利用DNA重组技术在柞蚕蛹中表达重组人促红细胞生成素(Recombinant human erythropoietin,rhEPO),经过亲和层析纯化后,用SDS-PAGE分离纯化各组分,并通过电喷雾电离串联质谱技术(ESI-MS/MS)检测其糖基化修饰.结果显示,在柞蚕蛹中表达的rh...利用DNA重组技术在柞蚕蛹中表达重组人促红细胞生成素(Recombinant human erythropoietin,rhEPO),经过亲和层析纯化后,用SDS-PAGE分离纯化各组分,并通过电喷雾电离串联质谱技术(ESI-MS/MS)检测其糖基化修饰.结果显示,在柞蚕蛹中表达的rhEPO比活性约为1190 U/μg,其糖基化修饰位点与人体表达的EPO一致,有3个N-糖基化位点和1个O-糖基化位点.凝集素杂交实验结合质谱结果表明,柞蚕蛹表达的rhEPO的糖链中缺乏唾液酸修饰,而缺少唾液酸修饰的EPO通过鼻腔给药后在多种神经系统疾病的治疗中发挥着重要的作用.所得结果为进一步研究外源蛋白在柞蚕蛹-柞蚕核型多角体病毒(Anthraea pernyi nucleopolyhedrorirus,ApNPV)宿主载体表达系统表达后的糖基化与生物活性提供了依据.展开更多
A cathepsin B-like proteinase was purified from eggs of Antheraea pernyi by 5 steps of purification. The molecular mass of the proteinase are estimated to be 47 kDa by using SDS-polyacrylamide gel electro-phoresis. Th...A cathepsin B-like proteinase was purified from eggs of Antheraea pernyi by 5 steps of purification. The molecular mass of the proteinase are estimated to be 47 kDa by using SDS-polyacrylamide gel electro-phoresis. The activity of the purified proteinase were strongly inhibited by E-64 and Leupeptin. The optimum pH is 3.5 as determined by using the bovine hemo-globin as substrate. The activity and quantity of the proteinase during embryo development were studied. The results suggested that the proteolytic activities during embryo development at least partially came from this proteinase.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
Exogenous DNA expressing green fluorescent protein( GFP) and labeled with fluorescein isothiocyanate( FITC) was used to transform the Chinese oak silkmoth Antheraea pernyi( A. pernyi)via sperm-mediated gene transfer( ...Exogenous DNA expressing green fluorescent protein( GFP) and labeled with fluorescein isothiocyanate( FITC) was used to transform the Chinese oak silkmoth Antheraea pernyi( A. pernyi)via sperm-mediated gene transfer( SMGT). Sperms entry into the female reproductive system and eggs were observed using fluorescence microscopy. The ability of A. pernyi sperms to uptake exogenous DNA was confirmed,and transfer of the exogenous DNA was shown by GFP expression in the transgenic eggs. Our result suggested that SMGT could also be used to directly generate transgenic A. pernyi expressing functional genes of interest.展开更多
Objective： Cancer patients undergoing large dose radiotherapy exhibit multifaceted defects in their immune capacity that are likely to contribute to an increased susceptibility to infections and disease progression. ...Objective： Cancer patients undergoing large dose radiotherapy exhibit multifaceted defects in their immune capacity that are likely to contribute to an increased susceptibility to infections and disease progression. The immune impairment may also constitute a barrier to effective immunotherapeutic interventions. Here, we evaluate whether supplementation with the male zooid of Antheraea pernyi extracts could enhance immune function in irradiated rats. Methods： Fifty male Wistar rats were randomly divided into a control group, a simple radiation group and a treatment group. The mice in the simple irradiation and treatment groups were given whole-chest irradiation with 6Gy. In the treatment group, the male zooid of Antheraea pernyi extracts was gavaged at the doses of 16.53mg/kg （large dose group）, 2.62mg/kg （medium dose group）, and 0.564mg/kg （small dose group） once a day for 14 days. The thymus and spleen indices were calculated. T cell subsets in peripheral blood were determined by flow cytometry and the expressions of IL-2, IFN-γ, IL-4 and IL-10 in sera were determined by ELISA on the 15th day. Results： The thymus index and spleen index of the high dose treatment group were statistically lower than that of the control group and higher than that of the radiation group （P〈0.01）. CD3＋ and CD4＋ T cells in the peripheral blood were increased in the high dose treatment group and decreased in the radiation group （P〈0.01）. Expression of IL-2 and INF-T in the radiation group was lower than that in control, and significantly increased during therapy. The production of IL-4 and IL-10 could be induced by radiation and was inhibited in the high dose treatment group （P〈0.01）. Conclusion： Our data indicate that the male zooid of Antheraea pernyi extracts may be administrated to improve immune function in irradiated rats and reverse the radial immune inhibition of rats by stimulating the proliferation of Th ceils and inducing the differentiation of Th2 to Th1.展开更多
