Hearing loss is the most frequent sensory disorder involving a multitude of factors, and at least 50% of cases are due to genetic etiology. To further characterize the molecular etiology of heating loss in the Chinese...Hearing loss is the most frequent sensory disorder involving a multitude of factors, and at least 50% of cases are due to genetic etiology. To further characterize the molecular etiology of heating loss in the Chinese population, we recruited a total of 135 unrelated patients with nonsyndromic sensorineural hearing loss (NSHL) for mutational screening of GJB2, GJB3, GJB6, SLC26A4, SLC26A5 IVS2-2A〉G and mitochondrial 12SrRNA, tRNASer(t/CN) by PCR amplification and direct DNA sequencing. The carrier frequencies of deafness-causing mutations in these patients were 35.55% in GJB2, 3.70% in GJB6, 15.56% in SLC26A4 and 8.14% in mitochondrial 12SrRNA, respectively. The results indicate the necessity of genetic screening for mutations of these causative genes in Chinese population with nonsyndromic heating loss.展开更多
Objective: To study the inhibition of proliferation of breast cancer by small interfering RNA(siRNA) targeting human prolactin (hPRLR) and the underlying mechanisms. Methods:The siRNA targeting hPRLR was chemica...Objective: To study the inhibition of proliferation of breast cancer by small interfering RNA(siRNA) targeting human prolactin (hPRLR) and the underlying mechanisms. Methods:The siRNA targeting hPRLR was chemically synthesized and transfected into MCF-7 cells, the expression of hPRLR was analyzed by real-time quantitive PCR, cell growth inhibition was measured with MTT assay, cell cycle of the transfected cells was examined by flow cytometry, meanwhile, expression of cyclin D1 was tested by semi-quantitative RT-PCR, Results:24 h after transfection with 100 nmol/L siRNA-PRLR, the expression of hPRLR mRNA was suppressed by 65%, cells in G1 phase increased, but cells in S phase decreased. Down regulated hPRLR expression exhibited significant inhibition in cell proliferation. And the expression of cyclin D 1 was down regulated. Conclusion:The results indicate that siRNA-hPRLR is a useful tool for silencing hPRLR expression and inhibiting cell proliferation in breast cancer MCF-7 cell line, and it may be a possible new approach for breast cancer gene therapy.展开更多
Objective: To investigate the frequency of mitochondrial 12S rRNA gene A1555G and 961 insC mutations among Chinese with sensorineural deafness. Methods: Blood samples from 78 sporadic cases with sensorineural deafne...Objective: To investigate the frequency of mitochondrial 12S rRNA gene A1555G and 961 insC mutations among Chinese with sensorineural deafness. Methods: Blood samples from 78 sporadic cases with sensorineural deafness were obtained and DNA was extracted from the leukocytes, then the mitochondrial DNA target fragments were amplified by polymerase chain reaction(PCR). The 1555G mutations were detected by BsmA 1 restriction endonuclease digestion, every fragment was analyzed by sequencing; All the 961 insC mutation were detected by direct sequencing. Results: The percent age of A1555G mutation and mt961C insertion were 6.4% and 2.6% in the hearing-impaired Chinese subjects respectively. Conclusion: A1555G and 961insC mutations in mitochondrial DNA 12S rRNA gene regions may play a role in the pathogenesis of hearing loss in the sporadic cases.展开更多
Overexpression of heat shock protein 27(HSP27)in gastric cancer is correlated with poor clinical prognosis.Melatonin,an endogenous hormone,shows promise in gastric cancer therapy.However,there is limited study on the ...Overexpression of heat shock protein 27(HSP27)in gastric cancer is correlated with poor clinical prognosis.Melatonin,an endogenous hormone,shows promise in gastric cancer therapy.However,there is limited study on the biological activity of HSP27 in response to melatonin treatment.In this study,we show an anti-proliferative action of melatonin on human gastric cancer cell lines BGC-823 and MGC-803.Biochemically,the inhibitory effect of melatonin is accompanied by the upregulation of HSP27 phosphorylation level.Transfection of gastric cancer cells with HSP27-specific siRNA effectively reduces HSP27 phosphorylation and potentiated melatonininduced inhibitory effect on cell proliferation.The reduction of cyclin D1 in melatonin-treated cells is also aggravated by HSP27 depletion.Moreover,melatonin stimulation increases p38 phosphorylation.Pretreatment with p38 inhibitor SB203580 not only remarkably suppresses melatonin-induced HSP27 phosphorylation,but also augment the inhibitory effect of melatonin on cyclin D1 expression as well as cell proliferation.Taken together,our study indicates the protective pathway of p38/HSP27 against melatonin-induced inhibitory effect on gastric cancer cell proliferation,suggesting that combined with p38/HSP27 pathway inhibitor,the therapeutic efficacy of melatonin on gastric cancer may be improved.展开更多
