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microRNA (miRNA)is a kind of non-protein-coding small RNA that is different from mRNA. miRNAs interact with the 3’ untranslated region (3’UTR )of the target mRNA,resulting in either complete degradation or translational inhibition of the target mRNA.miRNAs play a vital role in tumorigenesis,growth, migration and invasion.The regulation of miRNA is closely related to its function in cancer.Competing endogenous RNA (ceRNA),circular RNA (circRNA)and RNA binding protein (RBP)as regulatory molecules are involved in the expression regulation of target genes.It may be one reason why the miRNA has complicated and diversified functions in diverse cancer tissues or cancer cells.The future direction of miRNA research is to study the latest technology to find the network control mechanism of miRNA in the specific cells.
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<p><b>OBJECTIVE</b>To construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressing HKE-HERG-G572R.</p><p><b>METHODS</b>HERG-G572R mutant fragment was constructed by over-lap extension PCR and validated by DNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cell line stably expressing HKE-HERG-G572R.</p><p><b>RESULTS</b>The pcDNA3-HERG-G572R expression vector was successfully constructed and the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-fold HKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfected with pcDNA3 (P<0.01).</p><p><b>CONCLUSION</b>The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to provide a cell model for studying individualized therapy.</p>
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Humanos , Sequência de Bases , Canais de Potássio Éter-A-Go-Go , Genética , Expressão Gênica , Vetores Genéticos , Células HEK293 , Metabolismo , Mutação , TransfecçãoRESUMO
Objective Phosphatidylethanolamine (PE) is an important phospholipid component in the cell membrane and is involved in the formation of membrane asymmetry. PE is exposed on the cell surface with phosphatidylserine during apoptosis. However, the effects of PE on cell apoptosis are not clear. In this study, we investigated effects of PE on apoptosis in human cervical cancer HeLa cells. Methods HeLa cells were used as the experiment material, and were divided into five groups: blank PE, respectively. The cell growth was tested by MTT assay; the cell cycle and apoptosis were analyzed using flow cytometry. Results Compared with the control group, PE inhibited the growth of HeLa cells in all the treatment groups in dose- and time-dependent manners, and induced the apoptosis, but did not change the cell cycle. Conclusion PE inhibits the growth of HeLa cells by inducing the apoptosis.
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Objective: To investigate the mechanisms of apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase. Methods: Human leukemia cell line K562 were exposed to indole-3-acetic acid (IAA) at 20, 40, 60, 80 or 100 mol/L and horseradish peroxidase(HRP) at 1.2 g/mL for varying times. MTT assay was applied to detect the cell proliferation. Flow cytometry was performed to detect the arrest of cell cycle. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to measure apoptosis. 2, 7-dichlorofluorescin diacetate (DCFH-DA) uptake was measured to determine free radical by confocal microscope. Content of malondiadehyde (MDA) and activity of superoxide dismutase (SOD) were measured by biochemical methods. Results: IAA/HRP initiated growth inhibition of K562 cells in a dose- and time-dependent manner. Flow cytometry revealed that cell cycle arrested at G1/G0 after 24 hours treatment. After 72 hours treatment, apoptotic rate of 100 mol/L IAA group increased to 43.9%, which was 5 times that of control(P<0.01). Content of MDA and activity of SOD increased respectively in treatments compared with control. Meanwhile, IAA/HRP stimulated the formation of free radical, which was increased by IAA concentration-dependently. Conclusion: The combination of IAA and HRP can inhibit the growth of Human leukemia cell line K562 in vitro by inducing apoptosis which is associated with the increase of free radical. The combination of IAA and HRP might be a promising chemopreventive and chemotherapeutic agent against human leukemia.
