Background： Despite recent advances that have improved the pregnancy success rates that can be achieved via in vitro fertilization （IVF） therapy, it is not yet clear which blastocyst morphological paralneters best...Background： Despite recent advances that have improved the pregnancy success rates that can be achieved via in vitro fertilization （IVF） therapy, it is not yet clear which blastocyst morphological paralneters best predict the outcomes of single blastocyst transfer. In addition. most of the previous studies did not exclude the effect of embryo aneuploidy on blastocysts transfer. Thus, the present study investigated the predictive value of various parameters on the pregnancy outcomes achieved via the transfer of frozcn euploid blastocysts. Methods： The study retrospectively analyzed 914 single euploid blastocyst transfer cycles that were performed at the Peking U laivcrsity Third Hospital Reproductive Medical Center between June 2011 and May 2016. The expansion, trophectoderm （TE）. and inner cell mass （ICM） quality of the blastocysts were assessed based on blastocyst parameters, and used to differentiate between ＂excellent＂, ＂good＂, ＂average＂, and ＂poor＂-quality embryos. The relationship between these embryo grades and the achieved pregnancy outcomes was then analyzed via the Chi-square and logistic regression tests. Results： For embryo grades of excellent, good, average and poor, the clinical pregnancy rates were 65.0%. 50.3%, 50.3% and 33.3%. respectively; and the live-birth rates were 50.0%, 49.7%, 42.3% and 25.0%, respectively. Both the clinical pregnancy ratc （x2= 21.28. P = 0.001） and live-birth rate （x2 = 13.50, P 〈 0.001） increased with the overall blastocyst grade. Both rates were significanlly higher after the transfer era blastocyst that exhibited either an A-grade or B-grade TE, and similarly, an A-grade ICM. than after the transfer el a blastocyst that exhibited a C-grade TE and/or ICM. The degree of blastocysl expansion had no apparent effect on the clinical pregnancy or live-birth rate. All odds ratio were adjusted for patient age, body mass index, length （years） of infertility history, and infertility type. Conclusions： A higher overall euploid blastocyst quality is shown to correlate most strongly with optimal pregnancy outcomes. The study thus supports the use of the described TE and ICM morphological grades to augment current embryo selection criteria.展开更多
In the field of developmental biology and regenerative medicine,mammalian interspecific chimeras have been proved very useful for investigating early embryonic development and the immune system establishment,and exten...In the field of developmental biology and regenerative medicine,mammalian interspecific chimeras have been proved very useful for investigating early embryonic development and the immune system establishment,and extended to a promising potential for human organ generation（Rossant et al.,1982）.展开更多
Clinical assessment of fluid volume status in children during malaria can be taxing and often inaccurate. During malaria, changes in fluid volume are rather multifarious and estimating this parameter, especially in si...Clinical assessment of fluid volume status in children during malaria can be taxing and often inaccurate. During malaria, changes in fluid volume are rather multifarious and estimating this parameter, especially in sick children is very challenging for clinicians who frequently rely on indices such as long capillary refill times, tachycardia, central venous pressure and decreased urine volume as guides. Here, we present the UHAS-MIDA, an open-source software tool that calculates the red blood cell (RBC) concentration and blood volume during malaria in children determined using a stable isotope of chromium (<sup>53</sup>Cr as the label) by gas chromatography-mass spectrometry in selective ion monitoring (GC/MS-SIM) analysis. A key component involves the determination of the compositions of the most abundant naturally occurring isotopes of Cr (<sup>50</sup>Cr, <sup>52</sup>Cr, <sup>53</sup>Cr), and converting the proportions into a 3 × 3 matrix. To estimate unknown proportions of chromium isotopic mixtures from the measured abundances of three ions, an inverse matrix was calculated. The inverse together with several inputs is then used to calculate the corrected MS ion abundances. Thus, we constructed the software tool UHAS- MIDA using HTML, CSS/Bootstrap, JavaScript, and PHP scripting languages. The tool enables the user to efficiently determine RBC concentration and fluid volume. The source code, binary packages and associated materials for UHAS-MIDA are freely available at https://github.com/bentil078/Abaye-et-al_UHASmida展开更多
