In the present study we have found that proto-oncogene c-fos protein can expressin the noradrenergic neurons of rat hindbrain following peripheral electrical stimulation. Ratswere given peripheral electrical stimulati...In the present study we have found that proto-oncogene c-fos protein can expressin the noradrenergic neurons of rat hindbrain following peripheral electrical stimulation. Ratswere given peripheral electrical stimulation via thin stainless steel pins inserted into the pointsnear knee joint (S36) and ankle joint (Sp6) which mimic the manipulation of electroacupuncture(EA) performed in humans. Animals were perfused for double staining immunohistochemistry 2hafter the termination of EA. In rats subjected to EA stimulation Fos-like protein was found in thetyrosine hydroxylase (TH)-like immunoreactive neurons in rat hindbrain. The Fos and TH coex-isting neurons were distributed in the locus coeruleus, solitary tract nucleus, ventrolateral medul-la, periaqeductal gray, as well as superior colliculus. The percentage of the coexisting neuronscompared with the total number of neurons containing Fos-like protein in these nuclei rangedfrom 6% to 32%. The results suggest that the noradrenergic neurons in these regions may be ac-tivted by acupuncture stimulation.展开更多
AIM: To construct and highly express an epitope of hepatitis C virus (HCV) in a foreign epitope presenting vector based on an insect virus, and to study the antigenicity of the epitope. METHODS: The HCV epitope sequen...AIM: To construct and highly express an epitope of hepatitis C virus (HCV) in a foreign epitope presenting vector based on an insect virus, and to study the antigenicity of the epitope. METHODS: The HCV epitope sequence (amino acid residues 315 to 328: EGHRMAWDMMMNWS) of the El region was constructed at different positions of a foreign epitope presenting vector based on an insect virus, flock house virus (FHV) capsid protein encoding gene as a vector, and expressed in E. coli cells. Western blotting and ELISA were used to detect the immunoreactivity of these recombinant proteins. RESULTS: The gene encoding of the concerned B-cell epitope of HCV El envelope protein was expressed on FHV capsid carrier protein at positions I1 (aa 106), 12 (aa 153) and 13 (aa 305), respectively, on the surface of FHV capsid protein. The recombinant proteins in this system could be highly expressed in more than 40% of total cell protein of E. coli BL21. All the expressed recombinant proteins were in inclusion body form, and showed obvious immunoreactivity by Western blotting. Further purified recombinant proteins were detected by indirect ELISA as coating antigen respectively. All recombinant proteins could still show immunoreactivity. CONCLUSION: The epitope of HCV El envelope protein can be highly expressed in FHV carrier system as a chimeric protein with high immunoreactivity. This system has multiple entry sites conferring many possible conformations closer to the native one for a given sequence.展开更多
Rotavirus capsid protein Vp4 plays an important role in the virus adhering and entering the cells. In this study, a Vp4 gene cloned from a rotavirus strain TB-Chen was highly expressed in E.coli BL21 (DE3). The result...Rotavirus capsid protein Vp4 plays an important role in the virus adhering and entering the cells. In this study, a Vp4 gene cloned from a rotavirus strain TB-Chen was highly expressed in E.coli BL21 (DE3). The results of the Western blot showed that the protein possesses specific immuno-reactivities and can be specifically recognized by guinea pig antibodies against rotavirus strain SA11 or Wa. Some Vp4 dimers were formed during renaturation. These data obtained from this study provide a strong basis for further study on the structure and function of the Vp4.展开更多
