BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene ma...BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.展开更多
Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinom...Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinoma(G1 EAC)and the recurrence rate after treatment.Methods Sixty patients(aged 20-42 years)with AEH and/or grade 1 EAC limited to the endometrium were enrolled prospectively and randomized into two groups(n=30)to receive oral MA treatment at the daily dose of 160 mg(control)or MA plus oral metformin(850 mg,twice a day)for at least 6 months.The treatment could extend to 12 months until a complete response(CR)was achieved,and follow-up hysteroscopy and curettage were performed every 3 months.For all the patients who achieved CR,endometrial expressions of IGFBP-rP1,p-Akt and p-AMPK were detected immunohistochemically.Results A total of 58 patients completed the treatment.After 9 months of treatment,23(76.7%)patients in the combined treatment group and 20(71.4%)in the control group achieved CR;two patients in the control group achieved CR after converting to the combined treatment.The recurrence rate did not differ significantly between the control group and combined treatment group(30.0%vs 22.7%,P>0.05).Ten(35.7%)patients in the control group experienced significant weight gain of 5.7±6.1 kg,while none of the patients receiving the combined treatment exhibited significant body weight changes.Compared with the control group,the patients receiving the combined treatment showed enhanced endometrial expressions of IGFBP-rP1 and p-AMPK with lowered p-Akt expression.Conclusion Metformin combined with MA may provide an effective option for fertility-sparing treatment of AEH and grade 1 stage IA EAC,and the clinical benefits of metformin for controlling MA-induced weight gain and promoting endometrial expressions of IGFBP-rP1 and p-AMPK while inhibiting p-Akt expression warrants further study.展开更多
A recombinant inbred population (RI) was constructed from a cross between B5, an introgression. line from the wild rice Oryza officinalis Wall. ex Watt, and susceptible cultivar Minghui 63 ( O. sativa L.). The brown p...A recombinant inbred population (RI) was constructed from a cross between B5, an introgression. line from the wild rice Oryza officinalis Wall. ex Watt, and susceptible cultivar Minghui 63 ( O. sativa L.). The brown planthopper ( BPH) resistances of RI lines were evaluated. Based on bulked segregant analysis (BSA), two protein bulks were made by extracting proteins from equally mixed seedlings of extremely resistant and susceptible plants selected from the RI population, respectively. Two-dimensional electrophoresis was used to detect the changes of polypeptide pattern. Results showed that a protein P40 ( pI 6.3, Mw 40 kD) was significantly reduced or vanished after BPH infestation for 48 h in the susceptible bulk, while it remained uninfluenced in the resistant bulk. In connection with the physiological changes of the resistant and susceptible lines subjected to BPH sucking, we suppose that the protein P40 is related to the interaction responses of lice plants to BPH infestation.展开更多
Objective To explore the regulatory effect of fragile X mental retardation protein (FMRP) on the translation of microtubule associated protein 1B (MAP1B). Methods The expressions of MAP1B protein and MAP1B mRNA in...Objective To explore the regulatory effect of fragile X mental retardation protein (FMRP) on the translation of microtubule associated protein 1B (MAP1B). Methods The expressions of MAP1B protein and MAP1B mRNA in the brains of 1-week and 6-week old fragile X mental retardation-1 (FmrI) knockout (KO) mice were investigated by immunohistochemistry, Western blot, and in situ hybridization, with the age-matched wild type mice (WT) as controls. Results The mean optical density (MOD) of MAP1B was significantly decreased in each brain region in KO6W compared with WT6W, whereas in KO1W, this decrease was only found in the hippocampus and cerebellum. MAP1B in 6-week mice was much less than that in 1-week mice of the same genotype. The results of Western blot and in situ hybridization showed that MAP1B protein and MAP1B mRNA were significantly decreased in the hippocampus of both KO1W and KO6W. Conclusion The decreased MAP1B protein and MAP1B mRNA in the Fmrl knockout mice indicate that FMRP may positively regulate the expression of MAP1B.展开更多