Oxysterol binding protein like 2(OSBPL2), an important regulator in cellular lipid metabolism and transport, was identified as a novel deafness-causal gene in our previous work. To resemble the phenotypic features of ...Oxysterol binding protein like 2(OSBPL2), an important regulator in cellular lipid metabolism and transport, was identified as a novel deafness-causal gene in our previous work. To resemble the phenotypic features of OSBPL2 mutation in animal models and elucidate the potential genotypephenotype associations, the OSBPL2-disrupted Bama miniature(BM) pig model was constructed using CRISPR/Cas9-mediated gene editing, somatic cell nuclear transfer(SCNT) and embryo transplantation approaches, and then subjected to phenotypic characterization of auditory function and serum lipid profiles. The OSBPL2-disrupted pigs displayed progressive hearing loss(HL) with degeneration/apoptosis of cochlea hair cells(HCs) and morphological abnormalities in HC stereocilia, as well as hypercholesterolaemia. High-fat diet(HFD) feeding aggravated the development of HL and led to more severe hypercholesterolaemia. The dual phenotypes of progressive HL and hypercholesterolaemia resembled in OSBPL2-disrupted pigs confirmed the implication of OSBPL2 mutation in nonsydromic hearing loss(NSHL) and contributed to the potential linkage between auditory dysfunction and dyslipidaemia/hypercholesterolaemia.展开更多
基金supported by the Research Grant Award from the National Natural Science Foundation of China (No.31171217)the Open Research Grant of Medical Key Department (No.XF200719) from Jiangsu Province (No.KF200910)Technology Developmental Program from Nanjing Medical University (No.09NJMUM005)
文摘Hearing loss is the most frequent sensory disorder involving a multitude of factors, and at least 50% of cases are due to genetic etiology. To further characterize the molecular etiology of heating loss in the Chinese population, we recruited a total of 135 unrelated patients with nonsyndromic sensorineural hearing loss (NSHL) for mutational screening of GJB2, GJB3, GJB6, SLC26A4, SLC26A5 IVS2-2A〉G and mitochondrial 12SrRNA, tRNASer(t/CN) by PCR amplification and direct DNA sequencing. The carrier frequencies of deafness-causing mutations in these patients were 35.55% in GJB2, 3.70% in GJB6, 15.56% in SLC26A4 and 8.14% in mitochondrial 12SrRNA, respectively. The results indicate the necessity of genetic screening for mutations of these causative genes in Chinese population with nonsyndromic heating loss.
基金This project was supported by Scientific Foundation of Nanjing Medical University(CX2002004)
文摘Objective: To study the inhibition of proliferation of breast cancer by small interfering RNA(siRNA) targeting human prolactin (hPRLR) and the underlying mechanisms. Methods:The siRNA targeting hPRLR was chemically synthesized and transfected into MCF-7 cells, the expression of hPRLR was analyzed by real-time quantitive PCR, cell growth inhibition was measured with MTT assay, cell cycle of the transfected cells was examined by flow cytometry, meanwhile, expression of cyclin D1 was tested by semi-quantitative RT-PCR, Results:24 h after transfection with 100 nmol/L siRNA-PRLR, the expression of hPRLR mRNA was suppressed by 65%, cells in G1 phase increased, but cells in S phase decreased. Down regulated hPRLR expression exhibited significant inhibition in cell proliferation. And the expression of cyclin D 1 was down regulated. Conclusion:The results indicate that siRNA-hPRLR is a useful tool for silencing hPRLR expression and inhibiting cell proliferation in breast cancer MCF-7 cell line, and it may be a possible new approach for breast cancer gene therapy.
