Objective: To construct eukaryotic expression plasmid pcDNA3-fragile histidine triad(FHIT) and obtain its transient expression in COS-1 cells. Methods: FHIT gene was cloned from normal human thyroid tissue by RT-PCR a...Objective: To construct eukaryotic expression plasmid pcDNA3-fragile histidine triad(FHIT) and obtain its transient expression in COS-1 cells. Methods: FHIT gene was cloned from normal human thyroid tissue by RT-PCR and then inserted into eukaryotic expression vector pcDNA3. After the sequence was confirmed, the recombinant plasmid pcDNA3-FHIT was transfected into COS-1 cells by cation liposome. The transient expression in the cells was measured by immunocytochemistry. Results: The sequence of FHIT in pcDNA3 was correct and high expression was obtained in COS-1 cells. Conclusion: The eukaryotic expression plasmid pcDNA3-FHIT was constructed successfully and could highly express FHIT protein in COS-1 cells. This will be potentially useful for the research on gene therapy.展开更多
[Objective] The aims was to explore PTGMS line purification and multipli- cation technology. [Method] Three commercial PTGMS lines, Annong 810S, P88S and C815S, were used in this study. In the first year, two sterile ...[Objective] The aims was to explore PTGMS line purification and multipli- cation technology. [Method] Three commercial PTGMS lines, Annong 810S, P88S and C815S, were used in this study. In the first year, two sterile line populations with same genetic background were multiplied by dividing seedling; meanwhile, se- lection and multiplication of the core individuals were conducted; within the year, the core seeds harvested were multiplied under the isolated condition in Hainan, and at the same time, superior individuals were selected and used as the seeds for two populations with same genetic background in the next year. In the second year, seeds harvested in Hainan were planted in Changsha, while the core individuals were selected and multiplied; the core seeds harvested were continued to multiply in Hainan. The above procedures were repeated in the following years. [Result] The test results showed that in two years the proportion of core individuals in the popu- lation of Annong 810S, P88S and C815S increased from 40.3%, 0.7% and 0.3% in the first year to 71.67%, 66.00% and 68.67% in the second year respectively. [Conclusion] This suggested that this method of purification and multiplication of the PTGMS lines for consecutive years can effectively control the drift of the critical sterility inducing temperature, and at the same time, core seeds with high purity and controlled inducing temperature can be harvested.展开更多
An immunosuppressive animal model induced by physical stress that forced mice to swim in cold water(14±1℃)and the restorative effect of Tremella polysac charide(TP)on the suppressed immune function by stress wer...An immunosuppressive animal model induced by physical stress that forced mice to swim in cold water(14±1℃)and the restorative effect of Tremella polysac charide(TP)on the suppressed immune function by stress were studied in mice.It was found that the spleen plaque forming cell(PFC)response to sheep red blood cells,delayed cuta- neous hypersensitivity(DCH)induced by dinitrochlorobenzene and the lymphocyte prolifer- ation stimulated by concanavalin A(Con A)were significantly decreased in stressed mice. In addition.the maximal decrease of PFC was reached in 9-12 days after stress.A- drenolectomy could not affect the decrease of PFC in stressed mice.TP(200.400mg/kg) ig for 8-14 days significantly restored the PFC.DCH and lymphocyte proliferation to nor- mal level in stressed mice.展开更多
In field and laboratory conditions the reproductive potential (survival at different stages of development) of the Colorado potato beetle has been measured, the dependence of the parameters of insects fertility on v...In field and laboratory conditions the reproductive potential (survival at different stages of development) of the Colorado potato beetle has been measured, the dependence of the parameters of insects fertility on varietal features of the feed has been investigated, the food activity of the representatives of insects from four areas of the Southern Urals has been measured, the intensity and time of hypersensitivity reactions on potato leaves has been defined. In laboratory experiments, the level of activity of hydrolytic enzymes in insects and the level of activity of these enzymes inhibitors in the leaves of three potato varieties differing in their resistance to the Colorado potato beetle have been determined. The results show that the insects-representatives of different samples have different food activity in relation to different potato varieties. High mortality rates were revealed at feeding with leaves of potato of the Bashkir variety characterized by a high level of hydrolases inhibitors and relatively rapid development of necrotic reactions after the appearance of eggs laying on the leaves. We suppose that the neerogenetic barrier appearing in the form of necrosis is the major cause of the fetal mortality of insects. The level of the content of inhibitors inactivating larvae digestive enzymes affects the survival of insects at postembryonic stages. The speed of the necrosis emergence, the level of activity of digestive enzymes inhibitors in potato can be considered as criteria for the selection of breeding material for resistance to the Colorado potato beetle.展开更多
