Role of site-directed mutagenesis and adjuvants in the stability and potency of anthrax protective antigen

Anthrax is a zoonotic infection caused by the gram-positive, aerobic, spore-forming bacterium Bacillus anthracis. Depending on the origin of the infection, serious health problems or mortality is possible. The virulence of B. anthracis is reliant on three pathogenic factors, which are secreted upon infection: protective antigen (PA), lethal factor (LF), and edema factor (EF). Systemic illness results from LF and EF entering cells through the formation of a complex with the heptameric form of PA, bound to the membrane of infected cells through its receptor. The currently available anthrax vaccines have multiple drawbacks, and recombinant PA is considered a promising second-generation vaccine candidate. However, the inherent chemical instability of PA through Asn deamidation at multiple sites prevents its use after long-term storage owing to loss of potency. Moreover, there is a distinct possibility of B. anthracis being used as a bioweapon; thus, the developed vaccine should remain efficacious and stable over the long-term. Second-generation anthrax vaccines with appropriate adjuvant formulations for enhanced immunogenicity and safety are desired. In this article, using protein engineering approaches, we have reviewed the stabilization of anthrax vaccine candidates that are currently licensed or under preclinical and clinical trials. We have also proposed a formulation to enhance recombinant PA vaccine potency via adjuvant formulation.     

黄芩苷改善肿瘤缺氧微环境抑制乳腺癌移植瘤的生长＊

目的 通过观察黄芩苷对乳腺癌组织毛细血管通透性（Capillary Vessel Permeability，CVP）、赖氨酰氧化酶（Lysyl Oxidase，LOX）及血清丙二醛（Malondialdehyde，MDA）等相关指标的影响，探讨黄芩苷可能的抗乳腺癌作用机制。方法 通过在裸鼠皮下接种MDA-MB-231乳腺癌细胞株建立乳腺癌移植瘤模型；实验分为模型组、黄芩苷组、阿霉素组、黄芩苷+阿霉素组。于接种第7日开始：黄芩苷组每天灌胃黄芩苷水溶液（100 mg·kg^-1），连续14天；阿霉素组每3天腹腔注射一次阿霉素（5mg·kg^-1），共用药5次；模型组每天灌胃生理盐水（10 mL·kg^-1），连续14天；黄芩苷+阿霉素组每天灌胃黄芩苷水溶液（100 mg·kg^-1），连续14天，并每3天腹腔注射一次阿霉素5 mg·kg^-1，共5次。给药期间监测移植瘤体积；紫外可见分光光度计620 nm下测OD值来反映黄芩苷对肿瘤毛细血管通透性的影响；硫代巴比妥酸（Thiobarbituric acid，TBA）比色法检测黄芩苷对裸鼠血清中丙二醛（MDA）的影响；免疫组化染色法检测乳腺癌组织中赖氨酰氧化酶（LOX）的表达情况。结果 ①与模型组比较，黄芩苷组、阿霉素组、黄芩苷+阿霉素组瘤体重量均明显减轻（P<0.05）；黄芩苷+阿霉素组瘤体重量较阿霉素组明显减轻（P<0.05）。②与模型组比较，黄芩苷组降低乳腺癌组织毛细血管的通透性，而阿霉素组则增加肿瘤组织中毛细血管的通透性，差异均具有统计学意义（P<0.05）。③黄芩苷组能够明显抑制荷瘤裸鼠血清MDA的表达，与模型组比较差异有显著统计学意义（P<0.01）；阿霉素组促进MDA的表达，与模型组比较差异有统计学意义；黄芩苷+阿霉素组促进MDA的表达，与模型组比较差异无统计学意义。④与模型组比较，黄芩苷组裸鼠乳腺癌组织LOX的表达显著下调（P<0.01），阿霉素组肿瘤组织中LOX表达增加（P<0.05）；黄芩苷+阿霉素组能下调裸鼠乳腺癌组织LOX的表达，与模型组比较差异无统计学意义。结论 黄芩苷能够明显抑制乳腺癌移植瘤的生长，其机制可能与黄芩苷降低肿瘤组织毛细血管的通透性，抑制裸鼠血清中丙二醛、乳腺癌组织中赖氨酰氧化酶的表达，从而改变肿瘤缺氧微环境有关。     

Capillary leak syndrome: A rare cause of acute respiratory distress syndrome

Capillary leak syndrome (CLS) is a rare clinical syndrome associated with significant morbidity and mortality. Intensive care and supportive therapy constitute the mainstay of the treatment, along with judicious use of crystalloids and colloids such as dextran and starch during the leak phase. The advantages of proning, steroids, and intravenous immunoglobins are worth contemplating in patients with such a presentation. Extracorporeal membrane oxygenation appears to be an excellent strategy to surmount the impediments of the leak and post leak phase of CLS, especially in patients with severe or refractory hypoxemia.     