[ Objective ] This study aimed to identify the immune system-related TOLL-like receptor family gene of Aherea pernyi, to lay the foundation for further investigating the immune mechanism of Antherea perny/. [ Method] ...[ Objective ] This study aimed to identify the immune system-related TOLL-like receptor family gene of Aherea pernyi, to lay the foundation for further investigating the immune mechanism of Antherea perny/. [ Method] Immune system-related TOLL-like receptor family gene of Antherea perny/was cloned for se- quencing and phylogenetic analysis ; in addition, expression variations of TOLL-like receptor family gene in Antherea pernyi infected with Nosema pernyi and Nosema bombycis were detected to analyze the differences in immunological reactivity of Antherea pernyi to Nosema infection. [ Result] Based on cDNA cloning and sequen- cing, an immune system-related gene fragment was isolated from Antherea pernyi, which is the most homologous to the ToUl gene in Toll signaling pathway of Bom- byx mot/according to the sequencing result and phylogenetic analysis, which is named ApTolll gene. Subsequently, Antherea pernyi pupae were injected respective- ly with Nosema bombycis and Nosema pernyi, fluorescent real-time quantitative PCR analysis showed that ApTolll was abundantly expressed in Antherea pernyi pupa 2 h after Nosema bombycis injection, which began to express in Antherea pernyi pupa 11 h after Nosema pernyi injection, indicating that the immune response time induced by various Nosema strains varies in Toll signaling pathway of Antherea perny/. [ Conclusion ] ApTolll gene of Antherea perny/was first cloned in this study, which provides reference for further investigating the immune mechanism of Antherea pernyi.展开更多
The polyhedrin (polh) gene is often used to analyse evolution of baculovirus. In this report, the polh of Antheraea pernyi nucleopolyhedro-virus (AnpeNPV) was cloned and sequenced. The Open reading frame (ORF) of the ...The polyhedrin (polh) gene is often used to analyse evolution of baculovirus. In this report, the polh of Antheraea pernyi nucleopolyhedro-virus (AnpeNPV) was cloned and sequenced. The Open reading frame (ORF) of the AnpeNPV consists of 738 nucleotides encoding 245 amino acids with molecular masses of 29 kDa. The deduced amino acids were significant homol-ogy with other baculoviruses, such as Attacus ricini NPV (ArNPV) and Autographa californica NPV (AcNPV). A strongly hydrophilic region was predicted at positions from 30 to 50 of the An-peNPV Polh protein by bioinformatics analysis. Expression of the polh gene of AnpeNPV in E. coli was examined by SDS–PAGE, Western blot and Mass-spectrum analysis. The result showed that the bacterium expression system was suitable for the virus gene expression. It indi-cated that the products of the polh gene ex-pressed in this system can be easier to use for raising antibodies.展开更多
A new chlorine-containing neo-clerodane diterpene, teupernin D, was isolated from the aerial parts of Teucrium pernyi. The structure for teupernin D was established as (12S) -15, 16-epoxy-8β-hydroxy-17-chloro-19-nor1...A new chlorine-containing neo-clerodane diterpene, teupernin D, was isolated from the aerial parts of Teucrium pernyi. The structure for teupernin D was established as (12S) -15, 16-epoxy-8β-hydroxy-17-chloro-19-nor10α-neo-cleroda-4, 13(16), 14-triene-18,6β;20,12-diolide on the basis of spectral evidence. The absolute configuration was established by CD spectrum and confirmed by X-ray crystallographic analysis.展开更多
A 1.4Kb DNA fragment containing 3’ flanking sequence of fibroin gene of silkworm, Antheraea pernyi, was obtained from the silk gland’s mRNA of 5 th larva. Analysis of this sequence with another A.pernyi fibroin prot...A 1.4Kb DNA fragment containing 3’ flanking sequence of fibroin gene of silkworm, Antheraea pernyi, was obtained from the silk gland’s mRNA of 5 th larva. Analysis of this sequence with another A.pernyi fibroin protein (accession No. D83241) revealed that it consists of a completely open reading frame (ORF), which includes 14 polyalanine-containing units (motifs) and 100bp 3’-UTR. The sequence of the predicted amino acid reveals the highest level of overall identity (90%) with D83241. It was found that it loses a repeat region at the upstream of TAA codon and some mutations. A putative polyadenylation signal AATAAA tail was found in position 1300, which follows the termination codon.展开更多
Three furanoid ent-clerodane diterpenoids,teupernin A,B and C have been isolated from the acetone extract of Teucrium pernyi Franch.Their structures were established by spectroscopic means(UV,IR,MS,~1H NMR,^(13)C NMR,...Three furanoid ent-clerodane diterpenoids,teupernin A,B and C have been isolated from the acetone extract of Teucrium pernyi Franch.Their structures were established by spectroscopic means(UV,IR,MS,~1H NMR,^(13)C NMR,H-H COSY,C-H COSY and NOE)and by comparison with related compounds.展开更多
