1.Gene therapy of human ovarian carcinoma in vitro
Pu LI ; Dongtian LI ; Bingnan YIN ; Hua TANG ; Lirong YIN
Chinese Journal of Clinical Oncology 2014;(13):861-865
To construct a recombinant adenovirus containing CDglyES fusion gene, which can directly inhibit human ovarian cancer cell and indirectly inhibit vascular endothelial cell growth. Methods:We constructed prAdCDglyES using a homolo-gous recombination method in bacteria. The prAdCDglyES was transfected to 293 packaging cells using liposome, in which rAdCDgly-ES was packaged and amplified. MTT was used to observe the proliferation inhibition effect of rAdCDglyES on human ovarian cancer cells and the growth inhibition effect of expressing products of rAdCDglyES on ECV-304. Results:The titer of rAdCDglyES was 1 × 1013.3 TCID50/L, whereas the inhibition rate on human ovarian cancer cell SKOV-3 was (83.1±6.3)%. This result is significantly different from the control rAd-LacZ, which had an inhibition rate of (24.1 ± 13.2)% (P<0.01). The concentrated culture supernatant from cells transfected with rAdCDglyES can inhibit ECV-304 cell proliferation at a rate of (78.7 ± 1.6)%. This rate is significantly different com-pared with that of the control with the same concentration of culture supernatant from cells transfected with rAd-CD, with an effect on ECV-304 cell shown by an inhibition rate of (23.9 ± 9.7)%(P<0.01). Conclusion:The results showed that the recombinant adenovirus rAdCDglyES could inhibit human ovarian cancer cells directly and indirectly.
2.The Inhibitory Effect of CDES Gene on Mammary Cancer Mediated by Adenovirus
Qiouxiang LI ; Dongtian LI ; Huichun TONG ; Bingnan YIN ; Guangming LI
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective: To construct a recombinant adenovirus(rAdCDES) which is capable of both direct and indirect treatment to mammary cancer and enhancement to the antitumor effect of radiation. Methods: A method of homologous recombination in bacteria was used to construct prAdCDglyES. The recombination adenovirus was transfected to 293 cells by liposome, in which rAdCDES was packaged and generated. The growth curve and MTT methods was used to detect the growth inhibition effect of rAdCDES on MCF-7; rAdCDES was directly injected into established MA737 tumors-bearing mice for observing difference in tumor size and survival days of mice and enhancement of the antitumor effect of radiation. Results: The inhibiting rate of rAdCDES on MCF-7 cell was (83. 1?8. 1)% and had significant difference compared with control was (19.2 ? 7.8)% (P
3.Establishment of the method for simultaneous determination of levetiracetam and carbamazepine in human plasma
Min SHI ; Bingnan REN ; Huizhen WU ; Ping LIU ; Yin WU
China Pharmacy 2022;33(8):987-991
OBJECT IVE To e stablish the method for simultaneous determination of levetiracetam and carbamazepine concentrations in human plasma. METHODS After plasma samples were precipitated with methanol ,using carbamazepine-D 10 as the internal standard ,the concentrations of levetiracetam and carbamazepine were determined by high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The determination was performed on XBridge BEH C 18 column with methanol-0.1% formic acid as mobile phase (gradient elution )at the flow rate of 0.35 mL/min. The column temperature was set at 40℃,and sample size was 2 μL. With electrospray ion source,a multiple reaction monitoring mode was used for positive ion scanning;the detected ion pairs for quantitative analysis were m/z 171.3→126.3(levetiracetam),m/z 237.1→194.1(carbamazepine), 247.1→204.1(internal standard ). RESULTS The linear range of the concentrations of levetiracetam and carbamazepine were 0.5-50 and 0.2-20 μg/mL(r=0.997 3 and 0.998 5),respectively;the lower quantitative limits were 0.5 and 0.2 μg/mL,respectively. RSDs of intra-day and inter-day were all no more than 10.00%. RE of intra-day and inter-day were within ±4.00%;the average extraction recoveries rate were 95.60%-105.00%;the average internal standard correction matrix factors were 98.40%-110.00%; RSDs of stability tests were all not higher than 5.60%. The concentrations of levetiracetam and carbamazepine in the plasma of 22 patients measured by this method were 3.36-40.90 and 3.64-9.93 μg/mL,respectively. CONCLUSIONS The established method for determining the concentration of levetiracetam and carbamazepine in human plasma is f ast,sensitive,accurate and stable ,and can be used for the monitoring of plasma concentration and pharmacokinetic study in epilepsy patients.