Parasinilabeo longiventralis, a new species is here described from He jiang basin, a tributary of the Zhujiang River (Pearl River) drainage in Fuchuan County, Guangxi Province, southern China. It can be distinguishe...Parasinilabeo longiventralis, a new species is here described from He jiang basin, a tributary of the Zhujiang River (Pearl River) drainage in Fuchuan County, Guangxi Province, southern China. It can be distinguished from all other congeners by having the following combination of characteristics: maxillary barbels length is 78.3-90.4% of rostral barbels length; pelvic-fin length is 83.7-89.4% of the distance between the ventral-fin origin and to the anal-fin origin; a longitudinal wide black stripe running along the posterior lateral line and irregular brown pigments on the sides of the body.展开更多
[Objective] This study aimed to establish a real-time PCR method for de- tecting stx2 gene in Shiga toxin-producing E. coli (STEC). [Method] According to the known STEC stx2 gene sequences published in GenBank, PCR ...[Objective] This study aimed to establish a real-time PCR method for de- tecting stx2 gene in Shiga toxin-producing E. coli (STEC). [Method] According to the known STEC stx2 gene sequences published in GenBank, PCR primers and probes were designed based on the conserved region to construct recombinant plasmid as a positive template, thus optimizing the reaction conditions and establishing the real- time PCR method. [Result] A standard curve was established based on the opti- mized real-time PCR system, indicting a good linear correlation between the initial template concentration and Ct value, with the correlation coefficient F^e of above 0.995. The established method had a good specificity, without non-specific amplifica- tion for 10 non-STEC intestinal bacterial strains; the detection limit of initial template was 1.0x102 copies/μI, indicating a high sensitivity; furthermore, the coefficients of variation within and among batches were lower than 1% and 5% respectively, sug- gesting a good repeatability. [Conclusion] In this study, a real-time PCR method was successfully established for detecting STEC stx2 gene, which provided technical means for rapid detection of STEC in samples.展开更多
[Objective] The study aimed to discuss the influences of edge effect on orthopteran community in east slope of Helan Mountain.[Method] Samples are collected by using sample zone method.The different species of orthopt...[Objective] The study aimed to discuss the influences of edge effect on orthopteran community in east slope of Helan Mountain.[Method] Samples are collected by using sample zone method.The different species of orthopterans in different habitats are recorded.[Result] The percentage of Oedipodidae,Catantopidae and Pamphgidae in total are 42.65%,29.15% and 12.76% respectively.From scattered grassland interior to edge and then to desert grassland,abundance increase in turn,but diversity in the edge is the highe...展开更多
AIM: To screen the immunogenic membrane proteins of Shigella Aexneri 2a 2457T. METHODS: The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogeni...AIM: To screen the immunogenic membrane proteins of Shigella Aexneri 2a 2457T. METHODS: The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. Aexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides. RESULTS: A total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine. CONCLUSION: Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum.展开更多
Slope aspect has significant effect on the development and distribution of montane forest, especially in arid and semiarid regions. This paper, using SPOT5 images and 1:5o,ooo DEM, digitally extracts and analyzes the...Slope aspect has significant effect on the development and distribution of montane forest, especially in arid and semiarid regions. This paper, using SPOT5 images and 1:5o,ooo DEM, digitally extracts and analyzes the spatial information of montane coniferous forest (mainly Qinghai spruce), and thereby explores how the upper and lower limits and the altitudinal range of coniferous forest vary and how the area of coniferous forest is related with annual insolation with all aspects in the Helan Mountains. The results show that: 1) In the eastern flank, the lower limit of coniferous forest is between 1,6oo m and a,ooo m a.s.l., and the upper limit between 2,800 m and 3,ooo m a.s.1.; in the western flank, the lower limit of coniferous forest is between a,ooo m and 2,300 m a.s.l., and the upper limit between 9,80o m and 3,1oo m a.s.1.2) The altitudinal ranges of coniferous forest are 8o6-1,435 m, 438-1,14o m for eastern flank and western flank, respectively. 3) The area of coniferous forest takes on a normal distribution with aspect, and it has a close relationship with annual insolation. This distribution model developed in this paper quantitatively reveals the significant effect of slope aspect on the distribution of coniferous forest in arid and semi-arid land.展开更多
