Objecive To check and compare the expression levels of human androgen receptor (AR) mRNA in both normal adult prostate (NAP) and benign prostate hyperplasia (BPH) tissues, and to try to find if BPH results from the a...Objecive To check and compare the expression levels of human androgen receptor (AR) mRNA in both normal adult prostate (NAP) and benign prostate hyperplasia (BPH) tissues, and to try to find if BPH results from the abnormal transcription of ARmRNA in prostate. Methods Expression of the human ARmRNA in 14 paraffin-embedded prostate tissues (4 cases of NAP and 10 cases of BPH) was studied with the direct in situ reverse transcription-polymerase chain reaction (RT-PCR). Quantitative analysis of the ARmRNA products was performed using the image analysis system. Results ①Specific ARmRNA was detected in both NAP and BPH specimens and in both epithelia and interstitial cells. The positive products were relatively densely localized in the cytoplasm of perinuclear zone. ② The intensity of ARmRNA signals in epithelial cells was significantly stronger than that in interstitial cells (P<0.001). However, there was no statistically significant difference in ARmRNA level between NAP and BPH; ③ The heterogeneity of ARmRNA signal and the androgen-independent cells were observed in prostatic epithelia. Stronger positive signals of ARmRNA were shown in a few basal-cell layer (BCL) cells of BPH tissue, but were not found in that of NAP tissue. Conclusion Results of this study show that there is no significant difference in the ARmRNA expression between NAP and BPH groups in both epithelium and interstitial cells. It may indicate that BPH does not result from the ARmRNA transcription in the prostate.展开更多
本实验采用 RT- PCR法探讨大白鼠颌下腺是否存在 Gn RH受体 m RNA,并用原位杂交法对其细胞定位进行了研究。结果显示 RT- PCR可扩增出大白鼠颌下腺 Gn RH受体 m RNA的特异性片段 ,其碱基数与设计的一致。原位杂交发现颌下腺浆液性腺泡...本实验采用 RT- PCR法探讨大白鼠颌下腺是否存在 Gn RH受体 m RNA,并用原位杂交法对其细胞定位进行了研究。结果显示 RT- PCR可扩增出大白鼠颌下腺 Gn RH受体 m RNA的特异性片段 ,其碱基数与设计的一致。原位杂交发现颌下腺浆液性腺泡上皮细胞、颗粒曲管、排泄管及分泌管上皮细胞内有 Gn RH受体 m RNA的杂交信号。信号物质分布于胞质内 ,胞核阴性。上述结果表明大白鼠颌下腺能合成 Gn RH受体 ,颌下腺产生的 Gn RH可作用于颌下腺的靶细胞 ,参与颌下腺生理功能的调节。展开更多
The CCHC-type zinc finger motif has numerous biological activities （such as DNA binding and RNA binding） and can also mediate protein-protein interaction. This article gives a primary report about the human ZCCHC9 g...The CCHC-type zinc finger motif has numerous biological activities （such as DNA binding and RNA binding） and can also mediate protein-protein interaction. This article gives a primary report about the human ZCCHC9 gene. Protein ZCCHC9 contains four CCHC motifs and is highly conserved in humans, mice, and rats. The whole eDNA sequence of the ZCCHC9 gene has been amplified by PCR and a number of plasmids have been constructed for further study. The results show that ZCCHC9 is localized in the nucleus, and especially concentrated in the nueleolus. It is highly expressed in the brain and testicles of the mouse. This has been confirmed by real-time reverse transcription polymerase chain reaction （RT-PCR）. In situ hybridization of the mouse brain indicates that ZCCHC9 is mainly expressed in the cerebral cortex. Reporter gene assay shows that ZCCHC9 suppresses the transcription activities of NF-kappa B and SRE, and may play roles in the Mitogen-Aetivated Protein Kinase （MAPK） signaling transduetion pathway.展开更多
To detect the location of inducible nitric oxide synthetase (iNOS) protein and mRNA in lung during endotoxemia in rabbits Methods Northern blotting was performed before, 1 hour and 5 hours after the intravenous ...To detect the location of inducible nitric oxide synthetase (iNOS) protein and mRNA in lung during endotoxemia in rabbits Methods Northern blotting was performed before, 1 hour and 5 hours after the intravenous administration of lipopolysaccharide (LPS) in rabbits Immuno^histochemical analysis (IA), in situ hybridization and in situ reverse transcription polymerase chain reaction (in situ RT PCR) were also performed in lung sections Results iNOS mRNA expression was found using Northern blotting in lung 5 hours after LPS injection, while it was not found in control The positive stain was found only in macrophages in lung 5 hours after LPS injection by standard hybridization and IA; while by in situ RT PCR, the amplification products were found in macrophages, airway epithelial cells, vascular endothelial cells, smooth muscle cells and leukocytes, in addition to macrophages distributed abundantly throughout the lung The signal was absent in control or samples Conclusions Using an in situ RT PCR technique, iNOS expression was not only observed in macrophages but also in many other kinds of cells in lung during endotoxemia in rabbits This suggests that in situ RT PCR is much more sensitive than in situ hybridization, and can be used to examine genes with low expression展开更多
