A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Bo...A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Boom et al. (1990). By modifying the protocol, plant genome DNA could be extracted easily from dormant buds, mature leaves, and other parts of plant. Our results showed that the purified DNA was of high purity and could be analyzed by PCR. Furthermore, this CTAB-silica method took much less time for a successful DNA purification process compared to the traditional methods (CTAB and SDS). By our method, the suitable DNA can be extracted and purified from over 10 plant samples by one person in an hour.展开更多
Objective To clarify the influence on component and pharmacological action of Astragalus polysaccharides(APS) as complementary therapeutic agents prepared by different extraction and purification techniques.Methods Co...Objective To clarify the influence on component and pharmacological action of Astragalus polysaccharides(APS) as complementary therapeutic agents prepared by different extraction and purification techniques.Methods Components of APS prepared by different extraction and purification techniques were analyzed,and these APS were used for synergy and attenuation of chemotherapy,radiotherapy treatment with H22 liver cancer and Lewis lung cancer of tumor-bearing mice,and also used for the regulation of immune function to immunosuppression mice.Results Experimental data were analyzed by means of statistical method to get pharmaco-result:A3(extracted by microwave assistance and purified by membrane separation)>A4(extracted by refluxing and purified by membrane separation)>A1(extracted by refluxing and no purification)≈A2(extracted by microwave assistance and no purification).There were no significant differences on pharmacodynamic action between A1 and A2.However,compared with A1 and A2,it was worth noting that A3 and A4 exhibited good pharmacodynamic action.Then A3-in and A4-in,the samples in dialyzer after dialysis,were separated and purified to get homogeneous APS,which were the principal constituents of APS in dialyzer,with the molecular weight(Mw) of 7669 and 14142 determined by HPGPC,respectively.The average Mw of APS outside of the dialyzer,A3-out was 3102 and A4-out 3256,which were the main compositions of A3 and A4,accounted for 79.63% and 53.92%,respectively.Conclusion APS with Mw about 5000 Da exhibit better antitumor effect and immunological activity.Refluxing,microwave assistance extractions,and membrane enrichment techniques bring different cases on Mw distribution,components and pharmacodynamic action,and obviously exhibit relationship among component,Mw distribution,and pharmacological action.展开更多
The therapeutic efficacy of traditional Chinese medicine has been widely acknowledged due to its extensive history of clinical effectiveness.However,the precise active components underlying each prescription remain in...The therapeutic efficacy of traditional Chinese medicine has been widely acknowledged due to its extensive history of clinical effectiveness.However,the precise active components underlying each prescription remain incompletely understood.Polysaccharides,as a major constituent of water decoctions—the most common preparation method for Chinese medicinals—may provide a crucial avenue for deepening our understanding of the efficacy principles of Chinese medicine and establishing a framework for its modern development.The structural complexity and diversity of Chinese herbal polysaccharides present significant challenges in their separation and analysis compared to small molecules.This paper aims to explore the potential of Chinese herbal polysaccharides efficiently by briefly summarizing recent advancements in polysaccharide chemical research,focusing on methods of acquisition,structure elucidation,and quality control.展开更多
Purified lipopolysaccharide(LPS)was first obtained from fish-derived Aeromonas hydrophila(A.hydrophila)by hot phenol-water combined with enzymatic hydrolysis.Subsequently,different concentrations of LPS(0,0.2,0.4,0.8,...Purified lipopolysaccharide(LPS)was first obtained from fish-derived Aeromonas hydrophila(A.hydrophila)by hot phenol-water combined with enzymatic hydrolysis.Subsequently,different concentrations of LPS(0,0.2,0.4,0.8,and 1.2 mg/mL)were injected intraperitoneally into Qihe crucian carp(Carassius auratus var.Qihe)with a dose of 3μL/g body weight to evaluated their immunostimulatory activity by detecting the expression of immune-related genes in the spleen,head kidney,and hepatopancreas.The results showed that the average extraction rate of LPS was about 1.063%based on the precipitation weight of wet bacteria,of which the polysaccharide content was 9.39%.The silver staining band of LPS after SDS-PAGE electrophoresis showed a ladder