文摘Objective: To investigate the frequency of mitochondrial 12S rRNA gene A1555G and 961 insC mutations among Chinese with sensorineural deafness. Methods: Blood samples from 78 sporadic cases with sensorineural deafness were obtained and DNA was extracted from the leukocytes, then the mitochondrial DNA target fragments were amplified by polymerase chain reaction(PCR). The 1555G mutations were detected by BsmA 1 restriction endonuclease digestion, every fragment was analyzed by sequencing; All the 961 insC mutation were detected by direct sequencing. Results: The percent age of A1555G mutation and mt961C insertion were 6.4% and 2.6% in the hearing-impaired Chinese subjects respectively. Conclusion: A1555G and 961insC mutations in mitochondrial DNA 12S rRNA gene regions may play a role in the pathogenesis of hearing loss in the sporadic cases.
基金supported by the Training Programs of Innovation and Entrepreneurship for Undergraduates by Jiangsu Province(201810312018Z)to Jun Du,Chenchen Zhu,Kaiquan Lithe Training Programs of Innovation and Entrepreneurship for Undergraduates by Nanjing Medical University to Jun Du,Yuting Wang
文摘Overexpression of heat shock protein 27(HSP27)in gastric cancer is correlated with poor clinical prognosis.Melatonin,an endogenous hormone,shows promise in gastric cancer therapy.However,there is limited study on the biological activity of HSP27 in response to melatonin treatment.In this study,we show an anti-proliferative action of melatonin on human gastric cancer cell lines BGC-823 and MGC-803.Biochemically,the inhibitory effect of melatonin is accompanied by the upregulation of HSP27 phosphorylation level.Transfection of gastric cancer cells with HSP27-specific siRNA effectively reduces HSP27 phosphorylation and potentiated melatonininduced inhibitory effect on cell proliferation.The reduction of cyclin D1 in melatonin-treated cells is also aggravated by HSP27 depletion.Moreover,melatonin stimulation increases p38 phosphorylation.Pretreatment with p38 inhibitor SB203580 not only remarkably suppresses melatonin-induced HSP27 phosphorylation,but also augment the inhibitory effect of melatonin on cyclin D1 expression as well as cell proliferation.Taken together,our study indicates the protective pathway of p38/HSP27 against melatonin-induced inhibitory effect on gastric cancer cell proliferation,suggesting that combined with p38/HSP27 pathway inhibitor,the therapeutic efficacy of melatonin on gastric cancer may be improved.
基金supported by grants from the National Natural Science Foundation of China (81771000 and 31571302)the Key Research and Development Program of Jiangsu Province (Social Development: BE2016762)+2 种基金the Key Project of Science and Technology Innovation of Nanjing Medical University (2017NJMUCX001)grants from the China Postdoctoral Science Foundation (2016M600431)the Jiangsu Planned Projects for Postdoctoral Research Funds (1601071B)
文摘Oxysterol binding protein like 2(OSBPL2), an important regulator in cellular lipid metabolism and transport, was identified as a novel deafness-causal gene in our previous work. To resemble the phenotypic features of OSBPL2 mutation in animal models and elucidate the potential genotypephenotype associations, the OSBPL2-disrupted Bama miniature(BM) pig model was constructed using CRISPR/Cas9-mediated gene editing, somatic cell nuclear transfer(SCNT) and embryo transplantation approaches, and then subjected to phenotypic characterization of auditory function and serum lipid profiles. The OSBPL2-disrupted pigs displayed progressive hearing loss(HL) with degeneration/apoptosis of cochlea hair cells(HCs) and morphological abnormalities in HC stereocilia, as well as hypercholesterolaemia. High-fat diet(HFD) feeding aggravated the development of HL and led to more severe hypercholesterolaemia. The dual phenotypes of progressive HL and hypercholesterolaemia resembled in OSBPL2-disrupted pigs confirmed the implication of OSBPL2 mutation in nonsydromic hearing loss(NSHL) and contributed to the potential linkage between auditory dysfunction and dyslipidaemia/hypercholesterolaemia.