[Objective] To clone the actin gene of Cryptosporidium andersoni, and to study its eukaryotic expression in Hela cells. [Methed] Specific primers were designed for the partial encoding sequence of actin, which were ob...[Objective] To clone the actin gene of Cryptosporidium andersoni, and to study its eukaryotic expression in Hela cells. [Methed] Specific primers were designed for the partial encoding sequence of actin, which were obtained by screening the T7 phage display library of Cryptosporidium andersoni, and the actin gene CA42 was amplified by PCR. Recombinant eukaryotic expression plasmid pVAX1-CA42 was constructed and transfected to Hela cells with lipofection strategy. Indirect im- munofluorescence staining, SDS-PAGE and Western blotting analysis were used to detect the expression of recombinant protein in Hela cells. [Result] CA42 protein was successfully expressed in Hela cells, and the expression products had reactogenicity. [Conclusion] The partial encoding sequence of actin from Cryptosporidium andersoni has been successfully cloned, and it can be stably expressed in Hela Cells展开更多
AIM： To investigate whether supplementation of male zooid of Antheraea pernyi extracts （MZAPE） could enhance immune function of radiated tumor-bearing rats. METHODS： Eighty male Wistar rats were randomly divided i...AIM： To investigate whether supplementation of male zooid of Antheraea pernyi extracts （MZAPE） could enhance immune function of radiated tumor-bearing rats. METHODS： Eighty male Wistar rats were randomly divided into a control group, a simple radiation group, a MZAPE group, and a radiation plus MZAPE group. With the tumor model established by implanting Walker-256 ascites tumor cells, tumor weight and tumor control rate were calculated. The rats in the simple radiation and radiation plus MZAPE groups were underwent to radiation at 10 Gy within 2 d. In the MZAPE and radiation plus MZAPE groups, the MZAPE was gavaged at a dose of 16.53 mg/kg once a day for 7 d. T cell subsets in peripheral blood were determined by flow cytometry and the expression of IL-2, IFN-γ, IL-4 and IL-10 in sera were determined by ELISA on the 8th d. RESULTS： The tumor weight of simple radiation group, MZAPE group and radiation plus MZAPE group was lower than that of control group （P 〈 0.01） and tumor control rates were 63.08% ± 6.43%, 69.86%± 7.12% and 35.30% ± 7.67%, respectively. CD^4＋ T and CD^8＋ T cells in the peripheral blood of the simple radiation group were fewer than in control group. In the MZAPE and radiation plus MZAPE groups, the number of CD^4＋ T cells was higher while CD^8＋ T cells was lower than in the control and simple radiation groups. Expression of IL-2 and INF-y in the radiation group was lower than in control group, and significantly enhanced during MZAPE therapy （P 〈 0.05）. Expression of IL-4 and IL-10 in the radiation group had no significant changes compared with the control group, and decreased significantly after MZAPE treatment （P 〈 0.01）. CONCLUSION： MZAPE administration may help improve the immune function of the radiated tumor-bearing rats and reverse the radiation-induced immune inhibition by promoting the proliferation of T helper cells and inducing the transdifferentiation from Th2 to Th1.展开更多
AIM： To study the effects of concentrated liquor from male zooid of Antheraea pernyion immunological mice.METHODS： For each experiment, 40 mice were randomly divided into normal saline group （control group） and th...AIM： To study the effects of concentrated liquor from male zooid of Antheraea pernyion immunological mice.METHODS： For each experiment, 40 mice were randomly divided into normal saline group （control group） and three tested groups that were administered different dosages of concentrated liquor from male zooid of A. pernyiand food for 15 d. The typical FSR and HC50 value, monocybe-phagocytic exponent Kand emendated monocyte-phagocytic exponent were determined and calculated respectively.RESULTS： Alter 24 and 48 h, the FSR values of the three tested groups improved significantly in comparison to the conlyol group by variance analysis. The HCso values showed a significant difference between the high dosage group and the control group, as well as between the high dosage group and other two bested groups. The monocyte-phagocytic exponent Kand emendated exponent α showed rising tendencies, but no significant differences were found by variance analysis.CONCLUSION： The concentrated liquor from male zooid of A. pemyican significantly enhance cellular and humoral immune function in mice, but has no distinct influence on the monocyte-phagocytic system in mice.展开更多
