2019 Vol. 10, No. 2
2019, 10(2): 99-105.
doi: 10.15886/j.cnki.rdswxb.2019.02.001
Abstract
PDF 903KB
Abstract:
an attempt was made to study the cytotoxicity effect of α-conotoxin TxID in DMS114 s, a small cell lung cancer cell line. An α-conotoxin TxID was synthesized by solid phase peptide synthesis and two-step oxidation and identified by mass spectrometry and HPLC. The expression of α3 and β4 nAChR subunits in DMS114 cells was detected by PCR. MTT assay was used to detect the cytotoxicity of TxID or Adriamycin（ADM） or TxID combined with ADM in the DMS114 cells. The results showed that α-conotoxin TxID was successfully synthesized, and that α3 and α4 nAChR subunits were detected in DMS114 cells. The α-conotoxin TxID dependently inhibited the proliferation of small cell lung cancer DMS114 cells and HEL normal lung cells, and had higher cytotoxicity effect at some specific concentrations on DMS114 cells than on HEL cells. In addition, combination of TxID and ADM（1∶1） potentiated the cytotoxicity in DMS114 cells. In short, this study demonstrated for the first time that α-conotoxin TxID can inhibit the proliferation of small cell lung cancer DMS114 cells, which could provide guidance for the study of novel anticancer drugs.
an attempt was made to study the cytotoxicity effect of α-conotoxin TxID in DMS114 s, a small cell lung cancer cell line. An α-conotoxin TxID was synthesized by solid phase peptide synthesis and two-step oxidation and identified by mass spectrometry and HPLC. The expression of α3 and β4 nAChR subunits in DMS114 cells was detected by PCR. MTT assay was used to detect the cytotoxicity of TxID or Adriamycin（ADM） or TxID combined with ADM in the DMS114 cells. The results showed that α-conotoxin TxID was successfully synthesized, and that α3 and α4 nAChR subunits were detected in DMS114 cells. The α-conotoxin TxID dependently inhibited the proliferation of small cell lung cancer DMS114 cells and HEL normal lung cells, and had higher cytotoxicity effect at some specific concentrations on DMS114 cells than on HEL cells. In addition, combination of TxID and ADM（1∶1） potentiated the cytotoxicity in DMS114 cells. In short, this study demonstrated for the first time that α-conotoxin TxID can inhibit the proliferation of small cell lung cancer DMS114 cells, which could provide guidance for the study of novel anticancer drugs.
2019, 10(2): 106-110,134.
doi: 10.15886/j.cnki.rdswxb.2019.02.002
Abstract:
Salt stress suppresses growth and development of rice. Cloning salt tolerant related genes in rice for analysis of their functions might lay a foundation for research in rice salt tolerance mechanism and for salinity resistance breeding. A Na+/H+ antiporter gene OsNHX5 was cloned from Oryza sativa by polymerase chain reaction（PCR） method, and its localization expressing vector was constructed and observed through tobacco injection mediated by Agrobacterium tumefaciens.The expression patterns of OsNHX5 under salinity were analyzed by using real-time PCR. Phylogenetic analysis revealed that OsNHX5 was homologous to Arabidopsis AtNHX5 and AtNHX6. The results showed that OsNHX5 protein was localized in the intracellular membranes. The OsNHX5 gene was induced by KCl stress and its expression was upregulated in leaves but was not significantly different in roots under NaCl or KCl stress. Our results illustrated that OsNHX5 was an endosomal K+/H+ antiporter, and this finding would give a theoretical basis for molecular breeding of salt tolerant rice in the future.
Salt stress suppresses growth and development of rice. Cloning salt tolerant related genes in rice for analysis of their functions might lay a foundation for research in rice salt tolerance mechanism and for salinity resistance breeding. A Na+/H+ antiporter gene OsNHX5 was cloned from Oryza sativa by polymerase chain reaction（PCR） method, and its localization expressing vector was constructed and observed through tobacco injection mediated by Agrobacterium tumefaciens.The expression patterns of OsNHX5 under salinity were analyzed by using real-time PCR. Phylogenetic analysis revealed that OsNHX5 was homologous to Arabidopsis AtNHX5 and AtNHX6. The results showed that OsNHX5 protein was localized in the intracellular membranes. The OsNHX5 gene was induced by KCl stress and its expression was upregulated in leaves but was not significantly different in roots under NaCl or KCl stress. Our results illustrated that OsNHX5 was an endosomal K+/H+ antiporter, and this finding would give a theoretical basis for molecular breeding of salt tolerant rice in the future.
