Cytosine-substituted mildiomycin analogue （MIL-C） was produced effectively by supplementing cytosine into the culture of Streptoverticillium rimofaciens. In order to improve the yield of MIL-C, statistically-based e...Cytosine-substituted mildiomycin analogue （MIL-C） was produced effectively by supplementing cytosine into the culture of Streptoverticillium rimofaciens. In order to improve the yield of MIL-C, statistically-based experimental designs were applied to optimize the fermentation medium for S. rimofaciens ZJU 5119. Fifteen culture conditions were examined for their significances on MIL-C production using Plackett-Burman design. The Plackett-Burrnan design and one-variable-at-a-time design indicated that glucose and rice meal as the complex carbon sources, and peanut cake meal and NH4NO3 as the complex nitrogen sources were beneficial for MIL-C production in S. rimofaciens ZJU 5119. The results of further central composition design （CCD） showed that the optimal concentration of glucose, rice meal and peanut cake meal were 18.7 g/L, 64.8 g/L and 65.1 g/L, respectively. By using this optimal fermentation medium, the MIL-C concentration was increased up to 1336.5 mg/L, an approximate 3.8-fold improvement over the previous concentration （350.0 mg/L） with un-optimized medium. This work will be very helpful to the large-scale production of MIL-C in the future.展开更多
Fragment containing the whole riboflavin(rib)operons of B.cereus ATCC14579 was detected from GenBank and annotated.The rib operon of ATCC14579 was cloned with Pn,its native promoter,or with P43,the vegetative growth p...Fragment containing the whole riboflavin(rib)operons of B.cereus ATCC14579 was detected from GenBank and annotated.The rib operon of ATCC14579 was cloned with Pn,its native promoter,or with P43,the vegetative growth promoter,into the plasmid.Expression analysis showed that heterologous rib operon was operative in B.subtilis.Integrative plasmid with P43-rib fragment was integrated into the chromosome of B.subtilis RH33,yielding transformant B.subtilis PY.With optimized medium components,4.3 g·L -1 of riboflavin was achieved in batch culture of B.subtilis PY,which was 27%enhancement compared to the host strain.Real-time reverse transcription polymerase chain reaction(RT-PCR)analysis indicated that the transcriptional level of ribA maintained 2.8-fold higher with the expression of herterologous rib operon.Furthermore,the stability of B.subtilis PY was increased form 45%to 87%.The high transcriptional level of rib gene and higher stability of B.subtilis PY could explain the increased riboflavin production.展开更多
AIM： To enhance the differentiation of insulin producing cell （IPC） ability from embryonic stem （ES） cells in vitro. METHODS： Four-day embryoid body （EB）-formatted ES cells were dissociated as single cells for...AIM： To enhance the differentiation of insulin producing cell （IPC） ability from embryonic stem （ES） cells in vitro. METHODS： Four-day embryoid body （EB）-formatted ES cells were dissociated as single cells for the followed plasmid DNA delivery. The use of NucleofectorTM electroporator （Amaxa biosystems, Germany） in combination with medium-contained G418 provided a high efficiency of gene delivery for advanced selection. Neucleofected cells were plated on the top of fibronectincoated Petri dishes. Addition of Ly294002 and raised the glucose in medium at 24 h before examination.The differentiation status of these cells was monitored by semi-quantitative PCR （SQ-PCR） detection of the expression of relative genes, such as oct-4, sox-17, foxa2, mixll, pdx-1, insulin 1, glucagons and somatostatin. The percentage of IPC population on d 18 of the experiment was investigated by immunohistochemistry （IHC）, and the content/secretion of insulin was estimated by ELISA assay. The mice with severe combined immunodeficiency disease （SCID） pretreated with streptozotocin （STZ） were used to eliminate plasma glucose restoration after pax4^＋ ES implantation. RESULTS： A high efficiency of gene delivery was demonstrated when neucleofection was used in the present study; approximately 70% cells showed DsRed expression 2 d after neucleofection. By selection of medium-contained G418, the percentage of DsRed expressing cells kept high till the end of study. The pancreatic differentiation seemed to be accelerated by pax4 nucleofection. When compared to the group of cells with mock control, foxa2, mixll, pdxl, higher insulin and somatostatin levels were detected by SQ-PCR 4 d after nucleofection in the group of pax4 expressing plasmid delivery. Approximately 55% of neucleofected cells showed insulin expression 18 d after neucleofection, and only 18% of cells showed insulin expression in mock control. The disturbance was shown by nucleofected pax4 RNAi vector; only 8% of cells expressed insulin 18 d after nucleofection. A higher IPC population was also detected in the insulin content by ELISA assay, and the glucose dependency was demonstrated in insulin secretion level. In the animal model, improvement of average plasma glucose concentration was observed in the group of pax-4 expressed ES of SCID mice pretreated with STZ, but no significant difference was observed in the group of STZ-pretreated SCID mice who were transplanted ES with mock plasmid. CONCLUSION： Enhancement of IPC differentiation from EB-dissociated ES cells can be revealed by simply using pax4 expressing plasrnid delivery. Not only more IPCs but also pancreatic differentiation-related genes can be detected by SQ-PCR. Expression of relative genes, such as foxa 2, mixl 1, pdx-1, insulin 1 and somatostatin after nucleofection, suggests that pax4 accelerates the whole differentiation progress. The higher insulin production with glucose dependent modulation suggests that pax4 expression can drive more mature IPCs. Although further determination of the entire mechanism is required, the potential of pax-4-nucleofected cells in medical treatment is promising.展开更多
