From the fungus Trichoderma sp.,we isolated seven novel 18-residue peptaibols,neoatroviridins E-K(1−7),and six new 14-residue peptaibols,harzianins NPDG J-O(8−13).Additionally,four previously characterized 18-residue ...From the fungus Trichoderma sp.,we isolated seven novel 18-residue peptaibols,neoatroviridins E-K(1−7),and six new 14-residue peptaibols,harzianins NPDG J-O(8−13).Additionally,four previously characterized 18-residue peptaibols neoatroviridins A-D(14−17)were also identified.The structural configurations of the newly identified peptaibols(1−13)were determined by comprehensive nuclear magnetic resonance(NMR)and high-resolution electrospray ionization tandem mass spectrometry(HR-ESI-MS/MS)data.Their absolute configurations were further determined using Marfey’s method.Notably,compounds 12 and 13 represent the first 14-residue peptaibols containing an acidic amino acid residue.In antimicrobial assessments,all 18-residue peptaibols(1−7,14−17)exhibited moderate inhibitory activities against Staphylococcus aureus 209P,with minimum inhibitory concentration(MIC)values ranging from 8−32μg·mL^(−1).Moreover,compound 9 exhibited moderate inhibitory effect on Candida albicans FIM709,with a MIC value of 16μg·mL^(−1).展开更多
Objective: To investigate the law of age-related changes of insulin-like growth factor-1 (IGF-1) expression of rat thyroid cells cultured in vitro. Methods: Rat thyroid of different age (10, 45, 65, 100, 150 weeks) wa...Objective: To investigate the law of age-related changes of insulin-like growth factor-1 (IGF-1) expression of rat thyroid cells cultured in vitro. Methods: Rat thyroid of different age (10, 45, 65, 100, 150 weeks) was isolated and thyrocytes cultured. Total RNA was extracted in different rat age group when thyroid cells had been cultured for two weeks. mRNA IGF-1 expression was measured with reverse-transcription polymerase chain reaction( RT-PCR) in each group and compared. Results: Quantity of total RNA in thyroid cells decreased with ageing when the rat thyroid cells had been cultured for 2 weeks. There is significant difference among groups (P<0.05). Expression of IGF-1 mRNA could be detected in thyroid cells of different age cultured in vitro. Quantity of IGF-1 mRNA expression by RT-PCR analysis increased from 10 to 45 weeks old, and then decreased with ageing. Conclusion: Rat thyroid cells from different age cultured in vitro can express IGF-1 mRNA. Quantity of total RNA in thyroid cells cultured in vitro decreased with aging. IGF-1 mRNA expression was correlated to age (r=0.401, P<0.05).展开更多
Background:In traditional Chinese medicine,You-Gui-Wan(YGW)is typically used to treat osteoporosis associated with kidney-yang deficiency.However,there have been few mechanistic studies on the effectiveness of kidney-...Background:In traditional Chinese medicine,You-Gui-Wan(YGW)is typically used to treat osteoporosis associated with kidney-yang deficiency.However,there have been few mechanistic studies on the effectiveness of kidney-yang deficiency-type osteoporosis with YGW.To further clarify the role of YGW in the effect of osteoporosis with kidney-yang deficiency,the study analyzed the therapeutic advantages of YGW by comparing the therapeutic effects of YGW and alendronate(ALN)on osteoporosis with kidney-yang deficiency.Methods:SPF female SD rats were randomly divided into control,osteoporosis,osteoporosis with kidney-yang deficiency,osteoporosis with kidney-yang deficiency+YGW and osteoporosis with kidney-yang deficiency+ALN groups.Except for the control group,osteoporosis was induced by the removal of bilateral ovaries.After 12 weeks,rats with osteoporosis in the kidney-yang deficiency group had kidney-yang deficiency syndrome triggered by hydrocortisone for 14 days.Rats were treated with YGW or ALN for 12 weeks.The weights of rats were recorded.Hematoxylin-eosin staining staining was used to observe pathological changes in bone trabeculae,liver,spleen,and kidneys of rats.Depletion of the growth plate