The nucleotide sequence deduced from the amino acid sequence of the scorpion insectotoxin AaIT was chemically synthesized and was expressed in Escherichia coli. The authenticity of this in vitro expressed peptide was ...The nucleotide sequence deduced from the amino acid sequence of the scorpion insectotoxin AaIT was chemically synthesized and was expressed in Escherichia coli. The authenticity of this in vitro expressed peptide was confirmed by N-terminal peptide sequencing. Two groups of bioassays, artificial diet incorporation assay and contact insecticidal effect assay, were carried out separately to verify the toxicity of this recombinant toxin. At the end of a 24 h experimental period, more than 60% of the testing diamondback moth (Plutella xylostella) larvae were killed in both groups with LC50 value of 18.4 microM and 0.70 microM respectively. Cytotoxicity assay using cultured Sf9 insect cells and MCF-7 human cells demonstrated that the toxin AaIT had specific toxicity against insect cells but not human cells. Only 0.13 microM recombinant toxin was needed to kill 50% of cultured insect cells while as much as 1.3 microM toxin had absolutely no effect on human cells. Insect cells produced obvious intrusions from their plasma membrane before broken up. We infer that toxin AaIT bind to a putative sodium channel in these insect cells and open the channel persistently, which would result in Na+ influx and finally cause destruction of insect cells.展开更多
The management of chronic wounds in diabetes remains challenging due to the complexity of impaired wound healing,delayed healing,susceptibility to infection,and elevated risk of reopening,highlighting the need for eff...The management of chronic wounds in diabetes remains challenging due to the complexity of impaired wound healing,delayed healing,susceptibility to infection,and elevated risk of reopening,highlighting the need for effective chronic wound management with innovative approaches such as multifunctional hydrogels.Here,we have produced HA-DA@rhCol hydrogels consisting of dopamine-modified hyaluronic acid and recombinant human collagen type-III(rhCol)by oxidative coupling of the catechol group using the H_(2)O_(2)/HRP catalytic system.The post-reactive hydrogel has a good porous structure,swelling rate,reasonable degradation,rheological and mechanical properties,and the catechol group and dopamine impart to the hydrogel tissue adhesiveness,antioxidant capacity,and excellent photothermal effects leading to superior in vitro antimicrobial activity.In addition,the ability of rhCol to confer hydrogels to promote angiogenesis and wound repair has also been investigated.Cytotoxicity and hemolysis tests demonstrated the good biocompatibility of the hydrogel.Wound closure,collagen deposition and immunohistochemical examination confirmed the ability of the hydrogel to promote diabetic wound healing.In summary,the adhesive hemostatic antioxidative hydrogel with rhCol to promote wound healing in diabetic rat is an excellent chronic wound dressing.展开更多
Thr extract of E. containing recombinant human interferon- (rhIFN-) with 7.0 mol/L guanidine hydrochloride (Gu . HCl) was directly injected into a column of reverse phase liquid chromatography (RPLC) to separate and p...Thr extract of E. containing recombinant human interferon- (rhIFN-) with 7.0 mol/L guanidine hydrochloride (Gu . HCl) was directly injected into a column of reverse phase liquid chromatography (RPLC) to separate and purify rhIFN- with acetic acid-water as mobile phase. Gu I-ICI and most impure proteins can be separated by this way. Compared with the usual dilution method, the bioactivity recovery of the purified rhIFN- was found to be over 500%. In addition, compared to common organic solvents employed ill RPLC, acetic acid has higher freezing point, and therefore, it is easy to concentrate the aim-protein by freeze-drying when acetic acid-water is used as mobile phase ill RPLC.展开更多
目的综合分析抗人乳头瘤病毒(human papillomavirus,HPV)生物蛋白敷料联合藤甲酸氯化钠和重组人干扰素α2b(recombinant human interferonα-2b,rhIFN-α2b)治疗子宫颈鳞状上皮内病变HPV感染的效果。方法选取2022年9月—2023年3月牡丹...目的综合分析抗人乳头瘤病毒(human papillomavirus,HPV)生物蛋白敷料联合藤甲酸氯化钠和重组人干扰素α2b(recombinant human interferonα-2b,rhIFN-α2b)治疗子宫颈鳞状上皮内病变HPV感染的效果。方法选取2022年9月—2023年3月牡丹江医学院附属红旗医院妇科收治的100例子宫颈鳞状上皮内病变HPV感染患者为研究对象,采用随机数表法分为对照组和研究组,各50例。对照组采用rhIFN-α2b栓剂治疗,研究组在对照组的基础上采用抗HPV生物蛋白敷料以及藤甲酸氯化钠进行治疗。观察并比较两组HPV转阴情况、菌群恢复情况、不良反应发生情况以及治疗后白介素-4(interleukin-4,IL-4)、白介素-12(interleukin-12,IL-12)和干扰素-γ(interferon gamma,IFN-γ)水平。结果研究组的HPV转阴率(90.00%)高于对照组(72.00%),差异有统计学意义(χ^(2)=5.263,P<0.05)。研究组的菌群恢复情况、不良反应发生情况以及治疗后的IL-4、IL-12、IFN-γ水平均优于对照组,差异有统计学意义(P<0.05)。结论在子宫颈鳞状上皮内病变HPV感染中使用抗HPV生物蛋白敷料、rhIFN-α2b联合藤甲酸氯化钠具有较为积极的意义,能够提高患者预后。展开更多
基金This work was supported by a grant from 863High Technology Program,Chinese Ministry of Sci-ence and Technology
文摘The nucleotide sequence deduced from the amino acid sequence of the scorpion insectotoxin AaIT was chemically synthesized and was expressed in Escherichia coli. The authenticity of this in vitro expressed peptide was confirmed by N-terminal peptide sequencing. Two groups of bioassays, artificial diet incorporation assay and contact insecticidal effect assay, were carried out separately to verify the toxicity of this recombinant toxin. At the end of a 24 h experimental period, more than 60% of the testing diamondback moth (Plutella xylostella) larvae were killed in both groups with LC50 value of 18.4 microM and 0.70 microM respectively. Cytotoxicity assay using cultured Sf9 insect cells and MCF-7 human cells demonstrated that the toxin AaIT had specific toxicity against insect cells but not human cells. Only 0.13 microM recombinant toxin was needed to kill 50% of cultured insect cells while as much as 1.3 microM toxin had absolutely no effect on human cells. Insect cells produced obvious intrusions from their plasma membrane before broken up. We infer that toxin AaIT bind to a putative sodium channel in these insect cells and open the channel persistently, which would result in Na+ influx and finally cause destruction of insect cells.