A proteinase responsible for degradation of yolk proteins has been discovered in oocyte of Antheraea pernyi which was found to found to hydrolyze vitelluin effectively in acidic pH. Using bovine hemoglobin as substrat...A proteinase responsible for degradation of yolk proteins has been discovered in oocyte of Antheraea pernyi which was found to found to hydrolyze vitelluin effectively in acidic pH. Using bovine hemoglobin as substrate,the optimum pH was determine3d at 3.5.Unaffected by EDTA and DipF, the activity of the proteinase was strongly inhibited by E64 and other thiol blocking reagents. It is thus very likely to be a cysteine proteinase. Activity of the proteinas. was found to be increasing in developing oocyte.展开更多
文摘A cathepsin B-like proteinase was purified from eggs of Antheraea pernyi by 5 steps of purification. The molecular mass of the proteinase are estimated to be 47 kDa by using SDS-polyacrylamide gel electro-phoresis. The activity of the purified proteinase were strongly inhibited by E-64 and Leupeptin. The optimum pH is 3.5 as determined by using the bovine hemo-globin as substrate. The activity and quantity of the proteinase during embryo development were studied. The results suggested that the proteolytic activities during embryo development at least partially came from this proteinase.
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Scientific Research Project for High Schools of the Educational Department of Liaoning Province,China(No.2008643)
文摘Exogenous DNA expressing green fluorescent protein( GFP) and labeled with fluorescein isothiocyanate( FITC) was used to transform the Chinese oak silkmoth Antheraea pernyi( A. pernyi)via sperm-mediated gene transfer( SMGT). Sperms entry into the female reproductive system and eggs were observed using fluorescence microscopy. The ability of A. pernyi sperms to uptake exogenous DNA was confirmed,and transfer of the exogenous DNA was shown by GFP expression in the transgenic eggs. Our result suggested that SMGT could also be used to directly generate transgenic A. pernyi expressing functional genes of interest.
基金supported by the grants from the National Natural Science foundation of China(No.30472260)the Administration of Traditional Chinese Medicine of Shandong Province,China(No.2005-109)the Health Protection Committee of Shandong Province,China(No.2006059).
文摘Objective： Cancer patients undergoing large dose radiotherapy exhibit multifaceted defects in their immune capacity that are likely to contribute to an increased susceptibility to infections and disease progression. The immune impairment may also constitute a barrier to effective immunotherapeutic interventions. Here, we evaluate whether supplementation with the male zooid of Antheraea pernyi extracts could enhance immune function in irradiated rats. Methods： Fifty male Wistar rats were randomly divided into a control group, a simple radiation group and a treatment group. The mice in the simple irradiation and treatment groups were given whole-chest irradiation with 6Gy. In the treatment group, the male zooid of Antheraea pernyi extracts was gavaged at the doses of 16.53mg/kg （large dose group）, 2.62mg/kg （medium dose group）, and 0.564mg/kg （small dose group） once a day for 14 days. The thymus and spleen indices were calculated. T cell subsets in peripheral blood were determined by flow cytometry and the expressions of IL-2, IFN-γ, IL-4 and IL-10 in sera were determined by ELISA on the 15th day. Results： The thymus index and spleen index of the high dose treatment group were statistically lower than that of the control group and higher than that of the radiation group （P〈0.01）. CD3＋ and CD4＋ T cells in the peripheral blood were increased in the high dose treatment group and decreased in the radiation group （P〈0.01）. Expression of IL-2 and INF-T in the radiation group was lower than that in control, and significantly increased during therapy. The production of IL-4 and IL-10 could be induced by radiation and was inhibited in the high dose treatment group （P〈0.01）. Conclusion： Our data indicate that the male zooid of Antheraea pernyi extracts may be administrated to improve immune function in irradiated rats and reverse the radial immune inhibition of rats by stimulating the proliferation of Th ceils and inducing the differentiation of Th2 to Th1.
