Objective To investigate whether hypertonic saline (HS) can induce the synthesis and release of glutamate in cultured hypothalamic astrocytes or C6 cell line. Methods Astrocytes were isolated, cultured, purified and...Objective To investigate whether hypertonic saline (HS) can induce the synthesis and release of glutamate in cultured hypothalamic astrocytes or C6 cell line. Methods Astrocytes were isolated, cultured, purified and identified from the hypothalamus of newborn rat (1 day). The astrocytes were randomly divided into five groups: isotonic (IS) and HS groups, astrocytes were incubated by IS and HS (320 mosM NaCl) medium, respectively, for 1, 3, 5, 10 or 15 rain; carbenoxolone (CBX) +IS and CBX+HS groups, astrocytes were pre-treated with CBX (100 mmol/L) for 1 h at 37℃ in a 5% CO2 / 95% atmosphere, then removed to IS and HS medium, respectively, for 1, 3, 5, 10 or 15 min; Ca2++HS group, astrocytes were pre-incubated with Ca2+ (1 000 μmol/L) for 1 h at 37℃ in a 5% CO2 / 95% atmosphere, followed by a wash with isotonic FBS/DMEM, and then removed to hypertonic saline for 1, 3, 5, 10 or 15 min. The media of five groups were collected to analyze the medium glutamate concentration with high performance liquid chromatography. The astrocytes were fixed and double immunofluorescent stained with anti-glial fibrillary acidic protein (GFAP) and anti-glutamate. The C6 cells were divided into four groups: IS, HS, CBX+IS and CBX+HS groups, and used for quantitative measurement of glutamate in cells by flow cytometry (FCM). Results (1) Anti-GFAP immunofluorescent signal revealed no significant difference among various time points in each group, or among the five groups. (2) The anti-glutamate immunofluorescent signal was increased in HS group and peaked at 5 min, and decreased and returned to the level of IS group at 15 rain (P 〈 0.01 vs the 5 min of HS group). In CBX+HS group, the glutamate intensity was higher than that in CBX+IS and HS groups. (3) The medium glutamate concentration had no change after treatment with HS for 1 and 3 min, while increased markedly after treatment for 5 min to 15 min (P 〈 0.01 vs 1 min and 3 min). On the contrary, the medium glutamate concentrations in the CBX+HS or Ca2++HS group were significant lower than that in the HS group (P 〈 0.01). (4) FCM showed HS and CBX+HS induced glutamate increase in C6 cells. Conclusion HS induced cultured rat hypothalamic astrocytes or C6 cells to synthesize and release glutamate; CBX could block glutamate release, but could not disrupt glutamate synthesis.展开更多
A pot experiment was conducted to study the effect of nitrogen fertilizer on nitrogen and potassium uptake by four rice cultivars. Results showed that the quadatic parabola relationship between biomass of rice and nit...A pot experiment was conducted to study the effect of nitrogen fertilizer on nitrogen and potassium uptake by four rice cultivars. Results showed that the quadatic parabola relationship between biomass of rice and nitrogen levels was observed, with the maximum biomass at the nitrogen level of 150 mg kg-1.The rates of nitrogen and potassium uptake by the four rice cultivars depended on growth stage and rice cultivar with the maximum rate of N in Shanyou-63 and maximum rate of K in Kaiyou-5 (hybrid rice),respedively. The kinetics of nitrogen and potasssium uptake by rice plant could be quantitstively described by the following equations: y = a + blogt, y = ab + t1/2 and y = ae-bt. The b value in the equations was correlated significantly to the rates of nitrogen and potassium uptake (NR and KR, r=0.901**~0.990**),suggesting that the b value could be used to distinguish the index of nitrogen and potassium uptake capacity of rice. The maximum values of nitrogen uptake by plant (b value) and apparent recovery of fertilizer nitrogen were observed in Shanyou-63, and the minimum value in Eryou-6078. However, the capacity of potassium uptake (b value) by Kaiyou-5 ranked first and that by Shanyou-63 second. There was a significant linear relationship between nitrogen level and nitrogen uptake by rice, but a quadratic parabola relationship was found between nitrogen level and patassium uptake by rice. The application of nitrogen fertilizer decreased the ratios of potassium to nitrogen uptake by rice plant. The greatest reduction in the ratio was observed at high nitrogen level, and the least reduction was found in Kaiyou-5 and Shanyou-63 due to their greater ability to absorb potassium.展开更多