[Objective] The aim of this study was to determine the variation of phytic acid content (PAC) and its frequency distribution, as well as the relationship between PAC and protein content, kernel characteristics in 16...[Objective] The aim of this study was to determine the variation of phytic acid content (PAC) and its frequency distribution, as well as the relationship between PAC and protein content, kernel characteristics in 161 Chinese winter wheat cultivars from four regions. [Method] One hundred and sixty-one winter wheat varieties from China Wheat Zones I, II, III and IV (Table 1) were grown in a randomized block de- sign, in the 2009-2011 cropping season; and then the indexes for describing the grain morphological characteristics such as the thousand kernel weight (TKW), kernel length (KL), kernel width (KW) and kernel thickness (KT) were measured; the phytic acid content (PAC), protein content and sedimentation value were also determined; finally, the relationship between PAC and protein content, kernel characteristics were analyzed. [Result] The PAC in the cultivars tested ranged from 0.92% to 1.95% with a mean value of 1.41%. Protein content ranged from 12.60% to 19.20%, with a mean of 15.24%. Most (53.4%) of the wheat genotypes had a PAC value in the range of 1.25% to 1.55%. No significant correlation was found between PAC and protein content, sedimentation value, while protein content and SDS sedimentation value was significant correlated, which suggested the possibility of breeding wheat cultivars that have a low PAC but a high protein content and good gluten quality. There was a high correlation between TKW and KW (,.=0.79), KL (r=0.50) and KT (r=0.64). PAC was found having no significant correlation with TKW, KW, KL and KT. [Conclusion] The result suggests the possibility of breeding wheat cultivars that have a low PAC but high kernel weight.展开更多
The COP9 signalosome and the regulatory lid of the 26S proteasome are both eight-subunit protein complexes which are present in most eukaryotes. There is a one-to-one relationship between the corresponding subunits of...The COP9 signalosome and the regulatory lid of the 26S proteasome are both eight-subunit protein complexes which are present in most eukaryotes. There is a one-to-one relationship between the corresponding subunits of the two protein complexes in terms of their size and amino acid sequences. Eight groups of subunits from the COP9 signalosome and the proteasome lid complex of different organisms are collected from all the databases at the NCBI website. The corresponding subunits of COP9 signalosome and proteasome lid complex share at least 12% amino acid identity and some conserved regions, and the conserved sites spread evenly over the entire length of the subunits, suggesting that the two complexes have a common evolutionary ancestor. Phylogenetic analyses based on the amino acid sequences of the corresponding subunits of two protein complexes indicate that every tree consists of two clades. The subunits from one of the two protein complexes of different organisms are grouped into one of the two clades respectively. The sequences of single-cell organisms are always the basal groups to that of multi-cell animal and plant species. These results imply that the duplication/divergence events of COP9 signalosome and regulatory lid of the proteasome genes have occurred before the divergence of single-cell and multi-cell eukaryotes, and the genes of the two complexes are independently evolved. The analyses of dN/dS correlation show significant Pearson's correlations between 21 and 15 pairs of subunit-encoding sequences within the COP9 signalosome and the proteasome lid complex respectively, suggesting that those subunits pairs might have related functions and interacted with one another, and resulted in co-evolution.展开更多
Estrus-associated glycoprotein (EGP) is a special protein in oviductal fluid which appears before ovulation and fertilization and disappears after the embryo passes into the uterus. One-and two dimensional SDS-PAGE an...Estrus-associated glycoprotein (EGP) is a special protein in oviductal fluid which appears before ovulation and fertilization and disappears after the embryo passes into the uterus. One-and two dimensional SDS-PAGE and Western blotting with monoclonal antibody or peanut agglutinin were used to characterize the glycoprotein,The specificity of the marking agents enabled the study to proceed without purification. EGP was found to include two proteins: one is acidic with a pH of 4.5 while the other is basic with a pH of 8.0 The molecular weights of their subunits are the same, that is 110 kD. This study may shed light on the mechanism of reproductive Process and be helpful to the development of a more cffective culture medium for in vitro culture of early embryo.展开更多