Objective: To explore the effects of connective tissue growth factor (CTGF) on the pathogenesis of human keloid. Methods: CTGF antisense oligonucleotides (ASODN) conjugated with isothiocyananate fluorescence was encap...Objective: To explore the effects of connective tissue growth factor (CTGF) on the pathogenesis of human keloid. Methods: CTGF antisense oligonucleotides (ASODN) conjugated with isothiocyananate fluorescence was encapsulated by liposome, and then added into the human keloid fibroblasts (HKFs) culture media. The intracellular distribution of CTGF ASODN was observed by fluorescence microscopy in the fixed HKFs. The proliferation of HKFs was measured by MTT test. The collagen synthesis of HKFs was measured by 3H-proline incorporation method. Results: Compared with control group, the CTGF ASODN can inhibit the proliferation and collagen synthesis of the HKFs (P<0.01). Conclusion: CTGF ASODN has anti-fibrotic effects on keloid in vitro, and CTGF play an important role in promoting the fibrosis of keloid.展开更多
Objective To generate and characterize anti-idiotypic monoclonal antibody (Ab2) that bears the internal image of nasopharyngeal carcinoma (NPC) associated antigen. Methods Using NPC monoclonal antibody (Ab1) as immu...Objective To generate and characterize anti-idiotypic monoclonal antibody (Ab2) that bears the internal image of nasopharyngeal carcinoma (NPC) associated antigen. Methods Using NPC monoclonal antibody (Ab1) as immunogen, hybridoma cells were obtained by fusion of SP2/0 myeloma cells with immunized murine spleen cells. Positive clones were screened by Sandwich ELISA and a binding inhibition test. To determine whether Ab2 possess the internal image of the original antigen or not, mice were immunized with Ab2. ELISA and the competitive inhibition assay tested anti-anti-idiotypic antibodies (Ab3) in anti-sera. Cell-mediated immunity to tumors induced by Ab2 was investigated by a delayed-type hypersensitivity response and the mouse T-cell proliferation assay. Results Anti-idiotypic monoclonal antibodies against the monoclonal anti-NPC antibodies FC2 and HNL5 were generated that recognize NPC associated antigens. These Ab2, which were designated 2H4 and 5D3, could inhibit the binding of FC2 or HNL5 to NPC cell lines. Anti-sera from the immunized mice, which contained Ab3, could compete with FC2 or HNL5 for binding with NPC cell by a competitive inhibition assay. Mice immunized with 2H4 or 5D3 coupled with keyhole limpet hemocyanin (KLH), showed a positive and specific delayed-type hypersensitivity (DTH) reaction after stimulation by NPC cells. The mouse T cell proliferative assay indicated that there was a significantly higher proliferative response of the splenocytes in the experimental groups than that in control groups. Conclusions Anti-idiotypic antibodies 2H4 and 5D3 are Ab2 beta bearing the internal image of the epitope of NPC associated antigen. Either 2H4 or 5D3 expressing three-dimensional shapes that resemble the structure of natural antigens could induce humoral and cellular immune response.展开更多
Porcine reproductive and respiratory syndrome virus(PRRSV) shows characteristic antibody-dependent enhancement(ADE) of infection and causes porcine systemic inflammation, which is similar to a type I allergic reaction...Porcine reproductive and respiratory syndrome virus(PRRSV) shows characteristic antibody-dependent enhancement(ADE) of infection and causes porcine systemic inflammation, which is similar to a type I allergic reaction; however, the role of porcine FceεRI in ADE is still unclear. In this study, the expression of different Fc receptors(Fc Rs) on macrophages was investigated in a PRRSV 3D4/21 cell infection model in the presence or absence of PRRSV antibody. The transcription level of Fcc II and FceεRI was significantly up-regulated under PRRSV-antibody complex infection. Internalization and proliferation of PRRSV were promoted by the ADE mechanism when FceεRI was expressed in permissive 3D4/21 cells and the non-permissive cell line HEK 293T. Transcriptome sequencing data showed that the expression levels of AKT,ERK and other signal molecules in the anti-inflammatory pathway were significantly increased, especially in the cells infected with the PRRSV-antibody immune complex. Inflammatory regulatory molecules such as PLA2G6, LOX, TRPM8 and TRPM4 were significantly up-regulated following PRRSV infection but significantly down-regulated in the cells infected with the PRRSV-antibody immune complex. Our results demonstrated that FceεRI could be involved in PRRSV ADE, the antigen presenting process and regulation of the inflammatory response during PRRSV infection, which provides new insights into PRRSV infection mediated by FceεRI and the PRRSV-antibody immune complex.展开更多