Costaria costata(C.Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide;however,in China,no reports have been made on cultivation of the genus.To investigate potential cultivation of the...Costaria costata(C.Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide;however,in China,no reports have been made on cultivation of the genus.To investigate potential cultivation of the species in the northern part of China,trials on isolation and preservation of the gametophytes were conducted using C.costata from Korea;growth and development of the gametophytes were observed.We showed that at 10±1°C,60 μmol m-2s-1 and 12:12 h(L:D),freshly released zoospores settled down within 1 hour,and then developed into the primary cell during the following 2 days.After a vegetative growth phase lasting 6-8 days,female gametophytes became 3-4 times larger in diameter than that of the primary cell,but still remained at a unicellular stage,while male gametophytes divided into 4-10 cells with only a slight change in size.Fertilization occurred within 10 days after the zoospores were released from the sporangia,and the apical and basal tissues of the juvenile sporophyte divided and differentiated into the blade and stipe.Temperature and irradiance influenced gametophytic vegetative growth and developmental patterns.Generally,low irradiance(15 μmol m-2s-1 and 30 μmol m-2s-1) was unfavorable to the induction of fertility,but it enhanced female gametophyte division.The optimal conditions for vegetative growth were 15°C and 30 μmol m-2s-1.After transplantation of the juvenile seedlings and after eight months cultivation,the harvested mature blade reached 194 cm in length and 32.7 cm in width.Our study proves that it is feasible to implement propagation and large scale cultivation of C.costata in northern China.展开更多
Most studies on Haematococcus pluvialis have been focused on cell growth and astaxanthin accumulation;far less attention has been paid to cell cycles and proliferation patterns.The purpose of this study was to clarify...Most studies on Haematococcus pluvialis have been focused on cell growth and astaxanthin accumulation;far less attention has been paid to cell cycles and proliferation patterns.The purpose of this study was to clarify cell cycles and proliferation patterns in H.pluvialis microscopically using a camera and video recorder system.The complicated life history of H.pluvial i s can be divided into two stages:the motile stage and the non-motile stage.All the cells can be classifi ed into forms as follows:motile cell,nonmotile cell,zoospore and aplanospore.The main cell proliferation,both in the motile phase and non-motile phase in H.pluvialis,is by asexual reproduction.Under normal growth conditions,a motile cell usually produces two,sometimes four,and exceptionally eight zoospores.Under unfavorable conditions,the motile cell loses its fl agella and transforms into a non-motile cell,and the non-motile cell usually produces 2,4 or 8 aplanospores,and occasionally 20–32 aplanospores,which further develop into non-motile cells.Under suitable conditions,the non-motile cell is also able to release zoospores.The larger non-motile cells produce more than 16 zoospores,and the smaller ones produce 4 or 8 zoospores.Vegetative reproduction is by direct cell division in the motile phase and by occasional cell budding in the non-motile phase.There is,as yet,no convincing direct evidence for sexual reproduction.展开更多
The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived mono...The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived monoecious gametophyte isolate （designated as line 10-5-3） of U. pinnatifida was observed. To understand this phenomenon, a comprehensive screening of eighty-two previously identified male gametophyte cultures, isolated from three randomly selected cultivars （lines 10, 7, and 5） was performed. Thirty-six of the isolates developed both antheridia and oogonia on the same filamentous fragment in a standard gametogenesis test （SGT： 18℃, 60 umol photons/（m2.s））. Selfing of the monoecious gametophyte or crossing it with a normal male gametophyte both gave rise to morphologically normal sporophytic offspring. However, crossing resulted in a much higher fertilization rate （89.7%）. The hybrid and selfed