An isothermal amplification assay for the detection of Shigella based colloidal gold strip detection is developed. The primers designed corresponded to the ipaH gene of Shigella .The sensitivity of the colloidal gold ...An isothermal amplification assay for the detection of Shigella based colloidal gold strip detection is developed. The primers designed corresponded to the ipaH gene of Shigella .The sensitivity of the colloidal gold strip-NASBA method in this study was 6.3 x 101cfu/ml. The specificity of the detection system was detected and the result showed that the NASBA method could distinguish Shigella from other germs.展开更多
AIM: TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer's patch on the release of nitric oxide (NO) and IL-6 in response to Shigella lipopolysaccharide (LPS). METH...AIM: TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer's patch on the release of nitric oxide (NO) and IL-6 in response to Shigella lipopolysaccharide (LPS). METHODS: Human colonic epithelial cells (Caco-2) were mixed cocultured with lymphocytes of Peyer's patch from wild-type (C57 mice) and inducible NO synthase knockout mice, and challenged with Shigella F2a-12 LPS. Release of NO and raiL-6 was measured by Griess colorimetric assay and enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: In the absence of LPS challenge, NO was detected in the culture medium of Caco-2 epithelial cells but not in lymphocytes of Peyer's patch, and the NO release was further up-regulated in both cocultures with lymphocytes from either the wild-type or iNOS knockout mice, with a significantly higher level observed in the coculture with iNOS knockout lymphocytes. After Shigella F2a-12 LPS challenge for 24-h, NO production was significantly increased in both Caco-2 alone and the coculture with lymphocytes of Peyer's patch from the wild-type mice but not from iNOS knockout mice. LPS was found to stimulate the release of mIL-6 from lymphocytes, which was suppressed by coculture with Caco-2 epithelial cells. The LPS-induced mIL-6 production in lymphocytes from iNOS knockout mice was significantly greater than that from the wild-type mice. CONCLUSION: Lymphocytes of Peyer's patch maintain a constitutive basal level of NO production from the enteric epithelial cell Caco-2. LPS-induced mIL-6 release from lymphocytes of Peyer's patch is suppressed by the cocultured epithelial cells. While no changes are detectable in NO production in lymphocytes from both wild-type and iNOS knockout mice before and after LPS challenge, NO from lymphocytes appears to play an inhibitory role in epithelial NO release and their own mIL-6 release in response to LPS.展开更多
The genealogical relationship and agronomic characters of 13 peanut varieties,breeded by Hezhou Academy of Agricultural Sciences,which had been authorized were analyzed.The results showed that 20 direct parents were i...The genealogical relationship and agronomic characters of 13 peanut varieties,breeded by Hezhou Academy of Agricultural Sciences,which had been authorized were analyzed.The results showed that 20 direct parents were involved in 13 varieties,80%of which came from Guangxi and Guangdong Province.Shanyou 523 and Yueyou 193 were used for more times;The genealogical analysis of these certified varieties can be traced back to 30 ancestral parents,the proportion of which contained consanguinity of Fu Peanut and Shitouqi were 84.62%.This demonstrated that genetic basis was rather narrow and it was quite difficult to make a great breakthrough in breeding.Plant height,total number of branches and oil content changed very little,100-pod weight and pod yield increased,whereas there was a downward trend in shelling percentage.Therefore,it is quite necessary to further broaden the germplasm basis of parents,enrich breeding methods,and continue to strengthen disease-resistant breeding and specific peanut breeding.展开更多