Objective: To investigate the death mode of human hepatoma cells exposed to matrine and the role of glutathione (GSH) and cytochrome c. Methods: The MTT test and Cell Death Detection ELISA were used to identify cell d...Objective: To investigate the death mode of human hepatoma cells exposed to matrine and the role of glutathione (GSH) and cytochrome c. Methods: The MTT test and Cell Death Detection ELISA were used to identify cell death mode and viability of cells exposed to matrine. The volume of intracellular GSH was detected by GSH reductase. Finally Western blotting was chosen to analyze the expression of cytochrome c and Caspase-9 in HepG2 cells treated by matrine. Results: The apop-totic cell death induced by matrine in Hep G2 cells dramatically increased in the time-, dose-dependent manner. Matrine can exhaust intracellular GSH effectively to change the redox state in cells. Furthermore it affect the cytotoxicity of matrine. Re-sults of Western blotting showed that matrine induced the release of cytochrome c from mitochondria to cytoplasm, and then stimulate the cleavage of Caspase-9 in a time-dependent manner. Conclusion: Matrine induced apoptosis in Hep G2 cells through the mitochondrial pathway, and oxidative stress via depletion of GSH is directly involved in the apoptotic process.展开更多
Rice with low glutelin content is suitable as functional food for patients affected with diabetes and kidney failure. The fine mapping of the gene(s) responsible for low glutelin content will provide information regar...Rice with low glutelin content is suitable as functional food for patients affected with diabetes and kidney failure. The fine mapping of the gene(s) responsible for low glutelin content will provide information regarding the distribution of glutelin related genes in rice genome and will generate markers for the selection of low glutelin rice varieties. Following an SDS-PAGE screen of rice germplasm from Taihu Valley of China, Japonica selection W3660 is identified to be a novel mutant characterized with low glutelin content. For fine mapping the mutant gene for low glutelin content, F2 and F3 populations were derived from a cross between W3660 and Jingrennuo. SDS-PAGE analysis of the total endosperm protein showed that the low glutelin content trait was controlled by a single dominant nuclear gene. Genetic mapping, using SSRs, located this gene to chromosome 2, in the region between SSR2-001/SSR2-004 and RM1358. The dis- tances of the two markers to the target gene were 1.1 cM and 3.8 cM respectively. By semi-quantitative RT-PCR analysis, the transcripts of GluB4/GluB5 genes located within the region do not change. However, GluB5 gene located proximal to SSR2-001/SSR2-004 was specifically reduced. SSR profiles of seven Japonica varieties were compared with that of W3660 for loci in the relevant genetic region. The markers SSR2-004 and RM1358 were used for marker- assisted selection. The selection efficiencies of SSR2-004 and RM1358 were 96.8% and 92.7% respectively. This provides a standard starting point for the breeding of low glutelin content rice varieties in China.展开更多
Genetic comparison between Rhynchophorus ferrugineus (Coleoptera: Curculionidae: Rhynchophorinae) and other four Rhynchophorus spp. such as R. bilineatus, R. phoenicis, R. cruentatus and R. palmarum was investigat...Genetic comparison between Rhynchophorus ferrugineus (Coleoptera: Curculionidae: Rhynchophorinae) and other four Rhynchophorus spp. such as R. bilineatus, R. phoenicis, R. cruentatus and R. palmarum was investigated using partial sequences of Cytochrome c oxidase subunit I (COl) mitochondrial gene. The number ofpolymorphic sites ranged from 79 to 102 while the genetic distances ranged from 0.058 to 0.095. The number of polymorphic sites, genetic distances and phylogenetic relationships among R. ferrugineus and the other four tested species revealed that R.ferrugineus was more closely related to R. bilineatus while it was distantly related to R. palmarum. It is possible to rank the five tested species as follows: R.ferrugineus, R. bilineatus, R. phoenicis, R. cruentatus and R. palmarum. The result of this study is the first step of the revision of the uncertain taxonomic status and phylogenetic relationships among the Rhynchophorus genus. Further analysis should be performed using other Rhynchophorus species as well as other molecular markers.展开更多
Objective: The aim of this study was to study the effect of Ginkgo biloba extract (EGb761) on metabolism of afiatoxin B1 (AFB1) in Wistar rats. Methods: Seventy one Wistar rats were assigned at random to groups ...Objective: The aim of this study was to study the effect of Ginkgo biloba extract (EGb761) on metabolism of afiatoxin B1 (AFB1) in Wistar rats. Methods: Seventy one Wistar rats were assigned at random to groups A, B and C. Rats in groups A, B were injected with AFB1 (intraperitoneal, 100-200 ug/kg body weight, 1-3 times/week). Group C was normal control. Rats in group B were fed in food with EGb761, while rats in groups A, C were given normal food. Blood samples were collected and liver biopsies were performed on the 14th, 28th and 42nd week. All the rats were sacrificed on the 64th week. The incidence of hepatocarcinoma was investigated. The hepatic phase I drug-metabolizing enzyme Cytochrome-P450 (CYP450) and phase II metabolizing enzyme glutathione S-transferase (GST) were analyzed with spectrometry. Serum AFB1- lysine adduct levels were assessed with high performance liquid chromatography (HPLC). The expression of 