Threat-sensitive behavioral trade-offs allow prey animals to balance the conflicting demands of successful predator detection and avoidance and a suite of fitness-related activities such as forag- ing, mating, and ter...Threat-sensitive behavioral trade-offs allow prey animals to balance the conflicting demands of successful predator detection and avoidance and a suite of fitness-related activities such as forag- ing, mating, and territorial defense. Here, we test the hypothesis that background predation level and reproductive status interact to determine the form and intensity of threat-sensitive behavioral decisions of wild-caught female Trinidadian guppies Poecilia reticulata. Gravid and nongravid gup- pies collected from high- and low-predation pressure populations were exposed to serial dilutions of conspecific chemical alarm cues. Our results demonstrate that there was 'no effect of reproduct- ive status on the response of females originating from a low-predation population, with both gravid and nongravid guppies exhibiting strong anti-predator responses to the lowest concentration of alarm cues tested. Increasing cue concentrations did not result in increases in response intensity. Conversely, we found a significant effect of reproductive status among guppies from a high- predation population. Nongravid females from the high-predation population exhibited a strong graded (proportional) response to increasing concentrations of alarm cue. Gravid females from the same high-predation population, however, shifted to a nongraded response. Together, these re- sults demonstrate that accrued reproductive assets influence the threat-sensitive behavioral deci- sions of prey, but only under conditions of high-ambient predation risk.展开更多
Because antipredator behaviours are costly, the threat-sensitive predator avoidance hypothesis predicts that individual animals should express predator-avoidance behaviour proportionally to the perceived threat posed ...Because antipredator behaviours are costly, the threat-sensitive predator avoidance hypothesis predicts that individual animals should express predator-avoidance behaviour proportionally to the perceived threat posed by the predator. Here, we experimentally tested this hypothesis by providing wild passerine birds supplemental food (on a raised feeding platform) at either 1 or 4 m from the edge of forest cover (potential refuge), in either the presence or absence of a nearby simulated predation threat (a sharp-shinned hawk Accipiter striatus model). Compared with the control treatment, we observed proportionally fewer bird visits to the food patch, and the birds took longer to re-emerge from forest refuge and return to feed at the food patch, after the hawk presentation than before it. The observed threat-sensitive latency-to-return response was stronger when the food patch was further away from the nearest refuge. Overall, our results are consistent with the predictions of the threat-sensitive predator avoidance hypothesis in that wild passerine birds (primarily black-capped chickadees Poecile atricapillus) exhibited more intense antipre- dator behavioural responses with increasing level of apparent threat. The birds were thus sensitive to their local perceived threat of predation and traded-off safety from predation (by refuging) and foraging gains in open habitat in a graded, threat-sensitive manner [Current Zoology 60 (6): 719-728, 2014].展开更多
文摘Objective: To construct eukaryotic expression plasmid pcDNA3-fragile histidine triad(FHIT) and obtain its transient expression in COS-1 cells. Methods: FHIT gene was cloned from normal human thyroid tissue by RT-PCR and then inserted into eukaryotic expression vector pcDNA3. After the sequence was confirmed, the recombinant plasmid pcDNA3-FHIT was transfected into COS-1 cells by cation liposome. The transient expression in the cells was measured by immunocytochemistry. Results: The sequence of FHIT in pcDNA3 was correct and high expression was obtained in COS-1 cells. Conclusion: The eukaryotic expression plasmid pcDNA3-FHIT was constructed successfully and could highly express FHIT protein in COS-1 cells. This will be potentially useful for the research on gene therapy.
基金Supported by National High Technology Research and Development Program of China(2010AA101305,2011AA10A101)National Science and Technology Supporting Project(2012BAD48G01)~~
文摘[Objective] The aims was to explore PTGMS line purification and multipli- cation technology. [Method] Three commercial PTGMS lines, Annong 810S, P88S and C815S, were used in this study. In the first year, two sterile line populations with same genetic background were multiplied by dividing seedling; meanwhile, se- lection and multiplication of the core individuals were conducted; within the year, the core seeds harvested were multiplied under the isolated condition in Hainan, and at the same time, superior individuals were selected and used as the seeds for two populations with same genetic background in the next year. In the second year, seeds harvested in Hainan were planted in Changsha, while the core individuals were selected and multiplied; the core seeds harvested were continued to multiply in Hainan. The above procedures were repeated in the following years. [Result] The test results showed that in two years the proportion of core individuals in the popu- lation of Annong 810S, P88S and C815S increased from 40.3%, 0.7% and 0.3% in the first year to 71.67%, 66.00% and 68.67% in the second year respectively. [Conclusion] This suggested that this method of purification and multiplication of the PTGMS lines for consecutive years can effectively control the drift of the critical sterility inducing temperature, and at the same time, core seeds with high purity and controlled inducing temperature can be harvested.