The intra- or inter-strain reconstitutedblastocysts were produced by replacing the inner cell mass of Kunming mouse blastocysts with that of Kunming orC57BL/6 mouse strain blastocysts. A total of 192 intra-strain reco...The intra- or inter-strain reconstitutedblastocysts were produced by replacing the inner cell mass of Kunming mouse blastocysts with that of Kunming orC57BL/6 mouse strain blastocysts. A total of 192 intra-strain reconstituted blastocysts were transferred into 17pseudopregnant Kunming mice, and 2 reconstituted embryos were developed into term; while 115 inter-strain reconstituted blastocysts were produced, analysis of thereconstituted blastocysts showed that the morphology andcytoskeleton srtucture of the blastocysts were not differentfrom those of normal blastocysts, however, no viableoffspring was obtained after embryo transfer for these inter-strain reconstituted blastocysts. The resultsdemonstrated that the intra-strain reconstituted blastocystscould normally develop into term, whereas the inter-strain reconstituted blastocysts possessed less developmental potential as the intra-strain reconstituted blastocysts. Thisstudy may give light to solve the problem of low implantation rate and placenta abnormality in mammal cloning.展开更多
Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose ...Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose but immunoassays face limitation of both availability and qualities of antibody reagents that are necessary for development of immune assays. With genomics data base available, mass spectrometry (MS) can analyze protein tryptic peptides directly for quantitative determination of proteins. In this study, we report a method for detection of matrix metalloproteinase 1 (MMP1), an important extracellular matrix modulator, in human breast cancer cells by quadrupole time-of-flight (Q-TOF) MS. Absolute quantification of MMP1 was conducted using the selected reaction monitoring (SRM) on a triple quadrupole (Triple-Quad) MS via transitions selected from MMP1 tryptic peptides using non isotope labeled MMP1 protein as a titration standard. In comparison with immune based assay, this MS method showed picogram level sensitivity for quantitative determination of MMP1 intotal cell lysates. Our results demonstrated the feasibility of absolute quantification of low abundance proteins using label-free protein standard by mass spectrometry. Therefore, this method provides not only advantages of high sensitivity but also cost saving in comparison with the commonly used mass spectrometry that currently employs isotype labeled proteins for quantitative analysis.展开更多
AIM: To conduct the proteomic analysis of human colorectal carcinoma cell line, SW480 by using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption /ionization-time of flight mass spectrometry (...AIM: To conduct the proteomic analysis of human colorectal carcinoma cell line, SW480 by using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption /ionization-time of flight mass spectrometry (MALDITOFMS).METHODS: The total proteins of human colorectal carcinoma cell line, SW480 were separated with 2-DE by using immobilized pH gradient strips and visualized by staining with silver nitrate. The gel images were acquired by scanner and 2-DE analysis software, Image Master 2D Elite. Nineteen distinct protein spots were excised from gel randomly and digested in gel by TPCK-trypsin. Mass analysis ofthe tryptic digest peptides mixture was performed by using MALDI-TOF MS. Peptide mass fingerprints (PMFs) obtained by the MALDI-TOF analysis were used to search NCBI,SWISS-PROT and MSDB databases by using Mascot software.RESULTS: PMF maps of all spots were obtained by MALDI-TOF MS and thirteen proteins were preliminarily identified.CONCLUSION: The methods of analysis and identification of protein spots of tumor cells in 2-DE gel with silver staining by MALDI-TOF MS derived PMF have been established.Protein expression profile of SW480 has been obtained.It is demonstrated that a combination of proteomics and cell culture is a useful approach to comprehend the process of colon carcinogenesis.展开更多