With the methods of transganglionic transport of horseradish peroxidase(HRP) combined with postembedding colloidal gold stain immunoelectron microscopy, the shape and synaptic relationship of the trigeminal primary af...With the methods of transganglionic transport of horseradish peroxidase(HRP) combined with postembedding colloidal gold stain immunoelectron microscopy, the shape and synaptic relationship of the trigeminal primary afferent terminals containing substance展开更多
The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This repor...The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain).展开更多
An antigastric cancer monoclonal antibody, 3H11, and its Fab fragment, were labeled using p-(211At)-astatobenzoic acid (pAtBA) intermediate, in the yields of more than 30%. The results of in vitro experiments show tha...An antigastric cancer monoclonal antibody, 3H11, and its Fab fragment, were labeled using p-(211At)-astatobenzoic acid (pAtBA) intermediate, in the yields of more than 30%. The results of in vitro experiments show that 211At-3H11 and 211At-3H11Fab are stable, and have specific immunoreactivities and cytotoxic effects to human gastric cancer cell M85. The cytotoxic effects are dependent on the concentration of 211At-3H11 or 211At-3H11 Fab and obviously stronger than that of Na211At.展开更多
目的:制备粉尘螨变应原第27组分(Der f 27)重组蛋白,鉴定其免疫活性。方法:构建pET-28a(+)-Der f 27质粒,转化至E.coli BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 27。IgE-ELISA和IgE-Western blot检测rDer f 27与...目的:制备粉尘螨变应原第27组分(Der f 27)重组蛋白,鉴定其免疫活性。方法:构建pET-28a(+)-Der f 27质粒,转化至E.coli BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 27。IgE-ELISA和IgE-Western blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率,分别将变应性鼻炎患者PBMC、人支气管上皮细胞BESA-2B与rDer f 27共孵育24 h后测定细胞因子表达;生物信息学软件分析Der f 27的理化性质和结构。结果:成功构建pET-28a(+)-Der f 27质粒并转化至BL21(DE3)细胞,经IPTG诱导表达和纯化后,SDS-PAGE和Western blot在约48 kD处见特异性条带;IgE-ELISA和IgEWestern blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率分别为39.5%和45.5%;与对照组相比,变应性鼻炎患者PBMC与rDer f 27共孵育后,IL-6和IL-8表达升高(P<0.05);BESA-2B细胞与rDer f 27共孵育后,IL-10和TGF-β表达降低,IL-17A和IL-23A表达升高(P<0.05);生物信息学结果显示Der f 27属于丝氨酸蛋白酶抑制物家族,具有此家族通用结构及功能,二级结构主要为α螺旋(42.62%)和无规则卷曲(35.60%)。结论:成功制备了具有较好免疫反应性和免疫原性的重组变应原rDer f 27,并揭示其为变应性鼻炎的重要变应原之一。展开更多
基金Chinese postdoctoral foundationa grand from the National Institute of Drug Abuse, USA (DA 03983)
文摘In the present study we have found that proto-oncogene c-fos protein can expressin the noradrenergic neurons of rat hindbrain following peripheral electrical stimulation. Ratswere given peripheral electrical stimulation via thin stainless steel pins inserted into the pointsnear knee joint (S36) and ankle joint (Sp6) which mimic the manipulation of electroacupuncture(EA) performed in humans. Animals were perfused for double staining immunohistochemistry 2hafter the termination of EA. In rats subjected to EA stimulation Fos-like protein was found in thetyrosine hydroxylase (TH)-like immunoreactive neurons in rat hindbrain. The Fos and TH coex-isting neurons were distributed in the locus coeruleus, solitary tract nucleus, ventrolateral medul-la, periaqeductal gray, as well as superior colliculus. The percentage of the coexisting neuronscompared with the total number of neurons containing Fos-like protein in these nuclei rangedfrom 6% to 32%. The results suggest that the noradrenergic neurons in these regions may be ac-tivted by acupuncture stimulation.
