The small yellow croaker Larimichthys polyactis is an economically important marine fish in Northeast Asia.Currently,its natural resources are threatened by overfishing and environmental pollution.Therefore,research o...The small yellow croaker Larimichthys polyactis is an economically important marine fish in Northeast Asia.Currently,its natural resources are threatened by overfishing and environmental pollution.Therefore,research on the reproductive system of the fish is crucial.Here,we studied the testis development and ultrastructural features of spermatogenesis in cultured L.polyactis using anatomical,histological,and ultrastructural techniques.A pair of testes,consisting of a central sperm duct and radial seminiferous lobules,were observed.The reproduction cycle of testes can be divided into stages I–VI.March to May was confirmed as the breeding season for male L.polyactis,while April is the ideal period for artificial breeding.The male L.polyactis can attain sexual maturity within 1 year.The spermatogenesis of L.polyactis comprised spermatogonium,spermatocyte,spermatid,and mature spermatozoon.The morphology of spermatogenic cells changed obviously during spermiogenesis,including nuclear shaping,midpiece and flagellum formation.The mature sperms consist of an ellipsoidal head,a short midpiece,and a long flagellum.The anterior of the head with a kidney-shaped nucleus can be distinguished.The midpiece is located posterior to the head and includes four to six spherical mitochondria.The flagellum has irregular lateral fins.The testis of L.polyactis is an unrestricted lobular type,with cystic spermatogenesis,type II spermiogenesis,and type II spermatozoa.These features are highly similar to those of other Sciaenid species.Our findings provide useful insights into the mechanism underlying testis development and spermatogenesis of L.polyactis,which can facilitate the artificial breeding of this species.展开更多
BACKGROUND The advent of cutting-edge systemic therapies has driven advances in the treatment of hepatocellular carcinoma(HCC),and therapeutic strategies with multiple modes of delivery have been shown to be more effi...BACKGROUND The advent of cutting-edge systemic therapies has driven advances in the treatment of hepatocellular carcinoma(HCC),and therapeutic strategies with multiple modes of delivery have been shown to be more efficacious than mono-therapy.However,the mechanisms underlying this innovative treatment modality have not been elucidated.AIM To evaluate the clinical efficacy of targeted therapy plus immunotherapy combined with hepatic arterial infusion chemotherapy(HAIC)of FOLFOX in patients with unresectable HCC.METHODS We enrolled 53 patients with unresectable HCC who received a combination of targeted therapy,immunotherapy,and HAIC of FOLFOX between December 2020 and June 2021 and assessed the efficacy and safety of the treatment regimen.RESULTS The objective response rate was 60.4%(32/53),complete response was 24.5%(13/53),partial response was 35.9%(19/53),and stable disease was 39.6%(21/53).The median duration of response and median progression-free survival were 9.1 and 13.9 months,respectively.The surgical conversion rate was 34.0%(18/53),and 1-year overall survival was 83.0%without critical complicating diseases or adverse events(AEs).CONCLUSION The regimen of HAIC of FOLFOX,targeted therapy,and immunotherapy was curative for patients with unresectable HCC,with no serious AEs and a high rate of surgical conversion.展开更多
KIF3A and KIF3B are two N-terminal motor proteins belonging to the kinesin-II superfamily that play essential roles in spermiogenesis.To understand the roles played by KIF3 A/3B during spermatogenesis of large yellow ...KIF3A and KIF3B are two N-terminal motor proteins belonging to the kinesin-II superfamily that play essential roles in spermiogenesis.To understand the roles played by KIF3 A/3B during spermatogenesis of large yellow croaker Larimichthys crocea,we studied the testis characteristics at different developmental stages of L.crocea,and determined the spatiotemporal expression patterns of kif3a and kif3b during spermiogenesis.Quantitative real-time PCR(qR T-PCR)showed that the overall trends of kif3 a/3 b m RNA abundance during testis development are similar.From stage Ⅱ to stage V,kif3a/3b m RNA abundances first increased and then fell after reaching a peak at stage IV.Interestingly,the m RNA abundances of both genes at stage V were higher than those at stages Ⅱ and Ⅲ.In addition,it is worth of noting that kif3 b m RNA abundance was higher than that of kif3a at all stages.Fluorescence in situ hybridization results revealed that kif3a/3b m RNA abundance dynamics were consistent with the migration of mitochondria,the deformation of nucleus,and the formation of tail.The m RNA hybridization signals of both genes first appeared either around the nuclear periphery or on the side of the nuclei,then appeared at one side of nuclei,and finally were mainly on the tail during spermiogenesis.Our findings contributed to better understanding the molecular mechanisms of spermiogenesis in fish;and suggested that KIF3A and KIF3B may participate in the intracellular transport of mitochondria,nuclear deformation,and the formation of tail during the spermiogenesis in L.crocea.展开更多
Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during sp...Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during spermatogenesis is still un-known.In this study,the cytological features of spermatogenesis were investigated in Larimichthys crocea.In addition,the structure and function of prohibitin(PHB),which is associated with mitochondrial structure and dynamic,was also investigated.The full-length cDNA and protein(Lc-PHB)from the L.crocea phb gene(Lc-phb)contained 1625 base pairs and 271 amino acids,respec-tively.Lc-PHB had a conserved primary structure that resulted in a transmembrane,SPFH(the analogous region of proteins stomat-ins,prohibitins,flotillins and HflK/C),and coiled-coil domains.It was detected at high levels in the muscle,liver,and heart,and at intermediate levels in the testis,gill,and brain.Lc-phb mRNA expression was detected in spermatogenic cells by fluorescence in situ hybridization.An immunofluorescence assay revealed that PHB protein was localized in the mitochondria during spermatogenesis.Specifically,PHB expression was detected in the perinuclear cytoplasm of spermatogonia,spermatocytes,and spermatids in the early developmental stage,and mainly localized on one side of the nuclei in the cytoplasm of spermatids in a middle developmental stage,and finally on the sperm midpiece.Western blotting showed that PHB was located in the extracted mitochondria protein fraction but not in the cytoplasm protein fraction of testes.Conclusively,these results indicated that PHB was expressed in the mitochondria dur-ing spermatogenesis.In addition,the study explained the mitochondrial dynamic during fish spermatogenesis and proposed a possi-ble relationship among PHB,spermatogenesis,and male fertility.展开更多
The microtubule(MT)-associated proteins KIF3A and KIF3B are ubiquitously expressed in a wide range of taxa.This study investigated the functions of these proteins in spermiogenesis,which involves various MT-dependent ...The microtubule(MT)-associated proteins KIF3A and KIF3B are ubiquitously expressed in a wide range of taxa.This study investigated the functions of these proteins in spermiogenesis,which involves various MT-dependent processes,in Phasco-losoma esculenta.We cloned the complete cDNA of Pe-KIF3A/3B.Structural predictions showed that Pe-KIF3A/3B are composed of a highly conserved motor domain,a coiled-coil domain,and a tail domain.Real-time quantitative PCR showed that Pe-kif3a/3b are expressed in all tissues evaluated,with the highest levels in sperm masses.Fluorescence in situ hybridization and immunofluo-rescence were employed to analyze the dynamic expression patterns of KIF3A/3B during spermiogenesis.Pe-KIF3A/3B consistently co-localized with MTs at all stages of spermiogenesis,indicating their potential functions in cargo trafficking.Pe-KIF3A/3B co-localized with mitochondria,suggesting that they may mediate mitochondrial movement.In late-stage spermatids and mature sperm,co-localization was detected in the midpiece,suggesting that Pe-KIF3A/3B could facilitate midpiece formation.The co-localization of Pe-KIF3A and Pe-KIF3B at all stages of spermiogenesis suggests their function as the heterodimeric KIF3AB.Basing on the ob-served temporal and spatial expression patterns of Pe-KIF3A/3B,MTs,and mitochondria in our study,we suggest that heterodimer KIF3AB has a potential role in acrosome biogenesis,sperm head remodeling,enflagellation,and mitochondrial migration during spermiogenesis in P.esculenta.In addition,our study on the morphological characteristics of spermatogenic cells provided fundamental data on the reproductive biology of P.esculenta.展开更多
基金the NSFC-Zhejiang Joint Fund for the Integration of Industrialization and Informatization(No.U1809212)the Scientific and Technical Project of Zhejiang Province(Nos.2021C02055,2017C02013)+2 种基金the National Natural Science Foundation of China(No.31272642)Healthy Aquaculture,the K.C.Wong Magna Fund in Ningbo Universitythe Collaborative Innovation Center for Zhejiang Marine High-Efficiency。
文摘The small yellow croaker Larimichthys polyactis is an economically important marine fish in Northeast Asia.Currently,its natural resources are threatened by overfishing and environmental pollution.Therefore,research on the reproductive system of the fish is crucial.Here,we studied the testis development and ultrastructural features of spermatogenesis in cultured L.polyactis using anatomical,histological,and ultrastructural techniques.A pair of testes,consisting of a central sperm duct and radial seminiferous lobules,were observed.The reproduction cycle of testes can be divided into stages I–VI.March to May was confirmed as the breeding season for male L.polyactis,while April is the ideal period for artificial breeding.The male L.polyactis can attain sexual maturity within 1 year.The spermatogenesis of L.polyactis comprised spermatogonium,spermatocyte,spermatid,and mature spermatozoon.The morphology of spermatogenic cells changed obviously during spermiogenesis,including nuclear shaping,midpiece and flagellum formation.The mature sperms consist of an ellipsoidal head,a short midpiece,and a long flagellum.The anterior of the head with a kidney-shaped nucleus can be distinguished.The midpiece is located posterior to the head and includes four to six spherical mitochondria.The flagellum has irregular lateral fins.The testis of L.polyactis is an unrestricted lobular type,with cystic spermatogenesis,type II spermiogenesis,and type II spermatozoa.These features are highly similar to those of other Sciaenid species.Our findings provide useful insights into the mechanism underlying testis development and spermatogenesis of L.polyactis,which can facilitate the artificial breeding of this species.
