A vacuolar ATPase (V-ATPase.) B subunit gene has been cloned and characterized front a phosphorus starvation induced rice root subtractive cDNA library by suppression subtractive hybridization (SSH) method and RT-PCR ...A vacuolar ATPase (V-ATPase.) B subunit gene has been cloned and characterized front a phosphorus starvation induced rice root subtractive cDNA library by suppression subtractive hybridization (SSH) method and RT-PCR amplification. This gene encodes a polypeptide of 487 amino acid residues, containing a conservative ATP binding site and with a molecular weight of 54.06 kD and an isoelectric point of 4.99, southern analysis of the. genomic DNA indicates that V-ATPase B subunit is encoded by a single gene in rice genome. The amino acid homologies of V-ATPase B subunits among different organisms range from 76% to 97% and reveals that the evolution of V-ATPase B subunit is accompanied with the biological evolution. Expression pattern analysis indicated that the maximal expression of V-ATPase B subunit gene occurred at an early stage (6 - 12 h) after phosphorus starvation in roots, and lately stage (24 - 48 It) in leaves. Under phosphorus deficiency, the up-regulated expression of V-ATPase gene was presumed to strengthen the proton transport and provide the required energy to maintain an electrochemical gradient across the tonoplast to facilitate Phosphorus transport.展开更多
[Objective] The paper aimed to construct a new T vector based on plasmid pUC19 digested with Xcm Ⅰ. [Method] Two complementary oligonucleotide chains containing two Xcm Ⅰ sites were synthesized. After denaturation a...[Objective] The paper aimed to construct a new T vector based on plasmid pUC19 digested with Xcm Ⅰ. [Method] Two complementary oligonucleotide chains containing two Xcm Ⅰ sites were synthesized. After denaturation and renaturation,the adaptor was cloned into plasmid pUC19 between the Hind Ⅲ and BamH Ⅰ sites. The new plasmid,pUC19-HB-T vector,was digested with Xcm Ⅰ to derive a T-vector with 3′ end overhanging a T base. [Result] The constructed pUC19-HB-T vector was efficient in cloning PCR products,with an efficiency of 95% at least. [Conclusion] A new Xcm Ⅰ-based pUC19-HB-T vector was constructed,which could be applied to cloning of PCR products and other microbiology operations.展开更多
The main pathophysiological feature of sepsis is the uncontrollable activation of both pro-and anti-inflammatory responses arising from the overwhelming pro-duction of mediators such as pro-and anti-inflammatory cytok...The main pathophysiological feature of sepsis is the uncontrollable activation of both pro-and anti-inflammatory responses arising from the overwhelming pro-duction of mediators such as pro-and anti-inflammatory cytokines. Such an uncontrollable inflammatory response would cause many kinds of metabolic derangements. One such metabolic derangement is hyperglycemia. Accordingly, control of hyperglycemia in sepsis is considered to be a very effective therapeutic approach. However, despite the initial enthusiasm, recent studies reported that tight glycemic control with intensive insulin therapy failed to show a beneficial effect on mortality of patients with severe sepsis and septic shock. One of the main reasons for this disappointing result is the incidence of harmful hypoglycemia during intensive insulin therapy. Therefore, avoidance of hypoglycemia during intensive insulin therapy may be a key issue in effective tight glycemic control. It is generally accepted that glycemic control aimed at a blood glucose level of 80-100 mg/dL, as initially proposed by van den Berghe, seems to be too tight and that such a level of tight glycemic control puts septic patients at increased risk of hypoglycemia. Therefore, now many researchers suggest less strict glycemic control with a target blood glucose level of 140-180 mg/dL. Also specific targeting of glycemic control in diabetic patients should be considered. Since there is a significantcorrelation between success rate of glycemic control and the degree of hypercytokinemia in septic patients, some countermeasures to hypercytokinemia may be an important aspect of successful glycemic control. Thus, in future, use of an artificial pancreas to avoid hypoglycemia during insulin therapy, special consideration of septic diabetic patients, and control of hypercytokinemia should be considered for more effective glycemic control in patients with severe sepsis and septic shock.展开更多