sporophytic offspring were grown to maturity in flow tanks at an ambient temperature of 10-18℃ over a period of 69 days. Active zoospores were released from both types of mature sporophylls. The majority of these developed into male gametophytes, while 15%-20% developed into the observed monoecious structures on the same filament. Using PCR amplification it was found that all the monoecious gametophyte isolates and the sporophytic offspring resulting from the selfing and crossing lacked the femalelinked microsatellite sequence （a part of the locus Up-AC-2A8, GenBank accession No. AY738602.1）, indicating their male nature. U.pinnatifida is an invasive species in some regions and the implications of the above findings for this species in nature are briefly discussed.展开更多
The seasonal fluctuation of the plasmodia ofMyxobolus gibelioiWu et Wang, 1982 in the gill filaments of the allogynogenetic gibel carpCarassius auratus gibelio(Bloch) in a fish pond in Hubei Province of China was inve...The seasonal fluctuation of the plasmodia ofMyxobolus gibelioiWu et Wang, 1982 in the gill filaments of the allogynogenetic gibel carpCarassius auratus gibelio(Bloch) in a fish pond in Hubei Province of China was investigated from August 1999 to July 2000. A total of 445 fish was examined; the overall prevalence of the plasmodium infection in the fish was 64.94% and mean abundance of plasmodia was 11.65 ± 27.85 (mean ± SD). Significant seasonal changes in prevalence and mean abundance, with higher levels of the plasmodia infection from late spring to autumn, were observed.展开更多
The green algae of Order Chaetophorales distributed in Yellow-Bohai Sea of China are studied. More 3 species belonging respectively to 3 different genera are reported for the first time,viz. Entocladia viridis Reinke,...The green algae of Order Chaetophorales distributed in Yellow-Bohai Sea of China are studied. More 3 species belonging respectively to 3 different genera are reported for the first time,viz. Entocladia viridis Reinke, Pringsheimiella scutata (Reinke) Hhnel ex Marchewianka and Ulvella lens P.L. Crouan et H.M. Crouan, with a simple key. Entocladia viridis is different from other species in some features, such as disengaged branches in the external frond with concrescent midsection to distinct cells, irregular, slight swollen and sometime distorted at midsection, and column at external frond with 5～8 μm diameter and 1～5 times high to diameter. This species is found in Qingdao and Yantai, Shandong Province, and inner epiphytic to algae, especially to red algae. Pringsheimiella scutata (Reinke) Hhnel ex Marchewianka is different from other species in the shield frond of one cell layer, 1～2 mm diam., absent rhizoid, with ridgy center cells and flat marginal ones. This species is found in the Yellow Sea, and epiphytic to macroalgae, especially seen at the frond of genus Polysiphonia. Ulvella lens P.L. Crouan et H.M. Crouan is different from other species in the plate-like frond, ridgy midsection, with 1.5 mm diam. And 150～250 μm thick, small cells at midsection with usually 2～3 cells layers and 8～15 μm diameter, big cells at approximate margin of frond with 10～15 μm diameter and 20～30 μm long, cuneate cells at the margin of frond with 1 cell layer, 3.5～4.5 μm diameter and 15～25 μm long. This species is found in Qingdao, Shandong Province, and epiphytic to macroalgae, especially to the frond surface of red algae.展开更多
Models of circadian genetic oscillators involving interlinked feedback processes in molecular level genetic networks in Drosophila melanogaster and Neurospora crassa are studied, and mechanisms whereby synchronization...Models of circadian genetic oscillators involving interlinked feedback processes in molecular level genetic networks in Drosophila melanogaster and Neurospora crassa are studied, and mechanisms whereby synchronization can arise in an assembly of cells are examined. The individual subcellular circadian oscillatory processes are stochastic in nature due to the small numbers of molecules that are involved, and are subject to large fluctuations. The authors investigate and present the simulations of the stochastic dynamics of ensembles of clock-regulating proteins in different nuclei that communicate via ancillary small molecules, environmental parameters, additive cellular noise, or through diffusive processes. The results show that the emergence of collective oscillations is a macroscopic observable which has its origins in the microscopic coupling between distinct cellular oscillators.展开更多