Objective: Patients with head and neck cancer often suffer from malnutrition. This study aims to investigate the influence of body mass index(BMI) on the prognosis of laryngeal squamous cell carcinoma(LSCC).Methods: A...Objective: Patients with head and neck cancer often suffer from malnutrition. This study aims to investigate the influence of body mass index(BMI) on the prognosis of laryngeal squamous cell carcinoma(LSCC).Methods: A total of 473 patients with LSCC initially treated at Sun Yat-sen University Cancer Center between January 2005 and July 2009 were retrospectively reviewed. Survival analysis was performed by the Kaplan-Meier method and Cox regression model.Results: Low BMI before treatment was significantly associated with poor overall survival in patients with LSCC(P<0.001). BMI was an independent prognostic factor for patients with LSCC.Conclusion: Leanness before treatment was associated with poor prognosis in patients with LSCC. Good nutritional status is favorable to improve survival in patients with LSCC.展开更多
Carboxyl terminus of Hsp70-interacting protein（CHIP or STUB1） is an E3 ligase and regulates the stability of several proteins which are involved in different cellular functions. Our previous studies demonstrated tha...Carboxyl terminus of Hsp70-interacting protein（CHIP or STUB1） is an E3 ligase and regulates the stability of several proteins which are involved in different cellular functions. Our previous studies demonstrated that Chip deficient mice display bone loss phenotype due to increased osteoclast formation through enhancing TRAF6 activity in osteoclasts. In this study we provide novel evidence about the function of CHIP. We found that osteoblast differentiation and bone formation were also decreased in Chip KO mice. In bone marrow stromal（BMS） cells derived from Chip^-/- mice, expression of a panel of osteoblast marker genes was significantly decreased. ALP activity and mineralized bone matrix formation were also reduced in Chip-deficient BMS cells. We also found that in addition to the regulation of TRAF6, CHIP also inhibits TNFα-induced NF-κB signaling through promoting TRAF2 and TRAF5 degradation. Specific deletion of Chip in BMS cells downregulated expression of osteoblast marker genes which could be reversed by the addition of NF-κB inhibitor. These results demonstrate that the osteopenic phenotype observed in Chip^-/- mice was due to the combination of increased osteoclast formation and decreased osteoblast differentiation. Taken together, our findings indicate a significant role of CHIP in bone remodeling.展开更多
Bovine embryonic stem cell would be invaluable for researching the aspect of animal cloning, production transgenic animal and discussion of gene function in vitro. With the object of establishing an effective culture ...Bovine embryonic stem cell would be invaluable for researching the aspect of animal cloning, production transgenic animal and discussion of gene function in vitro. With the object of establishing an effective culture system for isolation and clone of bovine pluripotent stem cell, we cultured bovine embryos and mouse embryos including morula blastula and hatached blastula and obtained animal ICM on Primary murine embryonic fibroblast (Primary murine embryonic fibroblast, PMEF) feeder layer with tissue medium(DMEM supplemented with 15ml/100ml NBS,0.1μmol/L Na2SeO3, 0.1mmol/L p-mercaptoethanol, 1000ng/ml LIF, 10 ng/ml IGF, 1mmol/L necessary amino acid and 1mmol/L L-glutamine),then,we obtained mouse ICM and bovine ICM. Moreover, we isolated and cloned the 6 passage bovine ES like cells(12 cell lines) and 9 passage murine ES like cells (52 cell lines) deriving from bovine ICM and murine ICM respectively on the feeder layer of PMEF by disaggregating ICM and ES cell clones of bovine and murine into smaller clumps through digesting with 0.125g/100ml trypsin and 0.02g/100ml EDTA and scattering with a glass needle. The pluripotency of both murine and bovine ES like cells was identified with morphological character, histochemistry identification , karyotype analysis and differentiation of ES cells in vitro or in vivo. This result showed that bovine embryonic stem cell and murine embryonic stem cell had developmental pluripotency.展开更多