2019, 10(2): 111-118.
doi: 10.15886/j.cnki.rdswxb.2019.02.003
Abstract:
Cassava（Manihot esculenta Crantz） is characteristic of high yield, drought tolerance and poor soil tolerance. To probe into the molecular basis for tolerance of poor soils and enhance the utilization rate of nitrogen in cassava, two years old mature plants and 1 month-old tissue cultured plants of cassava cultivar "Huanan 8" were collected and cultured for 30 days in a medium as the experimental material, from which a high-affinity nitrate transporter gene NRT2 was isolated by using homologous cloning technique. Sequence analysis revealed that the open reading frame of this gene called MeNRT2.5 is 1 479 bp and encodes 492 amino acids. Bioinformatics analyses showed that the MeNRT2.5 protein had 10 transmembrane regions, and had a a high homology with the NRT2.5 protein of Hevea brasiliensis, Jatropha curcas and,Theobroma cacao, with its amino acid sequences being 94%, 89.84% and 87.4% in similarity, respectively. Semi-Quantitative real-time PCR analysis showed that the MeNRT2.5 gene was expressed in root, stem, leaf, flower and other organs, with its expression being higher in the roots of mature cassava plants and in the leaves of tissue cultured plants. Quantitative real-time PCR analysis showed that transcript of the MeNRT2.5 in root had a peak expression at 6 h under the treatment with low concentration NO3-(0.3 mmol·L-1) but its expression did not change significantly in the root, stem and leaf after the treatment with high concentration of NO3-(3 mmol·L-1), indicating the transcript of the MeNRT2.5 gene is inhibited by high concentrations of NO3-. All these results provide a theoretical foundation for further analysis of the functional verification of the MeNRT2.5 gene in cassava.
Cassava（Manihot esculenta Crantz） is characteristic of high yield, drought tolerance and poor soil tolerance. To probe into the molecular basis for tolerance of poor soils and enhance the utilization rate of nitrogen in cassava, two years old mature plants and 1 month-old tissue cultured plants of cassava cultivar "Huanan 8" were collected and cultured for 30 days in a medium as the experimental material, from which a high-affinity nitrate transporter gene NRT2 was isolated by using homologous cloning technique. Sequence analysis revealed that the open reading frame of this gene called MeNRT2.5 is 1 479 bp and encodes 492 amino acids. Bioinformatics analyses showed that the MeNRT2.5 protein had 10 transmembrane regions, and had a a high homology with the NRT2.5 protein of Hevea brasiliensis, Jatropha curcas and,Theobroma cacao, with its amino acid sequences being 94%, 89.84% and 87.4% in similarity, respectively. Semi-Quantitative real-time PCR analysis showed that the MeNRT2.5 gene was expressed in root, stem, leaf, flower and other organs, with its expression being higher in the roots of mature cassava plants and in the leaves of tissue cultured plants. Quantitative real-time PCR analysis showed that transcript of the MeNRT2.5 in root had a peak expression at 6 h under the treatment with low concentration NO3-(0.3 mmol·L-1) but its expression did not change significantly in the root, stem and leaf after the treatment with high concentration of NO3-(3 mmol·L-1), indicating the transcript of the MeNRT2.5 gene is inhibited by high concentrations of NO3-. All these results provide a theoretical foundation for further analysis of the functional verification of the MeNRT2.5 gene in cassava.
2019, 10(2): 119-126.
doi: 10.15886/j.cnki.rdswxb.2019.02.004
Abstract:
The mitogen-activated protein kinase（MAPK） cascade pathway is a signaling pathway widely present in eukaryotic organisms. Based on cassava genomic data, a comprehensive identification and phylogenetic tree analysis of mitogen-activated protein kinase kinase kinase（MKK） family members in cassava was carried out by bioinformatics methods. At the same time, the expression pattern of each gene of the MKK family was analyzed through hormone treatment and pathogen inoculation. The results showed that cassava encoded 11 MKK genes, which were distributed on chromosomes 3, 4, 6, 10, 12, 16, and 17 of cassava. The expression analysis showed that MKKs were responsive to ABA, JA and pathogen signal but not sensitive to ACC signal, of which MKK4, MKK5, MKK8, MKK9 and MKK11 may be involved in cassava-related pathogens defense pathways and hormone signaling pathways.
The mitogen-activated protein kinase（MAPK） cascade pathway is a signaling pathway widely present in eukaryotic organisms. Based on cassava genomic data, a comprehensive identification and phylogenetic tree analysis of mitogen-activated protein kinase kinase kinase（MKK） family members in cassava was carried out by bioinformatics methods. At the same time, the expression pattern of each gene of the MKK family was analyzed through hormone treatment and pathogen inoculation. The results showed that cassava encoded 11 MKK genes, which were distributed on chromosomes 3, 4, 6, 10, 12, 16, and 17 of cassava. The expression analysis showed that MKKs were responsive to ABA, JA and pathogen signal but not sensitive to ACC signal, of which MKK4, MKK5, MKK8, MKK9 and MKK11 may be involved in cassava-related pathogens defense pathways and hormone signaling pathways.