This work is devoted to obtaining radionuclide-labeled microspheres for radionuclide therapy as a therapeutic drug with a selective method of delivery. The nature of magnetic microspheres and quantitative elemental co...This work is devoted to obtaining radionuclide-labeled microspheres for radionuclide therapy as a therapeutic drug with a selective method of delivery. The nature of magnetic microspheres and quantitative elemental composition radiopreparats was considered. We have developed a new method of producing radionuclide-labeled microspheres （radiopharmaceutical ^140Nd-microspheres） for radionuclide therapy.展开更多
The feasibility of producing superheavy nuclei in proton evaporation channels was systematically studied within the dinuclear system(DNS)model.Due to the Z=114 proton-shell,one can synthesize Fl isotopes in proton eva...The feasibility of producing superheavy nuclei in proton evaporation channels was systematically studied within the dinuclear system(DNS)model.Due to the Z=114 proton-shell,one can synthesize Fl isotopes in proton evaporation channels.We only considered the case of evaporating one proton first and then n neutrons in this work,other cases were ignored due to the small cross-section.The production cross sections of unknown isotopes ^(290,291)Fl in ^(38)S+^(255)Es reaction are the highest compared with ^(50)Ti+^(243)Np and ^(54)Cr+^(239)Pa reactions,and the maximum cross sections are 1.1 and 15.1 pb,respectively.^(42)S+^(254)Es is a promising candidate to approach the island of stability as the radioactive beam facilities are upgraded in the future,and the production cross sections of ^(291−294)Fl in that reaction are estimated to be 3.2,6.0,4.0,and 0.1 pb,respectively.展开更多
Stars of～8-100 M_⊙end their lives as core-collapse supernovae(SNe). In the process they emit a powerful burst of neutrinos,produce a variety of elements, and leave behind either a neutron star or a black hole. The w...Stars of～8-100 M_⊙end their lives as core-collapse supernovae(SNe). In the process they emit a powerful burst of neutrinos,produce a variety of elements, and leave behind either a neutron star or a black hole. The wide mass range for SN progenitors results in diverse neutrino signals, explosion energies, and nucleosynthesis products. A major mechanism to produce nuclei heavier than iron is rapid neutron capture, or the r process. This process may be connected to SNe in several ways. A brief review is presented on current understanding of neutrino emission, explosion, and nucleosynthesis of SNe.展开更多
基金Project supported by the Ministry of Science and Technology, China (No. 2004BA308A22-14)the Department of Science and Tech- nology of Zhejiang Province, China (No. 011102543)
文摘Cytosine-substituted mildiomycin analogue （MIL-C） was produced effectively by supplementing cytosine into the culture of Streptoverticillium rimofaciens. In order to improve the yield of MIL-C, statistically-based experimental designs were applied to optimize the fermentation medium for S. rimofaciens ZJU 5119. Fifteen culture conditions were examined for their significances on MIL-C production using Plackett-Burman design. The Plackett-Burrnan design and one-variable-at-a-time design indicated that glucose and rice meal as the complex carbon sources, and peanut cake meal and NH4NO3 as the complex nitrogen sources were beneficial for MIL-C production in S. rimofaciens ZJU 5119. The results of further central composition design （CCD） showed that the optimal concentration of glucose, rice meal and peanut cake meal were 18.7 g/L, 64.8 g/L and 65.1 g/L, respectively. By using this optimal fermentation medium, the MIL-C concentration was increased up to 1336.5 mg/L, an approximate 3.8-fold improvement over the previous concentration （350.0 mg/L） with un-optimized medium. This work will be very helpful to the large-scale production of MIL-C in the future.
基金Supported by the National Natural Science Foundation of China（20536040） the State Key Development Program for Basic Research of China（2007CB707802） the Development Project of Science and Technology of Tianjin（05YFGZGX04500）
文摘Fragment containing the whole riboflavin(rib)operons of B.cereus ATCC14579 was detected from GenBank and annotated.The rib operon of ATCC14579 was cloned with Pn,its native promoter,or with P43,the vegetative growth promoter,into the plasmid.Expression analysis showed that heterologous rib operon was operative in B.subtilis.Integrative plasmid with P43-rib fragment was integrated into the chromosome of B.subtilis RH33,yielding transformant B.subtilis PY.With optimized medium components,4.3 g·L -1 of riboflavin was achieved in batch culture of B.subtilis PY,which was 27%enhancement compared to the host strain.Real-time reverse transcription polymerase chain reaction(RT-PCR)analysis indicated that the transcriptional level of ribA maintained 2.8-fold higher with the expression of herterologous rib operon.Furthermore,the stability of B.subtilis PY was increased form 45%to 87%.The high transcriptional level of rib gene and higher stability of B.subtilis PY could explain the increased riboflavin production.
