Objective: To examine the influence of Glyphaea (G.) brevis twig extract on the mitochondrial dehydrogenase activity, integrity of the tight junctions between adjacent cells, mitochondria, apoptosis, nucleus and expre...Objective: To examine the influence of Glyphaea (G.) brevis twig extract on the mitochondrial dehydrogenase activity, integrity of the tight junctions between adjacent cells, mitochondria, apoptosis, nucleus and expression of inhibin-β, stem cell factor, and androgen binding protein in TM4 Sertoli cells. Methods: TM4 cell line was used in this study as it exhibited properties similar to the Sertoli cells. TM4 Sertoli cells were exposed to G. brevis twig extract (0.1, 1.0, 10.0, 100.0, or 1000.0μg/mL) for 24, 48 and 72 h. Parameters studied included cell viability [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay], mitochondrial membrane potential (tetra methyl rhodamine ethyl ester dye), transepithelial electrical resistance, apoptosis (Annexin V Alexa Fluor?488/propidium iodide assay) and mRNA expression (quantitative reverse transcription polymerase chain reaction). Results: G. brevis twig extract had no cytotoxic impact on cell viability, thus, considerably increasing the activity of mitochondrial dehydrogenase enzyme after 24 and 72 h exposure. Transepithelial electrical resistance values revealed substantial (P<0.05) rise in treated groups, especially after 72 h of treatment. Moreover, there was a significant decrease in mitochondrial depolarization of TM4 Sertoli cells exposed to G. brevis twig extract when compared to controls. In addition, G. brevis twig extract significantly reduced necrosis and apoptosis of TM4 Sertoli cells when compared to control. Nevertheless, fluorescence microscopy revealed that the nuclei were egg-shaped and marked uniformly with consistent cell shape at the middle of the TM4 Sertoli cells. Significant stimulatory effects were observed on mRNA levels of inhibin-β, androgen binding protein and stem cell factor. Conclusions: G. brevis twig extract may increase the secretory roles of TM4 Sertoli cells, cells proliferation, as well as cell-cell tight junction integrity. Thus, G. brevis twig may enhance spermatogenesis.展开更多
AIM: To investigate the expression of lacrimal androgenbinding proteins(ABPs) in mice Pseudomonas aeruginosa(P. aeruginosa) keratitis.METHODS: P. aeruginosa mice model from different gender was developed by intra-stro...AIM: To investigate the expression of lacrimal androgenbinding proteins(ABPs) in mice Pseudomonas aeruginosa(P. aeruginosa) keratitis.METHODS: P. aeruginosa mice model from different gender was developed by intra-stromal injection. The expression of lacrimal ABPs in lacrimal gland specimens from P. aeruginosa keratitis mice was detected by the quantitative polymerase chain reaction(q RT-PCR). Corneal virulence was evaluated based on clinical scores. To study the mechanism of lacrimal ABPs' expression, experimental subjects were pre-treated with 4 E-BP1 inhibitor, and were used to evaluate the expression levels by q RT-PCR.RESULTS: Compared with control groups, the expression of ABPα, ABPη and ABPζ in lacrimal gland from P. aeruginosa keratitis mice had no meaningful changes, while ABPε and ABPδ were significantly higher at 1 d after infection. The expression of ABPδ in lacrimal gland of male mice was higher than female mice, regardless of whether or not P. aeruginosa keratitis occurred. After 4 E-BP1 inhibitor subconjunctival injection or lacrimal injection, the expression of ABPδ and ABPε has no significant change compared with the control group.CONCLUSION: ABPδ and ABPε secreted by mice lacrimal gland may involve in the progress of alleviating the severity of corneal damage in P. aeruginosa keratitis. The expression of ABPδ and ABPε upon P. aeruginosa infection is independent of cap-dependent m RNA translation activated by 4 E-BP1.展开更多