2019, 10(2): 127-134.
doi: 10.15886/j.cnki.rdswxb.2019.02.005
Abstract:
Based on the clues obtained from our previous proteomics studies, the mon1 gene from Fusarium oxysporum f.sp. cubense Race 4（Foc4） was found to play a role in pathogenic infection. To further characterize the functions of the mon1 in the infection of Foc4, the mon1 gene was knocked out through homologous recombination method and the phenotype and pathogenicity of the mutant strains analyzed. The results showed that the mutant strains of Foc4 grew slowly, produced less spores and thinner mycelia with fewer branches, and was lower in their resistance to exogenous oxygen stress, their capacity of cell wall penetration and their cellulose utilization, as compared with the wild strains. Pathogenicity test showed the mutant strains exhibited a significantly low infection against the banana seedlings. All the results suggested that the mon1 gene played a certain role in the growth, development, sporulation and pathogenicity of the Foc4.
Based on the clues obtained from our previous proteomics studies, the mon1 gene from Fusarium oxysporum f.sp. cubense Race 4（Foc4） was found to play a role in pathogenic infection. To further characterize the functions of the mon1 in the infection of Foc4, the mon1 gene was knocked out through homologous recombination method and the phenotype and pathogenicity of the mutant strains analyzed. The results showed that the mutant strains of Foc4 grew slowly, produced less spores and thinner mycelia with fewer branches, and was lower in their resistance to exogenous oxygen stress, their capacity of cell wall penetration and their cellulose utilization, as compared with the wild strains. Pathogenicity test showed the mutant strains exhibited a significantly low infection against the banana seedlings. All the results suggested that the mon1 gene played a certain role in the growth, development, sporulation and pathogenicity of the Foc4.
2019, 10(2): 135-139,164.
doi: 10.15886/j.cnki.rdswxb.2019.02.006
Abstract:
Environmental toxicology of pesticide is one of research hot topics at present. Selenastrum capricornutumwas was used as biological model to analyze acute toxicity of cyantraniliprole against Selenastrum capricornutum. S. capricornutum was exposed to cyantraniliprole at different concentrations to determine the contents of biomass, photosynthetic pigments and malonaldehyde（MDA）, and the activities of superoxide dismutase（SOD） and catalase（CAT）.The results showed that 72 h ErC50 and EyC50 of cyantraniliprole to S. capricornutum were 24.15 and 20.01 mg·L-1, respectively. Cyantraniliprole inhibited the growth of S. capricornutum and reduced the synthesis of photosynthetic pigments（chlorophyll a, chlorophyll b and carotenoid r）. S. capricornutum exposed to cyantraniliprole showed an oxidative stress response, leading to a decline in the SOD activity and an increase in the CAT activity and the MDA content.
Environmental toxicology of pesticide is one of research hot topics at present. Selenastrum capricornutumwas was used as biological model to analyze acute toxicity of cyantraniliprole against Selenastrum capricornutum. S. capricornutum was exposed to cyantraniliprole at different concentrations to determine the contents of biomass, photosynthetic pigments and malonaldehyde（MDA）, and the activities of superoxide dismutase（SOD） and catalase（CAT）.The results showed that 72 h ErC50 and EyC50 of cyantraniliprole to S. capricornutum were 24.15 and 20.01 mg·L-1, respectively. Cyantraniliprole inhibited the growth of S. capricornutum and reduced the synthesis of photosynthetic pigments（chlorophyll a, chlorophyll b and carotenoid r）. S. capricornutum exposed to cyantraniliprole showed an oxidative stress response, leading to a decline in the SOD activity and an increase in the CAT activity and the MDA content.
2019, 10(2): 140-144.
doi: 10.15886/j.cnki.rdswxb.2019.02.007
Abstract:
Flusilazole has a broad-spectrum germicidal efficacy to curb diseases caused by basidiomycetes and hemispheres.Chirocephalus is a salt tolerant small crustacean, which is used as live food for finfish and shellfish in aquaculture. In recent years it is used as a model organism to evaluate the environmental safety of pesticides. The effective concentration(EC50) of flusilazole against the Chirocephalus was determined by using an acute toxicity test after 38 h flusiazole exposure. Activities of superoxide dismutase（SOD） and peroxidase（POD） in the Chirocephalus were tested with an enzyme activity kit, and mamondialdehyde（MDA） content was tested by using the thiobarbituric acid（TBA） method after 38 h flusiazole exposure. The results showed that the EC50 of flusilazole to the Chirocephalus was 5.228 mg·L-1, which was of moderate toxicity. The hatching rate of the Chirocephalus was significantly decreased with an increase in concentration of flusilazole, as compared with that of the control. The toxic effects of flusilazole may cause oxidative stress in Chirocephalus. The activities of SOD and MDA content were increased, and the POD activity was decreased in all the treatment groups, compared with the control.
