1.Inhibition of human lung adenocarcinoma A549 cells by silencing survivin gene with RNAi
Meng RUI ; Changting LIU ; Xiaodan YU
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
0.05).The mRNA and protein expressions of survivin in cells transfected by 100-200nmol/L siRNA were significantly lower than those of untransfected cells(P0.05).Conclusion Silencing survivin gene by the RNAi technique can decrease effectively the expressions of survivin gene and protein,suppress significantly the growth and proliferation of A549 cells,and induce apoptosis of A549 cells.The inhibitory effect of RNAi is characterized by its specificity,high efficiency and durability.This may lay an experimental foundation for further study of gene therapy in lung cancer.
2.QuEChERS-Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry for Determination of 2,4,6-Trinitrophenol, 2,4,6-trinitroresorcinate and 2,4,6-Trinitrophloroglucinol in Soil
Yu LIU ; Tonglai ZHANG ; Li YANG ; Bidong WU ; Rui LIU
Chinese Journal of Analytical Chemistry 2014;(8):1184-1189
A simple, rapid sensitive and accurate ultra performance liquid chromatography tandem mass spectrometric ( UPLC-MS/MS ) method was developed for the simultaneous determination of 2 , 4 , 6-trinitrophenol, trinitroresorcinate, trinitrophloroglucinol residues in soil. The sample was pretreated by using the modified QuEChERS ( quick, easy, cheap, effective, rigged, and safe ) method that completed the extraction and clean-up steps in one procedure. In this none buffer QuEChERS method, samples were extracted with 1% formic acid ﹢ acetonitrile, cleaned up with primary graphitized carbon black ( GCB) and C18 sorbent, then centrifuged and filtrated before detection. The pretreatment method was simple, rapid and effective and can meet the detection requirements. The UPLC-MS/MS method was performed on Waters Accucore PFP (150 mm× 2. 1 mm, 2. 6 μm) and the column temperature was 30 ℃, the gradient elution with acetonitrile and ammonium acetate as the mobile phase and the flow rate was 0. 3 mL/min. The negative electrospray ionization ( ESI-) source under the multiple reaction monitoring ( MRM ) mode and external standard method were used for quantification. The results showed that the correlation coefficients up to 0. 9942 were obtained across a concentration range of 0. 005-5. 0 mg/L. The limits of detection ( LOD) ranged from 0 . 002 to 0 . 005 mg/kg ( S/N=3 ) . The method was validated with soil samples spiked at three fortification levels (0. 01, 0. 1 and 1 mg/kg) and recoveries were in the range of 79. 3%-94. 8% with relative standard deviations (RSD) of 3. 1%-6. 6%.
5.Antiviral Activity of Recombinant Cyanovirin-N against HSV-1
Hong YU ; Zongtao LIU ; Rui LV ; Wenqing ZHANG
Virologica Sinica 2010;25(6):432-439
In this study,a standard strain of HSV-1(strain SM44)was used to investigate the antiviral activity of the recombinant Cyanovirin-N(CV-N)against Herpes simplex virus type 1(HSV-1)in vitro and in vivo.Cytopathic effect(CPE)and MTT assays were used to evaluate the effect of CV-N on HSV-1 in Vero cells.The number of copies of HSV-DNA was detected by real-time fluorescence quantitative PCR(FQ-PCR).The results showed that CV-N had a low cytotoxicity on Vero cells with a CC50 of 359.03±0.56 μg/mL,and that it could not directly inactivate HSV-1 infectivity.CV-N not only reduced the CPE of HSV-1 when added before or after viral infection,with a 50% inhibitory concentration(IC50)with 2.26 and 30.16μg/mL respectively,but it also decreased the copies of HSV-1 DNA in infected host cells.The encephalitis model for HSV-1 infection was conducted in Kunming mice,and treated with three dosages of CV-N(0.5,5 & 10 mg/kg)which was administered intraperitoneally at 2h,3d,5d,7d post infection.The duration for the appearance of symptoms of encephalitis and the survival days were recorded and brain tissue samples were obtained for pathological examination(HEstaining).Compared with the untreated control group,in the 5mg/kg CV-N and 10mg/kg CV-N treated groups,the mice suffered light symptoms and the number of survival days were more than 9d and 14d respectively.HE staining also showed that in 5mg/kg CV-N and 10mg/kg CV-N treated groups,the brain cells did not show visible changes,except for a slight inflammation.Our results demonstrated that CV-N has pronounced antiviral activity against HSV-1 both in vitro and in vivo,and it would be a promising new candidate for anti-HSV therapeutics.