Pinus squamata X. W. Li, a recently-described species of the Pinaceae, is an extremely endangered pine with only 32 individuals in the wild. This species was thought to be intermediate between P. bungeana Zuec. ex End...Pinus squamata X. W. Li, a recently-described species of the Pinaceae, is an extremely endangered pine with only 32 individuals in the wild. This species was thought to be intermediate between P. bungeana Zuec. ex Endl. of subgen. Strobus and P. yunnanensis Franch. of subgen. Pinus by the original author. We made an effort to address the question of the phylogenetic relationship of this peculiar and important species in the context of the major groups of the genus Pinus by using sequences of the rbcL, matK,genes, rpl20-rps18 spacer, trnV intron of the chloroplast genome and the nuclear ribosomal ITS region. The results of the separated analysis and the combined analysis of the four cpDNA sequences and ITS sequence indicated that P. squamata was a stable member of subsect. Gerardianae and P. gerardiana Wall. was the closest species of P. squamata phylogenetically. As a result, the causes of the distribution pattern of subsect. Gerardianae were also discussed.展开更多
Zebrafish(Danio rerio) Z-OTU,containing OTU and TUDOR domains,was predicted to be a member of OTU-related protease,a family of the deubiquitylating enzymes(DUBs).A previous report from our laboratory clearly descr...Zebrafish(Danio rerio) Z-OTU,containing OTU and TUDOR domains,was predicted to be a member of OTU-related protease,a family of the deubiquitylating enzymes(DUBs).A previous report from our laboratory clearly describes the expression patterns of z-otu mRNA.Here,we characterized the Z-OTU protein during zebrafish oogenesis and early embryogenesis.After prokaryotic expression,the recombinant protein of the OTU domain and GST was purified and injected into rabbits to obtain the polyclonal antibody-anti-Z-OTU,which was used for immunohistochemistry in zebrafish ovaries and embryos.Interestingly,obvious differences existed between the expression patterns of z-otu mRNA and its protein during oogenesis and early embryogenesis.In stage I oocytes,z-otu mRNA was detected in cytoplasm while its protein existed in the germinal vesicle.In addition,its protein was distributed during entire oogenesis,while mRNA was not detected in oocytes at stage IV or mature oocytes.The z-otu mRNA disappeared after midblastula transition(MBT) and its protein gradually decreased after this stage.We inferred that Z-OTU protein,like other OTU-related protease with DUB activity,was required for germinal vesicle breakdown of oocytes during meiosis,germinal vesicle migration,and embryo cleavage maintenance.展开更多
文摘BACKGROUND:Adeno-associated virus(AAV)gene therapy has been proven to be reliable and safe for the treatment of osteoarthritis in recent years.However,given the complexity of osteoarthritis pathogenesis,single gene manipulation for the treatment of osteoarthritis may not produce satisfactory results.Previous studies have shown that nuclear factorκB could promote the inflammatory pathway in osteoarthritic chondrocytes,and bone morphogenetic protein 4(BMP4)could promote cartilage regeneration.OBJECTIVE:To test whether combined application of AAV-p65shRNA and AAV-BMP4 will yield the synergistic effect on chondrocytes regeneration and osteoarthritis treatment.METHODS:Viral particles containing AAV-p65-shRNA and AAV-BMP4 were prepared.Their efficacy in inhibiting inflammation in chondrocytes and promoting chondrogenesis was assessed in vitro and in vivo by transfecting AAV-p65-shRNA or AAV-BMP4 into cells.The experiments were divided into five groups:PBS group;osteoarthritis group;AAV-BMP4 group;AAV-p65shRNA group;and BMP4-p65shRNA 1:1 group.Samples were collected at 4,12,and 24 weeks postoperatively.Tissue staining,including safranin O and Alcian blue,was applied after collecting articular tissue.Then,the optimal ratio between the two types of transfected viral particles was further investigated to improve the chondrogenic potential of mixed cells in vivo.RESULTS AND CONCLUSION:The combined application of AAV-p65shRNA and AAV-BMP4 together showed a synergistic effect on cartilage regeneration and osteoarthritis treatment.Mixed cells transfected with AAV-p65shRNA and AAV-BMP4 at a 1:1 ratio produced the most extracellular matrix synthesis(P<0.05).In vivo results also revealed that the combination of the two viruses had the highest regenerative potential for osteoarthritic cartilage(P<0.05).In the present study,we also discovered that the combined therapy had the maximum effect when the two viruses were administered in equal proportions.Decreasing either p65shRNA or BMP4 transfected cells resulted in less collagen II synthesis.This implies that inhibiting inflammation by p65shRNA and promoting regeneration by BMP4 are equally important for osteoarthritis treatment.These findings provide a new strategy for the treatment of early osteoarthritis by simultaneously inhibiting cartilage inflammation and promoting cartilage repair.