cartilage of rats in different groups was observed by safranine-O staining.The expression of osteoclast key indices(ACP)and osteoblast key indices(ALP)in the bone tissue of rats in the different groups was observed by immunohistochemical staining.The expression of bone resorption-related indicators(TRAP and NXT-1),bone formation-related indicators(BALP,BGP,and P1NP),and major indicators of kidney-yang deficiency(ACTH,T3,T4,cAMP,and cGMP)were observed using an ELISA detection kit.The expression levels of the main indices of liver function(ALT and AST)were detected in different groups.Results:The differences between the osteoporosis with kidney-yang deficiency group and osteoporosis group were that the weight of rats and the expression of ACTH,T3,T4,and cAMP decreased significantly,and the expression of cGMP increased in the osteoporosis with kidney-yang deficiency group.Moreover,both YGW and ALN effectively improved the symptoms of osteoporosis,including the injury of bone trabeculae and growth plates,as well as the expression of bone metabolism-related indicators.However,unlike ALN,YGW simultaneously ameliorated the expression of key indicators of kidney-yang deficiency and prevented weight loss in rats.In addition,YGW caused no obvious damage to the liver,spleen,or kidney,whereas ALN led to liver cirrhosis.Conclusion:The results reveal that YGW plays a crucial part in osteoporosis with kidney-yang deficiency,increases bone mineral density,and improves bone metabolism indicators,and is safe and efficient for the efficacy of osteoporosis with kidney-yang deficiency.YGW might have a better therapeutic effect on osteoporosis in patients with kidney-yang deficiency.Therefore,alendronate should be used cautiously in patients with osteoporosis and poor liver function.展开更多
MHC/peptide tetramer technology has been widely used to study antigen-specific T cells, especially for identifying virus-specific CD8^＋ T cells in humans. The tetramer molecule is composed of HLA heavy chain, β2-mic...MHC/peptide tetramer technology has been widely used to study antigen-specific T cells, especially for identifying virus-specific CD8^＋ T cells in humans. The tetramer molecule is composed of HLA heavy chain, β2-microglobulin （β2m）, an antigenic peptide, and fluorescent-labeled streptavidin. To further investigate the HLA-A＊1101-restricted CD8^＋ T cell responses against human cytomegalovirus （HCMV）, we established an approach to prepare HLA-A＊1101 tetramer complexed with a peptide from HCMV. The cDNA encoding HLA-A＊1101 heavy chain was cloned and the prokaryotic expression vector for the ectodomain of HLA-A＊1101 fused with a BirA substrate peptide （HLA-A＊1101-BSP） at its carboxyl terminus was constructed. The fusion protein was highly expressed as inclusion bodies under optimized conditions in Escherichla coli. Moreover, HLA-A＊1101-BSP protein was refolded in the presence of β2m and an HCMV peptide pp6516.24 （GPISGHVLK, GPI）. Soluble HLA-A＊1101-GPI monomer was biotinylated and purified to a purity of 95%, which was subsequently combined with streptavidin to form tetramers at a yield of 〉 80%. The HLA-A＊1101-GPI tetramers could bind to virus-specific CD8^＋ T cells, suggesting soluble HLA-A＊1101-GPI tetramers were biologically functional. This study provides the basis for further evaluation of HLA-A＊1101-restricted CD8^＋ T cell responses against HCMV infection.展开更多
Major histocompatibility complex （MHC） class I tetramer technology has become the central technique for analyzing antigen-specific CD8^＋ T cell responses and it has been widely used to explore the differentiation a...Major histocompatibility complex （MHC） class I tetramer technology has become the central technique for analyzing antigen-specific CD8^＋ T cell responses and it has been widely used to explore the differentiation and formation of memory CD8^＋ T cells. Previously, a simplified and efficient procedure for preparing high quality HLA-A＊0201 tetramers has been established in our lab and the tetramers