基金Supported by the Natural Science Fund of Yunnan Province, No. 2003C0076M
文摘AIM: To construct and highly express an epitope of hepatitis C virus (HCV) in a foreign epitope presenting vector based on an insect virus, and to study the antigenicity of the epitope. METHODS: The HCV epitope sequence (amino acid residues 315 to 328: EGHRMAWDMMMNWS) of the El region was constructed at different positions of a foreign epitope presenting vector based on an insect virus, flock house virus (FHV) capsid protein encoding gene as a vector, and expressed in E. coli cells. Western blotting and ELISA were used to detect the immunoreactivity of these recombinant proteins. RESULTS: The gene encoding of the concerned B-cell epitope of HCV El envelope protein was expressed on FHV capsid carrier protein at positions I1 (aa 106), 12 (aa 153) and 13 (aa 305), respectively, on the surface of FHV capsid protein. The recombinant proteins in this system could be highly expressed in more than 40% of total cell protein of E. coli BL21. All the expressed recombinant proteins were in inclusion body form, and showed obvious immunoreactivity by Western blotting. Further purified recombinant proteins were detected by indirect ELISA as coating antigen respectively. All recombinant proteins could still show immunoreactivity. CONCLUSION: The epitope of HCV El envelope protein can be highly expressed in FHV carrier system as a chimeric protein with high immunoreactivity. This system has multiple entry sites conferring many possible conformations closer to the native one for a given sequence.
文摘Rotavirus capsid protein Vp4 plays an important role in the virus adhering and entering the cells. In this study, a Vp4 gene cloned from a rotavirus strain TB-Chen was highly expressed in E.coli BL21 (DE3). The results of the Western blot showed that the protein possesses specific immuno-reactivities and can be specifically recognized by guinea pig antibodies against rotavirus strain SA11 or Wa. Some Vp4 dimers were formed during renaturation. These data obtained from this study provide a strong basis for further study on the structure and function of the Vp4.
文摘With the methods of transganglionic transport of horseradish peroxidase(HRP) combined with postembedding colloidal gold stain immunoelectron microscopy, the shape and synaptic relationship of the trigeminal primary afferent terminals containing substance
基金Natural Science Foundation of China (30460098)China Postdoctoral Science Foundation funded project (2005038585)
文摘The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain).
文摘An antigastric cancer monoclonal antibody, 3H11, and its Fab fragment, were labeled using p-(211At)-astatobenzoic acid (pAtBA) intermediate, in the yields of more than 30%. The results of in vitro experiments show that 211At-3H11 and 211At-3H11Fab are stable, and have specific immunoreactivities and cytotoxic effects to human gastric cancer cell M85. The cytotoxic effects are dependent on the concentration of 211At-3H11 or 211At-3H11 Fab and obviously stronger than that of Na211At.
文摘目的:制备粉尘螨变应原第27组分(Der f 27)重组蛋白,鉴定其免疫活性。方法:构建pET-28a(+)-Der f 27质粒,转化至E.coli BL21(DE3)感受态细胞,经诱导表达和纯化后,获得重组变应原rDer f 27。IgE-ELISA和IgE-Western blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率,分别将变应性鼻炎患者PBMC、人支气管上皮细胞BESA-2B与rDer f 27共孵育24 h后测定细胞因子表达;生物信息学软件分析Der f 27的理化性质和结构。结果:成功构建pET-28a(+)-Der f 27质粒并转化至BL21(DE3)细胞,经IPTG诱导表达和纯化后,SDS-PAGE和Western blot在约48 kD处见特异性条带;IgE-ELISA和IgEWestern blot检测rDer f 27与粉尘螨变应性鼻炎患者血清IgE结合率分别为39.5%和45.5%;与对照组相比,变应性鼻炎患者PBMC与rDer f 27共孵育后,IL-6和IL-8表达升高(P<0.05);BESA-2B细胞与rDer f 27共孵育后,IL-10和TGF-β表达降低,IL-17A和IL-23A表达升高(P<0.05);生物信息学结果显示Der f 27属于丝氨酸蛋白酶抑制物家族,具有此家族通用结构及功能,二级结构主要为α螺旋(42.62%)和无规则卷曲(35.60%)。结论:成功制备了具有较好免疫反应性和免疫原性的重组变应原rDer f 27,并揭示其为变应性鼻炎的重要变应原之一。
