Quantitative analysis of pheromone‐binding protein specificity

Many pheromones have very low water solubility, posing experimental difficulties for quantitative binding measurements. A new method is presented for determining thermodynamically valid dissociation constants for ligands binding to pheromone‐binding proteins, using β‐cyclodextrin as a solubilizer and transfer agent. The method is applied to LUSH, a Drosophila odorant‐binding protein that binds the pheromone 11‐cis vaccenyl acetate (cVA). Refolding of LUSH expressed in Escherichia coli was assessed by measuring N‐phenyl‐1‐naphthylamine (NPN) binding and Förster resonance energy transfer between LUSH tryptophan 123 (W123) and NPN. Binding of cVA was measured from quenching of W123 fluorescence as a function of cVA concentration. The equilibrium constant for transfer of cVA between β‐cyclodextrin and LUSH was determined from a linked equilibria model. This constant, multiplied by the β‐cyclodextrin‐cVA dissociation constant, gives the LUSH‐cVA dissociation constant: ～100 nM. It was also found that other ligands quench W123 fluorescence. The LUSH‐ligand dissociation constants were determined to be ～200 nM for the silk moth pheromone bombykol and ～90 nM for methyl oleate. The results indicate that the ligand‐binding cavity of LUSH can accommodate a variety ligands with strong binding interactions. Implications of this for the Laughlin, Ha, Jones and Smith model of pheromone reception are discussed.     

芍药苷对醋酸铅诱导海马神经元凋亡及Bcl-2/Bax蛋白表达的影响

目的 探讨芍药苷(paeoniflorin,PF)对醋酸铅诱导海马神经元凋亡及Bcl-2/Bax蛋白表达的影响。方法 分离培养胎鼠海马神经元细胞,细胞免疫荧光染色鉴定纯度。MTT测定海马神经元细胞活力以确定醋酸铅最适造模浓度及时间,同时筛选合适剂量PF干预海马神经元凋亡。依据MTT测定结果,分为空白组、模型组和20,40,80 μmol·L^-1 PF组干预海马神经元细胞,作用24 h后,加醋酸铅染毒,检测细胞色素C (cytochrome C,Cyt-C)含量、线粒体膜电位及细胞内Ca²⁺浓度。流式细胞仪检测细胞凋亡情况,Western blotting测定海马神经元细胞中caspase-3、cleaved-caspase-3、caspase-8、cleaved-caspase-8、caspase-9、cleaved-caspase-9、Bax、Bcl-2蛋白表达水平。结果 细胞免疫荧光染色鉴定结果显示分离培养的细胞为海马神经元细胞,且纯度较高。MTT测定结果显示醋酸铅最适造模浓度及时间为25 μmol·L^-1染毒24 h;PF剂量为20,40,80 μmol·L-1可显著改善海马神经元细胞活性,呈剂量依赖性。与空白组相比,模型组Cyt-C含量、凋亡率、细胞内Ca²⁺浓度显著升高(P<0.01),线粒体膜电位显著降低(P<0.01)。与模型组相比,40 μmol·L^-1 PF组和80 μmol·L^-1 PF组可降低Cyt-C含量、凋亡率、细胞内Ca²⁺浓度(P<0.05或P<0.01),升高线粒体膜电位(P<0.01),20 μmol·L^-1PF组可显著升高线粒体膜电位(P<0.05)。此外,一定剂量的PF可下调cleaved-caspase-3、cleaved-caspase-8、cleaved-caspase-9、Bax蛋白表达,上调Bcl-2蛋白表达。结论 PF可抑制醋酸铅诱导的海马神经元凋亡,可通过调控Bcl-2/Bax蛋白表达发挥神经保护作用。     

醋酸戈舍瑞林缓释植入剂联合腹腔镜手术治疗子宫内膜异位症效果

目的:探讨醋酸戈舍瑞林缓释植入剂联合腹腔镜手术治疗子宫内膜异位症的临床疗效。方法:选择本院就诊的子宫内膜异位症患者80例,随机分为两组各40例,根据r-AFS分期进行腹腔镜手术,在术后1周内观察组醋酸戈舍瑞林缓释植入剂3.6mg皮下注射,每4周1次,连用6个月;对照组口服孕三烯酮2.5mg,每周2次,连用6个月。比较两组临床疗效,术前和术后6个月测定血清血管内皮生长因子(VEGF)、基质金属蛋白酶-9/组织金属蛋白酶抑制物-1(MMP-9/TIMP-1),评定患者视觉模拟评分法(VAS)和健康生活质量调查问卷(SF-36),随访3年术后不良反应和复发情况。结果:治疗有效率对照组(75.0%)低于观察组(92.5%)(P<0.05)。术后6个月时,对照组和观察组血清VEGF(158.41±56.28 ng/L、119.63±47.13ng/L),MMP-9/TIMP-1(1.35±0.25、1.14±0.23),生理健康评分(64.20±12.81分、78.53±13.23分),心理健康评分(69.14±13.26分、80.35±14.59分),VAS评分(1.21±0.46分、0.84±0.39分)比较均有差异(P<0.05);观察组无不良反应发生,术后3年复发率对照组(27.5%)高于观察组(10.0%)(P<0.05)。结论:醋酸戈舍瑞林缓释植入剂联合腹腔镜手术治疗子宫内膜异位症疗效确切,可降低术后血管侵袭指标及复发率,提高生活质量,安全性高。     

