在一些爬行动物中,个体的性别完全取决于胚胎发育过程中的环境温度,称之为温度依赖型性别决定(temperaturedependent sex determination,TSD).TSD的分子机制长期是个谜,特别是调控早期性腺分化的分子基础仍不清楚.本文通过表达分析和基...在一些爬行动物中,个体的性别完全取决于胚胎发育过程中的环境温度,称之为温度依赖型性别决定(temperaturedependent sex determination,TSD).TSD的分子机制长期是个谜,特别是调控早期性腺分化的分子基础仍不清楚.本文通过表达分析和基因敲低手段研究了Sox9基因在红耳龟雄性性腺分化中的生物学功能,为TSD动物的性别决定和性腺发育的分子机制的研究奠定了基础.qRT-PCR显示,从性腺分化前的17期起,Sox9呈现产雄温度(male-producing temperature,MPT)性腺特异性高表达,而在产雌温度(female-producing temperature,FPT)性腺中表达水平极低.免疫组化进一步证实了SOX9蛋白的MPT特异性表达趋势,其定位于Sertoli前体细胞核中.温度置换实验显示,与MPT性腺相比,MPT→FPT性腺中(16期置换)的Sox9表达量从17期起就显著降低,表明Sox9能快速响应温度变化.同时MPT性腺经过雌激素处理后,Sox9表达量亦快速下调.功能缺失研究显示,经过Sox9-RNAi处理后,90.9%(20/22)的MPT性腺结构明显雌性化,皮质区高度发育,髓质区退化,揭示Sox9的敲低能导致雄性向雌性性逆转.上述研究表明,Sox9是红耳龟早期睾丸分化的关键调控因子,参与TSD的雄性分化通路.展开更多
The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation （SSF） with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on ph...The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation （SSF） with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on phytic acid, crude protein, crude fat, and amino acid profile. Response surface methodology （RSM） with Box-Behnken design was used to optimize SSF. The optimal conditions derived from RSM for L. brevis fermentation were： pH=5. 1; inoculum size=10%; duration=72 h; substrate to water ratio=1.5. The minimum content of trypsin inhibitors was 6.4 mg g^-1 dry matter. The optimal conditions derived from RSM for A. oryzae fermentation were： substrate to water ratio= 0.8 1; inoculum size=4%; duration=120 h. The minimum content of trypsin inhibitors was 1.6 mg g^-1 dry matter. Both L. brevis and A. oryzae decreased trypsin inhibitors dramatically （57.1 and 89.2% respectively）. L. brevis fermentation did not affect phytic acid （0.4%） and crude fat （5.2%） considerably, whereas A. oryzae fermentation degraded phytic acid （34.8%） and crude fat （22.0%） contents to a certain extent. Crude protein content was increased after both fermentation （6.4 and 12.9% for L. brevis and A. oryzae respectively）. Urease activity was reduced greatly （83.3 and 58.3% for L. brevis and A. oryzae respectively）. In conclusion, SSF with A. oryzae and L. brevis reduced trypsin inhibitor content and modified major macronutrients in soybean meal.展开更多
This study aimed to investigate the effects of Sargassum fusiforme polysaccharide(SFPS I,II,and III)on the apoptosis and regulation of human erythroleukemia(HEL)cells.The effect of different doses of SFPS on HEL cell ...This study aimed to investigate the effects of Sargassum fusiforme polysaccharide(SFPS I,II,and III)on the apoptosis and regulation of human erythroleukemia(HEL)cells.The effect of different doses of SFPS on HEL cell growth was detected using the Cell Counting Kit-8 method,and apoptosis was detected by Hoechst staining.Cell cycle distribution and apoptosis were detected using flow cytometry.Expression of the cell cycle gene,p53,antiapoptotic genes,Bcl-xL and Bcl-2,and pro-apoptotic genes,Bax,Bad,and Caspase-3,as well as the expression of the corresponding proteins,were detected using real-time quantitative polymerase chain reaction(qPCR)and Western blot.The results showed that SFPS Ⅱ and Ⅲ decreased HEL cell viability and induced HEL cell