The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into fo...The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into four groups: control group, L-group (30 mg-kgt. d), M-group (60 mg·kg-1 ·d) and H-group (120 mg·kg-1· d) and were orally administrated with gossypol diluted by sodium carboxymethyl cellulose (SCC) or SCC (control group) for 20 days. On the 21st day, all the mice were killed and ultrastructure changes of testis were observed by TEM. mRNA expression of Bax and Bcl-2 in testis was measured by semiquantitative RT-PCR. The results showed that the testicular ultrastructure in three treated groups was gradually damaged, according to the dosage of gossypol and cellular structure disordered and organdie degenerated, manifesting vacuolation of mitochondria, expansion of endoplasmie reticulum, mRNA expression of Bcl-2 in testis significantly increased (p〈0.05) in L-group and then significantly decreased (p〈0.05, p〈0.01) in M-group and H-group compared with that in the control group; mRNA expression of Bcl-2 in M-group and H-group significantly decreased (p〈0.05, p〈0.01) than that in L-group and Bcl-2 mRNA expression in H-group showed a significant decrease (p〈0.05) compared with that in M-group. On the other hand, mRNA expression of Bax significant increased (p〈0.05,p〈0.01) in M-group and H-group than that in the control group. The ratio of Bcl-2/Bax significantly reduced 07〈0.05, p〈0.01) in the treated group than that in the control group and was found to be an obvious dose-dependent. It demonstrated that the gnssypol could induce the changes on ultrastructure of mice testis, down-regulate mRNA expression of Bcl-2 and up-regnlate mRNA expression of Bax, which indicated that sperm ceils were induced apoptosis.展开更多
This study objective was to express and characterize the catalytic domain of the human T cell protein tyrosine phosphatase(△TC-PTP) and to study immunohistochemically the expression of △TC-PTP in human non-small c...This study objective was to express and characterize the catalytic domain of the human T cell protein tyrosine phosphatase(△TC-PTP) and to study immunohistochemically the expression of △TC-PTP in human non-small cell lung cancers. △TC-PTP gene was PCR amplified with the cDNA of human TC-PTP as template, and cloned into the pT7 expression vector. The recombinant pT7-△TC-PTP was expressed in E. coli Rosetta ( DE3 ) host cells and puri- fied. The enzymatic characteristics of △TC-PTP including enzyme activity and kinetics assay were measured. The antiserum was prepared by immunizing rabbit with the purified recombinant △TC-PTP. Rabbit polyclonal antibody against △TC-PTP was purified by PVDF immobilized antigen affinity chromatography. Immunohistochemical staining of lung cancer tissues was performed with antibody against △TC-PTP protein. △TC-PTP gene was correctly cloned, expressed, and purified. The recombinant △TC-PTP had a highly catalytic activity of PTPase. Squamous cell lung carcinoma showed a significantly higher expression rate of △TC-PTP (76. 92%, 10/13 ) than adenocarcinoma (57.14%, 4/7) and normal lung tissue(20%, 1/5 ). This study represents the first demonstration that △TC-PTP is highly expressed in human squamous cell lung carcinomas. In addition, this study provides an important basis for further studying the biological function of TC-PTP and its relationship with lung carcinomas and other diseases.展开更多
A novel Schiff-base fluorescent probe 6,6'-((1E,1'E)-(ethane-1,2-diylbis(azaneylylidene))bis-(methaneylylidene))bis(3-(diethylamino)phenol)(L)was derived from the 2:1 M condensation of 4-(diethylamino)-2-hydro...A novel Schiff-base fluorescent probe 6,6'-((1E,1'E)-(ethane-1,2-diylbis(azaneylylidene))bis-(methaneylylidene))bis(3-(diethylamino)phenol)(L)was derived from the 2:1 M condensation of 4-(diethylamino)-2-hydroxybenzaldehyde with ethylenediamine and characterized by ^(1)H NMR,^(13)C NMR and FT-IR spectroscopies.The results of spectral analysis showed that the probe L is selective and sensitive to Cu^(2+).The detection limit of L is found to be 19 nmol·L^(-1).There is a good linear relationship between the fluorescence intensity of probe L and the concentration of Cu^(2+)in the range of 0 to 20μmol×L^(-1).X-ray crystal structure of the L-Cu^(2+)complex and the Job plot revealed a 1:1 L-Cu^(2+)identification.展开更多