To investigate feasibility of pathogen free industrial organics with higher agronomic value, the industrial organic wastes were subjected to vermistabilization. The body of earthworm work as "biofilter" and they can...To investigate feasibility of pathogen free industrial organics with higher agronomic value, the industrial organic wastes were subjected to vermistabilization. The body of earthworm work as "biofilter" and they can "purify" and also "disinfect" and "detoxify" municipal and industrial organics. The microbiomics of gut and cast of earthworm (Eisenia foetida Savigny) and their association with vermistabilization was studied to determine the microbial quantification in reactors (industrial organics). Worm were reared in five reactors viz. Sewage sludge (SS), Paper mill industry sludge (PS), Vegetable processing industry (VP), Tannery waste (TW) and Meat process industrial sludge (MP) for ninety days. The microbial load (Salmonella, Shigella, Escherichia, Mycobacterium, Streptococcus) in gut and cast, biomass, recovery of cast in different reactors were determined, periodically. The microbiomics of worms gut revealed the removal of Salmonella (12-18 × 10^8±0.02 to 0-4 × 10^3 + 0.05 CFU/g), Shigella (14-23 × 10^8 ± 0.04 to 0-4 × 10^3 ± 0.05 CFU/g), Escherichia (4-16 × 10^8 ± 0.02 to 0-4 × 10^2 ± 0.05 CFU/g), Mycobacterium (3-16 × 10^8 ± 0.02 to 0-3 × 10^2 ± 0.05 CFU/g), Streptococcus (6-16 × 10^8 ± 0.02 to 0-4 × 10^3 ± 0.05 CFU/g) during stabilization of industrial organics. Similarly, reduction in pathogens Salmonella (12-19 × 10^8 ± 0.02 to 0-8 ×10^3 ± 0.05 CFU/g), Shigella (7-20× 10^8 ± 0.04 to 0-2 ×10^3 ± 0.05 CFU/g), Escherichia (2-20 × 10^8 ± 0.02 to 0.0-2 × 10^3 ± 0.05), Mycobacterium (1-8 × 10^8 ±0.05 to 0.0-5 × 10^2 ± 0.05 CFU/g), Streptococcus (8-18 × 10^8 ± 0.02 to 0-7 × 10^3 ± 0.05 CFU/g) in castings of industrial organics indicates the selective nature of feeding of worm. This amply demonstrates that these pathogens have been eliminated as they entered in food chain of worms. However, it may not be possible to remove pathogens completely, but at least worms change the "microbial make-up" of industrial organics to make it harmless to the soil and enable its use as a nutritive organic fertilizer.展开更多
The mature Shiga toxin B (StxB) gene was optimized and generated by overlapping PCR. Recombinant expression vector pQE40-DHFR/StxB was constructed when the gene was cloned into pQE fusion expression vector. Induced ...The mature Shiga toxin B (StxB) gene was optimized and generated by overlapping PCR. Recombinant expression vector pQE40-DHFR/StxB was constructed when the gene was cloned into pQE fusion expression vector. Induced by Isopropyl β-D-thiogalactoside (IPTG), the DHFR/StxB fusion protein was highly expressed to the level of 41.36% in E. coli MI5 cells. The 35 kDa fusion protein with a 6 His-tag was one-step purified from inclusion bodies using Ni-NTA affinity chromatography column under denaturing conditions, and was refolded by dialyzing with a decreasing urea gradient. Purified DHFR/StxB fusion protein was used to immunize Kunming mice for generating the ascitic polyclonal antibody against recombinant StxB protein by injecting sarcoma 180 cells and the titer ofascitic polyclonal antibody is up to 1: 1× 10^6 detected by the indirect enzyme linked immunosorbent assy (ELISA). Western immunoblotting analysis revealed that the ascitic polyclonal antibody against StxB had a specific affinity for a 70 kDa shiga toxin protein of Shigella dysenteriae type 1. It is a new simple and quick method to produce a large amount of ascitic polyclonal antibody. The antibody is used to develop immunological method for detecting shiga toxin.展开更多
文摘Parasinilabeo longiventralis, a new species is here described from He jiang basin, a tributary of the Zhujiang River (Pearl River) drainage in Fuchuan County, Guangxi Province, southern China. It can be distinguished from all other congeners by having the following combination of characteristics: maxillary barbels length is 78.3-90.4% of rostral barbels length; pelvic-fin length is 83.7-89.4% of the distance between the ventral-fin origin and to the anal-fin origin; a longitudinal wide black stripe running along the posterior lateral line and irregular brown pigments on the sides of the body.