基金Supported by the National Key Technology R & D Program (2007BAD40B05),Chinathe National High Technology Research and Development Program of China(2006AA10A207)~~
文摘[Objective] To clone the actin gene of Cryptosporidium andersoni, and to study its eukaryotic expression in Hela cells. [Methed] Specific primers were designed for the partial encoding sequence of actin, which were obtained by screening the T7 phage display library of Cryptosporidium andersoni, and the actin gene CA42 was amplified by PCR. Recombinant eukaryotic expression plasmid pVAX1-CA42 was constructed and transfected to Hela cells with lipofection strategy. Indirect im- munofluorescence staining, SDS-PAGE and Western blotting analysis were used to detect the expression of recombinant protein in Hela cells. [Result] CA42 protein was successfully expressed in Hela cells, and the expression products had reactogenicity. [Conclusion] The partial encoding sequence of actin from Cryptosporidium andersoni has been successfully cloned, and it can be stably expressed in Hela Cells
基金National Natural Science Foundation of China, No. 30472260the Administration of Traditional Chinese Medicine of Shandong Province, No. 2005-109
文摘AIM： To investigate whether supplementation of male zooid of Antheraea pernyi extracts （MZAPE） could enhance immune function of radiated tumor-bearing rats. METHODS： Eighty male Wistar rats were randomly divided into a control group, a simple radiation group, a MZAPE group, and a radiation plus MZAPE group. With the tumor model established by implanting Walker-256 ascites tumor cells, tumor weight and tumor control rate were calculated. The rats in the simple radiation and radiation plus MZAPE groups were underwent to radiation at 10 Gy within 2 d. In the MZAPE and radiation plus MZAPE groups, the MZAPE was gavaged at a dose of 16.53 mg/kg once a day for 7 d. T cell subsets in peripheral blood were determined by flow cytometry and the expression of IL-2, IFN-γ, IL-4 and IL-10 in sera were determined by ELISA on the 8th d. RESULTS： The tumor weight of simple radiation group, MZAPE group and radiation plus MZAPE group was lower than that of control group （P 〈 0.01） and tumor control rates were 63.08% ± 6.43%, 69.86%± 7.12% and 35.30% ± 7.67%, respectively. CD^4＋ T and CD^8＋ T cells in the peripheral blood of the simple radiation group were fewer than in control group. In the MZAPE and radiation plus MZAPE groups, the number of CD^4＋ T cells was higher while CD^8＋ T cells was lower than in the control and simple radiation groups. Expression of IL-2 and INF-y in the radiation group was lower than in control group, and significantly enhanced during MZAPE therapy （P 〈 0.05）. Expression of IL-4 and IL-10 in the radiation group had no significant changes compared with the control group, and decreased significantly after MZAPE treatment （P 〈 0.01）. CONCLUSION： MZAPE administration may help improve the immune function of the radiated tumor-bearing rats and reverse the radiation-induced immune inhibition by promoting the proliferation of T helper cells and inducing the transdifferentiation from Th2 to Th1.
基金Supported by the National Natural Science Foundation of China, No. 30472260
文摘AIM： To study the effects of concentrated liquor from male zooid of Antheraea pernyion immunological mice.METHODS： For each experiment, 40 mice were randomly divided into normal saline group （control group） and three tested groups that were administered different dosages of concentrated liquor from male zooid of A. pernyiand food for 15 d. The typical FSR and HC50 value, monocybe-phagocytic exponent Kand emendated monocyte-phagocytic exponent were determined and calculated respectively.RESULTS： Alter 24 and 48 h, the FSR values of the three tested groups improved significantly in comparison to the conlyol group by variance analysis. The HCso values showed a significant difference between the high dosage group and the control group, as well as between the high dosage group and other two bested groups. The monocyte-phagocytic exponent Kand emendated exponent α showed rising tendencies, but no significant differences were found by variance analysis.CONCLUSION： The concentrated liquor from male zooid of A. pemyican significantly enhance cellular and humoral immune function in mice, but has no distinct influence on the monocyte-phagocytic system in mice.