文摘Objecive To check and compare the expression levels of human androgen receptor (AR) mRNA in both normal adult prostate (NAP) and benign prostate hyperplasia (BPH) tissues, and to try to find if BPH results from the abnormal transcription of ARmRNA in prostate. Methods Expression of the human ARmRNA in 14 paraffin-embedded prostate tissues (4 cases of NAP and 10 cases of BPH) was studied with the direct in situ reverse transcription-polymerase chain reaction (RT-PCR). Quantitative analysis of the ARmRNA products was performed using the image analysis system. Results ①Specific ARmRNA was detected in both NAP and BPH specimens and in both epithelia and interstitial cells. The positive products were relatively densely localized in the cytoplasm of perinuclear zone. ② The intensity of ARmRNA signals in epithelial cells was significantly stronger than that in interstitial cells (P<0.001). However, there was no statistically significant difference in ARmRNA level between NAP and BPH; ③ The heterogeneity of ARmRNA signal and the androgen-independent cells were observed in prostatic epithelia. Stronger positive signals of ARmRNA were shown in a few basal-cell layer (BCL) cells of BPH tissue, but were not found in that of NAP tissue. Conclusion Results of this study show that there is no significant difference in the ARmRNA expression between NAP and BPH groups in both epithelium and interstitial cells. It may indicate that BPH does not result from the ARmRNA transcription in the prostate.
文摘本实验采用 RT- PCR法探讨大白鼠颌下腺是否存在 Gn RH受体 m RNA,并用原位杂交法对其细胞定位进行了研究。结果显示 RT- PCR可扩增出大白鼠颌下腺 Gn RH受体 m RNA的特异性片段 ,其碱基数与设计的一致。原位杂交发现颌下腺浆液性腺泡上皮细胞、颗粒曲管、排泄管及分泌管上皮细胞内有 Gn RH受体 m RNA的杂交信号。信号物质分布于胞质内 ,胞核阴性。上述结果表明大白鼠颌下腺能合成 Gn RH受体 ,颌下腺产生的 Gn RH可作用于颌下腺的靶细胞 ,参与颌下腺生理功能的调节。
基金the National Natural Sci-ence Foundation of China (No. 30470945)Program for Changjiang Scholars and Innovative Research Team in University (No. IRT0445)
文摘The CCHC-type zinc finger motif has numerous biological activities （such as DNA binding and RNA binding） and can also mediate protein-protein interaction. This article gives a primary report about the human ZCCHC9 gene. Protein ZCCHC9 contains four CCHC motifs and is highly conserved in humans, mice, and rats. The whole eDNA sequence of the ZCCHC9 gene has been amplified by PCR and a number of plasmids have been constructed for further study. The results show that ZCCHC9 is localized in the nucleus, and especially concentrated in the nueleolus. It is highly expressed in the brain and testicles of the mouse. This has been confirmed by real-time reverse transcription polymerase chain reaction （RT-PCR）. In situ hybridization of the mouse brain indicates that ZCCHC9 is mainly expressed in the cerebral cortex. Reporter gene assay shows that ZCCHC9 suppresses the transcription activities of NF-kappa B and SRE, and may play roles in the Mitogen-Aetivated Protein Kinase （MAPK） signaling transduetion pathway.
基金ThisstudywassupportedbygrantsfromtheNationalNatural ScienceFoundationofChina (No 395 70 30 4)andHebeiNaturalScienceFoundation
文摘To detect the location of inducible nitric oxide synthetase (iNOS) protein and mRNA in lung during endotoxemia in rabbits Methods Northern blotting was performed before, 1 hour and 5 hours after the intravenous administration of lipopolysaccharide (LPS) in rabbits Immuno^histochemical analysis (IA), in situ hybridization and in situ reverse transcription polymerase chain reaction (in situ RT PCR) were also performed in lung sections Results iNOS mRNA expression was found using Northern blotting in lung 5 hours after LPS injection, while it was not found in control The positive stain was found only in macrophages in lung 5 hours after LPS injection by standard hybridization and IA; while by in situ RT PCR, the amplification products were found in macrophages, airway epithelial cells, vascular endothelial cells, smooth muscle cells and leukocytes, in addition to macrophages distributed abundantly throughout the lung The signal was absent in control or samples Conclusions Using an in situ RT PCR technique, iNOS expression was not only observed in macrophages but also in many other kinds of cells in lung during endotoxemia in rabbits This suggests that in situ RT PCR is much more sensitive than in situ hybridization, and can be used to examine genes with low expression
