Lipidomics approach by UPLC-Q-Exactive-MS was used for the identification,quantification,comparison,and characterization of sphingolipids in virus infected marine Emiliania huxleyi BOF92 cells.The results show that 16...Lipidomics approach by UPLC-Q-Exactive-MS was used for the identification,quantification,comparison,and characterization of sphingolipids in virus infected marine Emiliania huxleyi BOF92 cells.The results show that 16 significantly changed sphingolipids(including Cer,CerG1,and SPHm)were identified during viral infection.Our data confirmed previously recognized facts that viral infection led to a shift toward virus-specific sphingolipids,which is consistent with the down-regulation of genes involved in the host de novo sphingolipid biosynthesis.Moreover,we revealed the upregulation of virusencoded homologous genes participating in de novo sphingolipids biosynthesis and virus-specific hydroxylated long chain bases(LCBs)as phytoCer,suggesting the competitive inhibition of host sphingolipid synthesis to produce the required building blocks for viral production,replication,and assembly.Additionally,Cer 40꞉1;2,Cer 40꞉2;2 isomer,and CerG139꞉0;2,Cer 39꞉0;2 as novel metabolite markers might indicate the general dysfunctions in E.huxleyi in response to viral infection.Our results show that viral infection led to a profound remodeling of host sphingolipidome,by which viruses depend on the hijacking of host sphingolipid metabolism to support the viral life cycle.展开更多
Pollination biology studies of the endangered herbal medicines Dendrobium chrysotoxum were conducted in natural pollination conditions using flower observation,pollinator observation and artificial pollination experim...Pollination biology studies of the endangered herbal medicines Dendrobium chrysotoxum were conducted in natural pollination conditions using flower observation,pollinator observation and artificial pollination experiments.Populations of D.chrysotoxum with fragrance and nectar were pollinated by Ctenoplectra davidi Valhalla(Hymenoptera:Apidae)species.The floral structure of D.chrysotoxum adapted precisely to its pollinators.Flowers had a low capsule setting(0.17%)under natural conditions.However,compared to open pollination,artificial pollination experiments showed a significant increase in capsule setting,and D.chrysotoxum was cross-compatible and self-compatible,but there was pollinator limitation also.This study will provide important information for the preservation of this endangered species.展开更多
Emiliania huxleyi is the most prominent modern coccolithophore,a group of marine unicellular eukaryotes that play a critical role in ocean biogeochemistry.Coccolithoviruses are large double stranded DNA viruses,which ...Emiliania huxleyi is the most prominent modern coccolithophore,a group of marine unicellular eukaryotes that play a critical role in ocean biogeochemistry.Coccolithoviruses are large double stranded DNA viruses,which is responsible for the demise of large oceanic blooms formed by E.huxleyi.E.huxleyi virus(EhVs)acquired a series of enzyme-coding genes predicted to be involved in the sphingolipid biosynthesis by horizontal gene transfer between virus-host.Currently,there is limited experimental validation identifying the functions of these genes in EhV.Genetic transformation of eukaryotic cells is a powerful tool to get an insight into gene functions of the studied organisms.Serine palmitoyltransferase(SPT)catalyzes the first committed step in de novo sphingolipid biosynthetic pathway.Here,a novel vector system for the transformation of E.huxleyi was designed.It contained fragments of promoter and terminator sequences of E.huxleyi endogenic fucoxanthin chlorophyll a/c-binding protein gene“fcp”and harbored EhV-99B1 spt gene.The resultant recombinant transformation vectors pEhux-I-spt and pEhux-II were co-transferred into E.huxleyi BOF92 by electroporation.Transformants were obtained upon glufosinate-ammonium selection,and confirmed by Southern hybridization,genome PCR,qRT-PCR and Western blot screening of spt gene,which indicated that spt gene was integrated into the nuclear genome and was expressed at the mRNA and protein levels.The expression of the viral spt gene led to differences in lipid compositions analyzed using thin-layer chromatography(TLC).The results present the genetic transformation system for E.huxleyi,providing additional genetic resource with potential for exploring basic biological questions such as the virus-host interactions.展开更多