基金This study was reviewed and approved by the Ethics Committee of Zhongshan People’s Hospital(Approval No.2022-029).
文摘BACKGROUND The advent of cutting-edge systemic therapies has driven advances in the treatment of hepatocellular carcinoma(HCC),and therapeutic strategies with multiple modes of delivery have been shown to be more efficacious than mono-therapy.However,the mechanisms underlying this innovative treatment modality have not been elucidated.AIM To evaluate the clinical efficacy of targeted therapy plus immunotherapy combined with hepatic arterial infusion chemotherapy(HAIC)of FOLFOX in patients with unresectable HCC.METHODS We enrolled 53 patients with unresectable HCC who received a combination of targeted therapy,immunotherapy,and HAIC of FOLFOX between December 2020 and June 2021 and assessed the efficacy and safety of the treatment regimen.RESULTS The objective response rate was 60.4%(32/53),complete response was 24.5%(13/53),partial response was 35.9%(19/53),and stable disease was 39.6%(21/53).The median duration of response and median progression-free survival were 9.1 and 13.9 months,respectively.The surgical conversion rate was 34.0%(18/53),and 1-year overall survival was 83.0%without critical complicating diseases or adverse events(AEs).CONCLUSION The regimen of HAIC of FOLFOX,targeted therapy,and immunotherapy was curative for patients with unresectable HCC,with no serious AEs and a high rate of surgical conversion.
基金financially supported by the Scientific and Technical Project of Zhejiang Province (Nos. 2016C02055-7, LY18C190007)the Ningbo Natural Science Foundation (No. 2016A610081)+3 种基金the Scientific and Technical Project of Ningbo (No. 2015C110005)the National Natural Science Foundation of China (No. 31602140)the Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculturethe K. C. Wong Magna Fund in Ningbo University
文摘KIF3A and KIF3B are two N-terminal motor proteins belonging to the kinesin-II superfamily that play essential roles in spermiogenesis.To understand the roles played by KIF3 A/3B during spermatogenesis of large yellow croaker Larimichthys crocea,we studied the testis characteristics at different developmental stages of L.crocea,and determined the spatiotemporal expression patterns of kif3a and kif3b during spermiogenesis.Quantitative real-time PCR(qR T-PCR)showed that the overall trends of kif3 a/3 b m RNA abundance during testis development are similar.From stage Ⅱ to stage V,kif3a/3b m RNA abundances first increased and then fell after reaching a peak at stage IV.Interestingly,the m RNA abundances of both genes at stage V were higher than those at stages Ⅱ and Ⅲ.In addition,it is worth of noting that kif3 b m RNA abundance was higher than that of kif3a at all stages.Fluorescence in situ hybridization results revealed that kif3a/3b m RNA abundance dynamics were consistent with the migration of mitochondria,the deformation of nucleus,and the formation of tail.The m RNA hybridization signals of both genes first appeared either around the nuclear periphery or on the side of the nuclei,then appeared at one side of nuclei,and finally were mainly on the tail during spermiogenesis.Our findings contributed to better understanding the molecular mechanisms of spermiogenesis in fish;and suggested that KIF3A and KIF3B may participate in the intracellular transport of mitochondria,nuclear deformation,and the formation of tail during the spermiogenesis in L.crocea.