文摘The objective was to evaluate the toxicity effect of gossypol on ultrastructure of mouse testis and the expression of Bax mRNA and Bcl-2 mRNA of sperm cells in mice. Forty-eight male mice were randomly divided into four groups: control group, L-group (30 mg-kgt. d), M-group (60 mg·kg-1 ·d) and H-group (120 mg·kg-1· d) and were orally administrated with gossypol diluted by sodium carboxymethyl cellulose (SCC) or SCC (control group) for 20 days. On the 21st day, all the mice were killed and ultrastructure changes of testis were observed by TEM. mRNA expression of Bax and Bcl-2 in testis was measured by semiquantitative RT-PCR. The results showed that the testicular ultrastructure in three treated groups was gradually damaged, according to the dosage of gossypol and cellular structure disordered and organdie degenerated, manifesting vacuolation of mitochondria, expansion of endoplasmie reticulum, mRNA expression of Bcl-2 in testis significantly increased (p〈0.05) in L-group and then significantly decreased (p〈0.05, p〈0.01) in M-group and H-group compared with that in the control group; mRNA expression of Bcl-2 in M-group and H-group significantly decreased (p〈0.05, p〈0.01) than that in L-group and Bcl-2 mRNA expression in H-group showed a significant decrease (p〈0.05) compared with that in M-group. On the other hand, mRNA expression of Bax significant increased (p〈0.05,p〈0.01) in M-group and H-group than that in the control group. The ratio of Bcl-2/Bax significantly reduced 07〈0.05, p〈0.01) in the treated group than that in the control group and was found to be an obvious dose-dependent. It demonstrated that the gnssypol could induce the changes on ultrastructure of mice testis, down-regulate mRNA expression of Bcl-2 and up-regnlate mRNA expression of Bax, which indicated that sperm ceils were induced apoptosis.
基金Supported by the Fund of Science and Technology Department of Jilin Province(No20060563)in part by the Fund of Scienceand Technology Department of Jilin Province(No200505120) the Bureau of Science and Technology of Changchun City(No06GG110)
文摘This study objective was to express and characterize the catalytic domain of the human T cell protein tyrosine phosphatase(△TC-PTP) and to study immunohistochemically the expression of △TC-PTP in human non-small cell lung cancers. △TC-PTP gene was PCR amplified with the cDNA of human TC-PTP as template, and cloned into the pT7 expression vector. The recombinant pT7-△TC-PTP was expressed in E. coli Rosetta ( DE3 ) host cells and puri- fied. The enzymatic characteristics of △TC-PTP including enzyme activity and kinetics assay were measured. The antiserum was prepared by immunizing rabbit with the purified recombinant △TC-PTP. Rabbit polyclonal antibody against △TC-PTP was purified by PVDF immobilized antigen affinity chromatography. Immunohistochemical staining of lung cancer tissues was performed with antibody against △TC-PTP protein. △TC-PTP gene was correctly cloned, expressed, and purified. The recombinant △TC-PTP had a highly catalytic activity of PTPase. Squamous cell lung carcinoma showed a significantly higher expression rate of △TC-PTP (76. 92%, 10/13 ) than adenocarcinoma (57.14%, 4/7) and normal lung tissue(20%, 1/5 ). This study represents the first demonstration that △TC-PTP is highly expressed in human squamous cell lung carcinomas. In addition, this study provides an important basis for further studying the biological function of TC-PTP and its relationship with lung carcinomas and other diseases.
基金Financially supported by the Natural Science Foundation of Hunan Province (2021JJ30291)Undergraduate Training Program for Innovation and Entrepreneurship of Hunan Province ([2021]197-3585)+1 种基金Undergraduate Training Program for Innovation and Entrepreneurship of Hunan University of Science and Engineering (2021)the Construct Program of Applied Characteristic Discipline in Hunan University of Science and Engineering
文摘A novel Schiff-base fluorescent probe 6,6'-((1E,1'E)-(ethane-1,2-diylbis(azaneylylidene))bis-(methaneylylidene))bis(3-(diethylamino)phenol)(L)was derived from the 2:1 M condensation of 4-(diethylamino)-2-hydroxybenzaldehyde with ethylenediamine and characterized by ^(1)H NMR,^(13)C NMR and FT-IR spectroscopies.The results of spectral analysis showed that the probe L is selective and sensitive to Cu^(2+).The detection limit of L is found to be 19 nmol·L^(-1).There is a good linear relationship between the fluorescence intensity of probe L and the concentration of Cu^(2+)in the range of 0 to 20μmol×L^(-1).X-ray crystal structure of the L-Cu^(2+)complex and the Job plot revealed a 1:1 L-Cu^(2+)identification.