文摘Objective To evaluate the efficacy of medroxyprogesterone acetate(MA)plus metformin as the primary fertility-sparing treatment for atypical endometrial hyperplasia(AEH)and early-stage grade 1 endometrial adenocarcinoma(G1 EAC)and the recurrence rate after treatment.Methods Sixty patients(aged 20-42 years)with AEH and/or grade 1 EAC limited to the endometrium were enrolled prospectively and randomized into two groups(n=30)to receive oral MA treatment at the daily dose of 160 mg(control)or MA plus oral metformin(850 mg,twice a day)for at least 6 months.The treatment could extend to 12 months until a complete response(CR)was achieved,and follow-up hysteroscopy and curettage were performed every 3 months.For all the patients who achieved CR,endometrial expressions of IGFBP-rP1,p-Akt and p-AMPK were detected immunohistochemically.Results A total of 58 patients completed the treatment.After 9 months of treatment,23(76.7%)patients in the combined treatment group and 20(71.4%)in the control group achieved CR;two patients in the control group achieved CR after converting to the combined treatment.The recurrence rate did not differ significantly between the control group and combined treatment group(30.0%vs 22.7%,P>0.05).Ten(35.7%)patients in the control group experienced significant weight gain of 5.7±6.1 kg,while none of the patients receiving the combined treatment exhibited significant body weight changes.Compared with the control group,the patients receiving the combined treatment showed enhanced endometrial expressions of IGFBP-rP1 and p-AMPK with lowered p-Akt expression.Conclusion Metformin combined with MA may provide an effective option for fertility-sparing treatment of AEH and grade 1 stage IA EAC,and the clinical benefits of metformin for controlling MA-induced weight gain and promoting endometrial expressions of IGFBP-rP1 and p-AMPK while inhibiting p-Akt expression warrants further study.
文摘A recombinant inbred population (RI) was constructed from a cross between B5, an introgression. line from the wild rice Oryza officinalis Wall. ex Watt, and susceptible cultivar Minghui 63 ( O. sativa L.). The brown planthopper ( BPH) resistances of RI lines were evaluated. Based on bulked segregant analysis (BSA), two protein bulks were made by extracting proteins from equally mixed seedlings of extremely resistant and susceptible plants selected from the RI population, respectively. Two-dimensional electrophoresis was used to detect the changes of polypeptide pattern. Results showed that a protein P40 ( pI 6.3, Mw 40 kD) was significantly reduced or vanished after BPH infestation for 48 h in the susceptible bulk, while it remained uninfluenced in the resistant bulk. In connection with the physiological changes of the resistant and susceptible lines subjected to BPH sucking, we suppose that the protein P40 is related to the interaction responses of lice plants to BPH infestation.
文摘Objective To explore the regulatory effect of fragile X mental retardation protein (FMRP) on the translation of microtubule associated protein 1B (MAP1B). Methods The expressions of MAP1B protein and MAP1B mRNA in the brains of 1-week and 6-week old fragile X mental retardation-1 (FmrI) knockout (KO) mice were investigated by immunohistochemistry, Western blot, and in situ hybridization, with the age-matched wild type mice (WT) as controls. Results The mean optical density (MOD) of MAP1B was significantly decreased in each brain region in KO6W compared with WT6W, whereas in KO1W, this decrease was only found in the hippocampus and cerebellum. MAP1B in 6-week mice was much less than that in 1-week mice of the same genotype. The results of Western blot and in situ hybridization showed that MAP1B protein and MAP1B mRNA were significantly decreased in the hippocampus of both KO1W and KO6W. Conclusion The decreased MAP1B protein and MAP1B mRNA in the Fmrl knockout mice indicate that FMRP may positively regulate the expression of MAP1B.
基金Supported by the National Basic Research Program of China(2009CB118602)Young Backbone Teachers Program of Henan Province(2011)~~
文摘[Objective] The aim of this study was to determine the variation of phytic acid content (PAC) and its frequency distribution, as well as the relationship between PAC and protein content, kernel characteristics in 161 Chinese winter wheat cultivars from four regions. [Method] One hundred and sixty-one winter wheat varieties from China Wheat Zones I, II, III and IV (Table 1) were grown in a randomized block de- sign, in the 2009-2011 cropping season; and then the indexes for describing the grain morphological characteristics such as the thousand kernel weight (TKW), kernel length (KL), kernel width (KW) and kernel thickness (KT) were measured; the phytic acid content (PAC), protein content and sedimentation value were also determined; finally, the relationship between PAC and protein content, kernel characteristics were analyzed. [Result] The PAC in the cultivars tested ranged from 0.92% to 1.95% with a mean value of 1.41%. Protein content ranged from 12.60% to 19.20%, with a mean of 15.24%. Most (53.4%) of the wheat genotypes had a PAC value in the range of 1.25% to 1.55%. No significant correlation was found between PAC and protein content, sedimentation value, while protein content and SDS sedimentation value was significant correlated, which suggested the possibility of breeding wheat cultivars that have a low PAC but a high protein content and good gluten quality. There was a high correlation between TKW and KW (,.=0.79), KL (r=0.50) and KT (r=0.64). PAC was found having no significant correlation with TKW, KW, KL and KT. [Conclusion] The result suggests the possibility of breeding wheat cultivars that have a low PAC but high kernel weight.
