AIM: To explore the target relationship between microRNA-140-3p (miR-140-3p) and programmed cell death ligand 1 (PD-L1) and their effect on the viability, migration and invasion of non-small-cell lung cancer A549 cells.METHODS: RT-qPCR was used to detect the miR-140-3p expression in HLF-1, A549 and H1299 cells, and then the A549 cells with the most significant difference were selected as the subsequent research object. TargetScan software and dual-luciferase reporter assay were performed to predict and confirm the target relationship between miR-140-3p and PD-L1. RT-qPCR and Western blot were used to determine the effects of miR-140-3p mimic and inhibitor on PD-L1 expression level. MTT assay was used to detect the viability of A549 cells. Transwell assay was performed to detect the migration and invasion abilities of the A549 cells.RESULTS: miR-140-3p was significantly down-regulated in the A549 cells and H1299 cells (P<0.05). Transfection with miR-140-3p mimic decreased the expression of PD-L1 and inhibited the viability, migration and invasion of the A549 cells. Transfection with pcDNA3.0-PD-L1 reversed the inhibitory effect of miR-140-3p on the viability, migration and invasion of the A549 cells.CONCLUSION: miR-140-3p inhibits the viability, migration and invasion of A549 cells by targeting PD-L1. 相似文献
AIM: To investigate the effect of microRNA-204 (miR-204) on the proliferation of Hodgkin lymphoma cells and the underlying mechanism. METHODS: The expression of miR-204 and Sirt1 mRNA in Hodgkin lymphoma tissues was detected by RT-qPCR. After transfection with miR-204 mimic, Sirt1 siRNA and miR-204 mimic+pcDNA3.1-Sirt1 into the L428 cells, the cell viability and BrdU incorporation were measured by CCK-8 assay and BrdU assay, respectively. The protein levels of Sirt1 and acetylated p53 (ac-p53) were determined by Western blot.The targeting relationship between miR-204 and Sirt1 was verified by double luciferase reporter assay. RESULTS: The low expression of miR-204 and the high mRNA expression of Sirt1 were found in the Hodgkin lymphoma tissues. Compared with control group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were significantly decreased after L428 cells were transfected with miR-204 mimic or Sirt1 siRNA (P<0.05). Compared with miR-204 mimic alone group, the cell viability, BrdU incorporation and the protein levels of Sirt1 and ac-p53 were increased after L428 cells were co-transfected with miR-204 mimic and pcDNA3.1-Sirt1 (P<0.05). The results of double luciferase reporter assay confiermed that Sirt1 was the target gene of miR-204. CONCLUSION: The inhibitory effect of miR-204 on the proliferation of L428 cells may be achieved by inhibiting the expression of Sirt1 and promoting the up-regulation of ac-p53. 相似文献
Differential accumulation of [14C]imazalil and [14C]fenarimol by germlings of wild-type and DMI-resistant isolates ofPenicillium italicum was studied at various pH values. At pH 7 and 8 the low-resistant isolate E300–3 accumulated 22% and 35%, respectively, less imazalil than the wild-type isolate W5. Imazalil accumulation at pH 5 and 6 was similar. Isolate E300–3 also accumulated less fenarimol as compared with the wild-type isolate. This difference was much more obvious than for imazalil and was observed at all pH values tested. Differences in accumulation of both imazalil and fenarimol between low (E300–3), medium (H17) and high resistant (I33) isolates were not observed. These results suggest that decreased accumulation of DMIs is responsible for a low level of resistance only and that additional mechanisms of resistance might operate in isolates with a medium and high degree of resistance. With all isolates fenarimol accumulation was energy-dependent. This was not obvious for imazalil.The wild-type and DMI-resistant isolates had a similar plasma membrane potential as determined with the probe [14C]tetraphenylphosphonium bromide ([14C]TPP+). Various test compounds, among which ATPase inhibitors, ionophoric antibiotics and calmodulin antagonists, affected the accumulation of [14C]TPP+, [14C]imazalil and [14C]fenarimol. No obvious correlation between the effects of the test compounds on accumulation levels of the fungicides and [14C]TPP+ could be observed. These results indicate that the plasma membrane potential does not mediate the efflux of DMI fungicides byP. italicum. 相似文献
Chicks and chickens maintained under commercial conditions were vaccinated against Newcastle Disease via drinking water. Prior and after different times of vaccination blood samples were drawn from different numbers of birds and checked for HI antibodies. The modes of distribution of the antibody titers within the random sample were assayed. The following results were obtained:
1. 1. On the basis of the distribution of the serum titers can be concluded whether the antibody level within a flock is increasing or decreasing.
◦ —A right steep asymmetry can be observed up to 20 days post vaccination.
◦ —In the phase of maximal antibody levels an almost symmetric distribution of the titers is present.
◦ —In later times (more than three weeks p. vacc.) the distribution shows a left steep asymmetry (Poisson distribution).
2. 2. A poisson distribution is also observable during the elimination of maternal antibodies of chicks until complete elimination.
3. 3. The mode of distribution of the HI titers in sera of day-old chicks correlates with the mode of distribution of the dams. Therefore, conclusions are possible from the status of the chicks to the dams and reverse.
4. 4. Factors which interfere with the mode of distribution are:
◦ —Two or more peaks after vaccination of chickens. This indicates an uneven immune response within the flock.
◦ —Distributions with several peaks may also occur if flocks are composed of day-old chicks from parent flocks with different levels of antibody titers.
