AIM: TO explore the ability of recombinant toxin luteinizinghormone releasing hormone-Pseudomonas aeruginosaexotoxin 40 (LHRH-PE40) and LHRH binding to LHRH receptor(LHRHR) on the membrane surface of human liver cance...AIM: TO explore the ability of recombinant toxin luteinizinghormone releasing hormone-Pseudomonas aeruginosaexotoxin 40 (LHRH-PE40) and LHRH binding to LHRH receptor(LHRHR) on the membrane surface of human liver cancerHEPG cells.METHODS: LHRH was labeled by using 12sI with enzymaticreaction. The affinity and receptor volume of LHP, H-PE40and LHRH binding to LHRHR on the membrane surface ofhuman liver cancer cells were measured with radioligandreceptor assay.RESULTS: The specific activity of LHRH labeled with 12sIwas 2.7x 104 kBq/l^L, and its radiochemical purity reachedto 99.2-99.7%. The binding of 12sI to LHRH was maximalfor 240 min in the warm cultivation, and this binding wasstabilized. The inhibiting rates of ~2SI-LHRH and LHRH onthe proliferation of human liver cancer HEPG cells werenot significantly different. On the basis of the saturationcurve of ~2SI-LHRH binding to the membrane LHRHR of HEPGcells, ~2SI-LHRH of lx10s cpm was selected for radioligandreceptor assay. The affinity constants (Kd) of LHRH-PE40and LHRH binding to the membrane LHRHR of HEPG cellswere 0.43~0.12 nmol/L and 4.86~ 1.47 nmol/L, respectively,and their receptor volumes were 0.37~0.15 l^mol/g and0.42=I=0.13 l^mol/g, respectively. The binding of LHRH-PE40to the membrane protein of normal liver cells was notobserved.CONCLUSION: The recombinant toxin LHRH-PE40 binding tothe membrane surface of LHRHR of human liver cancer HEPGcells was very strong, while the specific binding of it to normalliver cells was not observed. The results provide an importantexperimental basis for the clinical application of LHRH-PE.展开更多
The effects of two intraperitoneal injections of human chorionic gonadotropin (HCG) or [ D-ALA6, pro9- N- ethylamide]-luteinizing hormone releasing hormone analogue (LHRH-A), respectively, and the combined effect of L...The effects of two intraperitoneal injections of human chorionic gonadotropin (HCG) or [ D-ALA6, pro9- N- ethylamide]-luteinizing hormone releasing hormone analogue (LHRH-A), respectively, and the combined effect of LHRH-A with pimozide (PIM), a dopamine antagonist, on ovulation in female mud skipper were investigated. A high rate of ovulation (83.3%)was observed after injection with HCG(30 I. U./g). Injection of LHRH-A at a total dosage of 2μg/g was 100% effective in inducing ovulation. No ovulations occurred in the group receiving a low dosage of LHRH-A (0.02μg/g). Injection of LHRH-A(0.2μg/g)with PIM (10 or 20 μg/g)induced greater ovulatory response than injection with LHRH-A (0.2μg/g)alone. PIM at total dosages of 0.2μg/g or 2μg/g did not enhance the potency of LHRH-A to stimulate ovulation.展开更多
Aim:To investigate the effect of Trx-LHRH, a new GnRH crasis protein, on antibody production and reproductive function in male rats. Methods: Trx-LHRH produced in vitro with a new crasis gene, which crasised Trx gene ...Aim:To investigate the effect of Trx-LHRH, a new GnRH crasis protein, on antibody production and reproductive function in male rats. Methods: Trx-LHRH produced in vitro with a new crasis gene, which crasised Trx gene and GnRH gene together, was used as a vaccine with hydroalaminum base as the adjuvant in adult male SD rats. A second inoculation was performed 5 weeks later. Antibody level was measured by ELISA and androgen level by RIA thereafter. Results: Trx-LHRH successfully induced polycolonal antibody at the level of 1:1 280 - 2 560 four weeks after the first inoculation and 1:2 000 SIX weeks after the enhanced treatment. Testosterone level was reduced significantly (P<0.01). Sperm count was also reduced by Trx-LHRH treatment. However, there was reasonable individual variations. Conclusion: Trx-LHRH can be used as an effective vaccine to induce antibody production; at the same time, it restrains the function of hypothatamas-pituitary-testis axis.展开更多
Using fluoromctric technique wc tcstcd ED50 ofhistaminc release by LHRH antagonists.There wcrc 9 LHRH antagonists synthesized by us and tested by this method. The range of ED50 is from 0.65±0.08/μg / ml, to 36.8...Using fluoromctric technique wc tcstcd ED50 ofhistaminc release by LHRH antagonists.There wcrc 9 LHRH antagonists synthesized by us and tested by this method. The range of ED50 is from 0.65±0.08/μg / ml, to 36.8±2.8 /μg / ml. We selected a LHRH展开更多
In the present study, we investigated the roles of TGF-β signaling pathway in a rat benign prostatic hyperplasia （BPH） model treated with cetrorelix. TGF-β1 and c-Myc expression were measured by qRT-PCR and Wester...In the present study, we investigated the roles of TGF-β signaling pathway in a rat benign prostatic hyperplasia （BPH） model treated with cetrorelix. TGF-β1 and c-Myc expression were measured by qRT-PCR and Western blotting in the proximal and distal region of ventral prostatic lobes, respectively. We observed that treatment with cetrorelix led to a significant reduction of ventral prostate weight in a dose-dependent manner. In the proximal region, after cetrorelix treatment, the expression of TGF-β1 was dramatically increased （P〈0.05）, while the expression of c-Myc was significantly decreased （P〈0.05）. In comparison with the control group, the cetrorelix groups had more TUNEL-positive cells. Our findings strongly suggest that the TGF-β signaling pathway may be one of the major causes responsible for prostate volume reduction in BPH rats after cetrorelix treatment.展开更多
