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
AIM: To study the effects of concentrated liquor from male zooid of Antheraea pernyi on immunological mice.METHODS: For each experiment, 40 mice were randomly divided into normal saline group (control group) and three...AIM: To study the effects of concentrated liquor from male zooid of Antheraea pernyi on 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. pernyi and food for 15 d. The typical FSR and HC50 value, monocyte-phagocytic exponent Kand emendated monocyte-phagocytic exponent α were determined and calculated respectively.RESULTS: After 24 and 48 h, the FSR values of the three tested groups improved significantly in comparison to the control group by variance analysis. The HC50 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 tested 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. pernyi can 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
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 ...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. CD4+ T and CD8+ 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 CD4+ T cells was higher while CD8+ T cells was lower than in the control and simple radiation groups. Expression of IL-2 and INF-γ 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
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
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
Gelatin(G)is a commonly used natural biomaterial owing to its good biocompatibility and easy availability.However,using pure gelatin as a bioink can barely achieve an ideal shape fidelity in 3D printing.In this study,...Gelatin(G)is a commonly used natural biomaterial owing to its good biocompatibility and easy availability.However,using pure gelatin as a bioink can barely achieve an ideal shape fidelity in 3D printing.In this study,Antheraea pernyi silk fibroin nanofibers(ASFNFs)with arginine-glycine-aspartic acid(RGD)peptide and partial natural silk structure are extracted and combined with pure gelatin bioink to simultaneously improve the shape fidelity and cytocompatibility of corresponding 3D printed scaffold.Results show that the optimum printing temperature is 30℃ for these bioinks.The printed filaments using 16G/4ASFNFs bioink(16wt%gelatin and 4wt%ASFNFs)demonstrate better morphology and larger pore size than those printed by pure gelatin bioink(20G,20wt%gelatin),thus successfully improve the shape fidelity and porosity of the 3D printed scaffold.The 16G/4ASFNFs scaffold also demonstrate higher swelling ratio and faster degradation rate than the pure gelatin scaffold.Moreover,the cell viability and proliferation ability of Schwann cells cultured on the 16G/4ASFNFs scaffold are significantly superior than those cultured on the pure 20G scaffold.The ASFNFs enhanced 16G/4ASFNFs scaffold reported here are expected to be a candidate with excellent potential for biomedical applications.展开更多
The sol-gel transition behavior of Antherae pernyi silk fibroin(Ap-SF) has not been systematically investigated.In this work,the influence of environmental temperature,pH,the concentration of Ap-SF,K+ and Ca2+ on the ...The sol-gel transition behavior of Antherae pernyi silk fibroin(Ap-SF) has not been systematically investigated.In this work,the influence of environmental temperature,pH,the concentration of Ap-SF,K+ and Ca2+ on the gelation time,and the structural changes of Ap-SF in sol-gel transformation were studied.The results indicated that the gelation time of the Ap-SF aqueous solution decreased with the increase of the Ap-SF concentration and environmental temperature.The sol-gel transformation of Ap-SF was much more rapid than that of Bombyx mori silk fibroin under the same conditions.The Ap-SF was sensitive to changes in the concentration of Ca2+ and K+.Upon gelation,the random coil structure of the Ap-SF was significantly transformed into the β-sheet structure.展开更多
The antibacterial peptide D (ABP-D) was induced from the haemolymph of the Chinese oak silkworm (Antheraea pemyi). It has a marked inhibition effect on some human, insect and plant pathogenic bacteria. In order to syn...The antibacterial peptide D (ABP-D) was induced from the haemolymph of the Chinese oak silkworm (Antheraea pemyi). It has a marked inhibition effect on some human, insect and plant pathogenic bacteria. In order to synthesize a large amount of ABP-D in microorganism by recombinant DNA technology, here we report the synthesis of the ABP-D gene.展开更多
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.
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 pernyi on 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. pernyi and food for 15 d. The typical FSR and HC50 value, monocyte-phagocytic exponent Kand emendated monocyte-phagocytic exponent α were determined and calculated respectively.RESULTS: After 24 and 48 h, the FSR values of the three tested groups improved significantly in comparison to the control group by variance analysis. The HC50 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 tested 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. pernyi can significantly enhance cellular and humoral immune function in mice, but has no distinct influence on the monocyte-phagocytic system in mice.
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. CD4+ T and CD8+ 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 CD4+ T cells was higher while CD8+ T cells was lower than in the control and simple radiation groups. Expression of IL-2 and INF-γ 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.
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.
基金This work was supported by the Natural Science Foundation of Shanghai(20ZR1402400)the National Natural Science Foundation of China(52173031,51903045,51703033)+4 种基金the Program of Shanghai Academic/Technology Research Leader(20XD1400100)the National Key Research and Development Program of China(2020YFC1910303,2018YFC1105800)the Basic Research Project of the Science and Technology Commission of Shanghai Municipality(21JC1400100)the Fundamental Research Funds for the Central Universities(2232020D-04,2232019A3-06,2232019D3-02)the Science and Technology Commission of Shanghai Municipality(20DZ2254900).
文摘Gelatin(G)is a commonly used natural biomaterial owing to its good biocompatibility and easy availability.However,using pure gelatin as a bioink can barely achieve an ideal shape fidelity in 3D printing.In this study,Antheraea pernyi silk fibroin nanofibers(ASFNFs)with arginine-glycine-aspartic acid(RGD)peptide and partial natural silk structure are extracted and combined with pure gelatin bioink to simultaneously improve the shape fidelity and cytocompatibility of corresponding 3D printed scaffold.Results show that the optimum printing temperature is 30℃ for these bioinks.The printed filaments using 16G/4ASFNFs bioink(16wt%gelatin and 4wt%ASFNFs)demonstrate better morphology and larger pore size than those printed by pure gelatin bioink(20G,20wt%gelatin),thus successfully improve the shape fidelity and porosity of the 3D printed scaffold.The 16G/4ASFNFs scaffold also demonstrate higher swelling ratio and faster degradation rate than the pure gelatin scaffold.Moreover,the cell viability and proliferation ability of Schwann cells cultured on the 16G/4ASFNFs scaffold are significantly superior than those cultured on the pure 20G scaffold.The ASFNFs enhanced 16G/4ASFNFs scaffold reported here are expected to be a candidate with excellent potential for biomedical applications.
基金supported by the National Basic Research Program of China (Grant No 2005CB623902)the National Natural Science Foundation of China (Grant No 30970714)College Natural Science Research Project of Jiangsu Province (Grant No 07KJA43010)
文摘The sol-gel transition behavior of Antherae pernyi silk fibroin(Ap-SF) has not been systematically investigated.In this work,the influence of environmental temperature,pH,the concentration of Ap-SF,K+ and Ca2+ on the gelation time,and the structural changes of Ap-SF in sol-gel transformation were studied.The results indicated that the gelation time of the Ap-SF aqueous solution decreased with the increase of the Ap-SF concentration and environmental temperature.The sol-gel transformation of Ap-SF was much more rapid than that of Bombyx mori silk fibroin under the same conditions.The Ap-SF was sensitive to changes in the concentration of Ca2+ and K+.Upon gelation,the random coil structure of the Ap-SF was significantly transformed into the β-sheet structure.
基金Project supported by the National Natural Science Foundation of China
文摘The antibacterial peptide D (ABP-D) was induced from the haemolymph of the Chinese oak silkworm (Antheraea pemyi). It has a marked inhibition effect on some human, insect and plant pathogenic bacteria. In order to synthesize a large amount of ABP-D in microorganism by recombinant DNA technology, here we report the synthesis of the ABP-D gene.