditional Chinese medicine. Therefore, metabonomics has comparatively strong advantages in the very respect of revealing the natural laws of syndrome differentiation. By reviewing and analyzing the current research on the concept of syndromes and the application of metabonomic technology to exploring the essential core of syndrome differentiation, the authors illustrated the potential commonalities. This would also show the issues requiring attention between the study of syndromes and the metabonomic technology. In the meantime this study reflected the core problems in detail and put forward suggestions with regard to reaching solutions.
5.The roles of RNA-editing enzyme ADAR1 in EV71 infection and virus mutation
Qingqing LIU ; Zhangmei CHANG ; Jinjin BAI ; Yan WANG ; Jianer LONG
Fudan University Journal of Medical Sciences 2017;44(3):253-260
Objective To identify the role of RNA-editing enzyme ADAR1 (adenosine deaminase acting on RNA) in EV71 infection and virus mutation.Methods RNAi technology was applied to establish ADAR1 knock-down stable cell lines.Then the cells were served to evaluate the role of ADAR1 in EV71 infection by MTT assay for detecting virus-induced cell viability,virus plaque assay for quantification of the virus titer and the cellular susceptibility to the virus,and Western blot for virus protein expressions.ADAR1-mediated RNA editing can result in the genetic A-G and T-C mutations.To further determine whether the effects of ADAR1 on EV71 infection were correlated with ADAR1-mediated EV71 RNA editing and therefore increased the viral mutations during the infection,the characteristics of EV71 mutation were analyzed based on the different full-length viral genomes from epidemic regions.The viral genome was also sequenced from the infected ADAR1 knock-down cells.Results After ADAR1 knock-down,the cell viability decreased quickly after the virus infection,and formed much more and larger sizes of plaques than the control cells.The virus capsid protein VP1 expressions and virus titer in the cells culture media were both increased in ADAR1 knockdown cells.Statistic analysis showed that A-G and T-C mutations were the major mutations of EV71,which were believed to be the hot sites for RNA-editing.However,the results of viral RNA genomic sequencing data indicated that ADAR1 did not edit EV71 genome directly.Conclusions ADAR1 was a restriction factor for controlling EV71.However,ADAR1 does not directly edit EV71 genome.
6.Analysis of eight cases with perinatal pulmonary embolism
Na LIU ; Yan LONG ; Li LIN ; Jie MENG
Chinese Journal of Perinatal Medicine 2017;20(8):618-622
Objective To analyze the incidence,risk factors,clinical characteristics and pregnant outcomes of perinatal pulmonary embolism(PPE).Methods Clinical data of eight patients who were admitted to Beijing Friendship Hospital of Capital Medical University for PPE from January 2006 to March 2016 were collected.General condition,symptoms,laboratory examinations,images,treatments and outcomes of these patients were analyzed retrospectively.Results The ten-year incidence of PPE was 0.029% (8/27 560) in this hospital.Among the eight cases,two cases were diagnosed in the first trimester,and treated successfully by thrombolytic therapy.But one of two cases stopped growth,while the other one was premature labor.There were one case in the third trimester who had successful anticoagulant therapy and five cases in the postpartum period after cesarean delivery.Among the five cases,three cases were recovered after anticoagulant therapy,one case was recovered after thrombolytic therapy and one case died.All of the eight patients were immobilized before the onset of PPE,and five of them were diagnosed after cesarean section.Four out of the eight patients were obese.Five patients had three or more high-risk factors for pulmonary embolism and the other three had two.Conclusions It is necessary to pay close attention to gravidas who have two or more high-risk factors of PPE due to its fatal outcome.
7.Study on Pharmacokinetics of Ferulic Acid in Ordinary Guipi Pills and Ultramicro Guipi Pills in Rabbits
Hui GUI ; Hong YAN ; Long ZOU ; Dongwen LIU ; Guangxian CAI
China Pharmacy 2005;0(23):-
OBJECTIVE: To study the pharmacokinetic parameter and the relative bioavailability of ferulic acid in ordinary Guipi pills and ultramicro Guipi pills in rabbit.METHODS: HPLC-MS/MS was used to identify the concentration of ferulic acid in the blood sample.RESULTS: Plasma concentration of ferulic acid in ordinary pills was different from that in ultramicro pills.The main pharmacokinetic parameters of ferulic acid in ordinary pills vs.that in ultramicro pills were as follows:t1/2a:(17.59?2.57) min vs.(23.51?4.49) min;t1/2?:(300.91?38.74) min vs.(167.17?45.66) min;tmax: (9.61?1.69) min vs.(12.33?2.27) min;Cmax:(58.92?8.42) ng?mL-1 vs.(112.35?31.29) ng?mL-1;AUC: (5 851.67?895.49) ng?min-1?mL-1 vs.(8 586.13?1 497.62) ng?min-1?mL-1.There were significant difference in Cmax and AUC between ordinary pills and ultramicro pills.CONCLUSION: The application of super fine crushing technique in the preparation of Guipi pills could improve the bioavailability of Guipi pills.