基金Supported by Key Project of Ministry of Education of China(210180)National Natural Science Foundation of China(31001037)
文摘[ Objective ] This study aimed to identify the immune system-related TOLL-like receptor family gene of Aherea pernyi, to lay the foundation for further investigating the immune mechanism of Antherea perny/. [ Method] Immune system-related TOLL-like receptor family gene of Antherea perny/was cloned for se- quencing and phylogenetic analysis ; in addition, expression variations of TOLL-like receptor family gene in Antherea pernyi infected with Nosema pernyi and Nosema bombycis were detected to analyze the differences in immunological reactivity of Antherea pernyi to Nosema infection. [ Result] Based on cDNA cloning and sequen- cing, an immune system-related gene fragment was isolated from Antherea pernyi, which is the most homologous to the ToUl gene in Toll signaling pathway of Bom- byx mot/according to the sequencing result and phylogenetic analysis, which is named ApTolll gene. Subsequently, Antherea pernyi pupae were injected respective- ly with Nosema bombycis and Nosema pernyi, fluorescent real-time quantitative PCR analysis showed that ApTolll was abundantly expressed in Antherea pernyi pupa 2 h after Nosema bombycis injection, which began to express in Antherea pernyi pupa 11 h after Nosema pernyi injection, indicating that the immune response time induced by various Nosema strains varies in Toll signaling pathway of Antherea perny/. [ Conclusion ] ApTolll gene of Antherea perny/was first cloned in this study, which provides reference for further investigating the immune mechanism of Antherea pernyi.
文摘The polyhedrin (polh) gene is often used to analyse evolution of baculovirus. In this report, the polh of Antheraea pernyi nucleopolyhedro-virus (AnpeNPV) was cloned and sequenced. The Open reading frame (ORF) of the AnpeNPV consists of 738 nucleotides encoding 245 amino acids with molecular masses of 29 kDa. The deduced amino acids were significant homol-ogy with other baculoviruses, such as Attacus ricini NPV (ArNPV) and Autographa californica NPV (AcNPV). A strongly hydrophilic region was predicted at positions from 30 to 50 of the An-peNPV Polh protein by bioinformatics analysis. Expression of the polh gene of AnpeNPV in E. coli was examined by SDS–PAGE, Western blot and Mass-spectrum analysis. The result showed that the bacterium expression system was suitable for the virus gene expression. It indi-cated that the products of the polh gene ex-pressed in this system can be easier to use for raising antibodies.
文摘A new chlorine-containing neo-clerodane diterpene, teupernin D, was isolated from the aerial parts of Teucrium pernyi. The structure for teupernin D was established as (12S) -15, 16-epoxy-8β-hydroxy-17-chloro-19-nor10α-neo-cleroda-4, 13(16), 14-triene-18,6β;20,12-diolide on the basis of spectral evidence. The absolute configuration was established by CD spectrum and confirmed by X-ray crystallographic analysis.
文摘A 1.4Kb DNA fragment containing 3’ flanking sequence of fibroin gene of silkworm, Antheraea pernyi, was obtained from the silk gland’s mRNA of 5 th larva. Analysis of this sequence with another A.pernyi fibroin protein (accession No. D83241) revealed that it consists of a completely open reading frame (ORF), which includes 14 polyalanine-containing units (motifs) and 100bp 3’-UTR. The sequence of the predicted amino acid reveals the highest level of overall identity (90%) with D83241. It was found that it loses a repeat region at the upstream of TAA codon and some mutations. A putative polyadenylation signal AATAAA tail was found in position 1300, which follows the termination codon.
文摘Three furanoid ent-clerodane diterpenoids,teupernin A,B and C have been isolated from the acetone extract of Teucrium pernyi Franch.Their structures were established by spectroscopic means(UV,IR,MS,~1H NMR,^(13)C NMR,H-H COSY,C-H COSY and NOE)and by comparison with related compounds.
文摘A proteinase responsible for degradation of yolk proteins has been discovered in oocyte of Antheraea pernyi which was found to found to hydrolyze vitelluin effectively in acidic pH. Using bovine hemoglobin as substrate,the optimum pH was determine3d at 3.5.Unaffected by EDTA and DipF, the activity of the proteinase was strongly inhibited by E64 and other thiol blocking reagents. It is thus very likely to be a cysteine proteinase. Activity of the proteinas. was found to be increasing in developing oocyte.
