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.
3.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%.
7.Mirror therapy combined with acupuncture can improve the upper extremity function of stroke survivors
Rui ZHANG ; Meilan ZHU ; Ying YU ; Bangliang LIU ; Lianhua WU
Chinese Journal of Physical Medicine and Rehabilitation 2017;39(8):588-593
Objective To explore the effects of mirror therapy combined with Tongdu Xingshen acupuncture on the upper extremity function of stroke survivors.Methods Sixty stroke survivors were randomly divided into a mirror group (n =20),an acupuncture group (n =20) and a combined group (n =20).In addition to routine rehabilitation treatment,those in each group received mirror therapy,acupuncture or both for 4 weeks.Before the treatment and after 2 and 4 weeks of treatment,all of the patients were assessed using the Fugl-Meyer motor function assessment (FMA),Brunnstrom upper limb and hand staging (BSULH),the Barthel Index (BI) and in terms of the active range of motion in the wrist contralateral to their hemiplegia (AROM).The combined group was also evaluated using functional magnetic resonance imaging (fMRI) before treatment and after the 4 weeks.Results After 2 and 4 weeks of the treatment,the average BI,AROM,FMA and BSULH results had improved significantly compared with before the treatment.The improvements in the combined group were significantly greater than in the other groups after 2 and 4 weeks.The fMRI demonstrated that there were high-intensity signals in the primary motor area,the premotor area and the supplementary motor area on the affected side after 4 weeks of treatment.Conclusions Mirror therapy combined with Tungdu Xingshen acupuncture can improve the functional performance of the upper extremities of stroke survivors.
8.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.
9.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.
10.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.