Heterotopic ossification(HO)is the abnormal formation of bone in extraskeletal sites.However,the mechanisms linking HO pathogenesis with bone mass dysfunction remain unclear.Here,we showed that mice harboring injury-i...Heterotopic ossification(HO)is the abnormal formation of bone in extraskeletal sites.However,the mechanisms linking HO pathogenesis with bone mass dysfunction remain unclear.Here,we showed that mice harboring injury-induced and BMP4-dependent HO exhibit bone mass loss similar to that presented by patients with HO.Moreover,we found that injury-induced hyperinflammatory responses at the injury site triggered HO initiation but did not result in bone mass loss at 1 day post-injury(dpi).In contrast,a suppressive immune response promoted HO propagation and bone mass loss by 7 dpi.Correcting immune dysregulation by PD1/PDL1 blockade dramatically alleviated HO propagation and bone mass loss.We further demonstrated that fetuin-A(FetA),which has been frequently detected in HO lesions but rarely observed in HO-adjacent normal bone,acts as an immunomodulator to promote PD1 expression and M2 macrophage polarization,leading to immunosuppression.Intervention with recombinant FetA inhibited hyperinflammation and prevented HO and associated bone mass loss.Collectively,our findings provide new insights into the osteoimmunological interactions that occur during HO formation and suggest that FetA is an immunosuppressor and a potential therapeutic option for the treatment of HO.展开更多
Objective: To establish angiogenesis model of xenografts of lung cancer cell in nude mouse and investigate the expression of the neuropilin-1 (NRP-1) protein in tumors and its role in progression and angiogenesis of l...Objective: To establish angiogenesis model of xenografts of lung cancer cell in nude mouse and investigate the expression of the neuropilin-1 (NRP-1) protein in tumors and its role in progression and angiogenesis of lung cancer. Methods: Human lung adenocarcinoma cells A549 were analyzed for the expression of vascular endothelial growth factor- 165 (VEGF165) mRNA using RT-PCR in vitro. Two groups of nude mice were subcutaneously inoculated with A549 at differ- ent tumor-loading time. Two groups of xenografts were identified by hematoxylin and eosin (HE) staining, their microvessel density (MVD) were analyzed meanwhile. Two groups were analyzed for the expression of NRP-1 protein and their mean absorbency by using immunohistochemistry and automatic image analysis system respectively. Results: A549 expressed VEGF165 mRNA, and xenografts of A549 in nude mice were successfully established and confirmed by HE staining. The atypia of cancer cells and angiogenesis were occurred in two groups. Two groups of MVD were 13.06 ± 1.58, 23.61 ± 3.11 (ves- sels/mm2) (P < 0.01). NRP-1 protein was expressed in cytoplasm of vascular endothelium cells and partial tumor cells. Two groups of mean absorbency of NRP-1 were 0.1095 ± 0.0228, 0.1784 ± 0.0151 (P < 0.01). Conclusion: The angiogenesis models of xenografts in nude mice with lung cancer cell A549 expressing VEGF165 mRNA at different tumor-loading times were established successfully. The expression of NRP-1 protein and MVD were increased with the tumor progression. Our results demonstrate that NRP-1 protein in lung cancer is related to angiogenesis.展开更多
Cervical cancer (CC) is the fourth most commonly diagnosed female malignancy and a leading cause of cancer-related mortality worldwide, especially in developing countries. Despite the use of advanced screening and pre...Cervical cancer (CC) is the fourth most commonly diagnosed female malignancy and a leading cause of cancer-related mortality worldwide, especially in developing countries. Despite the use of advanced screening and preventive vaccines, more than half of all CC cases are diagnosed at advanced stages, when therapeutic options are extremely limited and side effects are severe. Given these circumstances, new and effective treatments are needed. In recent years, exciting progress has been made in immunotherapies, including the rapid development of immune checkpoint inhibitors. Checkpoint blockades targeting the PD-1/PD-L1 axis have achieved effective clinical responses with acceptable toxicity by suppressing tumor progression and improving survival in several tumor types. In this review, we summarize recent advances in our understanding of the PD-1/PD-L1 signaling pathway, including the expression patterns of PD-1/PD-L1 and potential PD-l/PD-Ll-related therapeutic strategies for CC.展开更多