Flusilazole has a broad-spectrum germicidal efficacy to curb diseases caused by basidiomycetes and hemispheres.Chirocephalus is a salt tolerant small crustacean, which is used as live food for finfish and shellfish in aquaculture. In recent years it is used as a model organism to evaluate the environmental safety of pesticides. The effective concentration(EC50) of flusilazole against the Chirocephalus was determined by using an acute toxicity test after 38 h flusiazole exposure. Activities of superoxide dismutase（SOD） and peroxidase（POD） in the Chirocephalus were tested with an enzyme activity kit, and mamondialdehyde（MDA） content was tested by using the thiobarbituric acid（TBA） method after 38 h flusiazole exposure. The results showed that the EC50 of flusilazole to the Chirocephalus was 5.228 mg·L-1, which was of moderate toxicity. The hatching rate of the Chirocephalus was significantly decreased with an increase in concentration of flusilazole, as compared with that of the control. The toxic effects of flusilazole may cause oxidative stress in Chirocephalus. The activities of SOD and MDA content were increased, and the POD activity was decreased in all the treatment groups, compared with the control.
2019, 10(2): 145-150.
doi: 10.15886/j.cnki.rdswxb.2019.02.008
Abstract:
Insecticidal active ingredients were roughly extracted with methanol, petroleum ether, chloroform, ethyl acetate, N-butanol and azadirachtin from Capparis spinosa L. The antifeedant activity of each extract against Spodoptera litura was determined by leaf disc method. And the contact toxicity of chloroform extract against Lipaphis erysimi and Pierisrapae Linnaeus was determined by impregnation method. In addition, the field control effect of chloroform extract on Pierisrapae Linnaeus was determined by spray method. The results showed that all the extracts had good non-selective antifeedant activities against the 3 rd instar larvae. In particular, the chloroform extract had the highest activity, and each extract had lower antifeedant activity against the 5 th instar larvae. Moreover, the chloroform extract not only had good contact toxicity against L. erysimi and Pierisrapae Linnaeus, but also showed a good control effect on Pierisrapae Linnaeus infesting cabbage in the field.
Insecticidal active ingredients were roughly extracted with methanol, petroleum ether, chloroform, ethyl acetate, N-butanol and azadirachtin from Capparis spinosa L. The antifeedant activity of each extract against Spodoptera litura was determined by leaf disc method. And the contact toxicity of chloroform extract against Lipaphis erysimi and Pierisrapae Linnaeus was determined by impregnation method. In addition, the field control effect of chloroform extract on Pierisrapae Linnaeus was determined by spray method. The results showed that all the extracts had good non-selective antifeedant activities against the 3 rd instar larvae. In particular, the chloroform extract had the highest activity, and each extract had lower antifeedant activity against the 5 th instar larvae. Moreover, the chloroform extract not only had good contact toxicity against L. erysimi and Pierisrapae Linnaeus, but also showed a good control effect on Pierisrapae Linnaeus infesting cabbage in the field.
2019, 10(2): 151-158.
doi: 10.15886/j.cnki.rdswxb.2019.02.009
Abstract:
Pig manure separated from the dry and the wet was used as substrate, inoculated with earthworms（Eisenia fetida） and cultured indoors for 4 weeks. The contents of heavy metals（Cu, Zn, Cr, Pb, Cd, As） in the pig manure and earthworms were determined by the inductively coupled plasma mass spectrometry（ICP-MS）. The speciation changes in the pig manure and the enrichment of heavy metals in the earthworms were systematically analyzed under the action of the earthworms. The experimental results showed that the pig manure promoted the growth of earthworms, increased the body weight of the earthworms obviously after 4 weeks of treatment and had no effect on the reproduction of the earthworms. After 4 weeks of earthworm treatment, the pH value of the pig manure increased from 6.51 to 6.86. The total amount of Cu, Zn, Cr, Pb, Cd and As in the pig manure decreased by 13.69%, 24.47%, 30.70%, 39.17%, 21.91% and 9.36%, respectively, compared with the control group. Meanwhile, the proportion of Zn, Cr and Cd in the pig manure decreased by 15%, 27% and 27%, respectively compared with the control group. The earthworms had a certain effect on enrichment of heavy metals. The concentrations of Cu, Zn, Cd and As, except for Cr, in the earthworms increased by 0.49 mg·kg-1, 3.86 mg·kg-1, 0.012 mg·kg-1 and 0.064 mg·kg-1, respectively compared with those before the experiment, and showed a time-effect relationship. The biota-sediment accumulation factors increased in the order of Tr＞Cd＞Zn＞Cu＞As. This indicates the earthworms can enrich heavy metals and reduce the content of heavy metals in the pig manure and the proportion of heavy metals in the form of residue in the pig manure, which is conducive to the transfer and removal of heavy metals.