loaded with HCMV peptide pp6549s.50a has been successfully applied to investigate HCMV-specific CD8^＋ T cells in Chinese populations. Using similar procedure we reported here the construction of HLA-A＊0201 tetramer loaded with another dominant epitope derived from immediate early （IE）-1 316.324 （VLEETSVML, VLE） of HCMV （A2-VLE） and characterization of this tetramer. After A2-VLE monomer was prepared and purified, its tetramer was then formed at a yield of 83%. The optimized amount of A2-VLE tetramer for staining 100 μl whole blood was 0.5 μg with incubation at 4℃ for 1 h. Furthermore, the dissociation constant of the tetramer binding to the specific CD8^＋ T cells of one HLA-A2^＋ donor was estimated to be 32.7 nmol/L, which is markedly higher than that of MHC monomer. The construction of A2-VLE tetramer provides an alternative choice for investigating HCMV-specific CD8^＋ T cell responses and will deepen our understanding of the differentiation and formation of HCMV-specific memory CD8^＋ T cells. Cellular ＆ Molecular Immunology.展开更多
The high recurrence and low responsiveness to immunotherapy make ovarian cancer(OC)the most lethal gynecological malignancy.Tumor microenvironment is critical in risk stratification and the discovery of molecular targ...The high recurrence and low responsiveness to immunotherapy make ovarian cancer(OC)the most lethal gynecological malignancy.Tumor microenvironment is critical in risk stratification and the discovery of molecular targets.We developed a prognostic classification for OC,which could also predict the prognosis of other gynecological cancers including breast cancer,endometrial cancer,and cervical cancer.Somatic mutation,hallmark pathways,and immune landscapes were characterized.Integrative analysis of immune checkpoints and multiple immune signatures revealed the low-risk group responds better to immune checkpoint inhibitors,which was validated by an external immunotherapeutic cohort(IMvigor210).Singlecell RNA sequencing(scRNA-seq)confirmed the high expression of SERPINB1 and SERPINB9 in dendritic cells,and AlphaFold2 was used to infer their 3D protein structures.Putative molecular compounds binding to SERPINB1/SERPINB9 were predicted by virtual screening.展开更多
基金supported by the National Key Research and Development Program of China(No.2018YFA0903200//2018YFA0903201)the National Natural Science Foundation of China(Nos.81925037 and 81973213)+6 种基金National High-level Personnel of Special Support Program(No.2017RA2259)the 111 Project of Ministry of Education of the People's Republic of China(No.B13038)Local Innovative and Research Teams Project of Guangdong Pearl River Talents Program(No.2017BT01Y036)Guangdong International Science and Technology Cooperation Base(No.2021A0505020015)Innovative and Research Teams Project of Guangdong Higher Education Institution(No.2021KCXTD001)K.C.Wong Education Foundation(No.Guo-Dong Chen,2021,China)the Science and Technology Planning Project of Guangzhou(No.202201020048)。
文摘From the fungus Trichoderma sp.,we isolated seven novel 18-residue peptaibols,neoatroviridins E-K(1−7),and six new 14-residue peptaibols,harzianins NPDG J-O(8−13).Additionally,four previously characterized 18-residue peptaibols neoatroviridins A-D(14−17)were also identified.The structural configurations of the newly identified peptaibols(1−13)were determined by comprehensive nuclear magnetic resonance(NMR)and high-resolution electrospray ionization tandem mass spectrometry(HR-ESI-MS/MS)data.Their absolute configurations were further determined using Marfey’s method.Notably,compounds 12 and 13 represent the first 14-residue peptaibols containing an acidic amino acid residue.In antimicrobial assessments,all 18-residue peptaibols(1−7,14−17)exhibited moderate inhibitory activities against Staphylococcus aureus 209P,with minimum inhibitory concentration(MIC)values ranging from 8−32μg·mL^(−1).Moreover,compound 9 exhibited moderate inhibitory effect on Candida albicans FIM709,with a MIC value of 16μg·mL^(−1).