distribution typically characterized by smooth-type LPS.Protein and nucleic acid electrophoresis results showed very low amounts of residual protein and nucleic acid,and high-performance liquid chromatography(HPLC)results showed that the purity of the extracted A.hydrophila LPS was similar to that of high-purity commercial LPS.Pro-inflammatory cytokine genes such as IL-1,IL-8,TNF-α,and IL-6 were more sensitive to LPS stimulation than anti-inflammatory factors such as IL-10 and TGF-β.After 2 h of injection,their expression levels in the spleen,head kidney,and hepatopancreas were significantly increased(P<0.05),with the 0.8 and 1.2 mg/mL injection groups showing the most significant increase.The gene expression of IL-10 and TGF-βshowed slight fluctuations in response to A.hydrophila LPS stimulation(P<0.05).In conclusion,high-purity LPS was extracted from A.hydrophila derived from fish by modified hot phenol-water method,it demonstrating that LPS product could significantly regulate the innate immunity of Qihe crucian carp,with the most effective vaccination dose being 2 mg/kg/BW.These findings provide useful information for future research on endotoxin vaccines.展开更多
Objective To develop an efficient method to isolate and purify the main components isoaloeresin D and aloin from Aloe vera for its industrial production.Methods High-speed counter-current chromatography was used to is...Objective To develop an efficient method to isolate and purify the main components isoaloeresin D and aloin from Aloe vera for its industrial production.Methods High-speed counter-current chromatography was used to isolate isoaloeresin D and aloin in a one-step separation from dried crude extract of A.vera.The biphasic solvent system composed of hexane-ethyl acetate-acetone-water(0.2:5:1.5:5) was used at a flow rate of 1.0 mL/min,while the lipophilic phase was selected as the mobile phase and the apparatus was rotated at 840 r/min.The effluent was detected at 254 nm.Results Isoaloeresin D(53.1 mg) and aloin(106.9 mg) were separated from the crude extract(384.7 mg) with the purities of 98.6% and 99.5%,respectively.Conclusion HSCCC is a powerful technique for isolation and separation of chemical composition from aloe.展开更多
Rose is a kind of favorite ornamental plant. This article briefly introduced the cultivation and the use of rose around the world both in ancient time and nowadays. Today, volatile oil becomes the mainstream of the ro...Rose is a kind of favorite ornamental plant. This article briefly introduced the cultivation and the use of rose around the world both in ancient time and nowadays. Today, volatile oil becomes the mainstream of the rose industry. People pay attention to the effect of volatile oil; meanwhile, they speed up their research on extracting volatile oil and the ingredients.展开更多
Octacosanol is a widely distributed natural higher aliphatic alcohol that can be isolated and purified from rice bran,sugarcane,beeswax,insect wax,etc.Octacosanol exerts various biological effects,including anti-fatig...Octacosanol is a widely distributed natural higher aliphatic alcohol that can be isolated and purified from rice bran,sugarcane,beeswax,insect wax,etc.Octacosanol exerts various biological effects,including anti-fatigue,anti-hypoxia,antioxidant,anti-inflammatory,antitumor,etc.Meanwhile,it has the effects of regulating the body's immune function and energy metabolism and has potential benefits for cardiovascular disease,cerebrovascular disorders,diabetes,Parkinson's disease,and others.Octacosanol is primarily responsible for regulating multiple signaling pathways,such as AMPK,PI3K/Akt,and MAPK/NF-κB,to achieve different physiological functions.This review systemically summarized the progress in characterization,extraction and purification,biological functions,molecular mechanisms,and bioavailability of octacosanol.This study will provide a reference for many investigators to further explore the physiological functions of higher aliphatic alcohols and apply them as supplements in functional foods.展开更多
文摘A new method CTAB-silica for DNA extraction and purification from the leaves and buds of Castanea mollissima and Ginkgo biloba was tested. The method is based on the silica-based purification protocol developed by Boom et al. (1990). By modifying the protocol, plant genome DNA could be extracted easily from dormant buds, mature leaves, and other parts of plant. Our results showed that the purified DNA was of high purity and could be analyzed by PCR. Furthermore, this CTAB-silica method took much less time for a successful DNA purification process compared to the traditional methods (CTAB and SDS). By our method, the suitable DNA can be extracted and purified from over 10 plant samples by one person in an hour.