Pig manure separated from the dry and the wet was used as substrate, inoculated with earthworms（Eisenia fetida） and cultured indoors for 4 weeks. The contents of heavy metals（Cu, Zn, Cr, Pb, Cd, As） in the pig manure and earthworms were determined by the inductively coupled plasma mass spectrometry（ICP-MS）. The speciation changes in the pig manure and the enrichment of heavy metals in the earthworms were systematically analyzed under the action of the earthworms. The experimental results showed that the pig manure promoted the growth of earthworms, increased the body weight of the earthworms obviously after 4 weeks of treatment and had no effect on the reproduction of the earthworms. After 4 weeks of earthworm treatment, the pH value of the pig manure increased from 6.51 to 6.86. The total amount of Cu, Zn, Cr, Pb, Cd and As in the pig manure decreased by 13.69%, 24.47%, 30.70%, 39.17%, 21.91% and 9.36%, respectively, compared with the control group. Meanwhile, the proportion of Zn, Cr and Cd in the pig manure decreased by 15%, 27% and 27%, respectively compared with the control group. The earthworms had a certain effect on enrichment of heavy metals. The concentrations of Cu, Zn, Cd and As, except for Cr, in the earthworms increased by 0.49 mg·kg-1, 3.86 mg·kg-1, 0.012 mg·kg-1 and 0.064 mg·kg-1, respectively compared with those before the experiment, and showed a time-effect relationship. The biota-sediment accumulation factors increased in the order of Tr＞Cd＞Zn＞Cu＞As. This indicates the earthworms can enrich heavy metals and reduce the content of heavy metals in the pig manure and the proportion of heavy metals in the form of residue in the pig manure, which is conducive to the transfer and removal of heavy metals.
2019, 10(2): 159-164.
doi: 10.15886/j.cnki.rdswxb.2019.02.010
Abstract:
To evaluate analgesic and anti-inflammatory activities of artificially induced dragon’s blood of Dracaena cambodiana,the natural and artificially induced dragon’s bloods were evaluated and compared in terms of analgesic and anti-inflammatory activities by using the writhing model in mice, the mouse auricle swelling model and the capillary permeability in abdominal cavity of mice. Artificially induced dragon’s blood of D. cambodiana significantly increased the latency of mouse writhing induced by acetic acid, indicating that it had a higher analgesic activity than natural one. Both natural and artificially induced dragon’s blood markedly reduced auricle swelling caused by dimethylbenzene in mice, suggesting their similarity in anti-inflammatory activity. Artificially induced dragon’s blood of D. cambodiana had similar analgesic and anti-inflammatory activities with natural dragon’s blood.
To evaluate analgesic and anti-inflammatory activities of artificially induced dragon’s blood of Dracaena cambodiana,the natural and artificially induced dragon’s bloods were evaluated and compared in terms of analgesic and anti-inflammatory activities by using the writhing model in mice, the mouse auricle swelling model and the capillary permeability in abdominal cavity of mice. Artificially induced dragon’s blood of D. cambodiana significantly increased the latency of mouse writhing induced by acetic acid, indicating that it had a higher analgesic activity than natural one. Both natural and artificially induced dragon’s blood markedly reduced auricle swelling caused by dimethylbenzene in mice, suggesting their similarity in anti-inflammatory activity. Artificially induced dragon’s blood of D. cambodiana had similar analgesic and anti-inflammatory activities with natural dragon’s blood.
2019, 10(2): 165-171.
doi: 10.15886/j.cnki.rdswxb.2019.02.011
Abstract:
A floristic study of Yangshan Wetland in Haikou, Hainan Province, China was made from the aspects of plant species composition and geographical composition through field investigation, specimen textual research and literature consulting to have a good picture of the plant resources and floristic characteristics of Yangshan Wetland. The results showed thatYangshan wetland has a high species diversity, including 687 species of vascular plants belonging to 119 families, 413 genera and 687 species. Of the species5 are endemic species inHainan, accounting for 0.73% of the total tree species. Gramineae, Compositae, Moraceae and Rubiaceae are the main families of vascular plants in Yangshan Wetland. The main genera are Ficus, Cyperus, Polygonum and Syzygium. The main flora of wild vascular plants in Yangshan Wetland is tropical, and its tropical compositions accounted for 90.12% of the total families（exclusive of families worldwide distributed） and 90.5% of the total genera（exclusive of genera worldwide distributed）, respectively. The temperate floristic composition is low, accounting for 9.88% and 8.97% of the total families and genera, respectively, reflecting the tropical marginal nature and a weak tropical trend.
A floristic study of Yangshan Wetland in Haikou, Hainan Province, China was made from the aspects of plant species composition and geographical composition through field investigation, specimen textual research and literature consulting to have a good picture of the plant resources and floristic characteristics of Yangshan Wetland. The results showed thatYangshan wetland has a high species diversity, including 687 species of vascular plants belonging to 119 families, 413 genera and 687 species. Of the species5 are endemic species inHainan, accounting for 0.73% of the total tree species. Gramineae, Compositae, Moraceae and Rubiaceae are the main families of vascular plants in Yangshan Wetland. The main genera are Ficus, Cyperus, Polygonum and Syzygium. The main flora of wild vascular plants in Yangshan Wetland is tropical, and its tropical compositions accounted for 90.12% of the total families（exclusive of families worldwide distributed） and 90.5% of the total genera（exclusive of genera worldwide distributed）, respectively. The temperate floristic composition is low, accounting for 9.88% and 8.97% of the total families and genera, respectively, reflecting the tropical marginal nature and a weak tropical trend.