文摘Background： Despite recent advances that have improved the pregnancy success rates that can be achieved via in vitro fertilization （IVF） therapy, it is not yet clear which blastocyst morphological paralneters best predict the outcomes of single blastocyst transfer. In addition. most of the previous studies did not exclude the effect of embryo aneuploidy on blastocysts transfer. Thus, the present study investigated the predictive value of various parameters on the pregnancy outcomes achieved via the transfer of frozcn euploid blastocysts. Methods： The study retrospectively analyzed 914 single euploid blastocyst transfer cycles that were performed at the Peking U laivcrsity Third Hospital Reproductive Medical Center between June 2011 and May 2016. The expansion, trophectoderm （TE）. and inner cell mass （ICM） quality of the blastocysts were assessed based on blastocyst parameters, and used to differentiate between ＂excellent＂, ＂good＂, ＂average＂, and ＂poor＂-quality embryos. The relationship between these embryo grades and the achieved pregnancy outcomes was then analyzed via the Chi-square and logistic regression tests. Results： For embryo grades of excellent, good, average and poor, the clinical pregnancy rates were 65.0%. 50.3%, 50.3% and 33.3%. respectively; and the live-birth rates were 50.0%, 49.7%, 42.3% and 25.0%, respectively. Both the clinical pregnancy ratc （x2= 21.28. P = 0.001） and live-birth rate （x2 = 13.50, P 〈 0.001） increased with the overall blastocyst grade. Both rates were significanlly higher after the transfer era blastocyst that exhibited either an A-grade or B-grade TE, and similarly, an A-grade ICM. than after the transfer el a blastocyst that exhibited a C-grade TE and/or ICM. The degree of blastocysl expansion had no apparent effect on the clinical pregnancy or live-birth rate. All odds ratio were adjusted for patient age, body mass index, length （years） of infertility history, and infertility type. Conclusions： A higher overall euploid blastocyst quality is shown to correlate most strongly with optimal pregnancy outcomes. The study thus supports the use of the described TE and ICM morphological grades to augment current embryo selection criteria.
基金supported by the grants from the National Natural Science Foundation of China (Nos.31621004,31501188 and 31422038)the Key Research Projects of the Frontier Science of the Chinese Academy of Sciences (QYZDY-SSW-SMC002)the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA16000000)
文摘In the field of developmental biology and regenerative medicine,mammalian interspecific chimeras have been proved very useful for investigating early embryonic development and the immune system establishment,and extended to a promising potential for human organ generation（Rossant et al.,1982）.
文摘Clinical assessment of fluid volume status in children during malaria can be taxing and often inaccurate. During malaria, changes in fluid volume are rather multifarious and estimating this parameter, especially in sick children is very challenging for clinicians who frequently rely on indices such as long capillary refill times, tachycardia, central venous pressure and decreased urine volume as guides. Here, we present the UHAS-MIDA, an open-source software tool that calculates the red blood cell (RBC) concentration and blood volume during malaria in children determined using a stable isotope of chromium (<sup>53</sup>Cr as the label) by gas chromatography-mass spectrometry in selective ion monitoring (GC/MS-SIM) analysis. A key component involves the determination of the compositions of the most abundant naturally occurring isotopes of Cr (<sup>50</sup>Cr, <sup>52</sup>Cr, <sup>53</sup>Cr), and converting the proportions into a 3 × 3 matrix. To estimate unknown proportions of chromium isotopic mixtures from the measured abundances of three ions, an inverse matrix was calculated. The inverse together with several inputs is then used to calculate the corrected MS ion abundances. Thus, we constructed the software tool UHAS- MIDA using HTML, CSS/Bootstrap, JavaScript, and PHP scripting languages. The tool enables the user to efficiently determine RBC concentration and fluid volume. The source code, binary packages and associated materials for UHAS-MIDA are freely available at https://github.com/bentil078/Abaye-et-al_UHASmida
基金supported by the Climbing Project of the Ministry of Science and Technology of China(Climbing-special-08)the Knowledge Innovation Project of the Chinese Academy of Sciences(Grant No.KSCX1-05-01).