基金The National Key Scientific and Technological Project in 11th Five-year Plan (No:2009ZX09301-007)
文摘Objective To clarify the influence on component and pharmacological action of Astragalus polysaccharides(APS) as complementary therapeutic agents prepared by different extraction and purification techniques.Methods Components of APS prepared by different extraction and purification techniques were analyzed,and these APS were used for synergy and attenuation of chemotherapy,radiotherapy treatment with H22 liver cancer and Lewis lung cancer of tumor-bearing mice,and also used for the regulation of immune function to immunosuppression mice.Results Experimental data were analyzed by means of statistical method to get pharmaco-result:A3(extracted by microwave assistance and purified by membrane separation)>A4(extracted by refluxing and purified by membrane separation)>A1(extracted by refluxing and no purification)≈A2(extracted by microwave assistance and no purification).There were no significant differences on pharmacodynamic action between A1 and A2.However,compared with A1 and A2,it was worth noting that A3 and A4 exhibited good pharmacodynamic action.Then A3-in and A4-in,the samples in dialyzer after dialysis,were separated and purified to get homogeneous APS,which were the principal constituents of APS in dialyzer,with the molecular weight(Mw) of 7669 and 14142 determined by HPGPC,respectively.The average Mw of APS outside of the dialyzer,A3-out was 3102 and A4-out 3256,which were the main compositions of A3 and A4,accounted for 79.63% and 53.92%,respectively.Conclusion APS with Mw about 5000 Da exhibit better antitumor effect and immunological activity.Refluxing,microwave assistance extractions,and membrane enrichment techniques bring different cases on Mw distribution,components and pharmacodynamic action,and obviously exhibit relationship among component,Mw distribution,and pharmacological action.
基金supported by the Science and Technology Development Fund,Macao SAR (Nos.0075/2022/A and028/2022/ITP)the Zhuhai Science and Technology Plan Project in the Social Development Field (No.2220004000117)the University of Macao (Nos.MYRG-GRG2023-00082-ICMS-UMDF/CPG2024-00011-ICMS)。
文摘The therapeutic efficacy of traditional Chinese medicine has been widely acknowledged due to its extensive history of clinical effectiveness.However,the precise active components underlying each prescription remain incompletely understood.Polysaccharides,as a major constituent of water decoctions—the most common preparation method for Chinese medicinals—may provide a crucial avenue for deepening our understanding of the efficacy principles of Chinese medicine and establishing a framework for its modern development.The structural complexity and diversity of Chinese herbal polysaccharides present significant challenges in their separation and analysis compared to small molecules.This paper aims to explore the potential of Chinese herbal polysaccharides efficiently by briefly summarizing recent advancements in polysaccharide chemical research,focusing on methods of acquisition,structure elucidation,and quality control.
基金supported by the General Program of National Natural Science Foundation of China(32373149).