apoptosis.Different concentrations of SFPS(Ⅰ,Ⅱ,and Ⅲ)were detected that induced much less toxic effect in normal human embryonic lung(MRC-5)cells,and SFPS Ⅰ increased cell proliferation,indicating its favorable selectivity towards cancer cells.The mechanism by which SFPS induced apoptosis was also found to be related to the induction of cell cycle arrest in the G0/G1 phase and the increased expression of apoptosis-related genes and proteins.We concluded that SFPS induces HEL cell apoptosis,possibly via activation of the Caspase pathway,providing the theoretical basis for the development of SFPS-based anti-tumor drug products.展开更多
Sargassum fusiforme(S.fusiforme)has been used as an ingredient in Chinese herbal medicine for thousands of years.However,there are a limited number of studies concerning its therapeutic mechanism.High performance gel ...Sargassum fusiforme(S.fusiforme)has been used as an ingredient in Chinese herbal medicine for thousands of years.However,there are a limited number of studies concerning its therapeutic mechanism.High performance gel permeation chromatography(HPGPC)analysis showed that the average molecular weight of the S.fusiforme polysaccharide,SFPS 191212,is 43 kDa.SFPS 191212 is composed of mannose,rhamnose,galactose,xylose,glucose,and fucose(at a molar ratio:2.1:2.9:1.8:15.5:4.6:62.5)withα-andβ-configurations.The present research evaluated the anti-tumor potential of the S.fusiforme polysaccharide in human erythroleukemia(HEL)cells in vitro.To explore the SFPS 191212’s apoptosis mechanism in HEL cells,transcriptome analysis was performed on HEL cells that were incubated with SFPS 191212.The inhibitory effect of SFPS 191212 on HEL cell growth was also analyzed.It was found that SFPS 191212 inhibited HEL cell proliferation,reduced cell viability in a concentration-dependent manner,and induced an insignificant toxic effect on normal human embryonic lung(MRC-5)cells.Compared with the control group,transcriptome analysis identified a total of 598 differentially expressed genes(DEGs),including 243 up-regulated genes and 355 downregulated genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on all DEGs,and 900 GO terms and 52 pathways were found to be significantly enriched.Finally,23 DEGs were randomly selected and confirmed by quantitative real-time polymerase chain reaction(qRT-PCR).Moreover,SFPS 191212 down-regulated the PI3K/Akt signal transduction pathway.Our results provide a framework for understanding the effect of SFPS 191212 on cancer cells and can serve as a resource for delineating the anti-tumor mechanisms of S.fusiforme.展开更多
文摘在一些爬行动物中,个体的性别完全取决于胚胎发育过程中的环境温度,称之为温度依赖型性别决定(temperaturedependent sex determination,TSD).TSD的分子机制长期是个谜,特别是调控早期性腺分化的分子基础仍不清楚.本文通过表达分析和基因敲低手段研究了Sox9基因在红耳龟雄性性腺分化中的生物学功能,为TSD动物的性别决定和性腺发育的分子机制的研究奠定了基础.qRT-PCR显示,从性腺分化前的17期起,Sox9呈现产雄温度(male-producing temperature,MPT)性腺特异性高表达,而在产雌温度(female-producing temperature,FPT)性腺中表达水平极低.免疫组化进一步证实了SOX9蛋白的MPT特异性表达趋势,其定位于Sertoli前体细胞核中.温度置换实验显示,与MPT性腺相比,MPT→FPT性腺中(16期置换)的Sox9表达量从17期起就显著降低,表明Sox9能快速响应温度变化.同时MPT性腺经过雌激素处理后,Sox9表达量亦快速下调.功能缺失研究显示,经过Sox9-RNAi处理后,90.9%(20/22)的MPT性腺结构明显雌性化,皮质区高度发育,髓质区退化,揭示Sox9的敲低能导致雄性向雌性性逆转.上述研究表明,Sox9是红耳龟早期睾丸分化的关键调控因子,参与TSD的雄性分化通路.