基金funded by the Natural Science Foun dation of Zhejiang Province(No.LY18C190007)the Na tural Science Foundation of China-Zhejiang Joint Fund for the Integration of Industrialization and Informatization(No.U1809212)+3 种基金the Natural Science Foundation of Ningbo City(No.2018A610228)the Scientific and Technical Project of Zhejiang Province(No.2021C02069-1)the Scientific and Technical Project of Ningbo City(No.2021Z002),the Scientific Research Foundation of Ningbo University(No.XYL19023)the Collaborative Innovation Center for Zhejiang Marine High-efficiency and Healthy Aquaculture,the K.C.Wong Magna Fund in Ningbo University.
文摘Mitochondria are important for animals’fertility,and their morphologies and functions during spermatogenesis are un-der investigation.However,the molecular mechanism that regulates the mitochondrial dynamic during spermatogenesis is still un-known.In this study,the cytological features of spermatogenesis were investigated in Larimichthys crocea.In addition,the structure and function of prohibitin(PHB),which is associated with mitochondrial structure and dynamic,was also investigated.The full-length cDNA and protein(Lc-PHB)from the L.crocea phb gene(Lc-phb)contained 1625 base pairs and 271 amino acids,respec-tively.Lc-PHB had a conserved primary structure that resulted in a transmembrane,SPFH(the analogous region of proteins stomat-ins,prohibitins,flotillins and HflK/C),and coiled-coil domains.It was detected at high levels in the muscle,liver,and heart,and at intermediate levels in the testis,gill,and brain.Lc-phb mRNA expression was detected in spermatogenic cells by fluorescence in situ hybridization.An immunofluorescence assay revealed that PHB protein was localized in the mitochondria during spermatogenesis.Specifically,PHB expression was detected in the perinuclear cytoplasm of spermatogonia,spermatocytes,and spermatids in the early developmental stage,and mainly localized on one side of the nuclei in the cytoplasm of spermatids in a middle developmental stage,and finally on the sperm midpiece.Western blotting showed that PHB was located in the extracted mitochondria protein fraction but not in the cytoplasm protein fraction of testes.Conclusively,these results indicated that PHB was expressed in the mitochondria dur-ing spermatogenesis.In addition,the study explained the mitochondrial dynamic during fish spermatogenesis and proposed a possi-ble relationship among PHB,spermatogenesis,and male fertility.
基金supported by the Ningbo Science and Technology Plan Projects (Nos. 2019B10016, 2016C 10004)the Major Science and Technology Projects in Zhejiang Province (No. 2011C12013)+2 种基金the Natural Science Foundation of Zhejiang Province (No. LY18C190007)the National Natural Science Foundation of China (No. 312 72642)the Collaborative Innovation Center for Marine High-efficiency and Healthy Aquaculture of Zhejiang Province, and K.C.Wong Magna Fund in Ningbo University
文摘The microtubule(MT)-associated proteins KIF3A and KIF3B are ubiquitously expressed in a wide range of taxa.This study investigated the functions of these proteins in spermiogenesis,which involves various MT-dependent processes,in Phasco-losoma esculenta.We cloned the complete cDNA of Pe-KIF3A/3B.Structural predictions showed that Pe-KIF3A/3B are composed of a highly conserved motor domain,a coiled-coil domain,and a tail domain.Real-time quantitative PCR showed that Pe-kif3a/3b are expressed in all tissues evaluated,with the highest levels in sperm masses.Fluorescence in situ hybridization and immunofluo-rescence were employed to analyze the dynamic expression patterns of KIF3A/3B during spermiogenesis.Pe-KIF3A/3B consistently co-localized with MTs at all stages of spermiogenesis,indicating their potential functions in cargo trafficking.Pe-KIF3A/3B co-localized with mitochondria,suggesting that they may mediate mitochondrial movement.In late-stage spermatids and mature sperm,co-localization was detected in the midpiece,suggesting that Pe-KIF3A/3B could facilitate midpiece formation.The co-localization of Pe-KIF3A and Pe-KIF3B at all stages of spermiogenesis suggests their function as the heterodimeric KIF3AB.Basing on the ob-served temporal and spatial expression patterns of Pe-KIF3A/3B,MTs,and mitochondria in our study,we suggest that heterodimer KIF3AB has a potential role in acrosome biogenesis,sperm head remodeling,enflagellation,and mitochondrial migration during spermiogenesis in P.esculenta.In addition,our study on the morphological characteristics of spermatogenic cells provided fundamental data on the reproductive biology of P.esculenta.
