Characterization of proteolytic enzymes of the midgut and excreta of the biting fly Stomoxys calcitrans

Proteolytic enzymes were characterized in the midgut and the excreta of the stable fly Stomoxys calcitrans (L) with proteins, synthetic substrates, and inhibitors. Inhibition studies suggested trypsinlike activity in sugar-fed fly midguts, whereas excreta and blood-fed fly guts exhibited other proteases. Trypsinlike activity in midguts removed 20 and 30 h after a blood meal increased from 20% to 50% of the total proteolytic enzymes present. Trypsinlike activity was inhibited with human sera, trypsin-specific inhibitors, and a protein isolated from the stable fly thorax. When human albumin and globulin fractions were incubated with trypsinlike enzymes isolated from the midgut and excreta, the albumin fraction was less inhibitory than the globulin fractions and was readily hydrolyzed by the proteolytic enzymes. These results may indicate that the proteolytic enzymes produce an abortive complex with the globulin fractions of the sera. Such a complex may explain the temporary inhibition of proteolysis by the blood meal. Soybean trypsin inhibitor fed to stable flies caused 50% inhibition in proteolytic activity in the midguts of sugar-fed stable flies and 25% inhibition in the midguts of blood-fed stable flies. Complete inhibition of proteolytic enzyme activity was achieved only in vitro. pH profiles of proteolytic enzyme activity isolated from the excreta of blood-fed stable flies indicated that several proteolytic enzymes were excreted.     

皮下免疫治疗联合沙美特罗替卡松粉吸入剂对中度支气管哮喘缓解期患儿血清Th1/Th2相关细胞因子表达水平的影响

摘要 目的：探讨皮下免疫治疗联合应用沙美特罗替卡松粉吸入剂治疗中度支气管哮喘缓解期患儿的临床疗效及对Th1/Th2相关细胞因子表达水平的影响。方法：选取南方医科大学佛山市妇幼保健院2020年1月～2020年12月收治的中度支气管哮喘缓解期120例患儿,按随机分组原则分为观察组和对照组各60例。对照组予以沙美特罗替卡松粉吸入剂（舒利迭）治疗,早上及晚上各吸1次。观察组在对照组的基础上给予皮下免疫治疗,进行12个月疗程的治疗。对比两组患儿支气管哮喘发作次数以及临床控制率、肺功能变化。检测两组患儿血清Th1相关细胞因子[白细胞介素-2（IL-2）、γ干扰素（IFN-γ）]和Th2相关细胞因子[白细胞介素-4（IL-4）、白细胞介素-6（IL-6）]水平。结果：与治疗前相比,两组患儿治疗6月及治疗12个月后发作次数均显著降低（P＜0.05）,且观察组患儿哮喘发作的次数低于对照组（P＜0.01）;治疗12个月后,观察组患儿的临床控制率显示为90%,显著高于对照组的66.67％（P＜0.05）。治疗6个月后、治疗12个月后,观察组患儿的第1 秒呼气流速（FEV₁）占正常预计值百分比（FEV₁%）及呼气峰流速（PEF）占正常预计值百分比（PEF%）均显著高于对照组（P＜0.01）;而与治疗6个月后相比,两组治疗12个月后上述2项指标均显著升高,且观察组明显高于对照组（P＜0.01）。治疗12个月后,两组患儿血清IL-2、IFN-γ水平均显著升高,且观察组明显高于对照组（P＜0.01）;而两组血清IL-4、IL-6水平均显著下降,且观察组明显低于对照组（P＜0.01）。结论：皮下免疫治疗联合沙美特罗替卡松粉（舒利迭）治疗中度支气管哮喘缓解期患儿的临床疗效确切,患儿支气管哮喘发作次数明显减少,肺功能改善明显,其作用机制可能与调节Th1/Th2相关细胞因子平衡有关。     

Human urine: Epicatechin metabolites and antioxidant activity after cocoa beverage intake

Associations between cocoa consumption in humans, excreted metabolites and total antioxidant capacity (TAC) have been scarcely investigated. The aims of the study were to investigate the epicatechin (( ? )-Ec) metabolites excreted in urine samples after an intake of 40 g of cocoa powder along with the TAC of these urine samples and the relation between both the analyses. Each of the 21 volunteers received two interventions, one with a polyphenol-rich food (PRF) and one with a polyphenol-free food (PFF) in a randomized cross-over study. Urine samples were taken before and during 24 h at 0–6, 6–12 and 12–24 h periods after test intake. The excreted ( ? )-Ec metabolites and the TAC were determined in urine samples by LC-MS/MS and TEAC assay, respectively. The maximum excretion of ( ? )-Ec metabolites and the maximum TAC value were observed in urine samples excreted between 6 and 12 h after PRF consumption. Significance of TAC increase was found in urine samples excreted during 0–6 and 6–12 h (66.6 and 72.67%, respectively, with respect to the 0 h).     