2019, 10(2): 172-177.
doi: 10.15886/j.cnki.rdswxb.2019.02.012
Abstract:
Surveys were made of the weeds in the Arabica coffee plantations in Ph?ngsali province, Laos in 2016 and 2017. These surveys showed that there were 79 species, 66 genera and 23 families in the weeds in the Arabica coffee plantations in this province, of which Compositae and Gramineae were dominant weed populations, accounting for 39.24% of the total weed species. Dicotyledon weeds were dominant weed populations in the Arabica coffee plantations, accounting for 72.15% of the total number of weeds. Due to farming practices and human disturbance, perennial species of the weeds accounted for 51.90% of the total species in the Arabica coffee plantations.
Surveys were made of the weeds in the Arabica coffee plantations in Ph?ngsali province, Laos in 2016 and 2017. These surveys showed that there were 79 species, 66 genera and 23 families in the weeds in the Arabica coffee plantations in this province, of which Compositae and Gramineae were dominant weed populations, accounting for 39.24% of the total weed species. Dicotyledon weeds were dominant weed populations in the Arabica coffee plantations, accounting for 72.15% of the total number of weeds. Due to farming practices and human disturbance, perennial species of the weeds accounted for 51.90% of the total species in the Arabica coffee plantations.
2019, 10(2): 178-183.
doi: 10.15886/j.cnki.rdswxb.2019.02.013
Abstract:
In order to detect the expression of α7 nicotine acetylcholine receptor（nAChR） subunit gene（CHRNA7）, the primers were designed according to the sequence of human CHRNA7 gene, and then CHRNA7 gene was amplified by PCR. After being cloned into pMD-18 T vector, the recombinant plasmid was sequenced and prepared. The recombinant plasmids were gradient diluted and used as templates to establish standard curves for real-time PCR. The sensitivity and repeatability of the real-time PCR were carried out. A real-time PCR method for detecting CHRNA7 was successfully established. With this method CHRNA7 gene was detected 10 copies·μL-1 at the lowest. The variation coefficients of five replications of the three templates at 10~4, 105 and 106 copies·μL-1 were 1.22%, 1.90% and 2.63%, respectively, which indicated the real-time PCR detection had a good repeatability. Furthermore, the detection of α7 nAChR subunit gene expression in human cervical cancer cell line SiHa and human normal cervical cell line Ect1/E6 E7 showed that the expression of α7 nAChR subunit in SiHa cells was significantly lower than that in Ect1/E6 E7 cells（P=0.015）.
In order to detect the expression of α7 nicotine acetylcholine receptor（nAChR） subunit gene（CHRNA7）, the primers were designed according to the sequence of human CHRNA7 gene, and then CHRNA7 gene was amplified by PCR. After being cloned into pMD-18 T vector, the recombinant plasmid was sequenced and prepared. The recombinant plasmids were gradient diluted and used as templates to establish standard curves for real-time PCR. The sensitivity and repeatability of the real-time PCR were carried out. A real-time PCR method for detecting CHRNA7 was successfully established. With this method CHRNA7 gene was detected 10 copies·μL-1 at the lowest. The variation coefficients of five replications of the three templates at 10~4, 105 and 106 copies·μL-1 were 1.22%, 1.90% and 2.63%, respectively, which indicated the real-time PCR detection had a good repeatability. Furthermore, the detection of α7 nAChR subunit gene expression in human cervical cancer cell line SiHa and human normal cervical cell line Ect1/E6 E7 showed that the expression of α7 nAChR subunit in SiHa cells was significantly lower than that in Ect1/E6 E7 cells（P=0.015）.
2019, 10(2): 184-189.
doi: 10.15886/j.cnki.rdswxb.2019.02.014
Abstract:
In order to obtain the total DNA of the soil microbes in the rhizosphere of pitaya（Hylocereus undatus Britt.）, two methods, CTAB-SDS and SDS, were used to compare their extraction effect in extraction time, color of DNA supernatant in the extraction process, and total DNA extraction amount and purity after extraction. The results showed that both methods produced DNA fragments with about 23.1 kb. With the CTAB-SDS method the color of the supernatant was light yellow; the total amount of DNA extracted was（7.4 ～ 7.7） mg·kg-1; the OD260/280 value was between 1.81 and 1.95, and the OD260/230 value was between 1.26 and 2.03; the average DNA concentration was 0.51 g·L-1. With the SDS method the color of the supernatant was dark brown-yellow, and the total amount of DNA extracted was（3.5 ～ 4.15） mg·kg-1; the OD260/280 value was between 1.52 and 1.68, and the OD260/230 value was between 1.35 and 1.53; the average DNA concentration was 0.075 g·L-1. It was hence concluded that the fragment extracted by the CTAB-SDS method was relatively complete and highly pure, which could be directly used for subsequent experimental operation, and was more suitable for the extraction of total DNA from soil microbes in the rhyzosphere of pitaya.