TO evaluate the effects of abdominal trauma on hemorrhagic shock-induced acute lung injury in rats. METHODS： Five groups were allocated （n = 8） in the study. Group Ⅰ was taken as the control group, group Ⅱ as the...TO evaluate the effects of abdominal trauma on hemorrhagic shock-induced acute lung injury in rats. METHODS： Five groups were allocated （n = 8） in the study. Group Ⅰ was taken as the control group, group Ⅱ as the hemorrhagic shock group, group Ⅲ as hemorrhagic shock ＋ laparotomy, group Ⅳ as hemorrhagic shock ＋ splenectomy and group V as splenectomy ＋ omentectomy ＋ hemorrhagic shock group. Hemorrhagic shock was induced by drawing blood and reducing mean arterial pressure （MAP） to 40 mmHg within 10 min. After a hypotensive period of 1 h, animals were resuscitated. Bronchoalveolar lavage （BAL） was performed to recover cells from the alveolar space with 40 mL of BAL fluid after resuscitation malondialdehyde （MDA） and L-γ-glutamyl-L-cysteinylglycine （GSH） levels were measured in serum, erythrocytes and lung tissue. RESULTS： Serum, erythrocyte, lung tissue MDA and GSH levels were significantly increased in hemorrhagic shock groups Ⅱ-Ⅴ （P 〈 0.05）. Lymphocyte, neutrophil and alveolar macrophage counts in BAL fluid indicated a significant difference between control and shock groups （P 〈 0.05）. CONCLUSION： The degree of trauma increases hemorrhagic shock-induced acute lung injury.展开更多
Neurodegenerative diseases are characterized by the accumulation of intracellular or extracellular protein aggregates that result from conformational changes in proteins. These diseases may result from an imbalance be...Neurodegenerative diseases are characterized by the accumulation of intracellular or extracellular protein aggregates that result from conformational changes in proteins. These diseases may result from an imbalance between the produetion of misfolded proteins and normal chaperone capacity. Molecular chaperones provide a first line of defenee against misfolded, aggragation-prone proteins and are, therefore, promising therapeutic targets for neurodegenerative diseases.展开更多
As the title of this paper suggests, I intend to draw attention to what, in my opinion, lies at the core of the shared notion in both Kant and Habermas. This will be the concern of the first part of my paper. My focus...As the title of this paper suggests, I intend to draw attention to what, in my opinion, lies at the core of the shared notion in both Kant and Habermas. This will be the concern of the first part of my paper. My focus will then shift, in the second part, to Habermas＇s views on freedom. In due time, however, as the provocative question mark in the title suggests, the notion of freedom becomes questionable. I will conclude by examining Frankfurt＇s notion of coercion in order to show that Habermas＇s notion of freedom is not only questionable but can, at times, be coercive. Throughout this paper, the reader shall be encouraged to see and possibly appreciate that there is a degree of similarity between the two thinkers. Shall the reader be hard put to gauge this similarity or shall the reader promptly appreciate it and take it into consideration？ I will argue--and demonstrate--that a certain amount of similarity can readily be drawn between the two. I will leave it up to the reader to decide whether this similarity is a forced similarity or whether this is a similarity that one perceives at a first glance and thus ought to consider. This paper could better be appreciated if the reader has reasonable knowledge of the Hegelian critique of Kant＇s Categorical Imperative.展开更多
Environmental conditions, including ambient temperature, play important roles in survival growth development, and reproduction of the Japanese sea cucumber, Apostichopus japonicus. Low temperatures result in slowed gr...Environmental conditions, including ambient temperature, play important roles in survival growth development, and reproduction of the Japanese sea cucumber, Apostichopus japonicus. Low temperatures result in slowed growth and skin ulceration disease. In a previous study, we investigated the effect of low temperature on gene expression profiles inA.japonicus by suppression subtractive hybridization （SSH）. Genes encoding Ferritin, Lysozyme, Hsp70, gp96, and AjToll were selected from a subtracted cDNA library of A. japonicus under acute cold stress. The transcriptional expression profiles of these genes were investigated in different tissues （coelomocyte, respiratory tree, intestine, longitudinal muscle） after exposure to acute and mild temperature dropping treatments. The results show that （1） the five cold-tolerance-related genes were found in all four tissues and the highest mRNA levels were observed in coelomocyte and respiratory tree; （2） under the temperature dropping treatments, three types of transcriptional regulation patterns were observed： primary suppression followed by up-regulation at -2℃, suppressed expression throughout the two treatments, and more rarely an initial stimulation followed by suppression; and （3） gene expression suppression was more severe under acute temperature dropping than under mild temperature dropping treatment. The five cold-tolerance-related genes that were ,distributed mainly in coelomocyte and respiratory tissues were generally down-regulated by low temperature stress but an inverse up-regulation event was found at the extreme temperature （-2℃）.展开更多