基金supported by a research project of the Science and Technology Key Group in Zhejiang Provincethe research projects from the Science and Technology Department of Zhejiang Province,China (2009C12068)
文摘The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation （SSF） with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on phytic acid, crude protein, crude fat, and amino acid profile. Response surface methodology （RSM） with Box-Behnken design was used to optimize SSF. The optimal conditions derived from RSM for L. brevis fermentation were： pH=5. 1; inoculum size=10%; duration=72 h; substrate to water ratio=1.5. The minimum content of trypsin inhibitors was 6.4 mg g^-1 dry matter. The optimal conditions derived from RSM for A. oryzae fermentation were： substrate to water ratio= 0.8 1; inoculum size=4%; duration=120 h. The minimum content of trypsin inhibitors was 1.6 mg g^-1 dry matter. Both L. brevis and A. oryzae decreased trypsin inhibitors dramatically （57.1 and 89.2% respectively）. L. brevis fermentation did not affect phytic acid （0.4%） and crude fat （5.2%） considerably, whereas A. oryzae fermentation degraded phytic acid （34.8%） and crude fat （22.0%） contents to a certain extent. Crude protein content was increased after both fermentation （6.4 and 12.9% for L. brevis and A. oryzae respectively）. Urease activity was reduced greatly （83.3 and 58.3% for L. brevis and A. oryzae respectively）. In conclusion, SSF with A. oryzae and L. brevis reduced trypsin inhibitor content and modified major macronutrients in soybean meal.
基金Zhejiang Province Focuses on“Biological Engineering”Innovation Projects(No.CX2017001)the Autonomous Research Project of FSEKDNB(No.2020FSEKDNB001)。
文摘This study aimed to investigate the effects of Sargassum fusiforme polysaccharide(SFPS I,II,and III)on the apoptosis and regulation of human erythroleukemia(HEL)cells.The effect of different doses of SFPS on HEL cell growth was detected using the Cell Counting Kit-8 method,and apoptosis was detected by Hoechst staining.Cell cycle distribution and apoptosis were detected using flow cytometry.Expression of the cell cycle gene,p53,antiapoptotic genes,Bcl-xL and Bcl-2,and pro-apoptotic genes,Bax,Bad,and Caspase-3,as well as the expression of the corresponding proteins,were detected using real-time quantitative polymerase chain reaction(qPCR)and Western blot.The results showed that SFPS Ⅱ and Ⅲ decreased HEL cell viability and induced HEL cell apoptosis.Different concentrations of SFPS(Ⅰ,Ⅱ,and Ⅲ)were detected that induced much less toxic effect in normal human embryonic lung(MRC-5)cells,and SFPS Ⅰ increased cell proliferation,indicating its favorable selectivity towards cancer cells.The mechanism by which SFPS induced apoptosis was also found to be related to the induction of cell cycle arrest in the G0/G1 phase and the increased expression of apoptosis-related genes and proteins.We concluded that SFPS induces HEL cell apoptosis,possibly via activation of the Caspase pathway,providing the theoretical basis for the development of SFPS-based anti-tumor drug products.
基金partially funded by Zhejiang Wanli University Scientific Research and Innivation Team(No.SC1032110880210)Zhejiang Provincial Top Discipline of Biological Engineering(No.KF2021010)Ningbo Public Service Platform for High-Value Utilization of Marine Biological Resources(Nos.NBHY-2017-S5,NBHY-2017(1))。
文摘Sargassum fusiforme(S.fusiforme)has been used as an ingredient in Chinese herbal medicine for thousands of years.However,there are a limited number of studies concerning its therapeutic mechanism.High performance gel permeation chromatography(HPGPC)analysis showed that the average molecular weight of the S.fusiforme polysaccharide,SFPS 191212,is 43 kDa.SFPS 191212 is composed of mannose,rhamnose,galactose,xylose,glucose,and fucose(at a molar ratio:2.1:2.9:1.8:15.5:4.6:62.5)withα-andβ-configurations.The present research evaluated the anti-tumor potential of the S.fusiforme polysaccharide in human erythroleukemia(HEL)cells in vitro.To explore the SFPS 191212’s apoptosis mechanism in HEL cells,transcriptome analysis was performed on HEL cells that were incubated with SFPS 191212.The inhibitory effect of SFPS 191212 on HEL cell growth was also analyzed.It was found that SFPS 191212 inhibited HEL cell proliferation,reduced cell viability in a concentration-dependent manner,and induced an insignificant toxic effect on normal human embryonic lung(MRC-5)cells.Compared with the control group,transcriptome analysis identified a total of 598 differentially expressed genes(DEGs),including 243 up-regulated genes and 355 downregulated genes.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on all DEGs,and 900 GO terms and 52 pathways were found to be significantly enriched.Finally,23 DEGs were randomly selected and confirmed by quantitative real-time polymerase chain reaction(qRT-PCR).Moreover,SFPS 191212 down-regulated the PI3K/Akt signal transduction pathway.Our results provide a framework for understanding the effect of SFPS 191212 on cancer cells and can serve as a resource for delineating the anti-tumor mechanisms of S.fusiforme.
