OBJECTIVE: To evaluate the anti-tumor activity of ethyl acetate fraction(EFA), extracted with ethanol from the root of "Dai-Bai-Jie" in A549 cancer cells and its underlying mechanism.METHODS: "Dai-Bai-J...OBJECTIVE: To evaluate the anti-tumor activity of ethyl acetate fraction(EFA), extracted with ethanol from the root of "Dai-Bai-Jie" in A549 cancer cells and its underlying mechanism.METHODS: "Dai-Bai-Jie" was extracted with 95%ethanol-aqueous(DBJ-1), 50% ethanol-aqueous(DBJ-2), and water(DBJ-3) by reflux method. 95%ethanol-aqueous extract was separated byethyl acetate(EFA) and n-butyl alcohol(DBJ-5), consecutively. The SRB method was used to evaluate the cytotoxic activity. Annexin V-FITC staining was applied to observe the apoptosis and analyze the cell cycle activated by EFA in A549 tumor cell. Western blot was used to detect the apoptosis/related pro-teins expressions. A549 tumor cellsbearing nude mice model was employed to measure the tumor volume, mice weight, and tumor inhibition ratio in order to verify the antitumor activity in vivo.RESULTS: DBJ-1 and EFA showed better cytotoxic activity on A549 tumor cells with IC5025 and 3.5 μg/mL, respectively. EFA can exhibit the proliferation,arrest cell cycle at G0/G1 phase, and induce apoptosis in A549 tumor cells in vitro. The mechanisms of apoptosis induced by EFA may be associated with decreasing Bcl-2 protein expression and increasing p53, Bax, Caspase-3, and Caspase-8 proteins expression. EFA also possessed significant anti-tumor efficacy in nude mice, and little toxicity was observed in the host.CONCLUSION: EAF could induce A549 tumor cells apoptosis and G0/G1 cell cycle arrest. A549 tumor cells apoptosis induced by EAF may be associated with the decrease in the ratio of Bcl-2 and Bax mRNA levels, and increase in the expression of p53,Caspase-3, and Caspase-8 proteins.展开更多
OBJECTIVE:To observe the effects of New Dayuan powder(NDYP)on Methicillin-resistant Staphylococcus aureus(MRSA)biofilms and the embedded bacteria in vitro.METHODS:2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2 H-tetrazol...OBJECTIVE:To observe the effects of New Dayuan powder(NDYP)on Methicillin-resistant Staphylococcus aureus(MRSA)biofilms and the embedded bacteria in vitro.METHODS:2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2 H-tetrazolium-5-carboxanilide(XTT)assays were used to study the effects of NDYP on developing MRSA biofilms:100μL of bacterial culture and100μL drug solution were added to wells of96-well plates.After 24 h of incubation,the plates were washed and XTT-phenazine methyl sulfate(PMS)was added to enable counting of the number of live bacteria in biofilms using a microplate reader.XTT assays were also used to explore the effects of NDYP on mature MRSA biofilms:100μL of bacterial culture were added to wells of 96-well plates.Bacteria were cultured in the plates for 24 h,and then drug solution was added.The plates were cultured for another 24 h,and then XTT-PMS was added to detect the number of live bacteria in the biofilms.Scanning electron microscopy(SEM)was used to observe the effects of NDYP on mature MRSA biofilms:washed and sterilized glass coverslips were added to 24-well plates.Bacterial culture was added.After 24 h of incubation,drug solution was added.After another 24 h of incubation,the samples were observed by SEM.RESULTS:XTT assays showed that the number of live bacteria in both developing and mature MRSA biofilms decreased significantly(P<0.01)after the administration of NDYP.SEM images showed that NDYP could destroy the structure of the bacteria and resulted in uneven thickness of MRSA biofilms.CONCLUSION:In vitro,NDYP has obvious inhibitory effects on the formation of MRSA biofilms and on mature biofilms.展开更多
Functional synaptogenesis and network emergence are signature endpoints of neurogenesis. These behaviors provide higher-order confirmation that biochemical and cellular processes necessary for neurotransmitter release...Functional synaptogenesis and network emergence are signature endpoints of neurogenesis. These behaviors provide higher-order confirmation that biochemical and cellular processes necessary for neurotransmitter release, post-synaptic detection and network propagationof neuronal activity have been properly expressed and coordinated among cells. The development of synaptic neurotransmission can therefore be considered a defining property of neurons. Although dissociated primary neuron cultures readily form functioning synapses and network behaviors in vitro, continuously cultured neurogenic cell lines have historically failed to meet these criteria. Therefore, in vitro-derived neuron models that develop synaptic transmission are critically needed for a wide array of studies, including molecular neuroscience, developmental neurogenesis, disease research and neurotoxicology. Over the last decade, neurons derived from various stem cell lines have shown varying ability to develop into functionally mature neurons. In this review, we will discuss the neurogenic potential of various stem cells populations, addressing strengths and weaknesses of each, with particular attention to the emergence of functional behaviors. We will propose methods to functionally characterize new stem cell-derived neuron(SCN) platforms to improve their reliability as physiological relevant models. Finally, we will review how synaptically active SCNs can be applied to accelerate research in a variety of areas. Ultimately, emphasizing the critical importance of synaptic activity and network responses as a marker of neuronal maturation is anticipated to result in in vitro findings that better translate to efficacious clinical treatments.展开更多