In order to obtain the total DNA of the soil microbes in the rhizosphere of pitaya（Hylocereus undatus Britt.）, two methods, CTAB-SDS and SDS, were used to compare their extraction effect in extraction time, color of DNA supernatant in the extraction process, and total DNA extraction amount and purity after extraction. The results showed that both methods produced DNA fragments with about 23.1 kb. With the CTAB-SDS method the color of the supernatant was light yellow; the total amount of DNA extracted was（7.4 ～ 7.7） mg·kg-1; the OD260/280 value was between 1.81 and 1.95, and the OD260/230 value was between 1.26 and 2.03; the average DNA concentration was 0.51 g·L-1. With the SDS method the color of the supernatant was dark brown-yellow, and the total amount of DNA extracted was（3.5 ～ 4.15） mg·kg-1; the OD260/280 value was between 1.52 and 1.68, and the OD260/230 value was between 1.35 and 1.53; the average DNA concentration was 0.075 g·L-1. It was hence concluded that the fragment extracted by the CTAB-SDS method was relatively complete and highly pure, which could be directly used for subsequent experimental operation, and was more suitable for the extraction of total DNA from soil microbes in the rhyzosphere of pitaya.
2019, 10(2): 190-196.
doi: 10.15886/j.cnki.rdswxb.2019.02.015
Abstract:
The essential oil of agarwood induced by two methods, fungal inoculation and fire drilling, was extracted by the carbon dioxide supercritical fluid extraction（SFE-CO2） and analyzed with GC-MS, and its composition and relative contents were compared. A total of 59 compounds were identified from the agarwood induced by these two methods, of which 3 were chromones with their total relative contents of 54.59%（Fungal inoculation method） and 29.45%（Fire drilling method）, respectively. The results showed that the essential oil produced by the fungal inoculation method had a higher quality than that produced by the fire drilling method.
The essential oil of agarwood induced by two methods, fungal inoculation and fire drilling, was extracted by the carbon dioxide supercritical fluid extraction（SFE-CO2） and analyzed with GC-MS, and its composition and relative contents were compared. A total of 59 compounds were identified from the agarwood induced by these two methods, of which 3 were chromones with their total relative contents of 54.59%（Fungal inoculation method） and 29.45%（Fire drilling method）, respectively. The results showed that the essential oil produced by the fungal inoculation method had a higher quality than that produced by the fire drilling method.
2019, 10(2): 197-203.
doi: 10.15886/j.cnki.rdswxb.2019.02.016
Abstract:
A method for simultaneous detection of 13 sulfonamides drugs in seawater by using ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） was established and optimized, and used for analysis of the surface seawater samples collected from Dongzhai Harbour, Hainan. The results show that in case of the peak types of sulfonamides in the chromatogram are better and the correlation coefficient of the standard curve is high（R2 ≥ 0.996） when the sulfaonamides in the sea water were determined by using UPLC-MS/MS with mobile phase A: methanol-acetonitrile（V∶V=1∶1） and mobile phase B: 0.1% formic acid-10 mM ammonium acetate as the mobile phase and the initial mobile phase（5% mobile phase A and 95% mobile phase B） as the reconstituted solvent. Thirteen sulfonamides were separated well in 14 minutes and trheir recovery rates were from 70.4 to 103%, with a relative standard deviation（RSD） of between 1.2% and 12.8%. This method had high sensitivity and good reproducibility, and is suitable for simultaneous detection of sulfonamides in seawater. At the same time, five sulfonamides were detected in the water samples of the Dongzhai Harbour. The detection rates were from 60% to 100%. The highest detection frequency of sulfamethoxazole（SMX） was 100%, sulfameter（SME） and sulfachloropyridazine（SCP） had high detection rates（around 70%） in the Dongzhai Harbor, and the other two detected were sulfamethazine（SMA） and sulfadiazine（SDZ）. The highest concentration of sulfonamides detected was up to 3.35 ng·L-1. Compared with other areas, the concentrations of sulfonamides detected in Dongzhai Harbor was at a relatively low level.