中药栀子本草考证

栀子临床应用广泛,且历史悠久,为更好地利用栀子资源,通过梳理历代文献与现代研究,对栀子名称、品种、产地、炮制、功效等方面进行考证,发现栀子命名主要以形态、色泽为依据,一定程度上反映了栀子的特征,并且存在与水栀子混用"黄栀"之名的情况。古代文献中记载的栀子产地最早为南阳,但产量不高,受人口迁移以及生产力发展的影响,栀子的产地主要转移到南方,且以南方产者为佳。在历代本草文献中记载入药用的栀子为山栀子,水栀子不堪入药,但在《伤寒论》等古代方书中有"肥栀子""大栀子"入药用的记载,通过后世对相应方剂的记载,确定二者应为水栀子,为水栀子的临床应用提供了有力依据。之所以与本草文献记载有所差异,应与《雷公炮炙论》和《伤寒论》的年代不同,栀子的产量及采收能力有所差距,且本草书重规范,择优入药,方书注重实用等特点有关,并且根据现代实验研究表明山栀子与水栀子化学成分组成、含量相似,具有相同的利胆作用。此外,系统整理了历代本草与方书中栀子的炮制方法和功效,并进行了分析,为栀子属资源的进一步研究与利用提供了参考和借鉴。     

Interstitial lung diseases in children

Interstitial lung disease (ILD) in children (chILD) is a heterogeneous group of rare respiratory disorders that are mostly chronic and associated with high morbidity and mortality. The pathogenesis of the various chILD is complex and the diseases share common features of inflammatory and fibrotic changes of the lung parenchyma that impair gas exchanges. The etiologies of chILD are numerous. In this review, we chose to classify them as ILD related to exposure/environment insults, ILD related to systemic and immunological diseases, ILD related to primary lung parenchyma dysfunctions and ILD specific to infancy. A growing part of the etiologic spectrum of chILD is being attributed to molecular defects. Currently, the main genetic mutations associated with chILD are identified in the surfactant genes SFTPA1, SFTPA2, SFTPB, SFTPC, ABCA3 and NKX2-1. Other genetic contributors include mutations in MARS, CSF2RA and CSF2RB in pulmonary alveolar proteinosis, and mutations in TMEM173 and COPA in specific auto-inflammatory forms of chILD. However, only few genotype-phenotype correlations could be identified so far. Herein, information is provided about the clinical presentation and the diagnosis approach of chILD. Despite improvements in patient management, the therapeutic strategies are still relying mostly on corticosteroids although specific therapies are emerging. Larger longitudinal cohorts of patients are being gathered through ongoing international collaborations to improve disease knowledge and targeted therapies. Thus, it is expected that children with ILD will be able to reach the adulthood transition in a better condition.     

A validated capillary electrophoretic method for the determination of indacaterol and its application to a pharmaceutical preparation

Indacaterol is a new inhaled ultra-long acting ${\beta }_2$-agonist. It has been recently approved in the European Union for the treatment of chronic obstructive pulmonary disease. This paper reports, for the first time, a method for the determination and validation of Indacaterol (IND) using an internal standard in capsules. Capillary electrophoretic separation was performed on an uncoated fused-silica capillary (50 cm effective length, 75 μm i.d.) and background electrolyte composed of 20 mmol L^?1 of sodium tetraborate buffer, 15% (v/v) methanol (pH = 10.0) with the application of 20 kV of potential; 10 s at 5 × 10³ N m^?2 (50 mbar) of injection time; and wavelength of 200 nm and 25 °C of temperature. The linearity was evaluated in the range of 4.90 × 10^?6 mol L^?1 (2.50 μg mL^?1) and 3.94 × 10^?5 mol L^?1 (20.00 μg mL^?1), with R = 0.9993 for inter-day. LOD and LOQ values were 2.18 × 10^?8 mol L^?1 (0.011 μg mL^?1) and 7.25 × 10^?8 mol L^?1 (0.037 μg mL^?1) for inter-day, respectively. The precision values were 0.50–1.06% for intra-day and 2.12% for inter-day as RSD%. The accuracy was tested by the standard addition method with the recovery values being between 98.79 and 99.09 as percentages with RSD% interval of 0.01–0.80. The developed method was validated according to ICH guidelines. Indacaterol was successfully determined in Arcapta^® capsule dosage form by the validated CE method with a relative error of 0.28%. The result was within the requirements of the USP 34-NF29. Therefore, the validated method may be used for the determination of Indacaterol in its capsules in quality control laboratories.     