氮源对阿维菌素合成的影响及基于二氧化碳释放速率的阿维菌素发酵调控

中国是世界上最大的也是唯一的阿维菌素原料生产国,但在工业规模生产中与同类型大环内酯类抗生素相比其产量相对偏低。文中通过研究不同氮源对阿维链霉菌生长、代谢的影响,发现氮源在发酵中后期对菌丝活性、菌丝浓度以及阿维菌素B1a的合成都有较为显著的影响。在100 L生物反应器中,于发酵中后期基于二氧化碳释放速率(CER)控制补入酵母粉,效价达到8697mg/L,与原工艺相比,提高了26.9%。这一结论若在实际工业生产中应用,有望带来实际的经济效益。     

四黄散外敷对竹叶青蛇咬伤肿胀的疗效观察

目的探讨四黄散外敷治疗竹叶青蛇咬伤肿胀的临床疗效。方法选择2017年10月~2020年4月我院收治的112例竹叶青蛇咬伤肿胀患者为研究对象,采用随机数字表法将患者分为观察组和对照组,每组56例。对照组采用常规蛇咬伤处理方案,包括清创、引流、注射抗蛇毒血清及对症治疗等。观察组在对照组治疗基础上采用四黄散外敷治疗。观察比较两组患者的有效止痛时间、肿胀开始消退时间、肿胀完全消退时间,并对两组患者治疗7d时的临床疗效进行比较。结果观察组在有效止痛时间、肿胀开始消退时间、肿胀完全消退时间方面均显著低于对照组,两组比较差异具有统计学意义(均P<0.05)。两组治疗后的总体疗效比较,观察组的治愈率为89.3%(50/56),对照组的治愈率为69.6%(39/56),两组比较差异有统计学意义(P<0.05)。结论四黄散外敷治疗竹叶青蛇咬伤肿胀的疗效确切,能有效减轻患者的疼痛感,促进肿胀消退,减轻炎症反应,值得临床应用。     

蛹虫草菌粉胶囊抑制肺部炎症及缓解肺纤维化的研究

传统中药可治疗肺炎。新型急性呼吸综合征冠状病毒2型（SARS-CoV-2）外部囊膜上的S蛋白是决定病毒毒力的关键因素及主要抗原。本研究利用SARS-CoV-2病毒的S蛋白和细菌脂多糖（LPS）诱导的肺炎模型,初步探索了蛹虫草菌粉对肺炎模型的促炎性因子、单核/巨噬细胞和髓源抑制性细胞（MDSCs）以及纤维化的调节作用。研究发现蛹虫草菌粉可降低SARS-CoV-2病毒的S蛋白组和LPS组小鼠血清中TNF-α、IL-6和IL-10的表达,并降低肺组织、肺泡灌洗液和外周血中巨噬细胞的比例,降低外周血和脾脏中MDSC的数量。进一步研究发现,蛹虫草还可降低SARS-CoV-2病毒的S蛋白组和LPS组小鼠肺组织羟基脯氨酸的表达和成纤维细胞的分布,进而减轻小鼠的肺纤维化水平。qRT-PCR实验发现蛹虫草菌粉处理不仅可以降低SARS-CoV-2病毒的S蛋白组小鼠肺组织中TGF-β R1水平,还降低SARS-CoV-2病毒的S蛋白组和LPS组肺组织中smad2的表达。因此认为蛹虫草菌粉可缓解SARS-CoV-2病毒的S蛋白和LPS诱导的肺部炎症和纤维化症状,且纤维化缓解作用可能与TGF-β R1/smad2信号通路相关。     

Comparison of Rapid DNA Extraction Methods Applied to PCR Identification of Medicinal Mushroom Ganoderma spp.

Abstract

Four different DNA extraction methods were used to extract genomic DNA of the medicinal mushroom Lingzhi from its developing stage materials, such as mycelium, dry fruiting body, or sliced and spore powder or sporoderm‐broken spore powder. The DNA samples were analyzed using agarose gel electrophoresis, UV spectrophotometer, and PCR amplification. According to the average yields and purity of DNA, high salt concentrations and low pH methods were the best for DNA extraction. The mycelia and sporoderm‐broken spore powder yielded higher and purer DNA. The method developed could effectively eliminate the influence of the secondary metabolites to DNA extraction. The DNA samples extracted from the developed method could be successfully used for PCR applications.     