基金Supported by the Grant from the Basic Scientific Research Special of the Central Public Welfare Research Institutes of IMPLADChinese Academy of Medical Sciences:Study on anti-lung cancer chemical constituents and mechanism of"Dai-Bai-Jie"(No.YZYN1501)Study on the resources of"Dai-Bai-Jie"(No.YZYN-10-02)
文摘OBJECTIVE: To evaluate the anti-tumor activity of ethyl acetate fraction(EFA), extracted with ethanol from the root of "Dai-Bai-Jie" in A549 cancer cells and its underlying mechanism.METHODS: "Dai-Bai-Jie" was extracted with 95%ethanol-aqueous(DBJ-1), 50% ethanol-aqueous(DBJ-2), and water(DBJ-3) by reflux method. 95%ethanol-aqueous extract was separated byethyl acetate(EFA) and n-butyl alcohol(DBJ-5), consecutively. The SRB method was used to evaluate the cytotoxic activity. Annexin V-FITC staining was applied to observe the apoptosis and analyze the cell cycle activated by EFA in A549 tumor cell. Western blot was used to detect the apoptosis/related pro-teins expressions. A549 tumor cellsbearing nude mice model was employed to measure the tumor volume, mice weight, and tumor inhibition ratio in order to verify the antitumor activity in vivo.RESULTS: DBJ-1 and EFA showed better cytotoxic activity on A549 tumor cells with IC5025 and 3.5 μg/mL, respectively. EFA can exhibit the proliferation,arrest cell cycle at G0/G1 phase, and induce apoptosis in A549 tumor cells in vitro. The mechanisms of apoptosis induced by EFA may be associated with decreasing Bcl-2 protein expression and increasing p53, Bax, Caspase-3, and Caspase-8 proteins expression. EFA also possessed significant anti-tumor efficacy in nude mice, and little toxicity was observed in the host.CONCLUSION: EAF could induce A549 tumor cells apoptosis and G0/G1 cell cycle arrest. A549 tumor cells apoptosis induced by EAF may be associated with the decrease in the ratio of Bcl-2 and Bax mRNA levels, and increase in the expression of p53,Caspase-3, and Caspase-8 proteins.
基金Startup Fund for scientific researchFujian Medical University (Grant No. 2017XQ010)+1 种基金Beijing Medical and Health Foundation (Grant No. B183023)Natural Science Foundation of Fujian (Grant No. 2022J011011),China。
基金Support by the Fundamental Scientific Research Funds for China Academy of Chinese Medical Sciences(Independent Topic Selection Project)
文摘OBJECTIVE:To observe the effects of New Dayuan powder(NDYP)on Methicillin-resistant Staphylococcus aureus(MRSA)biofilms and the embedded bacteria in vitro.METHODS:2,3-Bis-(2-methoxy-4-nitro-5-sulfophenyl)-2 H-tetrazolium-5-carboxanilide(XTT)assays were used to study the effects of NDYP on developing MRSA biofilms:100μL of bacterial culture and100μL drug solution were added to wells of96-well plates.After 24 h of incubation,the plates were washed and XTT-phenazine methyl sulfate(PMS)was added to enable counting of the number of live bacteria in biofilms using a microplate reader.XTT assays were also used to explore the effects of NDYP on mature MRSA biofilms:100μL of bacterial culture were added to wells of 96-well plates.Bacteria were cultured in the plates for 24 h,and then drug solution was added.The plates were cultured for another 24 h,and then XTT-PMS was added to detect the number of live bacteria in the biofilms.Scanning electron microscopy(SEM)was used to observe the effects of NDYP on mature MRSA biofilms:washed and sterilized glass coverslips were added to 24-well plates.Bacterial culture was added.After 24 h of incubation,drug solution was added.After another 24 h of incubation,the samples were observed by SEM.RESULTS:XTT assays showed that the number of live bacteria in both developing and mature MRSA biofilms decreased significantly(P<0.01)after the administration of NDYP.SEM images showed that NDYP could destroy the structure of the bacteria and resulted in uneven thickness of MRSA biofilms.CONCLUSION:In vitro,NDYP has obvious inhibitory effects on the formation of MRSA biofilms and on mature biofilms.
文摘Functional synaptogenesis and network emergence are signature endpoints of neurogenesis. These behaviors provide higher-order confirmation that biochemical and cellular processes necessary for neurotransmitter release, post-synaptic detection and network propagationof neuronal activity have been properly expressed and coordinated among cells. The development of synaptic neurotransmission can therefore be considered a defining property of neurons. Although dissociated primary neuron cultures readily form functioning synapses and network behaviors in vitro, continuously cultured neurogenic cell lines have historically failed to meet these criteria. Therefore, in vitro-derived neuron models that develop synaptic transmission are critically needed for a wide array of studies, including molecular neuroscience, developmental neurogenesis, disease research and neurotoxicology. Over the last decade, neurons derived from various stem cell lines have shown varying ability to develop into functionally mature neurons. In this review, we will discuss the neurogenic potential of various stem cells populations, addressing strengths and weaknesses of each, with particular attention to the emergence of functional behaviors. We will propose methods to functionally characterize new stem cell-derived neuron(SCN) platforms to improve their reliability as physiological relevant models. Finally, we will review how synaptically active SCNs can be applied to accelerate research in a variety of areas. Ultimately, emphasizing the critical importance of synaptic activity and network responses as a marker of neuronal maturation is anticipated to result in in vitro findings that better translate to efficacious clinical treatments.