文摘Objective: To investigate the law of age-related changes of insulin-like growth factor-1 (IGF-1) expression of rat thyroid cells cultured in vitro. Methods: Rat thyroid of different age (10, 45, 65, 100, 150 weeks) was isolated and thyrocytes cultured. Total RNA was extracted in different rat age group when thyroid cells had been cultured for two weeks. mRNA IGF-1 expression was measured with reverse-transcription polymerase chain reaction( RT-PCR) in each group and compared. Results: Quantity of total RNA in thyroid cells decreased with ageing when the rat thyroid cells had been cultured for 2 weeks. There is significant difference among groups (P<0.05). Expression of IGF-1 mRNA could be detected in thyroid cells of different age cultured in vitro. Quantity of IGF-1 mRNA expression by RT-PCR analysis increased from 10 to 45 weeks old, and then decreased with ageing. Conclusion: Rat thyroid cells from different age cultured in vitro can express IGF-1 mRNA. Quantity of total RNA in thyroid cells cultured in vitro decreased with aging. IGF-1 mRNA expression was correlated to age (r=0.401, P<0.05).
基金supported by the National Natural Science Foundation of China(Grant No.81673996,81904220)the Jiangmen Association for Science and Technology-Youth science and technology talent lifting project(Grant No.2022-2023).
文摘Background:In traditional Chinese medicine,You-Gui-Wan(YGW)is typically used to treat osteoporosis associated with kidney-yang deficiency.However,there have been few mechanistic studies on the effectiveness of kidney-yang deficiency-type osteoporosis with YGW.To further clarify the role of YGW in the effect of osteoporosis with kidney-yang deficiency,the study analyzed the therapeutic advantages of YGW by comparing the therapeutic effects of YGW and alendronate(ALN)on osteoporosis with kidney-yang deficiency.Methods:SPF female SD rats were randomly divided into control,osteoporosis,osteoporosis with kidney-yang deficiency,osteoporosis with kidney-yang deficiency+YGW and osteoporosis with kidney-yang deficiency+ALN groups.Except for the control group,osteoporosis was induced by the removal of bilateral ovaries.After 12 weeks,rats with osteoporosis in the kidney-yang deficiency group had kidney-yang deficiency syndrome triggered by hydrocortisone for 14 days.Rats were treated with YGW or ALN for 12 weeks.The weights of rats were recorded.Hematoxylin-eosin staining staining was used to observe pathological changes in bone trabeculae,liver,spleen,and kidneys of rats.Depletion of the growth plate cartilage of rats in different groups was observed by safranine-O staining.The expression of osteoclast key indices(ACP)and osteoblast key indices(ALP)in the bone tissue of rats in the different groups was observed by immunohistochemical staining.The expression of bone resorption-related indicators(TRAP and NXT-1),bone formation-related indicators(BALP,BGP,and P1NP),and major indicators of kidney-yang deficiency(ACTH,T3,T4,cAMP,and cGMP)were observed using an ELISA detection kit.The expression levels of the main indices of liver function(ALT and AST)were detected in different groups.Results:The differences between the osteoporosis with kidney-yang deficiency group and osteoporosis group were that the weight of rats and the expression of ACTH,T3,T4,and cAMP decreased significantly,and the expression of cGMP increased in the osteoporosis with kidney-yang deficiency group.Moreover,both YGW and ALN effectively improved the symptoms of osteoporosis,including the injury of bone trabeculae and growth plates,as well as the expression of bone metabolism-related indicators.However,unlike ALN,YGW simultaneously ameliorated the expression of key indicators of kidney-yang deficiency and prevented weight loss in rats.In addition,YGW caused no obvious damage to the liver,spleen,or kidney,whereas ALN led to liver cirrhosis.Conclusion:The results reveal that YGW plays a crucial part in osteoporosis with kidney-yang deficiency,increases bone mineral density,and improves bone metabolism indicators,and is safe and efficient for the efficacy of osteoporosis with kidney-yang deficiency.YGW might have a better therapeutic effect on osteoporosis in patients with kidney-yang deficiency.Therefore,alendronate should be used cautiously in patients with osteoporosis and poor liver function.