基金supported by grants from National Natural Science Foundation of China No.82172206Zhejiang Provincial Medical and Healthy Science Foundation of China(No.2023RC183 and 2024KY110).
文摘The management of chronic wounds in diabetes remains challenging due to the complexity of impaired wound healing,delayed healing,susceptibility to infection,and elevated risk of reopening,highlighting the need for effective chronic wound management with innovative approaches such as multifunctional hydrogels.Here,we have produced HA-DA@rhCol hydrogels consisting of dopamine-modified hyaluronic acid and recombinant human collagen type-III(rhCol)by oxidative coupling of the catechol group using the H_(2)O_(2)/HRP catalytic system.The post-reactive hydrogel has a good porous structure,swelling rate,reasonable degradation,rheological and mechanical properties,and the catechol group and dopamine impart to the hydrogel tissue adhesiveness,antioxidant capacity,and excellent photothermal effects leading to superior in vitro antimicrobial activity.In addition,the ability of rhCol to confer hydrogels to promote angiogenesis and wound repair has also been investigated.Cytotoxicity and hemolysis tests demonstrated the good biocompatibility of the hydrogel.Wound closure,collagen deposition and immunohistochemical examination confirmed the ability of the hydrogel to promote diabetic wound healing.In summary,the adhesive hemostatic antioxidative hydrogel with rhCol to promote wound healing in diabetic rat is an excellent chronic wound dressing.
基金This work was supported by the National Natural Science Foundation of China!(29675017,39880003)
文摘Thr extract of E. containing recombinant human interferon- (rhIFN-) with 7.0 mol/L guanidine hydrochloride (Gu . HCl) was directly injected into a column of reverse phase liquid chromatography (RPLC) to separate and purify rhIFN- with acetic acid-water as mobile phase. Gu I-ICI and most impure proteins can be separated by this way. Compared with the usual dilution method, the bioactivity recovery of the purified rhIFN- was found to be over 500%. In addition, compared to common organic solvents employed ill RPLC, acetic acid has higher freezing point, and therefore, it is easy to concentrate the aim-protein by freeze-drying when acetic acid-water is used as mobile phase ill RPLC.
文摘目的综合分析抗人乳头瘤病毒(human papillomavirus,HPV)生物蛋白敷料联合藤甲酸氯化钠和重组人干扰素α2b(recombinant human interferonα-2b,rhIFN-α2b)治疗子宫颈鳞状上皮内病变HPV感染的效果。方法选取2022年9月—2023年3月牡丹江医学院附属红旗医院妇科收治的100例子宫颈鳞状上皮内病变HPV感染患者为研究对象,采用随机数表法分为对照组和研究组,各50例。对照组采用rhIFN-α2b栓剂治疗,研究组在对照组的基础上采用抗HPV生物蛋白敷料以及藤甲酸氯化钠进行治疗。观察并比较两组HPV转阴情况、菌群恢复情况、不良反应发生情况以及治疗后白介素-4(interleukin-4,IL-4)、白介素-12(interleukin-12,IL-12)和干扰素-γ(interferon gamma,IFN-γ)水平。结果研究组的HPV转阴率(90.00%)高于对照组(72.00%),差异有统计学意义(χ^(2)=5.263,P<0.05)。研究组的菌群恢复情况、不良反应发生情况以及治疗后的IL-4、IL-12、IFN-γ水平均优于对照组,差异有统计学意义(P<0.05)。结论在子宫颈鳞状上皮内病变HPV感染中使用抗HPV生物蛋白敷料、rhIFN-α2b联合藤甲酸氯化钠具有较为积极的意义,能够提高患者预后。