文摘The intra- or inter-strain reconstitutedblastocysts were produced by replacing the inner cell mass of Kunming mouse blastocysts with that of Kunming orC57BL/6 mouse strain blastocysts. A total of 192 intra-strain reconstituted blastocysts were transferred into 17pseudopregnant Kunming mice, and 2 reconstituted embryos were developed into term; while 115 inter-strain reconstituted blastocysts were produced, analysis of thereconstituted blastocysts showed that the morphology andcytoskeleton srtucture of the blastocysts were not differentfrom those of normal blastocysts, however, no viableoffspring was obtained after embryo transfer for these inter-strain reconstituted blastocysts. The resultsdemonstrated that the intra-strain reconstituted blastocystscould normally develop into term, whereas the inter-strain reconstituted blastocysts possessed less developmental potential as the intra-strain reconstituted blastocysts. Thisstudy may give light to solve the problem of low implantation rate and placenta abnormality in mammal cloning.
文摘Identification and quantification of low abundance growth factors and regulators in complex biological samples still present a challenging task in analytical biochemistry. Immunoassays are often used for such purpose but immunoassays face limitation of both availability and qualities of antibody reagents that are necessary for development of immune assays. With genomics data base available, mass spectrometry (MS) can analyze protein tryptic peptides directly for quantitative determination of proteins. In this study, we report a method for detection of matrix metalloproteinase 1 (MMP1), an important extracellular matrix modulator, in human breast cancer cells by quadrupole time-of-flight (Q-TOF) MS. Absolute quantification of MMP1 was conducted using the selected reaction monitoring (SRM) on a triple quadrupole (Triple-Quad) MS via transitions selected from MMP1 tryptic peptides using non isotope labeled MMP1 protein as a titration standard. In comparison with immune based assay, this MS method showed picogram level sensitivity for quantitative determination of MMP1 intotal cell lysates. Our results demonstrated the feasibility of absolute quantification of low abundance proteins using label-free protein standard by mass spectrometry. Therefore, this method provides not only advantages of high sensitivity but also cost saving in comparison with the commonly used mass spectrometry that currently employs isotype labeled proteins for quantitative analysis.
基金Supported by the Natural Science Foundation, Y100-573006Doctoral Foundation of Xi'an Jiaotong University, DFXJTU2002-11
文摘AIM: To conduct the proteomic analysis of human colorectal carcinoma cell line, SW480 by using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption /ionization-time of flight mass spectrometry (MALDITOFMS).METHODS: The total proteins of human colorectal carcinoma cell line, SW480 were separated with 2-DE by using immobilized pH gradient strips and visualized by staining with silver nitrate. The gel images were acquired by scanner and 2-DE analysis software, Image Master 2D Elite. Nineteen distinct protein spots were excised from gel randomly and digested in gel by TPCK-trypsin. Mass analysis ofthe tryptic digest peptides mixture was performed by using MALDI-TOF MS. Peptide mass fingerprints (PMFs) obtained by the MALDI-TOF analysis were used to search NCBI,SWISS-PROT and MSDB databases by using Mascot software.RESULTS: PMF maps of all spots were obtained by MALDI-TOF MS and thirteen proteins were preliminarily identified.CONCLUSION: The methods of analysis and identification of protein spots of tumor cells in 2-DE gel with silver staining by MALDI-TOF MS derived PMF have been established.Protein expression profile of SW480 has been obtained.It is demonstrated that a combination of proteomics and cell culture is a useful approach to comprehend the process of colon carcinogenesis.