基金Supported by Agricultural Science and Technology Support Program(Social Development)of Jiangsu Province(BE2011771)~~
文摘[Objective] This study aimed to establish a real-time PCR method for de- tecting stx2 gene in Shiga toxin-producing E. coli (STEC). [Method] According to the known STEC stx2 gene sequences published in GenBank, PCR primers and probes were designed based on the conserved region to construct recombinant plasmid as a positive template, thus optimizing the reaction conditions and establishing the real- time PCR method. [Result] A standard curve was established based on the opti- mized real-time PCR system, indicting a good linear correlation between the initial template concentration and Ct value, with the correlation coefficient F^e of above 0.995. The established method had a good specificity, without non-specific amplifica- tion for 10 non-STEC intestinal bacterial strains; the detection limit of initial template was 1.0x102 copies/μI, indicating a high sensitivity; furthermore, the coefficients of variation within and among batches were lower than 1% and 5% respectively, sug- gesting a good repeatability. [Conclusion] In this study, a real-time PCR method was successfully established for detecting STEC stx2 gene, which provided technical means for rapid detection of STEC in samples.
基金Supported by Ministry of Education,New Century Excellent Talents Scheme(NCET-07-0470)~~
文摘[Objective] The study aimed to discuss the influences of edge effect on orthopteran community in east slope of Helan Mountain.[Method] Samples are collected by using sample zone method.The different species of orthopterans in different habitats are recorded.[Result] The percentage of Oedipodidae,Catantopidae and Pamphgidae in total are 42.65%,29.15% and 12.76% respectively.From scattered grassland interior to edge and then to desert grassland,abundance increase in turn,but diversity in the edge is the highe...
基金Supported by the Capital "248" Key Innovation Project, No. H010210360119, State Basic Research Development Program of China No. 973 Program, G1999054103 and 2005CB22904 and National Natural Science Foundation of China No. 30470101
文摘AIM: To screen the immunogenic membrane proteins of Shigella Aexneri 2a 2457T. METHODS: The routine two-dimensional polyacrylamide gel electrophoresis (2-DE) and Western blotting were combined to screen immunogenic proteins of S. Aexneri 2a 2457T. Serum was gained from rabbits immunized with the same bacteria. Immunogenic spots were cut out from the polyacrylamide gel and digested by trypsin in-gel. Matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS) was performed to determine the molecular weight of peptides. Electrospray ionization (ESI-MS/MS) was performed to determine the sequences of the interesting peptides. RESULTS: A total of 20 spots were successfully identified from Coomassie brilliant blue stained gels representing 13 protein entries, 5 known antigens and 8 novel antigens. A hypothetical protein (YaeT) was detected, which might be a candidate target of vaccine. CONCLUSION: Membrane proteins of S. flexneri 2a 2457T were successfully observed by 2-DE. Several known and novel antigens were identified by mass spectrum.