文摘An immunosuppressive animal model induced by physical stress that forced mice to swim in cold water(14±1℃)and the restorative effect of Tremella polysac charide(TP)on the suppressed immune function by stress were studied in mice.It was found that the spleen plaque forming cell(PFC)response to sheep red blood cells,delayed cuta- neous hypersensitivity(DCH)induced by dinitrochlorobenzene and the lymphocyte prolifer- ation stimulated by concanavalin A(Con A)were significantly decreased in stressed mice. In addition.the maximal decrease of PFC was reached in 9-12 days after stress.A- drenolectomy could not affect the decrease of PFC in stressed mice.TP(200.400mg/kg) ig for 8-14 days significantly restored the PFC.DCH and lymphocyte proliferation to nor- mal level in stressed mice.
文摘In field and laboratory conditions the reproductive potential (survival at different stages of development) of the Colorado potato beetle has been measured, the dependence of the parameters of insects fertility on varietal features of the feed has been investigated, the food activity of the representatives of insects from four areas of the Southern Urals has been measured, the intensity and time of hypersensitivity reactions on potato leaves has been defined. In laboratory experiments, the level of activity of hydrolytic enzymes in insects and the level of activity of these enzymes inhibitors in the leaves of three potato varieties differing in their resistance to the Colorado potato beetle have been determined. The results show that the insects-representatives of different samples have different food activity in relation to different potato varieties. High mortality rates were revealed at feeding with leaves of potato of the Bashkir variety characterized by a high level of hydrolases inhibitors and relatively rapid development of necrotic reactions after the appearance of eggs laying on the leaves. We suppose that the neerogenetic barrier appearing in the form of necrosis is the major cause of the fetal mortality of insects. The level of the content of inhibitors inactivating larvae digestive enzymes affects the survival of insects at postembryonic stages. The speed of the necrosis emergence, the level of activity of digestive enzymes inhibitors in potato can be considered as criteria for the selection of breeding material for resistance to the Colorado potato beetle.
文摘Objective: To explore the effects of connective tissue growth factor (CTGF) on the pathogenesis of human keloid. Methods: CTGF antisense oligonucleotides (ASODN) conjugated with isothiocyananate fluorescence was encapsulated by liposome, and then added into the human keloid fibroblasts (HKFs) culture media. The intracellular distribution of CTGF ASODN was observed by fluorescence microscopy in the fixed HKFs. The proliferation of HKFs was measured by MTT test. The collagen synthesis of HKFs was measured by 3H-proline incorporation method. Results: Compared with control group, the CTGF ASODN can inhibit the proliferation and collagen synthesis of the HKFs (P<0.01). Conclusion: CTGF ASODN has anti-fibrotic effects on keloid in vitro, and CTGF play an important role in promoting the fibrosis of keloid.
基金ThisinvestigationwassupportedbygrantsfromtheNationalNaturalScienceFoundationofChina (No 392 70 716 )ChinaMedicalBoardinNewYork (No 90 5 2 9project 7)
文摘Objective To generate and characterize anti-idiotypic monoclonal antibody (Ab2) that bears the internal image of nasopharyngeal carcinoma (NPC) associated antigen. Methods Using NPC monoclonal antibody (Ab1) as immunogen, hybridoma cells were obtained by fusion of SP2/0 myeloma cells with immunized murine spleen cells. Positive clones were screened by Sandwich ELISA and a binding inhibition test. To determine whether Ab2 possess the internal image of the original antigen or not, mice were immunized with Ab2. ELISA and the competitive inhibition assay tested anti-anti-idiotypic antibodies (Ab3) in anti-sera. Cell-mediated immunity to tumors induced by Ab2 was investigated by a delayed-type hypersensitivity response and the mouse T-cell proliferation assay. Results Anti-idiotypic monoclonal antibodies against the monoclonal anti-NPC antibodies FC2 and HNL5 were generated that recognize NPC associated antigens. These Ab2, which were designated 2H4 and 5D3, could inhibit the binding of FC2 or HNL5 to NPC cell lines. Anti-sera from the immunized mice, which contained Ab3, could compete with FC2 or HNL5 for binding with NPC cell by a competitive inhibition assay. Mice immunized with 2H4 or 5D3 coupled with keyhole limpet hemocyanin (KLH), showed a positive and specific delayed-type hypersensitivity (DTH) reaction after stimulation by NPC cells. The mouse T cell proliferative assay indicated that there was a significantly higher proliferative response of the splenocytes in the experimental groups than that in control groups. Conclusions Anti-idiotypic antibodies 2H4 and 5D3 are Ab2 beta bearing the internal image of the epitope of NPC associated antigen. Either 2H4 or 5D3 expressing three-dimensional shapes that resemble the structure of natural antigens could induce humoral and cellular immune response.