8.Construction and expression of small interfering RNA expression vector of CXCR4
Yan LIU ; Long WANG ; Dawei ZHANG ; Deming ZHAO
Journal of Jilin University(Medicine Edition) 2006;0(06):-
Objective To construct the recombinant plasmid carrying small interfering RNA(siRNA) to CXCR4 and observe dumbness effect of chemokine receptor gene CXCR4 suppression by RNA interference(RNAi),and explore the new method of more efficacious therapeutic possibilities for HIV-1.Methods Small interfering RNAs targeting CXCR4 gene were designed by using software.The complement form was obtained by annealing and interted into vector Psilencer3.1-H1neo.After sequencing,MT4 cells were transfected using the plasmid vector.RT-PCR and flow cytometry detection were used to evaluate the suppression of CXCX4 expression in different groups.Results The recombinant plasmid had the correct special fragments and DNA sequence detected by PCR,electrophoresis and DNA sequencing;The siRNA obviously suppressed the expression of CXCR4.When transfected with plasmid vector for 48 h,the inhibitory rate was 70%,compared with control and negative groups,there were significant differences(P
9.The prognostic value of serum HE4 in lung cancer
Yigong ZHANG ; Yan ZHENG ; Long JIANG ; Zerui ZHAO ; Wanli LIU ; Hao LONG
Chinese Journal of Radiological Medicine and Protection 2014;34(6):423-426
Objective The study was designed to investigate the serum levels of HE4 in patients with lung cancer,and explore its prognostic value.Methods Blood samples of 106 healthy adults and 191 patients with lung cancer before treatment were measured by means of ELISA for HE4 levels in different stages and pathological types,for analyzing prognostic effect of HE4.Results The serum levels of HE4 in patients with lung cancer (median level 91.63 pmol/L) were significantly higher than control group (median level 56.42 pmol/L) (U =3 081.00,P < 0.05).AUC of serum HE4 was 0.85,with a cut-off value of 82.70 pmol/L (specificity =95.31%,sensitivity =62.32%).HE4 expression was not statistically related to clinical stage and pathological types of lung cancer.The median overall survival (mOS) was 36.87 months in the HE4-low group and 30.43 months in the HE4-high group (P <0.05),respectively.HE4 level was an independent prognostic factor for overall survival (HR =2.15,95% CI 1.49-3.12,P < 0.05).Conclusions The prognosis of patients with high HE4 expression is poor.Therefore,it might be necessary for patients with high level of HE4 to receive more aggressive adjuvant therapy.
10.Effect of miR-218 on biological functions of renal cell carcinoma cell line
Long HE ; Hongwei YANG ; Yan ZHANG ; Jun WANG ; Long LIU ; Lianhui FAN
Cancer Research and Clinic 2014;26(10):649-653
Objective To explore the effects of miR-218 on biological functions of renal cell carcinoma cell line through regulating the expression levels of miR-218.Methods The pcDNA3.1-miR-218 was transfected into renal carcinoma cell line A498 and 769-P and the expression of miR-218 was detected.The cell activity,invasion,apoptosis and proliferation of transfected renal carcinoma cell line were analyzed in vitro.Results After plasmid transfected A498/769-P renal carcinoma cell line,the expression level of miR-218 (1.99,1.64) was significantly higher than that of the control group (1.00) (t =60.82,10.89,P < 0.000 1) The cell viability (0.90±0.10,0.68±0.06) was lower than that of the control group (1.39±0.14,1.24±0.08) by CCK8 experiment (t =15.02,31.69,P < 0.000 1).The cell invasion was lower than that of control group by Transwell experiment (t =15.78,18.80,P < 0.000 1).The apoptosis ratio was higher by AnnxinV-FITC experiment,the apoptosis ration of control group and transfected group were respectively 0.25 %,45.77 % in A498 cell line and 0.11%,45.57 % in 769-P cell line.The proliferation was lower than that of the control group (P < 0.000 1).Conclusion Up-regulation of miR-218 expression can inhibit the growth of renal cell carcinoma cell line in vitro.