6.Therapeutic effect of umbilical cord-derived mesenchymal stem cell transplantation in systemic lupus erythematosus patients with different patterns of syndromes
Yu TANG ; Ruixia LIU ; Yingying QIU ; Jinbing RUI ; Jing LI
Chinese Journal of Tissue Engineering Research 2015;(45):7298-7303
BACKGROUND:Systemic lupus erythematosus (SLE) is classified into four types, and the major treatment is to tonify kidney and nourishyin, clear blood stasis and toxin by the traditional Chinese medicine (TCM). Even though, there are stil many patients with poor efficacy. Mesenchymal stem cels have the capacity of multiple differentiation, hematopoietic support and immune regulation, thus having been used for the treatment of refractory, recurrent SLE and achieving good effects. OBJECTIVE:To investigate the therapeutic effect of umbilical cord-derived mesenchymal stem cel transplantation on SLE patients with different patterns of syndromes. METHODS: Twenty-one SLE patients were clustered to four syndrome types of TCM, including heat-toxin,yin deficiency of liver and kidney,yang deficiency of spleen and kidney, andqi stagnation and blood stasis. The changes in clinical and laboratory indicators were analyzed statisticaly before and after cel transplantation. RESULTS AND CONCLUSION:The level of 24-hour proteinuria and SLE disease activity index scores in SLE patients were significantly decreased at 1, 3, 6 months after cel transplantation (P < 0.01). Umbilical cord-derived mesenchymal stem cel transplantation could significantly reduce the 24-hour proteinuria in SLE patients withyin deficiency of liver and kidney at 1, 3 and 6 months (P < 0.01), while slightly reduce the 24-hour proteinuria in SLE patients with heat-toxin andqi stagnation and blood stasis at 1, 3 months (P < 0.05) as wel as in SLE patients withyang deficiency of spleen and kidney at 1 month (P < 0.05). Additionaly, umbilical cord-derived mesenchymal stem cel transplantation could increase the serum albumin levels in al the SLE patients (P < 0.01), although the changes in patients with heat-toxin were moderate (P < 0.05). Al the SLE patients of four types had an increasing trend of their platelet counting after cel transplantation, but there was no statistical difference before and after cel transplantation. Taken together, umbilical cord-derived mesenchymal stem cel transplantation is effective for treatment of SLE, but has different therapeutic efficacy on SLE patients with different syndrome types of TCM.
7.Evaluation of the Vertebral Artery Anatomic Course with Multi-slice Spiral CT and Its Clinical Application
Rui HUA ; Jun LIU ; Jin ZHONG ; Yu ZHANG
Journal of Practical Radiology 2000;0(12):-
Objective To evaluate the value and the significance of 16-slice spiral computed tomography angiography(MSCTA) in identifying the vertebral artery course variation.Methods MSCTA data in 22 patients with anomalous path of vertebral artery were retrospectively analyzed,13 of them had undergone DSA examinations,and 5 of them had also undergone MRI examinations.Multi-planar reconstruction(MPR),curved MPR,thin-slab maximum intensity projection(MIP),volume rendering(VR) or slab VR(cut-plane VR) were used to assess the anomalous path of the vertebral artery and its relationship with the adjacent large vessels.Virtual endoscopy(VE) was used to evaluate the orifice of the vertebral arteries and its relationship with the aortic arch and the ostia of the adjacent large vessels.Different views of vertebral artery were analyzed on DSA.MSCTA and DSA images were analyzed in blinded fashion for the accuracy of anomalous vertebral artery path and origin detection.Results were compared in a secondary consensus evaluation.Results The anomalous path of the vertebral artery in all 22 patients were correctly displayed on MSCTA,7 of them had anomalous origin of vertebral artery,the anomalous of ostia showed clearly.The vertebral arteriae with course variation were all going up at the front of the foramen transversarium of C6,then bended into the C5,C4 or C3 foramen transversarium.DSA could only be detected the anomalous origin in 13 patients undergone DAS.5 patients indirectly showed having course variation of vertebral artery by MRI.Conclusion MSCTA is superior to DSA in showing the anomalous path and orifice of vertebral artery,it should be considered as a non-invasive imaging tool for these diseases.
9.Efftects of Tianma Xingnao Capsule on Brain Blood Circulation in Mice and Rats
Rui LIU ; Yu YUN ; Jindan ZHEN ; Jiashuo ZHANG ; Zhiqiang SHEN
Journal of Kunming Medical University 2006;0(06):-
Objective To study the protective effects of Tianma xingnao capsule on brain blood circulation in mice and rats.Methods The arteria carotis externa was ligated,and arteria carotis interna was kept.The left carotid artery was exposed and placed over an ultrasonic flow probe.The brain flow volume was recorded with an Ultrasonic Volume.The pursiness time of mice was observed to evaluate the effect of Tianma xingnao capsule on cerebral ischemia and anoxia.The permeability of normal blood-cerebral barrier was investigated in mice.Results Tianma xingnao capsule significantly increased brain blood flow and prolonged mouse pursiness time.Tianma xingnao capsule at a dose of 4 g/kg significantly elevated the content of Evans in mouse brain.Conclusions Tianma xingnao capsule can increase the brain blood flow in rats and improve brain blood circulation in mice.It is obviously advantageous to protect against cerebral ischemia and hypoxia.
10.Content Determination of Related Substances in Alprostadil for Injection by HPLC
Rui DING ; Hong JI ; Si CHEN ; Yiming LIU ; Li YU
China Pharmacy 2007;0(29):-
OBJECTIVE: To establish an HPLC method for the content determination of prostaglandin A1 and prostaglandin B1 in Alprostadil for injection.METHODS: The determination was performed on Alltech Alltima C18 column with mobile phase consisted of phosphate puffer(pH=6.3)-acetonitrile-methanol(70 ∶ 25 ∶ 5) at a flow rate of 1.5 mL? min-1.The detection wavelength was set at 196 nm.The column temperature was set at room temperature and the injection volume was 20 ?L.RESULTS: The prostaglandin A1 and prostaglandin B1 were well separated from main component and other impurities.The linear range of prostaglandin A1 and prostaglandin B1 were 0.175~19.00 ?g?mL-1(r=0.999 7) and 0.23~19.90 ?g?mL-1(r=0.999 2).The contents of prostaglandin A1 in 3 batches of samples were 4.7%,4.9% and 4.3%,and the contents of prostaglandin B1 in 3 batches of samples were 0.6%,0.8% and 0.5% respectively.CONCLUSIONS: This method is proved to be simple,specific and suitable for the content determination of related substances in Alprostadil for injection.