1.ATP1A1 gene silencing inhibits the invasion of human glioma cells U251 in vitro
Hong CHEN ; Song CHEN ; Gang HUO
Basic & Clinical Medicine 2017;37(4):506-511
Objective To investigate the effect and mechanism of silencing ATP1A1 gene on invasion ability of human U251 glioma cells.Methods The human U251 glioma cells were infected with lentivirus expressing shRNA-ATP1A1.The mRNA and protein expression of ATP1A1 in U251 glioma cells was detected by RT-qPCR and Western blot,respectively.The proliferation of U251 glioma cells was determined by MTT assay.The migration and invasion ability were detected by cell scratch test and Transwell chamber.The protein expression of matrix metallo proteinase 2 (MMP-2) and MMP-9 in U251 glioma cells were detected by Western blot.Results The mRNA and protein expression of ATP1A1 in the silence group were significantly inhibited,The ability of proliferation, migration and invasion were also significantly inhibited (P<0.05),The expression of MMP-2 and MMP-9 were also significantly reduced,there was a significant difference(P<0.05).Conclusions RNA interference targeting ATP1A1 gene can significantly inhibit the proliferation, migration and invasion of glioma U251 cells.The mechanism might be related to the down-reguLation of MMP-9 and MMP-2.ATP1A1 can be used as a potential target for the treatment of glioma.
2.Review of Studies on Azo Dyes Decolorization
Gang CHEN ; Liang CHEN ; Man-Hong HUANG ;
Microbiology 2008;0(07):-
Biological process is an important approach to treat the dye wastewater. The azo dyes decolouration by special azoreductase of different aerobic bacteria and fungi was summarized. Under anaerobic condition, reductive decolourization of azo dyes was carried in the presence of redox mediators which act as electron shuttle. It was also pointed out that azo dye reduction occurred mainly under anaerobic condition. Different electron donor resulted in different decolourization rate. Problems of current biotechnology were analyzed and corresponding measures were discussed.
3.Effect of different doses of propofol on cognitive function after chronic cerebral ischemia-induced injury in aged rats
Gang CHEN ; Weidong MI ; Hong ZHANG
Chinese Journal of Anesthesiology 2011;31(6):720-722
Objective To investigate the effects of different doses of propofol on cognitive function after chronic cerebral ischemia-induced injury in aged rats. Methods Eighty male SD rats, aged 18 months, weighing 400-500 g, were randomly divided into 4 groups ( n = 20 each): shame operation group (group S), chronic cerebral ischemia group (group I), two propofol groups (groups P1 and P2 ). The chronic cerebral ischemia was induced by permanent occlusion of bilateral common carotid arteries. On 1 day after operation, intraperitoneal normal saline 2.5 ml was injected twice a day for7 consecutive days in groups S and I, and intraperitoneal propofol 10 and 50 mg/kg in 2.5 ml of normal saline were injected twice a day for 7 consecutive days in groups P1 and P2 respectively. On 3rd and 33rd days after the last injection (T1.2), 10 rats in each group underwent Morris water maze test to assess the cognitive function. After the test was completed, the rats were sacrificed and the hippocampi were removed and sliced (450-500 μm thick). Schaffer lateral branch in CA1 region was stimulated to induce long-term potentiation (LTP). Results Compared with group S, the escape latency was significantly prolonged, the number of animals' swimming across the platform, the ratio of the swimming time spent in the forth quadrant to the total swimming time, and the success rate of LTP induction were significantly decreased at T1 and T2 in groups I, P1 and P2 (P < 0.05). Compared with group I, the escape latency was significantly prolonged, the number of animals' swimming across the platform, the ratio of the swimming time spent in the forth quadrant to the total swimming time, and the success rate of LTP induction were significantly decreased at T1 in groups P1 and P2, and at T2 in group P2 ( P < 0.05). Conclusion Propofol aggravates the damage to cognitive function while it attenuates the chronic cerebral ischemia-induced injury in aged rats, especially the high dose.
5.Application of Leucocyte Adherence Inhibition Test in Determining Immunological Activity of Ribonucleic Acid
Wang CAN ; Shao HONG ; Chen GANG
China Pharmacist 2015;(11):1848-1850
Objective:To establish a method for determining the immunological activity of ribonucleic acid. Methods: Leucocyte adherence inhibition test ( LAI) was applied, and the important parameters of LAI including the mouse strain, drug concentration, treatment time, content of buffer solution and cell density were researched. The immunological activity of RNAⅠ, Ⅱand Ⅲ was re-spectively determined by the method. Results:Stable and reliable parameters were obtained: the sample concentration was 10 mg· ml-1 , the treatment time was 2 hours, Ca2+ and Mg2+ were necessary for the buffer solution, and the cell density was about 4 × 107 cell·ml-1 . The strain of mouse showed no effect on the results. As a result, the determination method for immunological activity was established. Using the method, the immunological activity of RNA Ⅰ,Ⅱand Ⅲ was determined 3 times, and the results met the re-quirements with RSD below 20%. Conclusion:The method is suitable for determining the immunological activity of RNA.