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Objective To investigate the mechanisms of apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase. Methods Human leukemia cell line K562 were exposed to indole-3-acetic acid (IAA) at 20, 40, 60, 80 or 100 mol/L and horseradish peroxidase(HRP) at 1.2 g/mL for varying times. MTT assay was applied to detect the cell proliferation. Flow cytometry was performed to detect the arrest of cell cycle. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay was used to measure apoptosis. 2, 7-dichlorofluorescin diacetate (DCFH-DA) uptake was measured to determine free radical by confocal microscope. Content of malondiadehyde (MDA) and activity of superoxide dismutase (SOD) were measured by biochemical methods. Results IAA/HRP initiated growth inhibition of K562 cells in a dose- and time-dependent manner. Flow cytometry revealed that cell cycle arrested at G1/G0 after 24 hours treatment. After 72 hours treatment, apoptotic rate of 100 mol/L IAA group increased to 43.9%, which was 5 times that of control(P＜0.01). Content of MDA and activity of SOD increased respectively in treatments compared with control. Meanwhile, IAA/HRP stimulated the formation of free radical, which was increased by IAA concentration-dependently. Conclusion The combination of IAA and HRP can inhibit the growth of Human leukemia cell line K562 in vitro by inducing apoptosis which is associated with the increase of free radical. The combination of IAA and HRP might be a promising chemopreventive and chemotherapeutic agent against human leukemia.
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<p><b>OBJECTIVE</b>To analyze the genetic polymorphism of D16S539, D7S820 and D13S317 in Chinese Kazak ethnic population from Xinjiang.</p><p><b>METHODS</b>One hundred and two unrelated individuals and a sample of families (n=42) were investigated by multiplex amplification, 6% denaturing PAGE and silver staining. And, the obtained allele frequencies were compared with those of other populations.</p><p><b>RESULTS</b>Eight, seven, eight alleles were observed at the 3 STR loci respectively and the genotypes distributions were in accordance with Hardy-Weinberg equilibrium. The expected heterozygosities for these loci were 0.9439, 0.9356 and 0.9304; the calculated polymorphism formation content (PIC) was 0.9905; the discrimination power (DP), 0.9998; the paternity exclusion (PE), 0.9572. In addition, significant difference was found in comparison with other populations, and in the sample of families (n=42) no new mutations could be found.</p><p><b>CONCLUSION</b>The multiplex examination of 3 STR loci can be used in forensic identification and population genetics research.</p>
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Feminino , Humanos , Masculino , China , Etnologia , População Branca , Genética , Frequência do Gene , Genética Populacional , Genótipo , Polimorfismo Genético , Sequências de Repetição em Tandem , GenéticaRESUMO
Objective To study effects of temperature on learning and memory of Drosophila melanogaster.Methods Food was put in two centrifuge tubes.Drosophila can smell and eat food through pores in one centrifuge tube but Drosophila can only smell but not eat food through pores in another centrifuge tube with obstacle.After Drosophila were bred at different temperatures,the variation of the numbers of Drosophila in two tubes were observed.Results The number of Drosophila bred at high temperature entering the tube and eating the food was fewer than that at normal temperature and low temperature.Conclusion High temperature can decrease the learning and memory behavior of Drosophila.
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Corn meal was hydrolyzed in HCI ,and pH adjusted to 6.8 with (NH_4)OH. The substrate was inoculated with Cadida trapicalis ,then cultivated in a shaker at 30℃ and in 200r/min for 2 days. The Results indicated that the protein content in corn meal grew from 8.4 to 27.4%the lysine in the protein increased from 32.1 to 73.48/kg,the tryptophan from 6.8 to to 14.8g/kg. This shows that the nutritional value of corn meal can be improved with this method.
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10g corn meal was mixed with 200ml Na2HPO4/KH2PO4 buffer (1/15M, pH6.81, containtng ammonium oxalate 0.6%, MgSO4 0.01%, FeSO4 0.002%) in a 500ml flask, then was inoculated with Candids tropicalis, and 1.5g ?-amylase was added at the same time. The substrate was cultivated in a shaker at 30℃, 200r/min for 3 days. The protein content increased from 8.4% to 23.4% (P