2M NaCl-insoluble fraction of rat ventral prostatechromatin(residual proteins)contain proteins able tointeract specifically with androgen-receptor complex andis,therefore,a part of the acceptor complex.Amongresidual p...2M NaCl-insoluble fraction of rat ventral prostatechromatin(residual proteins)contain proteins able tointeract specifically with androgen-receptor complex andis,therefore,a part of the acceptor complex.Amongresidual proteins,a 97 KDa protein has been found whichbinds signifieantly to a genomic fragment containingan androgen-regulated gene coding for a 22 KDa protein.The biological significance of this binding in androgenaction need to be further studied. A mini-plasmid clone containing 22 KDa proteincoding sequence was cloned into Charon 4A genomiclibrary from which a 5.7 Kb genomic fragment wasisolated,identified by hybridization with a 5’ and a 3’cDNA probes,and shown to contain the 5’ flankingsequence.Restriction enzyme treatment of this fragmentyielded a 4.7 Kb restriction fragment representingthe 5’ upstream region and a 1.0 Kb containing part ofthe coding sequence.Deletion studies indicated that the97 KDa protein bound only to a subclone of about 300 bpsegment.Furthermore,gel shifting experiment supportedits DNA-prptein binding.展开更多
Aim: To study the DHT-binding ability of benign prostatic hypertrophy (BPH) and its etiological relationship with BPH. Methods: Cytosolic and nuclear fractions was obtained from 32 BPH tissues by superpubic prostatect...Aim: To study the DHT-binding ability of benign prostatic hypertrophy (BPH) and its etiological relationship with BPH. Methods: Cytosolic and nuclear fractions was obtained from 32 BPH tissues by superpubic prostatectomy and all the endogenous hormone were removed from the cytosolic and nuclear fractions by ether stripping. The content of the bound 3H-DHT was assayed after the addition of 3H-DHT. Results: The average DHT-binding capacity of the BPH is 0.024 nmol (protein)/g wet tissue. The DHT-binding capacity of the cytosolic and the nuclear fractions were (0.0128±0.0020) nmol/g and (0.0112±0.0059) nmol/g wet tissue, respectively, the difference of the two capacities being insignificant (P>0.05). Conclusion: The DHT-binding capacity of BPH is high and thus facilitates the effect of DHT on BPH.展开更多
目的:对腋臭患者和正常人腋部大汗腺进行形态学观察,探讨大汗腺分泌活动的周期。对两种人群大汗腺组织中雄激素受体(androgen receptor,AR)和大汗腺分泌物气味结合蛋白(apocri ne secreti on odor-bi ndi ng protei n,ASOB)的表达进行检...目的:对腋臭患者和正常人腋部大汗腺进行形态学观察,探讨大汗腺分泌活动的周期。对两种人群大汗腺组织中雄激素受体(androgen receptor,AR)和大汗腺分泌物气味结合蛋白(apocri ne secreti on odor-bi ndi ng protei n,ASOB)的表达进行检测,探讨二者的表达水平及其在腋臭发病中的作用。方法:对腋部皮肤组织进行HE染色,分析大汗腺不同分泌时期的形态和特点。利用免疫组织化学(i mmunohi stochemi stry)和免疫印迹法(eWstern bl otti ng)检测腋臭组与对照组大汗腺组织中AR和ASOB在大汗腺以及不同分泌时期中的表达情况,使用Image-Pro Pl us 5.0和Fl uorChemFC2分析系统对结果进行分析。结果:①大汗腺的分泌活动具有周期性,可分为活跃期和静息期;②腋臭组中AR和ASOB的表达明显高于对照组;③AR和ASOB在大汗腺不同分泌时期的表达差异有统计学意义;④AR和ASOB在两组人群相同分泌时期的表达差异有统计学意义;⑤AR和ASOB在腋臭大汗腺中的表达有正的直线相关关系。结论:AR和ASOB在腋臭患者大汗腺组织中的表达量同时增多,它们可能在腋臭发病的过程中起了重要作用。展开更多
文摘Objective: To examine the influence of Glyphaea (G.) brevis twig extract on the mitochondrial dehydrogenase activity, integrity of the tight junctions between adjacent cells, mitochondria, apoptosis, nucleus and expression of inhibin-β, stem cell factor, and androgen binding protein in TM4 Sertoli cells. Methods: TM4 cell line was used in this study as it exhibited properties similar to the Sertoli cells. TM4 Sertoli cells were exposed to G. brevis twig extract (0.1, 1.0, 10.0, 100.0, or 1000.0μg/mL) for 24, 48 and 72 h. Parameters studied included cell viability [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay], mitochondrial membrane potential (tetra methyl rhodamine ethyl ester dye), transepithelial electrical resistance, apoptosis (Annexin V Alexa Fluor?488/propidium iodide assay) and mRNA expression (quantitative reverse transcription polymerase chain reaction). Results: G. brevis twig extract had no cytotoxic impact on cell viability, thus, considerably increasing the activity of mitochondrial dehydrogenase enzyme after 24 and 72 h exposure. Transepithelial electrical resistance values revealed substantial (P<0.05) rise in treated groups, especially after 72 h of treatment. Moreover, there was a significant decrease in mitochondrial depolarization of TM4 Sertoli cells exposed to G. brevis twig extract when compared to controls. In addition, G. brevis twig extract significantly reduced necrosis and apoptosis of TM4 Sertoli cells when compared to control. Nevertheless, fluorescence microscopy revealed that the nuclei were egg-shaped and marked uniformly with consistent cell shape at the middle of the TM4 Sertoli cells. Significant stimulatory effects were observed on mRNA levels of inhibin-β, androgen binding protein and stem cell factor. Conclusions: G. brevis twig extract may increase the secretory roles of TM4 Sertoli cells, cells proliferation, as well as cell-cell tight junction integrity. Thus, G. brevis twig may enhance spermatogenesis.