基金Supported by the Shandong Agriculture Seedstocks Projectthe Knowledge Innovation Program of the Chinese Academy of Sciences(No.KSCX2-YW-N-247-02)
文摘Costaria costata(C.Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide;however,in China,no reports have been made on cultivation of the genus.To investigate potential cultivation of the species in the northern part of China,trials on isolation and preservation of the gametophytes were conducted using C.costata from Korea;growth and development of the gametophytes were observed.We showed that at 10±1°C,60 μmol m-2s-1 and 12:12 h(L:D),freshly released zoospores settled down within 1 hour,and then developed into the primary cell during the following 2 days.After a vegetative growth phase lasting 6-8 days,female gametophytes became 3-4 times larger in diameter than that of the primary cell,but still remained at a unicellular stage,while male gametophytes divided into 4-10 cells with only a slight change in size.Fertilization occurred within 10 days after the zoospores were released from the sporangia,and the apical and basal tissues of the juvenile sporophyte divided and differentiated into the blade and stipe.Temperature and irradiance influenced gametophytic vegetative growth and developmental patterns.Generally,low irradiance(15 μmol m-2s-1 and 30 μmol m-2s-1) was unfavorable to the induction of fertility,but it enhanced female gametophyte division.The optimal conditions for vegetative growth were 15°C and 30 μmol m-2s-1.After transplantation of the juvenile seedlings and after eight months cultivation,the harvested mature blade reached 194 cm in length and 32.7 cm in width.Our study proves that it is feasible to implement propagation and large scale cultivation of C.costata in northern China.
基金Supported by the National Natural Science Foundation of China(No.31572639)the Laboratory for Marine Biology and Biotechnology,Qingdao National Laboratory for Marine Science and Technology(No.Y62419101J)
文摘Most studies on Haematococcus pluvialis have been focused on cell growth and astaxanthin accumulation;far less attention has been paid to cell cycles and proliferation patterns.The purpose of this study was to clarify cell cycles and proliferation patterns in H.pluvialis microscopically using a camera and video recorder system.The complicated life history of H.pluvial i s can be divided into two stages:the motile stage and the non-motile stage.All the cells can be classifi ed into forms as follows:motile cell,nonmotile cell,zoospore and aplanospore.The main cell proliferation,both in the motile phase and non-motile phase in H.pluvialis,is by asexual reproduction.Under normal growth conditions,a motile cell usually produces two,sometimes four,and exceptionally eight zoospores.Under unfavorable conditions,the motile cell loses its fl agella and transforms into a non-motile cell,and the non-motile cell usually produces 2,4 or 8 aplanospores,and occasionally 20–32 aplanospores,which further develop into non-motile cells.Under suitable conditions,the non-motile cell is also able to release zoospores.The larger non-motile cells produce more than 16 zoospores,and the smaller ones produce 4 or 8 zoospores.Vegetative reproduction is by direct cell division in the motile phase and by occasional cell budding in the non-motile phase.There is,as yet,no convincing direct evidence for sexual reproduction.