The global burden caused by dengue has increased dramatically in recent decades. Dengue Fever and its severe clinical manifestation, Dengue Hemorrhagic Fever and Dengue Shock Syndrome, have become international public...The global burden caused by dengue has increased dramatically in recent decades. Dengue Fever and its severe clinical manifestation, Dengue Hemorrhagic Fever and Dengue Shock Syndrome, have become international public health problem endemic in more than 100 countries, risking 2.5 to 3 billion world populations in tropical and subtropical region. Envelope （E） protein of dengue virus has been proposed as the most important antigen that enables it as vaccine candidate or diagnostic materials. Recombinant protein E production is desirable for dengue vaccine and diagnostic development, especially in Indonesia, where dengue is epidemic. Cloning E gene in an expression vector is essential as an initial method to produce dengue E antigens. The purpose of this research was to clone E gene of dengue virus type 3 （Indonesia D3-1703 strain） into Saccharomyces cerevisiae expression vector pYES2/CT. The cloning method used was the in vitro ligation protocol. First, the cDNA from dengue virus type 3 strain （D3-1703） was generated. Then the polymerase chain reaction （PCR） amplification product of E gene cassette from this cDNA was obtained. The E gene cassette was ligated into linearized pYES2/CT resulting a recombinant vector named pYES2/CT-E. The cloned E gene was verified by restriction enzyme digestion and PCR. Sequencing analysis at its 5＇ end showed that the E gene was inserted at the right open reading frame. In conclusion, the results showed that for the first time the E gene originated from an Indonesian dengue virus type 3 strain was successfully cloned within the yeast expression vector pYES2/CT. In the future, this clone could be expressed and provided as materials for dengue vaccine and diagnostic kit, specific for Indonesian dengue virus strain.展开更多
Neutral perylene bisimides(PBI) are well-known n-type organic semiconductors, with number of challenging electronic properties in their neutral and reduced states. We report the characteristic electronic properties of...Neutral perylene bisimides(PBI) are well-known n-type organic semiconductors, with number of challenging electronic properties in their neutral and reduced states. We report the characteristic electronic properties of PBI anionic films. We unexpectedly discovered that pristine PBI dianion film showed p-type character, while oxidized dianion film(dominant neutral state with few radical anions) showed normal n-type semiconductor character based on Seebeck effect measurements. Both kinds of films exhibit high electrical conductivity with a potential for thermoelectric applications. The mechanism of polarity reversal is proposed.展开更多
文摘A vacuolar ATPase (V-ATPase.) B subunit gene has been cloned and characterized front a phosphorus starvation induced rice root subtractive cDNA library by suppression subtractive hybridization (SSH) method and RT-PCR amplification. This gene encodes a polypeptide of 487 amino acid residues, containing a conservative ATP binding site and with a molecular weight of 54.06 kD and an isoelectric point of 4.99, southern analysis of the. genomic DNA indicates that V-ATPase B subunit is encoded by a single gene in rice genome. The amino acid homologies of V-ATPase B subunits among different organisms range from 76% to 97% and reveals that the evolution of V-ATPase B subunit is accompanied with the biological evolution. Expression pattern analysis indicated that the maximal expression of V-ATPase B subunit gene occurred at an early stage (6 - 12 h) after phosphorus starvation in roots, and lately stage (24 - 48 It) in leaves. Under phosphorus deficiency, the up-regulated expression of V-ATPase gene was presumed to strengthen the proton transport and provide the required energy to maintain an electrochemical gradient across the tonoplast to facilitate Phosphorus transport.
文摘[Objective] The paper aimed to construct a new T vector based on plasmid pUC19 digested with Xcm Ⅰ. [Method] Two complementary oligonucleotide chains containing two Xcm Ⅰ sites were synthesized. After denaturation and renaturation,the adaptor was cloned into plasmid pUC19 between the Hind Ⅲ and BamH Ⅰ sites. The new plasmid,pUC19-HB-T vector,was digested with Xcm Ⅰ to derive a T-vector with 3′ end overhanging a T base. [Result] The constructed pUC19-HB-T vector was efficient in cloning PCR products,with an efficiency of 95% at least. [Conclusion] A new Xcm Ⅰ-based pUC19-HB-T vector was constructed,which could be applied to cloning of PCR products and other microbiology operations.
