青霉素杂质谱分析方法的优化与转换

目的建立青霉素杂质谱HPLC分析方法,并将其转换成UPLC/UHPLC方法。方法以青霉素混合降解溶液为样品;首先分析《中国药典》(2010年版,ChP2010)方法甲醇-缓冲盐二元流动相色谱系统的缺陷;再利用实验设计理念,以响应曲面法(response surface methodology,RSM)的中心组合设计(central composition design,CCD)对色谱系统进行优化,满意度函数法确定最优色谱条件,并优化梯度洗脱条件;最后,利用软件对HPLC方法的流速、进样体积和梯度时间进行几何缩放,并通过色谱柱的选择,将其分别转换为UPLC方法和UHPLC方法。结果新HPLC方法:色谱柱为Capcell Pak C18 MGII(4.6mm×250mm,5μm),流速:1.0m L/min,进样体积:20μL;UPLC方法:色谱柱为Cortecs C18(2.1mm×100mm,1.6μm),流速:0.35m L/min,进样体积:2.0μL;UHPLC方法:Cortecs C18(4.6mm×150mm,2.76μm),流速:0.8mL/min,进样体积:10μL。3种方法的检测波长均为225nm,柱温均为34℃;流动相A均为磷酸盐缓冲液(取磷酸二氢钾10.6g,加水至1000mL,用磷酸调pH至3.4)-甲醇(72:14,V/V),流动相B均为乙腈;均为梯度洗脱,但梯度洗脱表不同。结论 3个色谱系统的分离效果(出峰顺序和个数)相似。新HPLC方法可以分离出更多的降解杂质,并明显改善了青霉素峰的拖尾,缩短了分析时间。     

氦—氖激光照射对青霉素酶阳性金黄色葡萄球菌抑菌作用研究

研究氦－氖激光对青霉素酶阳性金黄色葡萄球菌的抑制作用。方法：采用２４ｍＷ氦－氖激光照射器直接照射青霉亲酶阳性金黄色葡萄球菌液或金葡菌化脓病灶区，再经活菌计数、组织病理、外周血白细胞检测等方法，观测激光照射后金葡菌活性、血清抗体、淋转率及白细胞变化。结果：氦－氖激光照射组金葡菌活性明显降低，化脓病灶缩小，血中白细胞减少，血清抗体及淋转率增高。结论：氦－氖激光照射具有抑制青霉素酶阳性金葡菌增强机体抗感染免疫作用。     

HPLC法测定固定化青霉素酰化酶合成头孢羟氨苄的活力

目的 建立一种HPLC法测定固定化青霉素酰化酶合成头孢羟氨苄活力的方法。方法 酶与其底物7-ADCA及D-对羟基苯甘氨酸甲酯，在20℃的溶液中，搅拌转速为250r/min条件下反应10min，通过测定反应液中头孢羟氨苄的含量，从而换算出合成酶活力的数值。采用十八烷基硅烷键合硅胶为填充剂；流动相：0.02mol/L磷酸二氢钾溶液(用lmol/L氢氧化钾溶液调节pH值至5.0)-甲醇(98:2)；流速：1.0mL/min；柱温：25℃；检测波长：230nm。结果 头孢羟氨苄在0.24~0.39mg/mL范围内线性关系良好(r=1.0000)，回收率均在99%~101%范围内，RSD为0.40%。结论 该测定方法专属性强、准确可靠、重现性好，适用于青霉素酰化酶合成酶活力的检测。     

Increase in penicillin-resistant invasive meningococcal serogroup W ST-11 complex isolates in England