基金supported by the Naional 973 project (2009CB421307) the National Natural Science Foundation of China (41030528 and 40971064)
文摘Slope aspect has significant effect on the development and distribution of montane forest, especially in arid and semiarid regions. This paper, using SPOT5 images and 1:5o,ooo DEM, digitally extracts and analyzes the spatial information of montane coniferous forest (mainly Qinghai spruce), and thereby explores how the upper and lower limits and the altitudinal range of coniferous forest vary and how the area of coniferous forest is related with annual insolation with all aspects in the Helan Mountains. The results show that: 1) In the eastern flank, the lower limit of coniferous forest is between 1,6oo m and a,ooo m a.s.l., and the upper limit between 2,800 m and 3,ooo m a.s.1.; in the western flank, the lower limit of coniferous forest is between a,ooo m and 2,300 m a.s.l., and the upper limit between 9,80o m and 3,1oo m a.s.1.2) The altitudinal ranges of coniferous forest are 8o6-1,435 m, 438-1,14o m for eastern flank and western flank, respectively. 3) The area of coniferous forest takes on a normal distribution with aspect, and it has a close relationship with annual insolation. This distribution model developed in this paper quantitatively reveals the significant effect of slope aspect on the distribution of coniferous forest in arid and semi-arid land.
文摘An isothermal amplification assay for the detection of Shigella based colloidal gold strip detection is developed. The primers designed corresponded to the ipaH gene of Shigella .The sensitivity of the colloidal gold strip-NASBA method in this study was 6.3 x 101cfu/ml. The specificity of the detection system was detected and the result showed that the NASBA method could distinguish Shigella from other germs.
基金Supported by Strategic Program of Chinese University of Hong KongDistinguished Young Investigator Fund of the National Natural Science Foundation of China, No. 30029002
文摘AIM: TO investigate the effect of interaction between enteric epithelial cells and lymphocytes of Peyer's patch on the release of nitric oxide (NO) and IL-6 in response to Shigella lipopolysaccharide (LPS). METHODS: Human colonic epithelial cells (Caco-2) were mixed cocultured with lymphocytes of Peyer's patch from wild-type (C57 mice) and inducible NO synthase knockout mice, and challenged with Shigella F2a-12 LPS. Release of NO and raiL-6 was measured by Griess colorimetric assay and enzyme-linked immunosorbent assay (ELISA), respectively. RESULTS: In the absence of LPS challenge, NO was detected in the culture medium of Caco-2 epithelial cells but not in lymphocytes of Peyer's patch, and the NO release was further up-regulated in both cocultures with lymphocytes from either the wild-type or iNOS knockout mice, with a significantly higher level observed in the coculture with iNOS knockout lymphocytes. After Shigella F2a-12 LPS challenge for 24-h, NO production was significantly increased in both Caco-2 alone and the coculture with lymphocytes of Peyer's patch from the wild-type mice but not from iNOS knockout mice. LPS was found to stimulate the release of mIL-6 from lymphocytes, which was suppressed by coculture with Caco-2 epithelial cells. The LPS-induced mIL-6 production in lymphocytes from iNOS knockout mice was significantly greater than that from the wild-type mice. CONCLUSION: Lymphocytes of Peyer's patch maintain a constitutive basal level of NO production from the enteric epithelial cell Caco-2. LPS-induced mIL-6 release from lymphocytes of Peyer's patch is suppressed by the cocultured epithelial cells. While no changes are detectable in NO production in lymphocytes from both wild-type and iNOS knockout mice before and after LPS challenge, NO from lymphocytes appears to play an inhibitory role in epithelial NO release and their own mIL-6 release in response to LPS.
文摘The genealogical relationship and agronomic characters of 13 peanut varieties,breeded by Hezhou Academy of Agricultural Sciences,which had been authorized were analyzed.The results showed that 20 direct parents were involved in 13 varieties,80%of which came from Guangxi and Guangdong Province.Shanyou 523 and Yueyou 193 were used for more times;The genealogical analysis of these certified varieties can be traced back to 30 ancestral parents,the proportion of which contained consanguinity of Fu Peanut and Shitouqi were 84.62%.This demonstrated that genetic basis was rather narrow and it was quite difficult to make a great breakthrough in breeding.Plant height,total number of branches and oil content changed very little,100-pod weight and pod yield increased,whereas there was a downward trend in shelling percentage.Therefore,it is quite necessary to further broaden the germplasm basis of parents,enrich breeding methods,and continue to strengthen disease-resistant breeding and specific peanut breeding.