基金Supported by the National Natural Science Foundation of China(Nos.42076086,41576166)the Natural Science Foundation of Fujian Province(No.2020J05138)+1 种基金the Education and Research Project for Young and Middle-aged Teachers of Fujian Province(No.JAT190343)the Cultivation Plan for Distinguished Young Scholars in Fujian Universities。
文摘Lipidomics approach by UPLC-Q-Exactive-MS was used for the identification,quantification,comparison,and characterization of sphingolipids in virus infected marine Emiliania huxleyi BOF92 cells.The results show that 16 significantly changed sphingolipids(including Cer,CerG1,and SPHm)were identified during viral infection.Our data confirmed previously recognized facts that viral infection led to a shift toward virus-specific sphingolipids,which is consistent with the down-regulation of genes involved in the host de novo sphingolipid biosynthesis.Moreover,we revealed the upregulation of virusencoded homologous genes participating in de novo sphingolipids biosynthesis and virus-specific hydroxylated long chain bases(LCBs)as phytoCer,suggesting the competitive inhibition of host sphingolipid synthesis to produce the required building blocks for viral production,replication,and assembly.Additionally,Cer 40꞉1;2,Cer 40꞉2;2 isomer,and CerG139꞉0;2,Cer 39꞉0;2 as novel metabolite markers might indicate the general dysfunctions in E.huxleyi in response to viral infection.Our results show that viral infection led to a profound remodeling of host sphingolipidome,by which viruses depend on the hijacking of host sphingolipid metabolism to support the viral life cycle.
基金funded by the Basic Research Priorities Program of Yunnan Provincethe Applied Basic Research Programs of Science and Technology Department for Young Scholars(No.2019FD005)Technological Innovation Talents Cultivation Project of Yunnan Province(No.202205AD160043)Technological Innovation Talents Cultivation Project of Dehong City(No.2021RC007).
文摘Pollination biology studies of the endangered herbal medicines Dendrobium chrysotoxum were conducted in natural pollination conditions using flower observation,pollinator observation and artificial pollination experiments.Populations of D.chrysotoxum with fragrance and nectar were pollinated by Ctenoplectra davidi Valhalla(Hymenoptera:Apidae)species.The floral structure of D.chrysotoxum adapted precisely to its pollinators.Flowers had a low capsule setting(0.17%)under natural conditions.However,compared to open pollination,artificial pollination experiments showed a significant increase in capsule setting,and D.chrysotoxum was cross-compatible and self-compatible,but there was pollinator limitation also.This study will provide important information for the preservation of this endangered species.
基金Supported by the National Natural Science Foundation of China(Nos.41576166,21707042,31771972)the Fujian Province Natural Science Foundation of China(Nos.2019J01696,2017J01447,2017J01636)。
文摘Emiliania huxleyi is the most prominent modern coccolithophore,a group of marine unicellular eukaryotes that play a critical role in ocean biogeochemistry.Coccolithoviruses are large double stranded DNA viruses,which is responsible for the demise of large oceanic blooms formed by E.huxleyi.E.huxleyi virus(EhVs)acquired a series of enzyme-coding genes predicted to be involved in the sphingolipid biosynthesis by horizontal gene transfer between virus-host.Currently,there is limited experimental validation identifying the functions of these genes in EhV.Genetic transformation of eukaryotic cells is a powerful tool to get an insight into gene functions of the studied organisms.Serine palmitoyltransferase(SPT)catalyzes the first committed step in de novo sphingolipid biosynthetic pathway.Here,a novel vector system for the transformation of E.huxleyi was designed.It contained fragments of promoter and terminator sequences of E.huxleyi endogenic fucoxanthin chlorophyll a/c-binding protein gene“fcp”and harbored EhV-99B1 spt gene.The resultant recombinant transformation vectors pEhux-I-spt and pEhux-II were co-transferred into E.huxleyi BOF92 by electroporation.Transformants were obtained upon glufosinate-ammonium selection,and confirmed by Southern hybridization,genome PCR,qRT-PCR and Western blot screening of spt gene,which indicated that spt gene was integrated into the nuclear genome and was expressed at the mRNA and protein levels.The expression of the viral spt gene led to differences in lipid compositions analyzed using thin-layer chromatography(TLC).The results present the genetic transformation system for E.huxleyi,providing additional genetic resource with potential for exploring basic biological questions such as the virus-host interactions.