8-hydroxydeoxy- guanosine (8-OHdG) was measured with immunohistochemistry. Results: The incidence of hepatocellular carcinoma (HCC) in group B was significantly lower than that in group A (26.92% vs 76.00%, P 〈 0.001). No HCC developed in group C. EGb761 showed no effects on the activities of CYP450 and GST in rat liver tissues. The level of AFB1-lysine adduct reached the peak (4356.01 pg/mg albumin) at the 14th week in group A. EGb761 significantly inhibited the formation of AFB1-lysine adduct in serum by 13.07% at the 14th week (P = 0.033), and 73.63% at the 42nd week (P = 0.002). The expression of 8-OHdG protein in rat liver tissues in group B was significantly lower than that in group A at the 28th, 42nd, and 64th week (P 〈 0.05). Conclusion: The main mechanism underlying the effect of EGb761 in blocking hepatocarcinogenesis induced by AFB1 may not be fully attributable to its influence on the activity of liver phase I and phase II metabolizing enzymes. EGb761 inhibits the production of AFB1-lysine adducts, decreases the expression of 8-OHdG protein, and finally alleviates the DNA oxidative injury, which may be one of the mechanisms for the effects of EGb761 in inhibiting or delaying AFB1-induced hepatocarcinogenesis.展开更多
[Objective] The aim was to explore an optimized method to measure the artificial pigment by means of high performance liquid chroma- tograph. [Method] The gradient elution technique of HPLC was used to isolate and ana...[Objective] The aim was to explore an optimized method to measure the artificial pigment by means of high performance liquid chroma- tograph. [Method] The gradient elution technique of HPLC was used to isolate and analyze tartrazine and sunset yellow in the cereals. The pre- treatment temperature, ultrasonic time, the column temperature and the elution conditions were studied, and the best expedmental conditions were confirmed. [ Result] At 40 ℃, the optimum chromatography analysis condition was moving phase, with pH (6.8), methanol/water of acetic acid- ammonium acetate (0.02 mol/L), and column temperature (35 ℃). Gradient elution condition was 20% -35% methanol, 3%/rain; 35% -98%, 6%/min; 98%, 8 min. The content of tartrazine and sunset yellow in cereal was tested precisely by the means of HPLC. The average recoveries of standard addition were 97.9% and 100.2%, and the detection limit were 0.02 mg/kg and 0.03 mg/kg. [ Conclusion] This method shortened the a- nalysis time, improved the detection sensitivity and the resolution of artificial color.展开更多
Because conditions for the grain production are not favorable and the input in the production factors ofmodern agriculture is not sufficient, the grain supply has been less than the demand for a long time. By means of...Because conditions for the grain production are not favorable and the input in the production factors ofmodern agriculture is not sufficient, the grain supply has been less than the demand for a long time. By means of greycorrelation analysis, this paper deals with the correlation situation among such input factors as the grain sown area, thetotal power of agricultural machinery, the fertilizer utilization volume, the actual irrigation area, the damage area, thenumber of farming labor force, in order to understand the major restricting and driving forces that have affected thegrain output in the recent 23 years. The analysis shows that the grain sown area and fertilizer utilization volume are thetwo most important leading factors that affect grain production in Guizhou Province, while the damage area and the totalpower of the farming-used machines are the following factors. Fertilizer utilization volume and the actual agriculturalmachinery and irrigation area are the two increasingly influential factors that affect grain production. The influence of thefarming labor force upon grain production became less strong. The main measures to increase grain production outputin Guizhou Province are to stabilize the arable land, increase the actual irrigation area, expedite the transfer of thefarming labor force and increase the input of modern agriculture.展开更多
We determined the sequence of mitochondrial Cytochrome oxidase subunit I (CO1) in two Drosophila melanogaster strains originating at "Evolution Canyon", Israel. CO1 nucleotide sequences from two iso-female strains...We determined the sequence of mitochondrial Cytochrome oxidase subunit I (CO1) in two Drosophila melanogaster strains originating at "Evolution Canyon", Israel. CO1 nucleotide sequences from two iso-female strains, 2-1 and 6-1, were 1,534 and 1,543 base-pairs, respectively. In each strain, ATAA was used in initiation of translation. Exchange rates for nucleotide and amino acid sequences were larger in the 6-1 strain than in the 2-1 strain when Oregon-R was used as the standard. Non-synonymous exchange rate was larger than synonymous exchange rate among the three strains.展开更多