基金Supported by the Key Programs of National Science and Technology,No.96-901-05-101
文摘AIM: TO explore the ability of recombinant toxin luteinizinghormone releasing hormone-Pseudomonas aeruginosaexotoxin 40 (LHRH-PE40) and LHRH binding to LHRH receptor(LHRHR) on the membrane surface of human liver cancerHEPG cells.METHODS: LHRH was labeled by using 12sI with enzymaticreaction. The affinity and receptor volume of LHP, H-PE40and LHRH binding to LHRHR on the membrane surface ofhuman liver cancer cells were measured with radioligandreceptor assay.RESULTS: The specific activity of LHRH labeled with 12sIwas 2.7x 104 kBq/l^L, and its radiochemical purity reachedto 99.2-99.7%. The binding of 12sI to LHRH was maximalfor 240 min in the warm cultivation, and this binding wasstabilized. The inhibiting rates of ~2SI-LHRH and LHRH onthe proliferation of human liver cancer HEPG cells werenot significantly different. On the basis of the saturationcurve of ~2SI-LHRH binding to the membrane LHRHR of HEPGcells, ~2SI-LHRH of lx10s cpm was selected for radioligandreceptor assay. The affinity constants (Kd) of LHRH-PE40and LHRH binding to the membrane LHRHR of HEPG cellswere 0.43~0.12 nmol/L and 4.86~ 1.47 nmol/L, respectively,and their receptor volumes were 0.37~0.15 l^mol/g and0.42=I=0.13 l^mol/g, respectively. The binding of LHRH-PE40to the membrane protein of normal liver cells was notobserved.CONCLUSION: The recombinant toxin LHRH-PE40 binding tothe membrane surface of LHRHR of human liver cancer HEPGcells was very strong, while the specific binding of it to normalliver cells was not observed. The results provide an importantexperimental basis for the clinical application of LHRH-PE.
文摘The effects of two intraperitoneal injections of human chorionic gonadotropin (HCG) or [ D-ALA6, pro9- N- ethylamide]-luteinizing hormone releasing hormone analogue (LHRH-A), respectively, and the combined effect of LHRH-A with pimozide (PIM), a dopamine antagonist, on ovulation in female mud skipper were investigated. A high rate of ovulation (83.3%)was observed after injection with HCG(30 I. U./g). Injection of LHRH-A at a total dosage of 2μg/g was 100% effective in inducing ovulation. No ovulations occurred in the group receiving a low dosage of LHRH-A (0.02μg/g). Injection of LHRH-A(0.2μg/g)with PIM (10 or 20 μg/g)induced greater ovulatory response than injection with LHRH-A (0.2μg/g)alone. PIM at total dosages of 0.2μg/g or 2μg/g did not enhance the potency of LHRH-A to stimulate ovulation.
文摘Aim:To investigate the effect of Trx-LHRH, a new GnRH crasis protein, on antibody production and reproductive function in male rats. Methods: Trx-LHRH produced in vitro with a new crasis gene, which crasised Trx gene and GnRH gene together, was used as a vaccine with hydroalaminum base as the adjuvant in adult male SD rats. A second inoculation was performed 5 weeks later. Antibody level was measured by ELISA and androgen level by RIA thereafter. Results: Trx-LHRH successfully induced polycolonal antibody at the level of 1:1 280 - 2 560 four weeks after the first inoculation and 1:2 000 SIX weeks after the enhanced treatment. Testosterone level was reduced significantly (P<0.01). Sperm count was also reduced by Trx-LHRH treatment. However, there was reasonable individual variations. Conclusion: Trx-LHRH can be used as an effective vaccine to induce antibody production; at the same time, it restrains the function of hypothatamas-pituitary-testis axis.
文摘Using fluoromctric technique wc tcstcd ED50 ofhistaminc release by LHRH antagonists.There wcrc 9 LHRH antagonists synthesized by us and tested by this method. The range of ED50 is from 0.65±0.08/μg / ml, to 36.8±2.8 /μg / ml. We selected a LHRH
基金supported by a grant from the National Basic Research Program of China(No.2003CB515304)a grant from the National Natural Science Foundation of China(No.30500535)
文摘In the present study, we investigated the roles of TGF-β signaling pathway in a rat benign prostatic hyperplasia （BPH） model treated with cetrorelix. TGF-β1 and c-Myc expression were measured by qRT-PCR and Western blotting in the proximal and distal region of ventral prostatic lobes, respectively. We observed that treatment with cetrorelix led to a significant reduction of ventral prostate weight in a dose-dependent manner. In the proximal region, after cetrorelix treatment, the expression of TGF-β1 was dramatically increased （P〈0.05）, while the expression of c-Myc was significantly decreased （P〈0.05）. In comparison with the control group, the cetrorelix groups had more TUNEL-positive cells. Our findings strongly suggest that the TGF-β signaling pathway may be one of the major causes responsible for prostate volume reduction in BPH rats after cetrorelix treatment.