文摘The main pathophysiological feature of sepsis is the uncontrollable activation of both pro-and anti-inflammatory responses arising from the overwhelming pro-duction of mediators such as pro-and anti-inflammatory cytokines. Such an uncontrollable inflammatory response would cause many kinds of metabolic derangements. One such metabolic derangement is hyperglycemia. Accordingly, control of hyperglycemia in sepsis is considered to be a very effective therapeutic approach. However, despite the initial enthusiasm, recent studies reported that tight glycemic control with intensive insulin therapy failed to show a beneficial effect on mortality of patients with severe sepsis and septic shock. One of the main reasons for this disappointing result is the incidence of harmful hypoglycemia during intensive insulin therapy. Therefore, avoidance of hypoglycemia during intensive insulin therapy may be a key issue in effective tight glycemic control. It is generally accepted that glycemic control aimed at a blood glucose level of 80-100 mg/dL, as initially proposed by van den Berghe, seems to be too tight and that such a level of tight glycemic control puts septic patients at increased risk of hypoglycemia. Therefore, now many researchers suggest less strict glycemic control with a target blood glucose level of 140-180 mg/dL. Also specific targeting of glycemic control in diabetic patients should be considered. Since there is a significantcorrelation between success rate of glycemic control and the degree of hypercytokinemia in septic patients, some countermeasures to hypercytokinemia may be an important aspect of successful glycemic control. Thus, in future, use of an artificial pancreas to avoid hypoglycemia during insulin therapy, special consideration of septic diabetic patients, and control of hypercytokinemia should be considered for more effective glycemic control in patients with severe sepsis and septic shock.
文摘TO evaluate the effects of abdominal trauma on hemorrhagic shock-induced acute lung injury in rats. METHODS： Five groups were allocated （n = 8） in the study. Group Ⅰ was taken as the control group, group Ⅱ as the hemorrhagic shock group, group Ⅲ as hemorrhagic shock ＋ laparotomy, group Ⅳ as hemorrhagic shock ＋ splenectomy and group V as splenectomy ＋ omentectomy ＋ hemorrhagic shock group. Hemorrhagic shock was induced by drawing blood and reducing mean arterial pressure （MAP） to 40 mmHg within 10 min. After a hypotensive period of 1 h, animals were resuscitated. Bronchoalveolar lavage （BAL） was performed to recover cells from the alveolar space with 40 mL of BAL fluid after resuscitation malondialdehyde （MDA） and L-γ-glutamyl-L-cysteinylglycine （GSH） levels were measured in serum, erythrocytes and lung tissue. RESULTS： Serum, erythrocyte, lung tissue MDA and GSH levels were significantly increased in hemorrhagic shock groups Ⅱ-Ⅴ （P 〈 0.05）. Lymphocyte, neutrophil and alveolar macrophage counts in BAL fluid indicated a significant difference between control and shock groups （P 〈 0.05）. CONCLUSION： The degree of trauma increases hemorrhagic shock-induced acute lung injury.
文摘Neurodegenerative diseases are characterized by the accumulation of intracellular or extracellular protein aggregates that result from conformational changes in proteins. These diseases may result from an imbalance between the produetion of misfolded proteins and normal chaperone capacity. Molecular chaperones provide a first line of defenee against misfolded, aggragation-prone proteins and are, therefore, promising therapeutic targets for neurodegenerative diseases.