The Red Palm Weevil (RPW), Rhynchophorus ferrugineus (Olivier), (Coleoptera, Curculionidae, Rhynchophorinae), is an invasive species that is originated from Southeast Asia. It has invaded Middle East, several co...The Red Palm Weevil (RPW), Rhynchophorus ferrugineus (Olivier), (Coleoptera, Curculionidae, Rhynchophorinae), is an invasive species that is originated from Southeast Asia. It has invaded Middle East, several countries of the Mediterranean Basin, Caribbean (Island of Curacao, Netherland Antilles), Lebanon and United States of America (Laguna Beach, Orange County, California) during the last three decades where it attacks palm trees. This study investigated the geographical variation among geographic populations of RPW collected from twelve invaded countries using cytochrome b (Cytb) and ITS2 partial sequences. The comparison among the Cytb sequences resulted in three different haplotypes designated as HB 1 to HB3. The three haplotypes were subdivided into two phylogenetic groups according to their geographic positions: (1) the "Multi-Continent" group containing HB 1 haplotype detected in eight populations belonging to three different Continents such as Africa-Egypt, Asia-Kingdom of Saudi Arabia and Turkey and Europe-Spain, Italy, Greece, Cyprus and France, and (2) the "Asian" group includes four populations from Iran (harboring HB2 haplotype), Pakistan, UAE, and Oman (harboring HB3 haplotype). This mitochondrial pattern of genetic diversity suggests that the tested RPW populations subdivided genetically into different sub-populations under the influence of genetic drift. According to these results we concluded that the twelve studied RPW populations are originating from different source populations in the RPW area of origin.展开更多
We summarized the discovery of liquid chromatography mass spectrometry (LC-MS) based glutathione (GSH) capture of reactive metabolites in traditional Chinese medicine (TCM), which provides scientific basis for ...We summarized the discovery of liquid chromatography mass spectrometry (LC-MS) based glutathione (GSH) capture of reactive metabolites in traditional Chinese medicine (TCM), which provides scientific basis for further research and clinical application of Chinese medicine toxicity discovery. This dissertation overviews relevant literatures of GSH capture of reactive metabolites in recent years. And then we review the principles and methods of LC-MS based GSH capture of reactive metabolites, as well as the research progress in the discovery of toxicity of TCM including pyrrolizidine alkaloids, furans and quinoid species. The review shows that the representative compounds of TCM includes adonifoline, lasiocarpine, diosbulbin B and safrol are well dctected by LC-MS based GSH capture technique. And the main analytic systems of LC-MS are triple quadrupole and Q-Trap mass spectrometer. Constant neutral loss scan (CNLS), precursor ion scan (PIS) and multiple reaction monitor (MRM) are main detection methods to monitor the characteristic GSH conjugate fragmentations. The approach of LC-MS based GSH-trapped metabolites has a good application prospect in the discovery of toxic components of TCM.展开更多
Recently much attention has been paid to the application of metal hybrid nanoparticles in industrial catalytic fields because of their super-efficient catalytic activity and attractive properties. We explored a novel ...Recently much attention has been paid to the application of metal hybrid nanoparticles in industrial catalytic fields because of their super-efficient catalytic activity and attractive properties. We explored a novel strategy to prepare GSH-capped Pt–Au–Ag-hybrid nanoclusters through the synergistic effect between ascorbic acid(VC) and glutathione(GSH) with chloroplatinic acid, chloroauric acid, and silver nitrate as precursors. The potential utilization of as-prepared GSH-capped Pt–Au–Aghybrid nanoclusters for catalytic applications has been evaluated through the reduction of 4-nitrophenol(4-NP) with NaBH4; we obtained the kinetic data by monitoring with UV-Vis spectroscopy. Our results illustrate that GSH-capped Pt–Au– Ag-hybrid nanoclusters could facilitate the process of reduction of 4-NP in a way that is unprecedented. This approach may offer a novel, non-cytotoxicity, efficient catalyst for industry.展开更多
Glutathione(GSH)plays a critical role in maintaining oxidation-reduction homeostasis in biological systems.Considering the detection of GSH by fluorescence sensors is limited by either the short wavelength emission or...Glutathione(GSH)plays a critical role in maintaining oxidation-reduction homeostasis in biological systems.Considering the detection of GSH by fluorescence sensors is limited by either the short wavelength emission or the poor photostability,a highly stable colorimetric and ratiometric NIR fluorescent sensor(DCM-S)for GSH detection has been constructed on the basis of dicyanomethylene-4H-pyran(DCM)chromophore.The specific disulfide bond is incorporated via a carbamate linker as the GSH responsive group,which simultaneously blue-shifts and quenches the fluorescence.Upon addition of GSH,DCM-S exhibits outstanding colorimetric(from yellow to red)and ratiometric fluorescent response with the 6-fold enhancement of NIR fluorescence at 665 nm in quantum yield.More importantly,the GSH-treated DCM-S(DCM-NH_2 actually)possesses 20-fold longer fluorescence half-life period as well as much better photostability than the FDA-approved ICG.Finally,the ratiometric detection of GSH is also successfully operated in the living cell imaging,exhibiting NIR fluorescence and large Stokes shift(215 nm)with nearly no background fluorescence interference.As a consequence,DCM-S can be utilized as colorimetric and ratiometric NIR fluorescent sensor for GSH,with a great potential in the development of GSH-induced drug delivery system.展开更多