基金supported by the National Natural Science Foundation of China (31272540)the underprop project of Tianjin Science and Technology Committee in China (16YFZCNC00640)
文摘Porcine reproductive and respiratory syndrome virus(PRRSV) shows characteristic antibody-dependent enhancement(ADE) of infection and causes porcine systemic inflammation, which is similar to a type I allergic reaction; however, the role of porcine FceεRI in ADE is still unclear. In this study, the expression of different Fc receptors(Fc Rs) on macrophages was investigated in a PRRSV 3D4/21 cell infection model in the presence or absence of PRRSV antibody. The transcription level of Fcc II and FceεRI was significantly up-regulated under PRRSV-antibody complex infection. Internalization and proliferation of PRRSV were promoted by the ADE mechanism when FceεRI was expressed in permissive 3D4/21 cells and the non-permissive cell line HEK 293T. Transcriptome sequencing data showed that the expression levels of AKT,ERK and other signal molecules in the anti-inflammatory pathway were significantly increased, especially in the cells infected with the PRRSV-antibody immune complex. Inflammatory regulatory molecules such as PLA2G6, LOX, TRPM8 and TRPM4 were significantly up-regulated following PRRSV infection but significantly down-regulated in the cells infected with the PRRSV-antibody immune complex. Our results demonstrated that FceεRI could be involved in PRRSV ADE, the antigen presenting process and regulation of the inflammatory response during PRRSV infection, which provides new insights into PRRSV infection mediated by FceεRI and the PRRSV-antibody immune complex.
文摘Threat-sensitive behavioral trade-offs allow prey animals to balance the conflicting demands of successful predator detection and avoidance and a suite of fitness-related activities such as forag- ing, mating, and territorial defense. Here, we test the hypothesis that background predation level and reproductive status interact to determine the form and intensity of threat-sensitive behavioral decisions of wild-caught female Trinidadian guppies Poecilia reticulata. Gravid and nongravid gup- pies collected from high- and low-predation pressure populations were exposed to serial dilutions of conspecific chemical alarm cues. Our results demonstrate that there was 'no effect of reproduct- ive status on the response of females originating from a low-predation population, with both gravid and nongravid guppies exhibiting strong anti-predator responses to the lowest concentration of alarm cues tested. Increasing cue concentrations did not result in increases in response intensity. Conversely, we found a significant effect of reproductive status among guppies from a high- predation population. Nongravid females from the high-predation population exhibited a strong graded (proportional) response to increasing concentrations of alarm cue. Gravid females from the same high-predation population, however, shifted to a nongraded response. Together, these re- sults demonstrate that accrued reproductive assets influence the threat-sensitive behavioral deci- sions of prey, but only under conditions of high-ambient predation risk.
文摘Because antipredator behaviours are costly, the threat-sensitive predator avoidance hypothesis predicts that individual animals should express predator-avoidance behaviour proportionally to the perceived threat posed by the predator. Here, we experimentally tested this hypothesis by providing wild passerine birds supplemental food (on a raised feeding platform) at either 1 or 4 m from the edge of forest cover (potential refuge), in either the presence or absence of a nearby simulated predation threat (a sharp-shinned hawk Accipiter striatus model). Compared with the control treatment, we observed proportionally fewer bird visits to the food patch, and the birds took longer to re-emerge from forest refuge and return to feed at the food patch, after the hawk presentation than before it. The observed threat-sensitive latency-to-return response was stronger when the food patch was further away from the nearest refuge. Overall, our results are consistent with the predictions of the threat-sensitive predator avoidance hypothesis in that wild passerine birds (primarily black-capped chickadees Poecile atricapillus) exhibited more intense antipre- dator behavioural responses with increasing level of apparent threat. The birds were thus sensitive to their local perceived threat of predation and traded-off safety from predation (by refuging) and foraging gains in open habitat in a graded, threat-sensitive manner [Current Zoology 60 (6): 719-728, 2014].