文摘The COP9 signalosome and the regulatory lid of the 26S proteasome are both eight-subunit protein complexes which are present in most eukaryotes. There is a one-to-one relationship between the corresponding subunits of the two protein complexes in terms of their size and amino acid sequences. Eight groups of subunits from the COP9 signalosome and the proteasome lid complex of different organisms are collected from all the databases at the NCBI website. The corresponding subunits of COP9 signalosome and proteasome lid complex share at least 12% amino acid identity and some conserved regions, and the conserved sites spread evenly over the entire length of the subunits, suggesting that the two complexes have a common evolutionary ancestor. Phylogenetic analyses based on the amino acid sequences of the corresponding subunits of two protein complexes indicate that every tree consists of two clades. The subunits from one of the two protein complexes of different organisms are grouped into one of the two clades respectively. The sequences of single-cell organisms are always the basal groups to that of multi-cell animal and plant species. These results imply that the duplication/divergence events of COP9 signalosome and regulatory lid of the proteasome genes have occurred before the divergence of single-cell and multi-cell eukaryotes, and the genes of the two complexes are independently evolved. The analyses of dN/dS correlation show significant Pearson's correlations between 21 and 15 pairs of subunit-encoding sequences within the COP9 signalosome and the proteasome lid complex respectively, suggesting that those subunits pairs might have related functions and interacted with one another, and resulted in co-evolution.
文摘Estrus-associated glycoprotein (EGP) is a special protein in oviductal fluid which appears before ovulation and fertilization and disappears after the embryo passes into the uterus. One-and two dimensional SDS-PAGE and Western blotting with monoclonal antibody or peanut agglutinin were used to characterize the glycoprotein,The specificity of the marking agents enabled the study to proceed without purification. EGP was found to include two proteins: one is acidic with a pH of 4.5 while the other is basic with a pH of 8.0 The molecular weights of their subunits are the same, that is 110 kD. This study may shed light on the mechanism of reproductive Process and be helpful to the development of a more cffective culture medium for in vitro culture of early embryo.
文摘Pinus squamata X. W. Li, a recently-described species of the Pinaceae, is an extremely endangered pine with only 32 individuals in the wild. This species was thought to be intermediate between P. bungeana Zuec. ex Endl. of subgen. Strobus and P. yunnanensis Franch. of subgen. Pinus by the original author. We made an effort to address the question of the phylogenetic relationship of this peculiar and important species in the context of the major groups of the genus Pinus by using sequences of the rbcL, matK,genes, rpl20-rps18 spacer, trnV intron of the chloroplast genome and the nuclear ribosomal ITS region. The results of the separated analysis and the combined analysis of the four cpDNA sequences and ITS sequence indicated that P. squamata was a stable member of subsect. Gerardianae and P. gerardiana Wall. was the closest species of P. squamata phylogenetically. As a result, the causes of the distribution pattern of subsect. Gerardianae were also discussed.
基金supported by grants from the ICGEB (International Center for Genetic Engineering and Biotechnology) (CRP/CHN02-01) (SONG Ping)the National Basic Research Program of China (2004CB117400) (SONG Ping)+1 种基金the National Natural Science Foundation of China (30150005 30270675) (SONG Ping)
文摘Zebrafish(Danio rerio) Z-OTU,containing OTU and TUDOR domains,was predicted to be a member of OTU-related protease,a family of the deubiquitylating enzymes(DUBs).A previous report from our laboratory clearly describes the expression patterns of z-otu mRNA.Here,we characterized the Z-OTU protein during zebrafish oogenesis and early embryogenesis.After prokaryotic expression,the recombinant protein of the OTU domain and GST was purified and injected into rabbits to obtain the polyclonal antibody-anti-Z-OTU,which was used for immunohistochemistry in zebrafish ovaries and embryos.Interestingly,obvious differences existed between the expression patterns of z-otu mRNA and its protein during oogenesis and early embryogenesis.In stage I oocytes,z-otu mRNA was detected in cytoplasm while its protein existed in the germinal vesicle.In addition,its protein was distributed during entire oogenesis,while mRNA was not detected in oocytes at stage IV or mature oocytes.The z-otu mRNA disappeared after midblastula transition(MBT) and its protein gradually decreased after this stage.We inferred that Z-OTU protein,like other OTU-related protease with DUB activity,was required for germinal vesicle breakdown of oocytes during meiosis,germinal vesicle migration,and embryo cleavage maintenance.