A method for simultaneous detection of 13 sulfonamides drugs in seawater by using ultra performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS） was established and optimized, and used for analysis of the surface seawater samples collected from Dongzhai Harbour, Hainan. The results show that in case of the peak types of sulfonamides in the chromatogram are better and the correlation coefficient of the standard curve is high（R2 ≥ 0.996） when the sulfaonamides in the sea water were determined by using UPLC-MS/MS with mobile phase A: methanol-acetonitrile（V∶V=1∶1） and mobile phase B: 0.1% formic acid-10 mM ammonium acetate as the mobile phase and the initial mobile phase（5% mobile phase A and 95% mobile phase B） as the reconstituted solvent. Thirteen sulfonamides were separated well in 14 minutes and trheir recovery rates were from 70.4 to 103%, with a relative standard deviation（RSD） of between 1.2% and 12.8%. This method had high sensitivity and good reproducibility, and is suitable for simultaneous detection of sulfonamides in seawater. At the same time, five sulfonamides were detected in the water samples of the Dongzhai Harbour. The detection rates were from 60% to 100%. The highest detection frequency of sulfamethoxazole（SMX） was 100%, sulfameter（SME） and sulfachloropyridazine（SCP） had high detection rates（around 70%） in the Dongzhai Harbor, and the other two detected were sulfamethazine（SMA） and sulfadiazine（SDZ）. The highest concentration of sulfonamides detected was up to 3.35 ng·L-1. Compared with other areas, the concentrations of sulfonamides detected in Dongzhai Harbor was at a relatively low level.
2019, 10(2): 204-207.
doi: 10.15886/j.cnki.rdswxb.2019.02.017
Abstract:
To study the anticoagulant activity and purify the anticoagulation components from P. manillensis in Hainan. Poecilobdella manillensis Lession collected in Hainan was extracted with different extraction methods and purified with molecular weight ultra membranes to determine its anticoagulant components and activities by using the thrombin titration method. The relative molecular weight of the anticoagulant components was calculated by using the SDS-PAGE test. The results showed that the sodium chloride solution was better than the water extraction, the water extraction and alcohol precipitation, and the acetone extract in extraction of anticoagulant components. The anticoagulant activity of the extract from the head of P. manillensis extracted by the sodium chloride solution was 1 811.11 U·g-1, the highest among the other extraction methods. The ultra-membranes with molecular weights of 10×103 and 30×103 were selected for purification, and the molecular weight of the purified protein was 15.96×103.
To study the anticoagulant activity and purify the anticoagulation components from P. manillensis in Hainan. Poecilobdella manillensis Lession collected in Hainan was extracted with different extraction methods and purified with molecular weight ultra membranes to determine its anticoagulant components and activities by using the thrombin titration method. The relative molecular weight of the anticoagulant components was calculated by using the SDS-PAGE test. The results showed that the sodium chloride solution was better than the water extraction, the water extraction and alcohol precipitation, and the acetone extract in extraction of anticoagulant components. The anticoagulant activity of the extract from the head of P. manillensis extracted by the sodium chloride solution was 1 811.11 U·g-1, the highest among the other extraction methods. The ultra-membranes with molecular weights of 10×103 and 30×103 were selected for purification, and the molecular weight of the purified protein was 15.96×103.
2019, 10(2): 208-214.
doi: 10.15886/j.cnki.rdswxb.2019.02.018
Abstract:
Hydrogen sulfide（H2S） is an endogenous gas signaling molecule, often synergistic with carbon monoxide and nitric oxide, and plays an important role in the regulation of angiogenesis, neural activity, glucose metabolism, antioxidant and inflammatory reactions. However, endogenous H2S metabolic abnormalities are associated with many diseases, including cancer, hypertension, diabetes, and neurodegenerative diseases. Therefore, the regulation of endogenous H2S is of great significance for the treatment of the H2S related diseases and clinical studies. In the present study, the main research method is to evaluate the effect of endogenous H2S in human pathology, supplemented with exogenous H2S so as to seek breakthroughs in pathology and find therapeutic targets of H2S and its products in diseases by relying on this research method. In this paper, we will focus on the recent progress of H2S donor and H2S in inflammation and autophagy, two of the most popular pathology-related studies, and the potential research methods of endogenous H2S quantifying.The research prospect and development direction of H2S are prospected.
Hydrogen sulfide（H2S） is an endogenous gas signaling molecule, often synergistic with carbon monoxide and nitric oxide, and plays an important role in the regulation of angiogenesis, neural activity, glucose metabolism, antioxidant and inflammatory reactions. However, endogenous H2S metabolic abnormalities are associated with many diseases, including cancer, hypertension, diabetes, and neurodegenerative diseases. Therefore, the regulation of endogenous H2S is of great significance for the treatment of the H2S related diseases and clinical studies. In the present study, the main research method is to evaluate the effect of endogenous H2S in human pathology, supplemented with exogenous H2S so as to seek breakthroughs in pathology and find therapeutic targets of H2S and its products in diseases by relying on this research method. In this paper, we will focus on the recent progress of H2S donor and H2S in inflammation and autophagy, two of the most popular pathology-related studies, and the potential research methods of endogenous H2S quantifying.The research prospect and development direction of H2S are prospected.
Email alert
RSS