6.Alleviating Japanese encephalitis virus infection with multiple receptor tyrosine kinase inhibitor sunitinib
ZHANG Chen ; AI Jun-hong ; AN Jing ; WANG Pei-gang
China Tropical Medicine 2022;22(10):947-
Abstract: Objective To evaluate the effects of sunitinib on Japanese encephalitis virus (JEV) infection in vitro and vivo. Methods The 4-week-old BALB/c mice infected with JEV by intraperitoneal injection. The infected mice were treated with sunitinib for 5 days and 10 days respectively. After that, the change of weight and survival rate were evaluated continuously. The viral load variation in mice brain were detected by qRT-PCR. Indirect immunohistochemical staining assay (IFA) was used to detect the number and distribution of CD4+/CD8+T cells in mouse brain. IFA was also used to observe the expression of virus E protein in the brain of mice. Vero cells were infected with JEV in vitro and given a certain concentration of sunitinib to observe the cell survival status. The expression of virus E protein in cells was detected by IFA. Results Continuous administration of sunitinib significantly improved the survival rate of infected mice. Survival rate and body weight changes showed that the 5-day's administration strategy was significantly better than the 10-day's administration strategy. The treatment of sunitinib decreased the infiltration of CD4+/CD8+T cells in the brain and reduced the changes of vascular sleeve. However, the variation of viral load and E protein expression in brain were not obvious. The cytopathic effect (CPE) of infected Vero cells were slightly relieved after giving sunitinib, and the expression of E protein was also slightly changed. Conclusion Sunitinib can significantly reduce the mortality of infected mice, and the 5-day's administration strategy is significantly better than the 10-day's administration strategy. Sunitinib decrease T lymphocyte infiltration in brain of mice, relieve the encephalitis symptoms ,and prolonged the life of mice.
7.Characteristics of HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou
ZHAO Gang ; ZHANG Xingliang ; CHEN Junfang ; XU Ke ; WU Hong
Journal of Preventive Medicine 2021;33(12):1209-1214
Objective :
To analyze the characteristics of HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou, and the influencing factors for new infection and local infection, so as to provide the evidence for AIDS prevention and control.
Methods :
From 2017 to 2019, the newly reported HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou were recruited, and their demographic information, previous sexual behaviors and history of HIV testing were collected in the questionnaire survey. The multivariate logistic regression model was used to analyze the influencing factors for new infections and local infections.
Results :
A total of 522 participants from 668 newly reported HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou during this period were surveyed.Among 522 cases, 263 ( 50.38% ) were aged 40 years or above, 218 ( 41.76% ) were married, 326 ( 62.45% ) had an educational level of junior high school or below, and 340 ( 65.13% ) were not local. Among 504 cases whose infection time could be determined, 72 ( 14.29% ) were newly infected within one year; age of 40 years below ( OR=4.148, 95%CI: 1.956-8.795 ), history of HIV testing ( OR=2.049, 95%CI: 1.163-3.609 ) and history of sexually transmitted diseases ( OR=2.169, 95%CI: 1.076-4.374 ) were risk factors for new infection. Among 454 cases whose infection location could be determined, 267 ( 58.81% ) were infected in Hangzhou; educational level of high school or below ( OR=2.538, 95%CI: 1.252-5.145 ) , Hangzhou residence ( OR=7.835, 95%CI: 4.227-14.353 ), living in Hangzhou for a year or over ( OR=18.960, 95%CI: 8.755-41.060 ) and monthly income of 3 000 yuan or over ( OR=2.630, 95%CI: 1.546-4.474 ) were risk factors for local infection.
Conclusions
The HIV/AIDS cases with non-marital or non-commercial heterosexual transmission in Hangzhou are mainly floating population and less educated. The newly infected cases are more likely to be young and middle-aged people and patients with sexually transmitted diseases, the locally infected cases are more likely to be people with permanent residence, less educated and high income.
10.Effect of GABA on expression of VEGF and MMP-9 in monocrotaline-induced pulmonary hypertension rats
Gang CHEN ; Jun SONG ; Hong XIONG ; Lujie ZHANG ; Jinquan LV
Chongqing Medicine 2014;(30):4043-4045
Objective To study the therapeutical effect of γ-aminobutyric acid on monocrotaline(MCT )induced pulmonary hy-pertension rats ,and to elucidate the expression of VEGF and MMP-9 .Methods Thirty SD rats were randomly divided into 3 groups :a normal control group(control group) ,a MCT-induced pulmonary hypertension group(model control group) ,and anγ-ami-nobutyric acid treatment group(treatment group) .The mean right ventricular pressure(mRVP)were detected ,the right ventricular hypertrophy index(RVHI)were measured ,WT% and WA% were evaluated ,and the expression of VEGF mRNA in the lung tissue and MMP-9 were detected wtih FQ-PCR and immunohistochemical staining method respectively .Results mRVP ,RVHl ,WT% , WA% and the expression of MMP-9 and VEGF mRNA of treatment group were lower than those in the model control group(P<0 .05) ,but higher than the control group(P<0 .05) .Conclusion GABA has a therapeutic effect on pulmonary hypertension rats through regulating the expression of VEGF mRNA and MMP-9 protein .