中药配方颗粒一种新调剂模式的探索与实践

目的：探索门诊中药房中药配方颗粒调剂新模式,提升调剂工作质量,保障调剂药品安全。方法：按PDCA循环管理计划、实施、检查、处理4个步骤对中药配方颗粒调剂模式进行探索与实践。结果：将袋装配方颗粒或散装配方颗粒现有通用调剂模式改进为新的一种智能配发模式,利用智能发药设备实现机器调配辅以药品信息码扫描再复核为主的调剂模式,实施后调配一张常见数量、帖数处方的平均时间由原先的每张3.5 min缩短为0.6 min,平均复核时间由每张1.5 min缩短为0.5 min,显著提高中药配方颗粒的调剂效率和准确率,同时避免散装配方颗粒机器调剂后设备有残余粉末的现象,保障用药安全。结论：新的智能调剂模式可行性强,尤其适用于中药配方颗粒处方量大的医院门诊中药房。     

低渗丙酮酸钠口服补液盐对50%总体表面积烫伤大鼠血管通透性、胃肠及脏器功能的影响

目的探讨低渗丙酮酸钠口服补液盐对严重烧伤大鼠血管通透性、胃肠及脏器功能的影响。 方法选择雄性SD大鼠80只,建立50%总体表面积(TBSA)Ⅲ度烫伤模型,按随机数字表法将大鼠分为4组：烫伤不补液组(NR组)、低渗丙酮酸钠口服补液盐组(PR组)、低渗枸橼酸钠口服补液盐组(CR组)、假烫伤对照组(SR组),每组20只。NR组、PR组及CR组采用96 ℃水浴浸泡大鼠背部15 s、双下肢15 s、腹部8 s,造成50%TBSAⅢ度烫伤模型(经病理切片证实,为严重烧伤);SR组采用37 ℃水浴浸泡相同时间。PR组、CR组于伤后即刻开始采用灌胃输入的方法口服补液,补液量和速度均依据Parkland公式,即每1%TBSA补液4 mL/kg,伤后第1个8 h补补液量的一半,之后16 h补另一半,每0.5 h均严格按照计算所得补液量灌注。SR组自由饮水,NR组不予口服补液。观察烫伤后8、24 h各组大鼠各脏器组织含水率、脏器功能指标变化及胃肠功能变化。数据比较采用单因素方差分析、t检验及χ²检验。 结果(1)伤后8、24 h,4组大鼠的脏器组织(心脏、肝脏、肺、肾脏和肠)含水率比较,差异均有统计学意义(P值均小于0.05);与NR比较,其余3组大鼠脏器组织含水率均降低,差异均有统计学意义(P值均小于0.05)。伤后8 h,PR组心脏、肝脏、肺、肾脏和肠的组织含水率分别为(75.66±1.21)%、(72.83±1.12)%、(75.91±1.24)%、(77.67±1.17)%、(79.16±1.01)%,均低于CR组(79.48±1.25) %、(74.16±1.12) %、(78.87±0.88) %、(79.39±1.19)%、(81.23±0.86)%,差异均有统计学意义(t＝3.698、4.368、5.112、5.287、4.257,P值均小于0.05);伤后24 h,PR组心脏、肝脏、肺、肾脏和肠的组织含水率分别为(71.78±1.08)%、(66.89±1.11)%、(71.42±1.18)%、(71.64±1.17)%、(73.91±1.03)%,均低于CR组(77.12±1.22)%、(71.13±1.09)%、(75.81±1.14)%、(76.78±1.15)%、(78.42±0.94)%,差异均有统计学意义(t＝4.165、4.572、4.981、4.653、5.017,P值均小于0.05)。(2)伤后8、24 h 4组大鼠各脏器功能指标[磷酸肌酸激酶同工酶(CK-MB)、血清谷丙转氨酶(ALT)、肌酐]比较,差异均有统计学意义(P值均小于0.05);与NR组比较,PR组和CR组大鼠伤后8、24 h CK－MB、ALT、肌酐均明显下降,差异均有统计学意义(P值均小于0.05); 伤后8 h,PR组CK-MB、ALT和肌酐分别为(2575.6±165.1)U/L、(270.3±61.2)U/L、(46.1±6.4)μmol/L,均低于CR组(3949.4±165.5)U/L、(542.6±60.1)U/L、(66.7±6.8)μmol/L,差异均有统计学意义(t＝2.396、5.465、5.146,P值均小于0.05);伤后24 h,PR组CK-MB、ALT和肌酐分别为(1652.8±167.8)U/L、(226.9±12.1)U/L、(38.2±4.8)μmol/L,均低于CR组(3247.2±121.2)U/L、(418.1±10.9)U/L、(51.1±5.4)μmol/L,差异均有统计学意义(t＝2.382、4.957、4.060,P值均小于0.05)。(3)伤后8、24 h,PR组胃排空率(85.1±1.4)%、(91.2±1.8)%,均高于CR组(45.7±1.8)%、(66.1±1.4)%,差异均有统计学意义(χ²＝37.327,38.421,P值均小于0.05)。(4)伤后8、24 h,PR组肠吸收总量(20.07±0.78) 、(44.07±2.54) mL,均高于CR组(14.81±0.69)、(31.53±1.62) mL,差异均有统计学意义(χ²＝4.716、5.217,P值均小于0.05)。 结论低渗丙酮酸钠口服补液盐能显著改善严重烫伤大鼠血管通透性、胃肠及脏器功能,可能对严重烫伤大鼠的复苏提供一定的治疗作用。