IntroductionInvasive meningococcal disease (IMD) caused by serogroup W meningococci belonging to the ST-11 complex (MenW:cc11) has been increasing globally since the early 2000s. Penicillin resistance among meningococci due to the production of beta-lactamase remains relatively rare. Isolates displaying resistance and reduced susceptibility to penicillin due to alterations in the penA gene (encoding Penicillin Binding Protein 2) are increasingly reported. In 2016, a penicillin-resistant clade of MenW:cc11 isolates with altered penA genes was identified in Australia. More recently, an increase in penicillin-resistant invasive MenW:cc11 isolates was observed in England. Here, we investigate the distribution of penicillin resistance among English invasive MenW:cc11 isolates.MethodsIsolates from IMD cases in England from July 2010 to August 2019 underwent whole genome sequencing and antibiotic susceptibility testing as part of routine surveillance. The PubMLST Neisseria database was used to determine the distribution of penicillin resistance among English MenW:cc11 isolates and to identify other closely related isolates.ResultsTwenty-five out of 897 English invasive MenW:cc11 isolates were resistant to penicillin; identified among six distinct sublineages and a singleton. Expansion of the Australian penicillin-resistant clade included isolates from several new countries as well as 20 English isolates. A newly identified penicillin resistance-associated lineage was also identified among several countries.ConclusionPenicillin resistance among diverse MenW:cc11 isolates is increasing. Surveillance of antibiotic resistance among meningococci is essential to ensure continued effective use.     

青霉素与妇科千金片联合治疗急性盆腔炎临床分析

目的探讨青霉素联合妇科千金片治疗急性盆腔炎的临床疗效。方法 136例急性盆腔炎患者随机分为观察组和对照组各68例,对照组在常规治疗的基础上给予青霉素400×104U静脉滴注,每日2次;观察组在对照组治疗的基础上给予妇科千金片6片口服,每日3次;10~15 d为1个疗程。比较2组的疗效、症状消除时间和血清中肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）和C反应蛋白（CRP）水平。结果观察组总有效率为94.1%,高于对照组的80.9%（P〈0.05）;观察组的体温恢复正常时间、腹痛消失时间和包块消除时间均显著短于对照组,差异有统计学意义（P〈0.01）;2组治疗后血清中TNF-α、IL-6、CRP水平均明显下降（均P〈0.01）,组间比较,差异有统计学意义（P〈0.01）;2组不良反应发生率无明显差别（P〉0.05）。结论青霉素联合妇科千金片治疗急性盆腔炎疗效显著,可以改善患者的免疫功能,安全性好,值得在临床推广。     

Tamoxifen enhances erlotinib-induced cytotoxicity through down-regulating AKT-mediated thymidine phosphorylase expression in human non-small-cell lung cancer cells

Tamoxifen is a triphenylethylene nonsteroidal estrogen receptor (ER) antagonist used worldwide as an adjuvant hormone therapeutic agent in the treatment of breast cancer. However, the molecular mechanism of tamoxifen-induced cytotoxicity in non-small cell lung cancer (NSCLC) cells has not been identified. Thymidine phosphorylase (TP) is an enzyme of the pyrimidine salvage pathway which is upregulated in cancers. In this study, tamoxifen treatment inhibited cell survival in two NSCLC cells, H520 and H1975. Treatment with tamoxifen decreased TP mRNA and protein levels through AKT inactivation. Furthermore, expression of constitutively active AKT (AKT-CA) vectors significantly rescued the decreased TP protein and mRNA levels in tamoxifen-treated NSCLC cells. In contrast, combination treatment with PI3K inhibitors (LY294002 or wortmannin) and tamoxifen further decreased the TP expression and cell viability of NSCLC cells. Knocking down TP expression by transfection with small interfering RNA of TP enhanced the cytotoxicity and cell growth inhibition of tamoxifen. Erlotinib (Tarceva, OSI-774), an orally available small molecular inhibitor of epidermal growth factor receptor (EGFR) tyrosine kinase, is approved for clinical treatment of NSCLC. Compared to a single agent alone, tamoxifen combined with erlotinib resulted in cytotoxicity and cell growth inhibition synergistically in NSCLC cells, accompanied with reduced activation of phospho-AKT and phospho-ERK1/2, and reduced TP protein levels. These findings may have implications for the rational design of future drug regimens incorporating tamoxifen and erlotinib for the treatment of NSCLC.