雄蚕蛾精液中氨基酸成分的提取分离及鉴定

雄蚕蛾乙醇提取液经离子交换树脂分离，氨基酸分析仪测定，确定含有１６种人体必需的氨基酸．     

HCV CE2融合蛋白在家蚕培养细胞、蚕幼虫中的表达及其初步应用

目的:高效表达HCV CE2融合蛋白并进行初步应用. 方法:将2a型HCV全长CE2基因的cDNA重组于质粒pBacPAK8,构建重组转移载体pBacPAK-CE2,与经线性化修饰的家蚕核型多角体病毒(BmNPV)DNA共转染家蚕培养细胞株,构建重组病毒. 双酶切及PCR鉴定重组病毒基因组中目的片段的表达. 重组病毒感染BmN细胞株及五龄家蚕幼虫后,SDS-PAGE分析细胞培养上清、细胞抽提物及幼虫体液样品中,HCV CE2融合蛋白的特异性条带. 并用间接ELISA法初步检测表达产物的生物活性. 结果:双酶切及PCR鉴定重组病毒基因组含有约1.6 kb的目的片段,重组病毒感染后的BmN细胞培养上清、细胞抽提物及幼虫体液样品中,均可见一Mr约90×103的特异性条带;用间接ELISA检测证明表达产物具有较好的免疫原性. 结论:HCV CE2融合基因在家蚕培养细胞及蚕幼虫中获得了高效表达,并具有生物活性,为进一步进行疫苗的研究及临床诊断试剂的开发奠定基础.     

Familial Anaphylaxis after Silkworm Ingestion

Herein, we present a case of anaphylaxis in multiple family members after ingesting silkworms, an Asian delicacy. While food allergies, including anaphylaxis are unfortunately common, there are no previous reports of multiple family members suffering an anaphylactic reaction after eating silkworms. In addition, both family members required multiple doses of epinephrine and eventually an epinephrine infusion to improve their blood pressures. All interventions, including the epinephrine infusions, were started by emergency medical services (EMS) with on-line medical direction. Both the reaction and the required treatment are not extensively documented in the medical literature.     

基因工程人丁酰胆碱酯酶的纯化及其在体外对琥珀酰胆碱的解毒作用(英文)

目的　开发丰富又安全的人丁酰胆碱酯酶来源 ,并检测其解毒效率。方法　家蚕血淋巴液中重组人丁酰胆碱酯酶 (rhBChE)的纯化采用批量硫酸铵分段分离和普鲁卡因酰胺 脂糖凝胶 4B亲和层析技术。结果　聚丙烯酰胺凝胶电泳纯的rhBChE从血淋巴液中的总收率为 6 6 % ,酶比活性达 715mmol·min- 1·g- 1蛋白。rhBChE在 - 2 0℃含 2 0 %甘油及0 .0 2 %NaN3 的 10mmol·L- 1磷酸盐缓冲液 (pH 7.4 )中存放 1年 ,酶活性无明显变化。 18只小鼠ip事先与rhBChE孵温的 1.5LD的琥珀酰胆碱 ,不出现任何中毒症状和体征 ,全部存活 ;而琥珀酰胆碱对照组 ,18只小鼠均剧烈抽搐并在 2～ 5min内窒息死亡。结论　实验结果说明 ,从感染家蚕幼虫血淋巴液中纯化rhBChE的方法可靠 ,基因工程rhBChE在琥珀酰胆碱中毒危象的治疗中有潜在应用前景。     

Complementation of the exoS gene in the pvdE pyoverdine synthesis gene-deficient mutant of Pseudomonas aeruginosa results in recovery of the pvdE gene-mediated penetration through the intestinal epithelial cell barrier but not the pvdE-mediated virulence in silkworms

Translocation of endogenous Pseudomonas aeruginosa from the colonized intestinal tract is an important pathogenic phenomenon. Comparative genome hybridization analysis of high virulent and low virulent strains allowed us to identify bacterial genes that are associated with bacterial translocation from gut in infected hosts. Here we focused on the pvdE pyoverdine synthesis gene among the identified bacterial genes, showing that the pvdE gene is required for bacterial penetration through epithelial cell monolayers and for bacterial translocation from gut to hemolymph in infected silkworms. We next revealed that mRNA expression level of the exoS gene in a pvdE-deficient mutant (ΔpvdE) after incubation with Caco-2 cells was greatly reduced as compared with that in the wild-type strain. The pvdE- and exoS-complemented ΔpvdE strains (ΔpvdE/pvdE and ΔpvdE/exoS) showed recovery of the ability of bacterial penetration through Caco-2 cell monolayers and of the ability of bacterial translocation from gut to hemolymph in infected silkworms. However, there were differences between the ability of ΔpvdE/pvdE and ΔpvdE/exoS to kill silkworms after intestinal infection and to replicate in hemolymph following direct injection into the hemolymph: ΔpvdE/pvdE could kill silkworms after intestinal infection and could replicate in hemolymph to levels similar to those of the wild-type strain, but ΔpvdE/exoS could not. Taken together, our results suggest that the virulence of the wild-strain mediated by the pvdE gene is the result of the ability to both penetrate through the intestinal epithelial cell barrier depending on ExoS and to replicate in hemolymph independently of ExoS.     

保留卧蚕双平面眼袋整复术和传统眼袋整复手术的临床效果

分析保留卧蚕双平面眼袋整复术治疗眼袋的临床效果。方法 选取南通市康美整形医院整 形科2022年6月-2023年6月收治的106例眼袋患者为研究对象,采用随机数字表法分为传统组和新式组, 各53例。传统组予以传统眼袋整复手术治疗,新式组予以保留卧蚕双平面眼袋整复术治疗,比较两组临床疗 效、满意度、并发症发生情况及保留显露卧蚕比例。结果 新式组治疗总有效率高于传统组（P＜0.05）;新 式组满意度高于传统组（P＜0.05）;新式组术后并发症发生率低于传统组（P ＜0.05）;新式组术后保留显 露卧蚕比例为81.13%,高于传统组的20.75%（P ＜0.05）。结论 对眼袋患者应用保留卧蚕的双平面眼袋整 复术效果较佳,可提高患者满意度,且安全性较高,值得临床应用。     

Encapsulation of Sesbania grandiflora extract in polymeric micelles to enhance its solubility,stability, and antibacterial activity

Clinical applications of Sesbania grandiflora bark extract (SGE) are limited because of its poor water solubility and stability. SGE was loaded in micelles of Pluronics. In vitro and in vivo antibacterial and toxicity tests were investigated using broth dilution and silkworm model. Aqueous solubility of SGE was improved by these micelles. Activity and toxicity of SGE loaded micelles were dependent on type and concentration of Pluronics. The micelles composed of 1:3 SGE to Pluronic F68 (SGE-PF68-13) showed small size (24.95?±?0.34?nm), narrow PdI (<0.2), high entrapment efficiency (99.63?±?0.19%) and negative zeta potential (?41.53?±?0.15?mV). Stability of SGE in SGE-PF68-13 was 10 times higher than the unentrapped SGE. SGE-PF68-13 showed a dose dependent activity and significantly higher therapeutic effect than the unentrapped SGE. It is concluded that encapsulation of SGE in Pluronic micelles can enhance SGE solubility, stability, and antibacterial activity. SGE-PF68-13 is suitable for further study in mammalian animals.