基金supported by grants from the National Natural Science Foundation of China(No.30230350,No.30572199 and No.30371651).
文摘MHC/peptide tetramer technology has been widely used to study antigen-specific T cells, especially for identifying virus-specific CD8^＋ T cells in humans. The tetramer molecule is composed of HLA heavy chain, β2-microglobulin （β2m）, an antigenic peptide, and fluorescent-labeled streptavidin. To further investigate the HLA-A＊1101-restricted CD8^＋ T cell responses against human cytomegalovirus （HCMV）, we established an approach to prepare HLA-A＊1101 tetramer complexed with a peptide from HCMV. The cDNA encoding HLA-A＊1101 heavy chain was cloned and the prokaryotic expression vector for the ectodomain of HLA-A＊1101 fused with a BirA substrate peptide （HLA-A＊1101-BSP） at its carboxyl terminus was constructed. The fusion protein was highly expressed as inclusion bodies under optimized conditions in Escherichla coli. Moreover, HLA-A＊1101-BSP protein was refolded in the presence of β2m and an HCMV peptide pp6516.24 （GPISGHVLK, GPI）. Soluble HLA-A＊1101-GPI monomer was biotinylated and purified to a purity of 95%, which was subsequently combined with streptavidin to form tetramers at a yield of 〉 80%. The HLA-A＊1101-GPI tetramers could bind to virus-specific CD8^＋ T cells, suggesting soluble HLA-A＊1101-GPI tetramers were biologically functional. This study provides the basis for further evaluation of HLA-A＊1101-restricted CD8^＋ T cell responses against HCMV infection.
基金This work was supported by Key Project of the National Natural Science Foundation of China （No.30230350）.
文摘Major histocompatibility complex （MHC） class I tetramer technology has become the central technique for analyzing antigen-specific CD8^＋ T cell responses and it has been widely used to explore the differentiation and formation of memory CD8^＋ T cells. Previously, a simplified and efficient procedure for preparing high quality HLA-A＊0201 tetramers has been established in our lab and the tetramers loaded with HCMV peptide pp6549s.50a has been successfully applied to investigate HCMV-specific CD8^＋ T cells in Chinese populations. Using similar procedure we reported here the construction of HLA-A＊0201 tetramer loaded with another dominant epitope derived from immediate early （IE）-1 316.324 （VLEETSVML, VLE） of HCMV （A2-VLE） and characterization of this tetramer. After A2-VLE monomer was prepared and purified, its tetramer was then formed at a yield of 83%. The optimized amount of A2-VLE tetramer for staining 100 μl whole blood was 0.5 μg with incubation at 4℃ for 1 h. Furthermore, the dissociation constant of the tetramer binding to the specific CD8^＋ T cells of one HLA-A2^＋ donor was estimated to be 32.7 nmol/L, which is markedly higher than that of MHC monomer. The construction of A2-VLE tetramer provides an alternative choice for investigating HCMV-specific CD8^＋ T cell responses and will deepen our understanding of the differentiation and formation of HCMV-specific memory CD8^＋ T cells. Cellular ＆ Molecular Immunology.
基金the China Postdoctoral Science Foundation(No.2022M720896).
文摘The high recurrence and low responsiveness to immunotherapy make ovarian cancer(OC)the most lethal gynecological malignancy.Tumor microenvironment is critical in risk stratification and the discovery of molecular targets.We developed a prognostic classification for OC,which could also predict the prognosis of other gynecological cancers including breast cancer,endometrial cancer,and cervical cancer.Somatic mutation,hallmark pathways,and immune landscapes were characterized.Integrative analysis of immune checkpoints and multiple immune signatures revealed the low-risk group responds better to immune checkpoint inhibitors,which was validated by an external immunotherapeutic cohort(IMvigor210).Singlecell RNA sequencing(scRNA-seq)confirmed the high expression of SERPINB1 and SERPINB9 in dendritic cells,and AlphaFold2 was used to infer their 3D protein structures.Putative molecular compounds binding to SERPINB1/SERPINB9 were predicted by virtual screening.