基金supported by the National Nature Science Foundation of China(No.39770251)the Medical Foundation of the People's Liberation Army,China(No.01Z082,06MA234)the Foundation of the Fourth Military Medical University(No.05ZXJM001)
文摘Objective To investigate whether hypertonic saline (HS) can induce the synthesis and release of glutamate in cultured hypothalamic astrocytes or C6 cell line. Methods Astrocytes were isolated, cultured, purified and identified from the hypothalamus of newborn rat (1 day). The astrocytes were randomly divided into five groups: isotonic (IS) and HS groups, astrocytes were incubated by IS and HS (320 mosM NaCl) medium, respectively, for 1, 3, 5, 10 or 15 rain; carbenoxolone (CBX) +IS and CBX+HS groups, astrocytes were pre-treated with CBX (100 mmol/L) for 1 h at 37℃ in a 5% CO2 / 95% atmosphere, then removed to IS and HS medium, respectively, for 1, 3, 5, 10 or 15 min; Ca2++HS group, astrocytes were pre-incubated with Ca2+ (1 000 μmol/L) for 1 h at 37℃ in a 5% CO2 / 95% atmosphere, followed by a wash with isotonic FBS/DMEM, and then removed to hypertonic saline for 1, 3, 5, 10 or 15 min. The media of five groups were collected to analyze the medium glutamate concentration with high performance liquid chromatography. The astrocytes were fixed and double immunofluorescent stained with anti-glial fibrillary acidic protein (GFAP) and anti-glutamate. The C6 cells were divided into four groups: IS, HS, CBX+IS and CBX+HS groups, and used for quantitative measurement of glutamate in cells by flow cytometry (FCM). Results (1) Anti-GFAP immunofluorescent signal revealed no significant difference among various time points in each group, or among the five groups. (2) The anti-glutamate immunofluorescent signal was increased in HS group and peaked at 5 min, and decreased and returned to the level of IS group at 15 rain (P 〈 0.01 vs the 5 min of HS group). In CBX+HS group, the glutamate intensity was higher than that in CBX+IS and HS groups. (3) The medium glutamate concentration had no change after treatment with HS for 1 and 3 min, while increased markedly after treatment for 5 min to 15 min (P 〈 0.01 vs 1 min and 3 min). On the contrary, the medium glutamate concentrations in the CBX+HS or Ca2++HS group were significant lower than that in the HS group (P 〈 0.01). (4) FCM showed HS and CBX+HS induced glutamate increase in C6 cells. Conclusion HS induced cultured rat hypothalamic astrocytes or C6 cells to synthesize and release glutamate; CBX could block glutamate release, but could not disrupt glutamate synthesis.
文摘A pot experiment was conducted to study the effect of nitrogen fertilizer on nitrogen and potassium uptake by four rice cultivars. Results showed that the quadatic parabola relationship between biomass of rice and nitrogen levels was observed, with the maximum biomass at the nitrogen level of 150 mg kg-1.The rates of nitrogen and potassium uptake by the four rice cultivars depended on growth stage and rice cultivar with the maximum rate of N in Shanyou-63 and maximum rate of K in Kaiyou-5 (hybrid rice),respedively. The kinetics of nitrogen and potasssium uptake by rice plant could be quantitstively described by the following equations: y = a + blogt, y = ab + t1/2 and y = ae-bt. The b value in the equations was correlated significantly to the rates of nitrogen and potassium uptake (NR and KR, r=0.901**~0.990**),suggesting that the b value could be used to distinguish the index of nitrogen and potassium uptake capacity of rice. The maximum values of nitrogen uptake by plant (b value) and apparent recovery of fertilizer nitrogen were observed in Shanyou-63, and the minimum value in Eryou-6078. However, the capacity of potassium uptake (b value) by Kaiyou-5 ranked first and that by Shanyou-63 second. There was a significant linear relationship between nitrogen level and nitrogen uptake by rice, but a quadratic parabola relationship was found between nitrogen level and patassium uptake by rice. The application of nitrogen fertilizer decreased the ratios of potassium to nitrogen uptake by rice plant. The greatest reduction in the ratio was observed at high nitrogen level, and the least reduction was found in Kaiyou-5 and Shanyou-63 due to their greater ability to absorb potassium.