基金Supported by the National Natural Science Foundation of China(No.81300730)China Postdoctoral Science Foundation(No.2018M630482)+2 种基金Key Research Project of Shandong Province(No.2018GSF118193)Science and Technology Project of Qingdao(No.19-6-1-39-nsh)Qingdao Outstanding Health Professional Development Fund
文摘AIM: To investigate the expression of lacrimal androgenbinding proteins(ABPs) in mice Pseudomonas aeruginosa(P. aeruginosa) keratitis.METHODS: P. aeruginosa mice model from different gender was developed by intra-stromal injection. The expression of lacrimal ABPs in lacrimal gland specimens from P. aeruginosa keratitis mice was detected by the quantitative polymerase chain reaction(q RT-PCR). Corneal virulence was evaluated based on clinical scores. To study the mechanism of lacrimal ABPs' expression, experimental subjects were pre-treated with 4 E-BP1 inhibitor, and were used to evaluate the expression levels by q RT-PCR.RESULTS: Compared with control groups, the expression of ABPα, ABPη and ABPζ in lacrimal gland from P. aeruginosa keratitis mice had no meaningful changes, while ABPε and ABPδ were significantly higher at 1 d after infection. The expression of ABPδ in lacrimal gland of male mice was higher than female mice, regardless of whether or not P. aeruginosa keratitis occurred. After 4 E-BP1 inhibitor subconjunctival injection or lacrimal injection, the expression of ABPδ and ABPε has no significant change compared with the control group.CONCLUSION: ABPδ and ABPε secreted by mice lacrimal gland may involve in the progress of alleviating the severity of corneal damage in P. aeruginosa keratitis. The expression of ABPδ and ABPε upon P. aeruginosa infection is independent of cap-dependent m RNA translation activated by 4 E-BP1.
文摘2M NaCl-insoluble fraction of rat ventral prostatechromatin(residual proteins)contain proteins able tointeract specifically with androgen-receptor complex andis,therefore,a part of the acceptor complex.Amongresidual proteins,a 97 KDa protein has been found whichbinds signifieantly to a genomic fragment containingan androgen-regulated gene coding for a 22 KDa protein.The biological significance of this binding in androgenaction need to be further studied. A mini-plasmid clone containing 22 KDa proteincoding sequence was cloned into Charon 4A genomiclibrary from which a 5.7 Kb genomic fragment wasisolated,identified by hybridization with a 5’ and a 3’cDNA probes,and shown to contain the 5’ flankingsequence.Restriction enzyme treatment of this fragmentyielded a 4.7 Kb restriction fragment representingthe 5’ upstream region and a 1.0 Kb containing part ofthe coding sequence.Deletion studies indicated that the97 KDa protein bound only to a subclone of about 300 bpsegment.Furthermore,gel shifting experiment supportedits DNA-prptein binding.
文摘Aim: To study the DHT-binding ability of benign prostatic hypertrophy (BPH) and its etiological relationship with BPH. Methods: Cytosolic and nuclear fractions was obtained from 32 BPH tissues by superpubic prostatectomy and all the endogenous hormone were removed from the cytosolic and nuclear fractions by ether stripping. The content of the bound 3H-DHT was assayed after the addition of 3H-DHT. Results: The average DHT-binding capacity of the BPH is 0.024 nmol (protein)/g wet tissue. The DHT-binding capacity of the cytosolic and the nuclear fractions were (0.0128±0.0020) nmol/g and (0.0112±0.0059) nmol/g wet tissue, respectively, the difference of the two capacities being insignificant (P>0.05). Conclusion: The DHT-binding capacity of BPH is high and thus facilitates the effect of DHT on BPH.
文摘目的:对腋臭患者和正常人腋部大汗腺进行形态学观察,探讨大汗腺分泌活动的周期。对两种人群大汗腺组织中雄激素受体(androgen receptor,AR)和大汗腺分泌物气味结合蛋白(apocri ne secreti on odor-bi ndi ng protei n,ASOB)的表达进行检测,探讨二者的表达水平及其在腋臭发病中的作用。方法:对腋部皮肤组织进行HE染色,分析大汗腺不同分泌时期的形态和特点。利用免疫组织化学(i mmunohi stochemi stry)和免疫印迹法(eWstern bl otti ng)检测腋臭组与对照组大汗腺组织中AR和ASOB在大汗腺以及不同分泌时期中的表达情况,使用Image-Pro Pl us 5.0和Fl uorChemFC2分析系统对结果进行分析。结果:①大汗腺的分泌活动具有周期性,可分为活跃期和静息期;②腋臭组中AR和ASOB的表达明显高于对照组;③AR和ASOB在大汗腺不同分泌时期的表达差异有统计学意义;④AR和ASOB在两组人群相同分泌时期的表达差异有统计学意义;⑤AR和ASOB在腋臭大汗腺中的表达有正的直线相关关系。结论:AR和ASOB在腋臭患者大汗腺组织中的表达量同时增多,它们可能在腋臭发病的过程中起了重要作用。