基金supported by the Science and Technology Planning Project of Guangdong Province, China (Grant No. 2010B031600237)
文摘Objective: Patients with head and neck cancer often suffer from malnutrition. This study aims to investigate the influence of body mass index(BMI) on the prognosis of laryngeal squamous cell carcinoma(LSCC).Methods: A total of 473 patients with LSCC initially treated at Sun Yat-sen University Cancer Center between January 2005 and July 2009 were retrospectively reviewed. Survival analysis was performed by the Kaplan-Meier method and Cox regression model.Results: Low BMI before treatment was significantly associated with poor overall survival in patients with LSCC(P<0.001). BMI was an independent prognostic factor for patients with LSCC.Conclusion: Leanness before treatment was associated with poor prognosis in patients with LSCC. Good nutritional status is favorable to improve survival in patients with LSCC.
基金supported by National Institutes of Health Grants, R01 AR054465, R01 AR070222, and R01 AR070222supported by the grants of Natural Science Foundation of China (NSFC) to TW (grants No. 81301531 and 81572104)+1 种基金supported by the grant from Shenzhen Science and Technology Innovation Committee, China (grant No. JCYJ20160331114205502)the grant from Shenzhen Development and Reform Committee, China for Shenzhen Engineering Laboratory of Orthopedic Regenerative Technologies
文摘Carboxyl terminus of Hsp70-interacting protein（CHIP or STUB1） is an E3 ligase and regulates the stability of several proteins which are involved in different cellular functions. Our previous studies demonstrated that Chip deficient mice display bone loss phenotype due to increased osteoclast formation through enhancing TRAF6 activity in osteoclasts. In this study we provide novel evidence about the function of CHIP. We found that osteoblast differentiation and bone formation were also decreased in Chip KO mice. In bone marrow stromal（BMS） cells derived from Chip^-/- mice, expression of a panel of osteoblast marker genes was significantly decreased. ALP activity and mineralized bone matrix formation were also reduced in Chip-deficient BMS cells. We also found that in addition to the regulation of TRAF6, CHIP also inhibits TNFα-induced NF-κB signaling through promoting TRAF2 and TRAF5 degradation. Specific deletion of Chip in BMS cells downregulated expression of osteoblast marker genes which could be reversed by the addition of NF-κB inhibitor. These results demonstrate that the osteopenic phenotype observed in Chip^-/- mice was due to the combination of increased osteoclast formation and decreased osteoblast differentiation. Taken together, our findings indicate a significant role of CHIP in bone remodeling.
基金Supported by the National Natural Science Foundation of China （20573045, 20843001）, the ＂Tenth Five-Year＂ National Key Scientific and Technological Project （2004BA313B-17） and the National High Technology Research and Development Pro gram of China （2006AA06Z321）.
基金supported by the National Natural Science Foundation(C36970359).
文摘Bovine embryonic stem cell would be invaluable for researching the aspect of animal cloning, production transgenic animal and discussion of gene function in vitro. With the object of establishing an effective culture system for isolation and clone of bovine pluripotent stem cell, we cultured bovine embryos and mouse embryos including morula blastula and hatached blastula and obtained animal ICM on Primary murine embryonic fibroblast (Primary murine embryonic fibroblast, PMEF) feeder layer with tissue medium(DMEM supplemented with 15ml/100ml NBS,0.1μmol/L Na2SeO3, 0.1mmol/L p-mercaptoethanol, 1000ng/ml LIF, 10 ng/ml IGF, 1mmol/L necessary amino acid and 1mmol/L L-glutamine),then,we obtained mouse ICM and bovine ICM. Moreover, we isolated and cloned the 6 passage bovine ES like cells(12 cell lines) and 9 passage murine ES like cells (52 cell lines) deriving from bovine ICM and murine ICM respectively on the feeder layer of PMEF by disaggregating ICM and ES cell clones of bovine and murine into smaller clumps through digesting with 0.125g/100ml trypsin and 0.02g/100ml EDTA and scattering with a glass needle. The pluripotency of both murine and bovine ES like cells was identified with morphological character, histochemistry identification , karyotype analysis and differentiation of ES cells in vitro or in vivo. This result showed that bovine embryonic stem cell and murine embryonic stem cell had developmental pluripotency.