基金the Chinese Traditional Medicine Founda-tion of Zhejiang (No. 2007CB184)
文摘Objective: To investigate the death mode of human hepatoma cells exposed to matrine and the role of glutathione (GSH) and cytochrome c. Methods: The MTT test and Cell Death Detection ELISA were used to identify cell death mode and viability of cells exposed to matrine. The volume of intracellular GSH was detected by GSH reductase. Finally Western blotting was chosen to analyze the expression of cytochrome c and Caspase-9 in HepG2 cells treated by matrine. Results: The apop-totic cell death induced by matrine in Hep G2 cells dramatically increased in the time-, dose-dependent manner. Matrine can exhaust intracellular GSH effectively to change the redox state in cells. Furthermore it affect the cytotoxicity of matrine. Re-sults of Western blotting showed that matrine induced the release of cytochrome c from mitochondria to cytoplasm, and then stimulate the cleavage of Caspase-9 in a time-dependent manner. Conclusion: Matrine induced apoptosis in Hep G2 cells through the mitochondrial pathway, and oxidative stress via depletion of GSH is directly involved in the apoptotic process.
基金supported by the grants from Hi-Tech Research and Development Program of China("863"Program,No.2003AA222131,2003AA207020)the National Natural Science Foundation of China(No.30170570)Special Program for gene-transfering(No.JY03-B-07,JY03-A-07-02)
文摘Rice with low glutelin content is suitable as functional food for patients affected with diabetes and kidney failure. The fine mapping of the gene(s) responsible for low glutelin content will provide information regarding the distribution of glutelin related genes in rice genome and will generate markers for the selection of low glutelin rice varieties. Following an SDS-PAGE screen of rice germplasm from Taihu Valley of China, Japonica selection W3660 is identified to be a novel mutant characterized with low glutelin content. For fine mapping the mutant gene for low glutelin content, F2 and F3 populations were derived from a cross between W3660 and Jingrennuo. SDS-PAGE analysis of the total endosperm protein showed that the low glutelin content trait was controlled by a single dominant nuclear gene. Genetic mapping, using SSRs, located this gene to chromosome 2, in the region between SSR2-001/SSR2-004 and RM1358. The dis- tances of the two markers to the target gene were 1.1 cM and 3.8 cM respectively. By semi-quantitative RT-PCR analysis, the transcripts of GluB4/GluB5 genes located within the region do not change. However, GluB5 gene located proximal to SSR2-001/SSR2-004 was specifically reduced. SSR profiles of seven Japonica varieties were compared with that of W3660 for loci in the relevant genetic region. The markers SSR2-004 and RM1358 were used for marker- assisted selection. The selection efficiencies of SSR2-004 and RM1358 were 96.8% and 92.7% respectively. This provides a standard starting point for the breeding of low glutelin content rice varieties in China.
文摘Genetic comparison between Rhynchophorus ferrugineus (Coleoptera: Curculionidae: Rhynchophorinae) and other four Rhynchophorus spp. such as R. bilineatus, R. phoenicis, R. cruentatus and R. palmarum was investigated using partial sequences of Cytochrome c oxidase subunit I (COl) mitochondrial gene. The number ofpolymorphic sites ranged from 79 to 102 while the genetic distances ranged from 0.058 to 0.095. The number of polymorphic sites, genetic distances and phylogenetic relationships among R. ferrugineus and the other four tested species revealed that R.ferrugineus was more closely related to R. bilineatus while it was distantly related to R. palmarum. It is possible to rank the five tested species as follows: R.ferrugineus, R. bilineatus, R. phoenicis, R. cruentatus and R. palmarum. The result of this study is the first step of the revision of the uncertain taxonomic status and phylogenetic relationships among the Rhynchophorus genus. Further analysis should be performed using other Rhynchophorus species as well as other molecular markers.