Résumé
Des poulets et des poussins maintenus dans un élevage conventionnel furent vaccinés contre la maladie de Newcastle par un vaccin administré dans l'eau de boisson. Avant et à différents temps après vaccination, des échantillons de sang furent prélevés sur un certain nombre de poussins et les anticorps inhibants de l'hémagglutination furent recherchés. Les modes de la distribution des titres en anticorps pour les échantillons pris au hasard furent recherchés. Les résultats suivants furent obtenus:
1. 1. Sur la base de la distribution des titres sériques, on peut conclure si le taux en anticorps à l'intérieur d'une population a augmenté ou diminué.
◦ —Une courbe asymétrique avec un pic déplacé vers la droite peut être observée 20 jours après la vaccination.
◦ —La phase correspondant au taux maximal en anticorps présente une distribution presque normale.
◦ —Plus tard, (au-delà de 3 semaines après la vaccination), la distribution apparaît asymétrique avec un déplacement vers la gauche (distribution de Poisson).
2. 2. Une distribution de Poisson peut aussi être observée au moment de l'élimination des anticorps d'origine maternelle chez des poussins jusqu'à complète élimination.
3. 3. Le mode de distribution des titres en anticorps inhibant l'hémagglutination dans des sérums de poussins d'un jour correspond au mode de distribution observé chez les mères. Des conclusions peuvent donc être faites à partir de l'état immunologique des poussins vis-à-vis des mères et viceversa.
4. 4. Les facteurs qui peuvent intervenir dans le mode de distribution sont:
◦ —Deux pics ou plus après la vaccination des poussins. Ceci indique qu'il existe une réponse immunitaire inégale dans la population.
◦ —Des distributions avec plusieurs pics peuvent être également observées si les populations sont composées de poussins de un jour provenant de populations parentales ayant des taux différents de titres en anticorps.
AIM and METHODS:To analysis the factor that involved in renal carcinogenesis, we used the bait gene AK001518 to screen GenBank. To understand the relationship between cell cycle related gene(CCRG) and p15, we did RT-PCR and Northern Blot experiments. Then we examined CCRG expression level in renal carcinogenesis. RESULTS:Gained a function unknown gene CCRG that was 67% a mino acid identical with the gene AK001518 that was regulated by p15. It was shown that the CCRG mRNA was dramatically decreased when p15 gene was over-expressed. CCRG expression level was much higher in tumor tissues and cells than normal tissues and cells. CONCLUSION:The novel gene CCRG expressed highly in the renal carcinoma, which might play a significant role in the renal carcinogenesis. 相似文献
AIM:To investigate the protein expression of cyclin D2 and p16 in proliferation and differentiation of cultured cardiac myocytes.METHODS:One-day-old Sparague-Dawley rats were used. Cardiac myocytes(CM) were collected by a trypsin-dispersal method and cultured. Cell growth line and fluorescence activated cell sorting (FACS) were used to investigate the proliferation of CM. Ultra-thin sections were made to observe the ultrastructure of CM under transmission electron microscope. The expression of cyclin D2 and p16 in CM were measured using immunocytochemistry and image analysis.RESULTS:①Results of cell growth line and FACS analysis showed that cultured CM could proliferate in the first 3 cultured days, but the ability decreased quickly, concomitant with differentiation. CM was obseved quiescent in cell cycle three days later. The ultrastructure of CM showed the large amount of myofilaments and mitochondrion. ②The protein expression of cyclin D2 in 3,4,5 day CM group was 0.89 times(P＜0.05),0.80 times (P＜0.05) and 0.56 times (P＜0.01) of that in 1 day group, respectively. The expression of p16 in CM was increased during the culture process, 2,3,4,5 day group were 1.63 times, 1.72 times, 1.99 times and 2.84 times (P＜0.01) of that in 1 day group, respectively.CONCLUSION:Cultured neonatal rat cardiac myocytes could proliferate during the first 3 days after incubation, but the ability of proliferation decreased, from the fourth day, concomitant with differentiation. Cyclin D2 and p16 play the key roles in CM postnatal development. Downregulation of cyclin D2 and upregulation of p16 may induce CM differentiation. 相似文献
AIM: In order to study the relationship between the ERK and p38 MAPK activation and the protection of 11, 12-epoxyeicosatrienoic acid (11, 12-EET) and ischemia preconditioning (IP), the effects of 11, 12-EET and ischemic preconditioning on phosphorylated ERK and p38 MAPK during ischemia and reperfusion in rat myocardium were examined. METHODS: The rat heart was subjected to ischemia for 5 min by ligating the left anterior descending coronary artery followed by reperfusion for 5 min (two times) to undergo ischemia preconditioning. The rats were divided into 5 groups: (1) control; (2) sham group; (3) ischemia/reperfusion (I/R) group, in which the rat heart suffered from 60 min ischemia followed by 30 min reperfusion; (4) IP plus I/R group; (5) EET plus I/R group, in which 6.28×10-8 mol/L 11, 12-EET was injected intravenously 20 min before I/R. The heart function was examined, and phosphorylated ERK and p38 MAPK were detected by Western blot. RESULTS: At 30 min reperfusion, +dp/dtmax, -dp/dtmax and LVDP decreased significantly in I/R group compared with sham group, IP plus I/R group and EET plus I/R group; Phosphorylated ERK1/2 level was higher in I/R group than sham group, but was lower in I/R group than IP plus I/R group and EET plus I/R group; Phosphorylated p38 MAPK level was lower in control, sham, IP plus I/R and EET plus I/R group than I/R group. CONCLUSION: 11,12-EET protects rat heart against ischemia/reperfusion injury, the mechanism may be related to activation of ERK1/2 and inhibition of p38 MAPK. 相似文献