文摘To investigate feasibility of pathogen free industrial organics with higher agronomic value, the industrial organic wastes were subjected to vermistabilization. The body of earthworm work as "biofilter" and they can "purify" and also "disinfect" and "detoxify" municipal and industrial organics. The microbiomics of gut and cast of earthworm (Eisenia foetida Savigny) and their association with vermistabilization was studied to determine the microbial quantification in reactors (industrial organics). Worm were reared in five reactors viz. Sewage sludge (SS), Paper mill industry sludge (PS), Vegetable processing industry (VP), Tannery waste (TW) and Meat process industrial sludge (MP) for ninety days. The microbial load (Salmonella, Shigella, Escherichia, Mycobacterium, Streptococcus) in gut and cast, biomass, recovery of cast in different reactors were determined, periodically. The microbiomics of worms gut revealed the removal of Salmonella (12-18 × 10^8±0.02 to 0-4 × 10^3 + 0.05 CFU/g), Shigella (14-23 × 10^8 ± 0.04 to 0-4 × 10^3 ± 0.05 CFU/g), Escherichia (4-16 × 10^8 ± 0.02 to 0-4 × 10^2 ± 0.05 CFU/g), Mycobacterium (3-16 × 10^8 ± 0.02 to 0-3 × 10^2 ± 0.05 CFU/g), Streptococcus (6-16 × 10^8 ± 0.02 to 0-4 × 10^3 ± 0.05 CFU/g) during stabilization of industrial organics. Similarly, reduction in pathogens Salmonella (12-19 × 10^8 ± 0.02 to 0-8 ×10^3 ± 0.05 CFU/g), Shigella (7-20× 10^8 ± 0.04 to 0-2 ×10^3 ± 0.05 CFU/g), Escherichia (2-20 × 10^8 ± 0.02 to 0.0-2 × 10^3 ± 0.05), Mycobacterium (1-8 × 10^8 ±0.05 to 0.0-5 × 10^2 ± 0.05 CFU/g), Streptococcus (8-18 × 10^8 ± 0.02 to 0-7 × 10^3 ± 0.05 CFU/g) in castings of industrial organics indicates the selective nature of feeding of worm. This amply demonstrates that these pathogens have been eliminated as they entered in food chain of worms. However, it may not be possible to remove pathogens completely, but at least worms change the "microbial make-up" of industrial organics to make it harmless to the soil and enable its use as a nutritive organic fertilizer.
文摘The mature Shiga toxin B (StxB) gene was optimized and generated by overlapping PCR. Recombinant expression vector pQE40-DHFR/StxB was constructed when the gene was cloned into pQE fusion expression vector. Induced by Isopropyl β-D-thiogalactoside (IPTG), the DHFR/StxB fusion protein was highly expressed to the level of 41.36% in E. coli MI5 cells. The 35 kDa fusion protein with a 6 His-tag was one-step purified from inclusion bodies using Ni-NTA affinity chromatography column under denaturing conditions, and was refolded by dialyzing with a decreasing urea gradient. Purified DHFR/StxB fusion protein was used to immunize Kunming mice for generating the ascitic polyclonal antibody against recombinant StxB protein by injecting sarcoma 180 cells and the titer ofascitic polyclonal antibody is up to 1: 1× 10^6 detected by the indirect enzyme linked immunosorbent assy (ELISA). Western immunoblotting analysis revealed that the ascitic polyclonal antibody against StxB had a specific affinity for a 70 kDa shiga toxin protein of Shigella dysenteriae type 1. It is a new simple and quick method to produce a large amount of ascitic polyclonal antibody. The antibody is used to develop immunological method for detecting shiga toxin.