文摘Objective: The aim of this study was to study the effect of Ginkgo biloba extract (EGb761) on metabolism of afiatoxin B1 (AFB1) in Wistar rats. Methods: Seventy one Wistar rats were assigned at random to groups A, B and C. Rats in groups A, B were injected with AFB1 (intraperitoneal, 100-200 ug/kg body weight, 1-3 times/week). Group C was normal control. Rats in group B were fed in food with EGb761, while rats in groups A, C were given normal food. Blood samples were collected and liver biopsies were performed on the 14th, 28th and 42nd week. All the rats were sacrificed on the 64th week. The incidence of hepatocarcinoma was investigated. The hepatic phase I drug-metabolizing enzyme Cytochrome-P450 (CYP450) and phase II metabolizing enzyme glutathione S-transferase (GST) were analyzed with spectrometry. Serum AFB1- lysine adduct levels were assessed with high performance liquid chromatography (HPLC). The expression of 8-hydroxydeoxy- guanosine (8-OHdG) was measured with immunohistochemistry. Results: The incidence of hepatocellular carcinoma (HCC) in group B was significantly lower than that in group A (26.92% vs 76.00%, P 〈 0.001). No HCC developed in group C. EGb761 showed no effects on the activities of CYP450 and GST in rat liver tissues. The level of AFB1-lysine adduct reached the peak (4356.01 pg/mg albumin) at the 14th week in group A. EGb761 significantly inhibited the formation of AFB1-lysine adduct in serum by 13.07% at the 14th week (P = 0.033), and 73.63% at the 42nd week (P = 0.002). The expression of 8-OHdG protein in rat liver tissues in group B was significantly lower than that in group A at the 28th, 42nd, and 64th week (P 〈 0.05). Conclusion: The main mechanism underlying the effect of EGb761 in blocking hepatocarcinogenesis induced by AFB1 may not be fully attributable to its influence on the activity of liver phase I and phase II metabolizing enzymes. EGb761 inhibits the production of AFB1-lysine adducts, decreases the expression of 8-OHdG protein, and finally alleviates the DNA oxidative injury, which may be one of the mechanisms for the effects of EGb761 in inhibiting or delaying AFB1-induced hepatocarcinogenesis.
基金Supported by Heilongjiang Natural Science Fund(B200911)Young Teacher' Scientific Research Support Program in Qiqihaer University(2010k-M17)Qiqihaer City Industrial Research and Development Program(GYGG2010-06-1)
文摘[Objective] The aim was to explore an optimized method to measure the artificial pigment by means of high performance liquid chroma- tograph. [Method] The gradient elution technique of HPLC was used to isolate and analyze tartrazine and sunset yellow in the cereals. The pre- treatment temperature, ultrasonic time, the column temperature and the elution conditions were studied, and the best expedmental conditions were confirmed. [ Result] At 40 ℃, the optimum chromatography analysis condition was moving phase, with pH (6.8), methanol/water of acetic acid- ammonium acetate (0.02 mol/L), and column temperature (35 ℃). Gradient elution condition was 20% -35% methanol, 3%/rain; 35% -98%, 6%/min; 98%, 8 min. The content of tartrazine and sunset yellow in cereal was tested precisely by the means of HPLC. The average recoveries of standard addition were 97.9% and 100.2%, and the detection limit were 0.02 mg/kg and 0.03 mg/kg. [ Conclusion] This method shortened the a- nalysis time, improved the detection sensitivity and the resolution of artificial color.
文摘Because conditions for the grain production are not favorable and the input in the production factors ofmodern agriculture is not sufficient, the grain supply has been less than the demand for a long time. By means of greycorrelation analysis, this paper deals with the correlation situation among such input factors as the grain sown area, thetotal power of agricultural machinery, the fertilizer utilization volume, the actual irrigation area, the damage area, thenumber of farming labor force, in order to understand the major restricting and driving forces that have affected thegrain output in the recent 23 years. The analysis shows that the grain sown area and fertilizer utilization volume are thetwo most important leading factors that affect grain production in Guizhou Province, while the damage area and the totalpower of the farming-used machines are the following factors. Fertilizer utilization volume and the actual agriculturalmachinery and irrigation area are the two increasingly influential factors that affect grain production. The influence of thefarming labor force upon grain production became less strong. The main measures to increase grain production outputin Guizhou Province are to stabilize the arable land, increase the actual irrigation area, expedite the transfer of thefarming labor force and increase the input of modern agriculture.
文摘We determined the sequence of mitochondrial Cytochrome oxidase subunit I (CO1) in two Drosophila melanogaster strains originating at "Evolution Canyon", Israel. CO1 nucleotide sequences from two iso-female strains, 2-1 and 6-1, were 1,534 and 1,543 base-pairs, respectively. In each strain, ATAA was used in initiation of translation. Exchange rates for nucleotide and amino acid sequences were larger in the 6-1 strain than in the 2-1 strain when Oregon-R was used as the standard. Non-synonymous exchange rate was larger than synonymous exchange rate among the three strains.