Bioactivity of Piper guineense Schum. & Thonn seed and Moringa oleifera Lam. leaf powder against Trogoderma granarium Everts (Coleoptera: Dermestidae)

Laboratory studies were carried out to investigate bioactivity of Piper guineense seeds and Moringa oleifera leaf powders applied singly or in a mixture against larvae and adult Trogoderma granarium Everts in airtight containers. Three levels (0.0 g, 0.5 g, and 1.0 g/20 g groundnut seeds) of the plant powders were used and pirimiphos-methyl was applied at 0.01 g/20 g seeds (recommended dose). Another control consisting of untreated seeds with aerated lids was included in the bioassay. Both larvae and adults were not killed in control with aerated lids throughout the experimental period and larvae were also tolerant to airtight storage conditions. Adults were more susceptible to plant powders than larvae and adult mortality recorded in P. guineense at 1.0 g, 0.5 g and M. oleifera at 1.0 g/20 g seeds were not significantly different from the mortality observed with the recommended dose of Pirimiphos methyl at five days after treatment (DAT). Larval mortality observed in a mixture of both plants (1:1; w/w) caused significantly higher mortality (77.5%) than other treatments at 5 DAT. All treatments (P. guineense and M. oleifera applied singly or in a mixture) were repellant to larvae T. granarium with 60% repellency recorded in the mixture of plants, 50% repellency in P. guineense and 30% repellency in M. oleifera slurry. The water absorption capacity of treated seeds was not affected by treatment with plant powders and ranged from 31.98% to 37.59%.     

The Piperaceae Amides I: Structure of Pipercide,A New Insecticidal Amide from Piper nigrum L.

It was evidenced that mutagenic principles in tryptophan pyrolysate, 3-amino-1,4-dimethyl-5H pyrido(4,3-b) indole and 3-amino-1-methyl-5H pyrido(4,3-b) indole (abbreviated as Trp-P-1 and Trp-P-2, respectively) bind to DNA without activation by rat liver microsomes. The bindings of Trp-P-1 and Trp-P-2 were not random and did not introduce strand scissions into DNA. Trp-P-1 bound more easily than Trp-P-2. The bindings of these mutagenic principles to DNA were concluded by using negatively superhelical simian virus 40 (SV40) DNA from following experimental data. (1) The intensity of ethidium bromide (EtBr)-DNA fluorescence by illumination with UV light and the electrophoretic mobility of superhelical DNA in agarose gel decreased as a function of the amounts of Trp-P-1 and Trp-P-2. (2) In vitro RNA synthesis catalyzed by Escherichia coli DNA-dependent RNA polymerase and nick-translation catalyzed by Escherichia coli DNA polymerase I (Kornberg enzyme) were inhibited significantly on DNA treated with Trp-P-1 and Trp-P-2. (3) The negative superhelicity of SV40 DNA introduces unpaired regions into DNA. These regions can be cleaved by single-strand-specific S1 endonuclease to generate unit length linear duplex molecules. It was found that this S1-sensitivity of DNA decreased by treatment with Trp-P-1. (4) The cleavage patterns of Trp-P-1 treated DNA with five restriction endonucleases were investigated. The protection of the cleavage site by the drug was observed against HincII, HindIII and EcoRII, whereas not against HaeIII and HinfI. These results show that the binding of Trp-P-1 to DNA is not random. Identical results were also obtained in Trp-P-2.

However, the bindings of Trp-P-1 and Trp-P-2 were not so tight, and phenol extraction of the complex dissociated these drugs from DNA.     

粉末包衣技术在药物制剂领域的应用研究进展

粉末包衣技术是薄膜包衣技术发展至今的一个重要分支,其在药物制剂领域的应用优势突出,近年来受到药剂学研究者的广泛关注。分类综述目前应用于药物制剂的几种主要粉末包衣技术,包括属于物理化学法中的凝聚法、溶剂蒸发法和熔融分散法以及物理机械法中的喷雾干燥法、喷雾冷凝法、干法包衣技术和气流悬浮包衣技术,并探讨粉末包衣技术的主要功用,如用于制备缓控释制剂、药物粉末表面改性、改善口服制剂感官效果和提高药物及制剂稳定性等。