基金grants of Stem Cell Project of TVGHthe Joint Projects of UTVGH, No. 94-P1-04/06/10+1 种基金Yen Tjing-Ling Medical FoundationNational Yang-Ming University, Taiwan, China
文摘AIM： To enhance the differentiation of insulin producing cell （IPC） ability from embryonic stem （ES） cells in vitro. METHODS： Four-day embryoid body （EB）-formatted ES cells were dissociated as single cells for the followed plasmid DNA delivery. The use of NucleofectorTM electroporator （Amaxa biosystems, Germany） in combination with medium-contained G418 provided a high efficiency of gene delivery for advanced selection. Neucleofected cells were plated on the top of fibronectincoated Petri dishes. Addition of Ly294002 and raised the glucose in medium at 24 h before examination.The differentiation status of these cells was monitored by semi-quantitative PCR （SQ-PCR） detection of the expression of relative genes, such as oct-4, sox-17, foxa2, mixll, pdx-1, insulin 1, glucagons and somatostatin. The percentage of IPC population on d 18 of the experiment was investigated by immunohistochemistry （IHC）, and the content/secretion of insulin was estimated by ELISA assay. The mice with severe combined immunodeficiency disease （SCID） pretreated with streptozotocin （STZ） were used to eliminate plasma glucose restoration after pax4^＋ ES implantation. RESULTS： A high efficiency of gene delivery was demonstrated when neucleofection was used in the present study; approximately 70% cells showed DsRed expression 2 d after neucleofection. By selection of medium-contained G418, the percentage of DsRed expressing cells kept high till the end of study. The pancreatic differentiation seemed to be accelerated by pax4 nucleofection. When compared to the group of cells with mock control, foxa2, mixll, pdxl, higher insulin and somatostatin levels were detected by SQ-PCR 4 d after nucleofection in the group of pax4 expressing plasmid delivery. Approximately 55% of neucleofected cells showed insulin expression 18 d after neucleofection, and only 18% of cells showed insulin expression in mock control. The disturbance was shown by nucleofected pax4 RNAi vector; only 8% of cells expressed insulin 18 d after nucleofection. A higher IPC population was also detected in the insulin content by ELISA assay, and the glucose dependency was demonstrated in insulin secretion level. In the animal model, improvement of average plasma glucose concentration was observed in the group of pax-4 expressed ES of SCID mice pretreated with STZ, but no significant difference was observed in the group of STZ-pretreated SCID mice who were transplanted ES with mock plasmid. CONCLUSION： Enhancement of IPC differentiation from EB-dissociated ES cells can be revealed by simply using pax4 expressing plasrnid delivery. Not only more IPCs but also pancreatic differentiation-related genes can be detected by SQ-PCR. Expression of relative genes, such as foxa 2, mixl 1, pdx-1, insulin 1 and somatostatin after nucleofection, suggests that pax4 accelerates the whole differentiation progress. The higher insulin production with glucose dependent modulation suggests that pax4 expression can drive more mature IPCs. Although further determination of the entire mechanism is required, the potential of pax-4-nucleofected cells in medical treatment is promising.
文摘This work is devoted to obtaining radionuclide-labeled microspheres for radionuclide therapy as a therapeutic drug with a selective method of delivery. The nature of magnetic microspheres and quantitative elemental composition radiopreparats was considered. We have developed a new method of producing radionuclide-labeled microspheres （radiopharmaceutical ^140Nd-microspheres） for radionuclide therapy.
文摘The feasibility of producing superheavy nuclei in proton evaporation channels was systematically studied within the dinuclear system(DNS)model.Due to the Z=114 proton-shell,one can synthesize Fl isotopes in proton evaporation channels.We only considered the case of evaporating one proton first and then n neutrons in this work,other cases were ignored due to the small cross-section.The production cross sections of unknown isotopes ^(290,291)Fl in ^(38)S+^(255)Es reaction are the highest compared with ^(50)Ti+^(243)Np and ^(54)Cr+^(239)Pa reactions,and the maximum cross sections are 1.1 and 15.1 pb,respectively.^(42)S+^(254)Es is a promising candidate to approach the island of stability as the radioactive beam facilities are upgraded in the future,and the production cross sections of ^(291−294)Fl in that reaction are estimated to be 3.2,6.0,4.0,and 0.1 pb,respectively.
基金supported by the US Department of Energy(Grant No.DEFG02-87ER40328)
文摘Stars of～8-100 M_⊙end their lives as core-collapse supernovae(SNe). In the process they emit a powerful burst of neutrinos,produce a variety of elements, and leave behind either a neutron star or a black hole. The wide mass range for SN progenitors results in diverse neutrino signals, explosion energies, and nucleosynthesis products. A major mechanism to produce nuclei heavier than iron is rapid neutron capture, or the r process. This process may be connected to SNe in several ways. A brief review is presented on current understanding of neutrino emission, explosion, and nucleosynthesis of SNe.