文摘The Red Palm Weevil (RPW), Rhynchophorus ferrugineus (Olivier), (Coleoptera, Curculionidae, Rhynchophorinae), is an invasive species that is originated from Southeast Asia. It has invaded Middle East, several countries of the Mediterranean Basin, Caribbean (Island of Curacao, Netherland Antilles), Lebanon and United States of America (Laguna Beach, Orange County, California) during the last three decades where it attacks palm trees. This study investigated the geographical variation among geographic populations of RPW collected from twelve invaded countries using cytochrome b (Cytb) and ITS2 partial sequences. The comparison among the Cytb sequences resulted in three different haplotypes designated as HB 1 to HB3. The three haplotypes were subdivided into two phylogenetic groups according to their geographic positions: (1) the "Multi-Continent" group containing HB 1 haplotype detected in eight populations belonging to three different Continents such as Africa-Egypt, Asia-Kingdom of Saudi Arabia and Turkey and Europe-Spain, Italy, Greece, Cyprus and France, and (2) the "Asian" group includes four populations from Iran (harboring HB2 haplotype), Pakistan, UAE, and Oman (harboring HB3 haplotype). This mitochondrial pattern of genetic diversity suggests that the tested RPW populations subdivided genetically into different sub-populations under the influence of genetic drift. According to these results we concluded that the twelve studied RPW populations are originating from different source populations in the RPW area of origin.
文摘We summarized the discovery of liquid chromatography mass spectrometry (LC-MS) based glutathione (GSH) capture of reactive metabolites in traditional Chinese medicine (TCM), which provides scientific basis for further research and clinical application of Chinese medicine toxicity discovery. This dissertation overviews relevant literatures of GSH capture of reactive metabolites in recent years. And then we review the principles and methods of LC-MS based GSH capture of reactive metabolites, as well as the research progress in the discovery of toxicity of TCM including pyrrolizidine alkaloids, furans and quinoid species. The review shows that the representative compounds of TCM includes adonifoline, lasiocarpine, diosbulbin B and safrol are well dctected by LC-MS based GSH capture technique. And the main analytic systems of LC-MS are triple quadrupole and Q-Trap mass spectrometer. Constant neutral loss scan (CNLS), precursor ion scan (PIS) and multiple reaction monitor (MRM) are main detection methods to monitor the characteristic GSH conjugate fragmentations. The approach of LC-MS based GSH-trapped metabolites has a good application prospect in the discovery of toxic components of TCM.
基金supported by the National Basic Research Program of China(2010CB732404)the National Natural Science Foundation of China(81325011,21327902,21175020)+1 种基金the National High Technology Research and Development Program of China(2012AA022703)the Major Science&Technology Project of Suzhou(ZXY2012028)
文摘Recently much attention has been paid to the application of metal hybrid nanoparticles in industrial catalytic fields because of their super-efficient catalytic activity and attractive properties. We explored a novel strategy to prepare GSH-capped Pt–Au–Ag-hybrid nanoclusters through the synergistic effect between ascorbic acid(VC) and glutathione(GSH) with chloroplatinic acid, chloroauric acid, and silver nitrate as precursors. The potential utilization of as-prepared GSH-capped Pt–Au–Aghybrid nanoclusters for catalytic applications has been evaluated through the reduction of 4-nitrophenol(4-NP) with NaBH4; we obtained the kinetic data by monitoring with UV-Vis spectroscopy. Our results illustrate that GSH-capped Pt–Au– Ag-hybrid nanoclusters could facilitate the process of reduction of 4-NP in a way that is unprecedented. This approach may offer a novel, non-cytotoxicity, efficient catalyst for industry.
基金the National Basic Research Program(2013CB733700)the National Natural Science Foundation of China for Creative Research Groups(21421004)+2 种基金Distinguished Young Scholars(21325625)the Oriental Scholarship,the Fundamental Research Funds for the Central Universities(WJ1416005)Science and Technology Commission of Shanghai Municipality(15XD1501400)
文摘Glutathione(GSH)plays a critical role in maintaining oxidation-reduction homeostasis in biological systems.Considering the detection of GSH by fluorescence sensors is limited by either the short wavelength emission or the poor photostability,a highly stable colorimetric and ratiometric NIR fluorescent sensor(DCM-S)for GSH detection has been constructed on the basis of dicyanomethylene-4H-pyran(DCM)chromophore.The specific disulfide bond is incorporated via a carbamate linker as the GSH responsive group,which simultaneously blue-shifts and quenches the fluorescence.Upon addition of GSH,DCM-S exhibits outstanding colorimetric(from yellow to red)and ratiometric fluorescent response with the 6-fold enhancement of NIR fluorescence at 665 nm in quantum yield.More importantly,the GSH-treated DCM-S(DCM-NH_2 actually)possesses 20-fold longer fluorescence half-life period as well as much better photostability than the FDA-approved ICG.Finally,the ratiometric detection of GSH is also successfully operated in the living cell imaging,exhibiting NIR fluorescence and large Stokes shift(215 nm)with nearly no background fluorescence interference.As a consequence,DCM-S can be utilized as colorimetric and ratiometric NIR fluorescent sensor for GSH,with a great potential in the development of GSH-induced drug delivery system.