基金supported by the National Key Research and Development Program of China(reference number 2019YFA0801800)the Natural Science Foundation of China(reference numbers 32070916,82102573,8157152,81472087,81670097 and 81870085)the Natural Science Foundation of Anhui Province(reference numbers 1508085MC45 and 1908085QH359).
文摘Heterotopic ossification(HO)is the abnormal formation of bone in extraskeletal sites.However,the mechanisms linking HO pathogenesis with bone mass dysfunction remain unclear.Here,we showed that mice harboring injury-induced and BMP4-dependent HO exhibit bone mass loss similar to that presented by patients with HO.Moreover,we found that injury-induced hyperinflammatory responses at the injury site triggered HO initiation but did not result in bone mass loss at 1 day post-injury(dpi).In contrast,a suppressive immune response promoted HO propagation and bone mass loss by 7 dpi.Correcting immune dysregulation by PD1/PDL1 blockade dramatically alleviated HO propagation and bone mass loss.We further demonstrated that fetuin-A(FetA),which has been frequently detected in HO lesions but rarely observed in HO-adjacent normal bone,acts as an immunomodulator to promote PD1 expression and M2 macrophage polarization,leading to immunosuppression.Intervention with recombinant FetA inhibited hyperinflammation and prevented HO and associated bone mass loss.Collectively,our findings provide new insights into the osteoimmunological interactions that occur during HO formation and suggest that FetA is an immunosuppressor and a potential therapeutic option for the treatment of HO.
基金Province Science and Technology of Hubei Key Program Foundation (No. 2003AA301C05)
文摘Objective: To establish angiogenesis model of xenografts of lung cancer cell in nude mouse and investigate the expression of the neuropilin-1 (NRP-1) protein in tumors and its role in progression and angiogenesis of lung cancer. Methods: Human lung adenocarcinoma cells A549 were analyzed for the expression of vascular endothelial growth factor- 165 (VEGF165) mRNA using RT-PCR in vitro. Two groups of nude mice were subcutaneously inoculated with A549 at differ- ent tumor-loading time. Two groups of xenografts were identified by hematoxylin and eosin (HE) staining, their microvessel density (MVD) were analyzed meanwhile. Two groups were analyzed for the expression of NRP-1 protein and their mean absorbency by using immunohistochemistry and automatic image analysis system respectively. Results: A549 expressed VEGF165 mRNA, and xenografts of A549 in nude mice were successfully established and confirmed by HE staining. The atypia of cancer cells and angiogenesis were occurred in two groups. Two groups of MVD were 13.06 ± 1.58, 23.61 ± 3.11 (ves- sels/mm2) (P < 0.01). NRP-1 protein was expressed in cytoplasm of vascular endothelium cells and partial tumor cells. Two groups of mean absorbency of NRP-1 were 0.1095 ± 0.0228, 0.1784 ± 0.0151 (P < 0.01). Conclusion: The angiogenesis models of xenografts in nude mice with lung cancer cell A549 expressing VEGF165 mRNA at different tumor-loading times were established successfully. The expression of NRP-1 protein and MVD were increased with the tumor progression. Our results demonstrate that NRP-1 protein in lung cancer is related to angiogenesis.
基金This work was supported by the National Natural Science Foundation of China (No. 81373867).
文摘Cervical cancer (CC) is the fourth most commonly diagnosed female malignancy and a leading cause of cancer-related mortality worldwide, especially in developing countries. Despite the use of advanced screening and preventive vaccines, more than half of all CC cases are diagnosed at advanced stages, when therapeutic options are extremely limited and side effects are severe. Given these circumstances, new and effective treatments are needed. In recent years, exciting progress has been made in immunotherapies, including the rapid development of immune checkpoint inhibitors. Checkpoint blockades targeting the PD-1/PD-L1 axis have achieved effective clinical responses with acceptable toxicity by suppressing tumor progression and improving survival in several tumor types. In this review, we summarize recent advances in our understanding of the PD-1/PD-L1 signaling pathway, including the expression patterns of PD-1/PD-L1 and potential PD-l/PD-Ll-related therapeutic strategies for CC.