基金Supported by the National Natural Science Foundation of China(Nos.41176135,41206142)the Innovation Program of Chinese Academy of Sciences(No.KSCX2-EW-B-3)the Ministry of Agriculture of China(No.200903030)
文摘The annual life cycle of the brown seaweed Undariapinnatifida （Harvey） Suringer comprises a macroscopic diploid sporophyte stage and a microscopic haploid gametophyte stage. In 2011, an unusual zoospore-derived monoecious gametophyte isolate （designated as line 10-5-3） of U. pinnatifida was observed. To understand this phenomenon, a comprehensive screening of eighty-two previously identified male gametophyte cultures, isolated from three randomly selected cultivars （lines 10, 7, and 5） was performed. Thirty-six of the isolates developed both antheridia and oogonia on the same filamentous fragment in a standard gametogenesis test （SGT： 18℃, 60 umol photons/（m2.s））. Selfing of the monoecious gametophyte or crossing it with a normal male gametophyte both gave rise to morphologically normal sporophytic offspring. However, crossing resulted in a much higher fertilization rate （89.7%）. The hybrid and selfed sporophytic offspring were grown to maturity in flow tanks at an ambient temperature of 10-18℃ over a period of 69 days. Active zoospores were released from both types of mature sporophylls. The majority of these developed into male gametophytes, while 15%-20% developed into the observed monoecious structures on the same filament. Using PCR amplification it was found that all the monoecious gametophyte isolates and the sporophytic offspring resulting from the selfing and crossing lacked the femalelinked microsatellite sequence （a part of the locus Up-AC-2A8, GenBank accession No. AY738602.1）, indicating their male nature. U.pinnatifida is an invasive species in some regions and the implications of the above findings for this species in nature are briefly discussed.
文摘The seasonal fluctuation of the plasmodia ofMyxobolus gibelioiWu et Wang, 1982 in the gill filaments of the allogynogenetic gibel carpCarassius auratus gibelio(Bloch) in a fish pond in Hubei Province of China was investigated from August 1999 to July 2000. A total of 445 fish was examined; the overall prevalence of the plasmodium infection in the fish was 64.94% and mean abundance of plasmodia was 11.65 ± 27.85 (mean ± SD). Significant seasonal changes in prevalence and mean abundance, with higher levels of the plasmodia infection from late spring to autumn, were observed.
文摘The green algae of Order Chaetophorales distributed in Yellow-Bohai Sea of China are studied. More 3 species belonging respectively to 3 different genera are reported for the first time,viz. Entocladia viridis Reinke, Pringsheimiella scutata (Reinke) Hhnel ex Marchewianka and Ulvella lens P.L. Crouan et H.M. Crouan, with a simple key. Entocladia viridis is different from other species in some features, such as disengaged branches in the external frond with concrescent midsection to distinct cells, irregular, slight swollen and sometime distorted at midsection, and column at external frond with 5～8 μm diameter and 1～5 times high to diameter. This species is found in Qingdao and Yantai, Shandong Province, and inner epiphytic to algae, especially to red algae. Pringsheimiella scutata (Reinke) Hhnel ex Marchewianka is different from other species in the shield frond of one cell layer, 1～2 mm diam., absent rhizoid, with ridgy center cells and flat marginal ones. This species is found in the Yellow Sea, and epiphytic to macroalgae, especially seen at the frond of genus Polysiphonia. Ulvella lens P.L. Crouan et H.M. Crouan is different from other species in the plate-like frond, ridgy midsection, with 1.5 mm diam. And 150～250 μm thick, small cells at midsection with usually 2～3 cells layers and 8～15 μm diameter, big cells at approximate margin of frond with 10～15 μm diameter and 20～30 μm long, cuneate cells at the margin of frond with 1 cell layer, 3.5～4.5 μm diameter and 15～25 μm long. This species is found in Qingdao, Shandong Province, and epiphytic to macroalgae, especially to the frond surface of red algae.
文摘Models of circadian genetic oscillators involving interlinked feedback processes in molecular level genetic networks in Drosophila melanogaster and Neurospora crassa are studied, and mechanisms whereby synchronization can arise in an assembly of cells are examined. The individual subcellular circadian oscillatory processes are stochastic in nature due to the small numbers of molecules that are involved, and are subject to large fluctuations. The authors investigate and present the simulations of the stochastic dynamics of ensembles of clock-regulating proteins in different nuclei that communicate via ancillary small molecules, environmental parameters, additive cellular noise, or through diffusive processes. The results show that the emergence of collective oscillations is a macroscopic observable which has its origins in the microscopic coupling between distinct cellular oscillators.