文摘Purified lipopolysaccharide(LPS)was first obtained from fish-derived Aeromonas hydrophila(A.hydrophila)by hot phenol-water combined with enzymatic hydrolysis.Subsequently,different concentrations of LPS(0,0.2,0.4,0.8,and 1.2 mg/mL)were injected intraperitoneally into Qihe crucian carp(Carassius auratus var.Qihe)with a dose of 3μL/g body weight to evaluated their immunostimulatory activity by detecting the expression of immune-related genes in the spleen,head kidney,and hepatopancreas.The results showed that the average extraction rate of LPS was about 1.063%based on the precipitation weight of wet bacteria,of which the polysaccharide content was 9.39%.The silver staining band of LPS after SDS-PAGE electrophoresis showed a ladder distribution typically characterized by smooth-type LPS.Protein and nucleic acid electrophoresis results showed very low amounts of residual protein and nucleic acid,and high-performance liquid chromatography(HPLC)results showed that the purity of the extracted A.hydrophila LPS was similar to that of high-purity commercial LPS.Pro-inflammatory cytokine genes such as IL-1,IL-8,TNF-α,and IL-6 were more sensitive to LPS stimulation than anti-inflammatory factors such as IL-10 and TGF-β.After 2 h of injection,their expression levels in the spleen,head kidney,and hepatopancreas were significantly increased(P<0.05),with the 0.8 and 1.2 mg/mL injection groups showing the most significant increase.The gene expression of IL-10 and TGF-βshowed slight fluctuations in response to A.hydrophila LPS stimulation(P<0.05).In conclusion,high-purity LPS was extracted from A.hydrophila derived from fish by modified hot phenol-water method,it demonstrating that LPS product could significantly regulate the innate immunity of Qihe crucian carp,with the most effective vaccination dose being 2 mg/kg/BW.These findings provide useful information for future research on endotoxin vaccines.
基金Science and Technology Project of Zhuhai (PC20051072),2005
文摘Objective To develop an efficient method to isolate and purify the main components isoaloeresin D and aloin from Aloe vera for its industrial production.Methods High-speed counter-current chromatography was used to isolate isoaloeresin D and aloin in a one-step separation from dried crude extract of A.vera.The biphasic solvent system composed of hexane-ethyl acetate-acetone-water(0.2:5:1.5:5) was used at a flow rate of 1.0 mL/min,while the lipophilic phase was selected as the mobile phase and the apparatus was rotated at 840 r/min.The effluent was detected at 254 nm.Results Isoaloeresin D(53.1 mg) and aloin(106.9 mg) were separated from the crude extract(384.7 mg) with the purities of 98.6% and 99.5%,respectively.Conclusion HSCCC is a powerful technique for isolation and separation of chemical composition from aloe.
文摘Rose is a kind of favorite ornamental plant. This article briefly introduced the cultivation and the use of rose around the world both in ancient time and nowadays. Today, volatile oil becomes the mainstream of the rose industry. People pay attention to the effect of volatile oil; meanwhile, they speed up their research on extracting volatile oil and the ingredients.
基金supported by the[National Natural Science Foundation of China]under Grant[number 31571874][the Grain-oil Process and Quality Control 2011 Collaborative and Innovative Grant from Hunan Province]under Grant[2013,number 448]+1 种基金[the Key Project of the Education Department of Hunan Province]under Grant[number 16A228,13A124][the Graduate Innovative Research Project of Hunan province and Central South University of Forestry and Technology]under Grant[number CX20200699,CX20201018].
文摘Octacosanol is a widely distributed natural higher aliphatic alcohol that can be isolated and purified from rice bran,sugarcane,beeswax,insect wax,etc.Octacosanol exerts various biological effects,including anti-fatigue,anti-hypoxia,antioxidant,anti-inflammatory,antitumor,etc.Meanwhile,it has the effects of regulating the body's immune function and energy metabolism and has potential benefits for cardiovascular disease,cerebrovascular disorders,diabetes,Parkinson's disease,and others.Octacosanol is primarily responsible for regulating multiple signaling pathways,such as AMPK,PI3K/Akt,and MAPK/NF-κB,to achieve different physiological functions.This review systemically summarized the progress in characterization,extraction and purification,biological functions,molecular mechanisms,and bioavailability of octacosanol.This study will provide a reference for many investigators to further explore the physiological functions of higher aliphatic alcohols and apply them as supplements in functional foods.