文摘As the title of this paper suggests, I intend to draw attention to what, in my opinion, lies at the core of the shared notion in both Kant and Habermas. This will be the concern of the first part of my paper. My focus will then shift, in the second part, to Habermas＇s views on freedom. In due time, however, as the provocative question mark in the title suggests, the notion of freedom becomes questionable. I will conclude by examining Frankfurt＇s notion of coercion in order to show that Habermas＇s notion of freedom is not only questionable but can, at times, be coercive. Throughout this paper, the reader shall be encouraged to see and possibly appreciate that there is a degree of similarity between the two thinkers. Shall the reader be hard put to gauge this similarity or shall the reader promptly appreciate it and take it into consideration？ I will argue--and demonstrate--that a certain amount of similarity can readily be drawn between the two. I will leave it up to the reader to decide whether this similarity is a forced similarity or whether this is a similarity that one perceives at a first glance and thus ought to consider. This paper could better be appreciated if the reader has reasonable knowledge of the Hegelian critique of Kant＇s Categorical Imperative.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2012AA10A412)the National Natural Science Foundation of China(No.31072230)
文摘Environmental conditions, including ambient temperature, play important roles in survival growth development, and reproduction of the Japanese sea cucumber, Apostichopus japonicus. Low temperatures result in slowed growth and skin ulceration disease. In a previous study, we investigated the effect of low temperature on gene expression profiles inA.japonicus by suppression subtractive hybridization （SSH）. Genes encoding Ferritin, Lysozyme, Hsp70, gp96, and AjToll were selected from a subtracted cDNA library of A. japonicus under acute cold stress. The transcriptional expression profiles of these genes were investigated in different tissues （coelomocyte, respiratory tree, intestine, longitudinal muscle） after exposure to acute and mild temperature dropping treatments. The results show that （1） the five cold-tolerance-related genes were found in all four tissues and the highest mRNA levels were observed in coelomocyte and respiratory tree; （2） under the temperature dropping treatments, three types of transcriptional regulation patterns were observed： primary suppression followed by up-regulation at -2℃, suppressed expression throughout the two treatments, and more rarely an initial stimulation followed by suppression; and （3） gene expression suppression was more severe under acute temperature dropping than under mild temperature dropping treatment. The five cold-tolerance-related genes that were ,distributed mainly in coelomocyte and respiratory tissues were generally down-regulated by low temperature stress but an inverse up-regulation event was found at the extreme temperature （-2℃）.
文摘The global burden caused by dengue has increased dramatically in recent decades. Dengue Fever and its severe clinical manifestation, Dengue Hemorrhagic Fever and Dengue Shock Syndrome, have become international public health problem endemic in more than 100 countries, risking 2.5 to 3 billion world populations in tropical and subtropical region. Envelope （E） protein of dengue virus has been proposed as the most important antigen that enables it as vaccine candidate or diagnostic materials. Recombinant protein E production is desirable for dengue vaccine and diagnostic development, especially in Indonesia, where dengue is epidemic. Cloning E gene in an expression vector is essential as an initial method to produce dengue E antigens. The purpose of this research was to clone E gene of dengue virus type 3 （Indonesia D3-1703 strain） into Saccharomyces cerevisiae expression vector pYES2/CT. The cloning method used was the in vitro ligation protocol. First, the cDNA from dengue virus type 3 strain （D3-1703） was generated. Then the polymerase chain reaction （PCR） amplification product of E gene cassette from this cDNA was obtained. The E gene cassette was ligated into linearized pYES2/CT resulting a recombinant vector named pYES2/CT-E. The cloned E gene was verified by restriction enzyme digestion and PCR. Sequencing analysis at its 5＇ end showed that the E gene was inserted at the right open reading frame. In conclusion, the results showed that for the first time the E gene originated from an Indonesian dengue virus type 3 strain was successfully cloned within the yeast expression vector pYES2/CT. In the future, this clone could be expressed and provided as materials for dengue vaccine and diagnostic kit, specific for Indonesian dengue virus strain.
基金supported by the National Natural Science Foundation of China (51573055, 51373054, 21334002, 51403063)National Basic Research Program of China (2014CB643504)+2 种基金Fundamental Research Funds for the Central UniversitiesChina Postdoctoral Science Fund (2014M562174)Introduced Innovative Research & Development Team of Guangdong (201101C0105067115)
文摘Neutral perylene bisimides(PBI) are well-known n-type organic semiconductors, with number of challenging electronic properties in their neutral and reduced states. We report the characteristic electronic properties of PBI anionic films. We unexpectedly discovered that pristine PBI dianion film showed p-type character, while oxidized dianion film(dominant neutral state with few radical anions) showed normal n-type semiconductor character based on Seebeck effect measurements. Both kinds of films exhibit high electrical conductivity with a potential for thermoelectric applications. The mechanism of polarity reversal is proposed.
