Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assist...Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.展开更多
Objective： To determine the role of intensity of plum-blossom needle tapping in treating alopecia areata. Methods： The BALB/c mice were randomized into a normal group, a control group, a roller-needle（RN） group, a...Objective： To determine the role of intensity of plum-blossom needle tapping in treating alopecia areata. Methods： The BALB/c mice were randomized into a normal group, a control group, a roller-needle（RN） group, a mild plum-blossom needle（MP） group, and a heavy plum-blossom needle（HP） group. An area of hair was removed by external application of 8% sodium sulfide on BALB/c mice. The hair regrowth, hair follicle changes, and local inflammatory factor changes after cutaneous acupuncture were observed. Results： After treated with sodium sulfide, the hair was completely removed, the local hair follicles reached the catagen phase, and the expressions of interleukin（IL）-1α, IL-1β, IL-2, tumor necrosis factor（TNF）-α, interferon（IFN）-γ, and IL-17 were increased. Mice intervened by RN achieved the same hair growth rating as the controls but with thicker hair shafts; mice in the MP group had incomplete and uneven hair growth but thicker hair shafts; mice in the HP group didn＇t show hair growth. Pathological analysis revealed significant inflammatory infiltration into the local follicle bulbs and increased catagen-phase follicles in the control group, while RN and MP groups showed significantly increased anagen-phase follicles, coarser individual hairs, and obvious hair shafts. Meanwhile, most of the hair follicles in the HP group were in telogen phase and showed obvious surrounding inflammatory infiltration. RN, MP, and HP significantly down-regulated the increased IL-2, IL-1α, IL-1β, TNF-α, and IFN-γ levels（P〈0.05）, but didn＇t notably affect the increased CD34 expression（P〈0.05）. Conclusion： Cutaneous acupuncture with heavy stimulation intensity can inhibit hair growth in hair removal mice, while RN, with the lightest stimulation intensity, is unlikely to affect hair growth but may make hair shafts thicker and follicles larger.展开更多
文摘Objective The detection of RNA single nucleotide polymorphism(SNP)is of great importance due to their association with protein expression related to various diseases and drug responses.At present,splintR ligase-assisted methods are important approaches for RNA direct detection,but its specificity will be limited when the fidelity of ligases is not ideal.The aim of this study was to create a method to improve the specificity of splintR ligase for RNA detection.Methods In this study,a dualcompetitive-padlock-probe(DCPLP)assay without the need for additional enzymes or reactions is proposed to improve specificity of splintR ligase ligation.To verify the method,we employed dual competitive padlock probe-mediated rolling circle amplification(DCPLP-RCA)to genotype the CYP2C9 gene.Results The specificity was well improved through the competition and strand displacement of dual padlock probe,with an 83.26%reduction in nonspecific signal.By detecting synthetic RNA samples,the method demonstrated a dynamic detection range of 10 pmol/L-1 nmol/L.Furthermore,clinical samples were applied to the method to evaluate its performance,and the genotyping results were consistent with those obtained using the qPCR method.Conclusion This study has successfully established a highly specific direct RNA SNP detection method,and provided a novel avenue for accurate identification of various types of RNAs.
基金supported by Shanghai Baoshan Hospital of Integrated Traditional Chinese and Western MedicineNo.201405Excellent TCM Doctor Training Project in the Three-year Traditional Chinese Medicine Action Plan from Shanghai Health and Family Planning Commission,No.ZY3-RCPY-2-2041Youth Medical Talents Training Project from Baoshan District Health and Family Planning Commission of Shanghai,BSWSYQ-2014-A05~~
文摘Objective： To determine the role of intensity of plum-blossom needle tapping in treating alopecia areata. Methods： The BALB/c mice were randomized into a normal group, a control group, a roller-needle（RN） group, a mild plum-blossom needle（MP） group, and a heavy plum-blossom needle（HP） group. An area of hair was removed by external application of 8% sodium sulfide on BALB/c mice. The hair regrowth, hair follicle changes, and local inflammatory factor changes after cutaneous acupuncture were observed. Results： After treated with sodium sulfide, the hair was completely removed, the local hair follicles reached the catagen phase, and the expressions of interleukin（IL）-1α, IL-1β, IL-2, tumor necrosis factor（TNF）-α, interferon（IFN）-γ, and IL-17 were increased. Mice intervened by RN achieved the same hair growth rating as the controls but with thicker hair shafts; mice in the MP group had incomplete and uneven hair growth but thicker hair shafts; mice in the HP group didn＇t show hair growth. Pathological analysis revealed significant inflammatory infiltration into the local follicle bulbs and increased catagen-phase follicles in the control group, while RN and MP groups showed significantly increased anagen-phase follicles, coarser individual hairs, and obvious hair shafts. Meanwhile, most of the hair follicles in the HP group were in telogen phase and showed obvious surrounding inflammatory infiltration. RN, MP, and HP significantly down-regulated the increased IL-2, IL-1α, IL-1β, TNF-α, and IFN-γ levels（P〈0.05）, but didn＇t notably affect the increased CD34 expression（P〈0.05）. Conclusion： Cutaneous acupuncture with heavy stimulation intensity can inhibit hair growth in hair removal mice, while RN, with the lightest stimulation intensity, is unlikely to affect hair growth but may make hair shafts thicker and follicles larger.