中药方剂抗肺纤维化方治疗肺纤维化患者的临床研究

〔摘 要〕 目的：研究中药方剂抗肺纤维化方治疗肺纤维化（PF）患者的疗效。方法：选取福建中医药大学附属 第二人民医院 2020 年 3 月至 2021 年 10 月期间收治的 70 例 PF 患者，采用随机数字表法分为对照组与观察组，各 35 例。对照组患者采用常规西药治疗，观察组患者在常规西药治疗基础上采用中药方剂抗肺纤维化方治疗。比较两 组患者的临床疗效、治疗前及治疗 3 个月后中医证候积分、肺功能指标、肺纤维化指标、圣乔治呼吸疾病问卷（SGRQ）、 高分辨计算机断层扫描（HRCT）评分。结果：观察组患者治疗总有效率为 88.67 %，高于对照组的 62.86 %，差异具 有统计学意义（P ＜ 0.05）。治疗后观察组患者的乏力、呼吸气短、自汗、咳嗽评分均低于对照组，差异具有统计学 意义（P ＜ 0.05）。治疗后观察组患者的第 1 秒用力呼气容积（FEV1）、用力肺活量（FVC）、最大呼气流量（PEF） 均高于对照组，差异具有统计学意义（P ＜ 0.05）。治疗后观察组患者的透明质酸（HA）、层粘连蛋白（LN）、 Ⅲ型胶原（Ⅲ –Col）均低于对照组，差异具有统计学意义（P ＜ 0.05）。治疗后观察组患者的 SGRQ、HRCT 评分均 低于对照组，差异具有统计学意义（P ＜ 0.05）。结论：采用中药方剂抗肺纤维化方与常规西药联合治疗 PF 患者， 能进一步改善患者临床症状，降低肺纤维化程度，增强肺功能，提高临床疗效。     

Investigation of Solid Dosage Form Components Interaction Using Orthogonal Techniques- Discovery of New Forms of Sodium Stearyl Fumarate

Physical or chemical interactions between drug product (DP) components can occur during manufacturing and/or upon storage; and may alter DP shelf life and performance. In this work a new Powder X-ray Diffraction (PXRD) peak was observed in DP under accelerated storage conditions. Due to the complex drug product matrix (including API, polymer, fillers, super disintegrant and lubricant), it was challenging to pinpoint the component(s) responsible for the new peak. In addition to PXRD, other orthogonal techniques including Differential Scanning Calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor sorption (DVS), Solid State Nuclear Magnetic Resonance (SSNMR) and Infrared (IR) spectroscopy were employed in this investigation to understand the root cause mechanistically. Specifically, multi nuclei SSNMR (¹H, ²³Na, ¹³C) was instrumental in delineating the components of the matrix. We identified the root cause to be an acid base reaction occurring in the DP, whereby sodium ion in sodium stearyl fumarate (SSF) is replaced by proton leading to SSF form conversion. We also identified commercially available SSF to be a hydrate that can dehydrate to an anhydrous form upon heating. In general, the same techniques can be used to investigate interactions of any multi component solid dosage forms.     

Phase Ib dose-finding study of abiraterone acetate plus buparlisib (BKM120) or dactolisib (BEZ235) in patients with castration-resistant prostate cancer

BackgroundThe phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) signalling axis and androgen receptor (AR) pathways exhibit reciprocal feedback regulation in phosphatase and tensin homologue (PTEN)-deficient metastatic castration-resistant prostate cancer (CRPC) in preclinical models. This phase Ib study evaluated the pan-PI3K inhibitor buparlisib (BKM120) and the dual pan-PI3K/ mammalian target of rapamycin (mTOR) inhibitor dactolisib (BEZ235) in combination with abiraterone acetate (AA) in patients with CRPC.Materials and methodsPatients with CRPC who had progressed on AA therapy received escalating doses of either buparlisib or dactolisib, along with fixed doses of AA (1000 mg once daily (qd)) and prednisone (5 mg twice daily (bid)). The primary objective was to define the maximum tolerated dose (MTD) and/or the recommended dose for expansion (RDE) of either buparlisib or dactolisib in combination with AA. Secondary objectives included safety, antitumour activity (Prostate Cancer Working Group 2 (PCWG2) criteria; 30% of prostate-specific antigen (PSA) decline at ≥week 12) and pharmacokinetic (PK) profile.ResultsIn buparlisib + AA arm, 25 patients received buparlisib + AA (median age, 67 years; Eastern Cooperative Oncology Group performance status (ECOG PS) of 0/1/2 for 7/17/1 patients, respectively). At 100 mg qd; two patients experienced dose-limiting toxicities (DLTs) (grade 3 hyperglycaemia; grade 2 asthenia), and this was the maximum buparlisib dose explored. Buparlisib + AA showed a 26% lower median area under the curve from time zero to 24°h (AUC_0–24) and 48% lower median maximum serum concentration (Cmax) versus the single-agent buparlisib assessed in first-in-human study. No objective response and few PSA decreases were reported.In dactolisib + AA arm, 18 patients (median age, 71 years; ECOG PS of 0/1 for 6/12 patients, respectively) received dactolisib + AA at the first dose level (200 mg bid). Five patients had 9 DLTs (grades 2&3 stomatitis; grade 3 hyperglycaemia; grades 2& 3 diarrhoea; grades 1& 2 pyrexia, grade 2 vomiting, and grade 2 chills).ConclusionsBased on the assessment of available pharmacokinetics, safety, and efficacy data, no further study is planned for either buparlisib or dactolisib in combination with AA in CRPC.     

Treatment of Patients with Metastatic Hormone-Sensitive Prostate Cancer: A Systematic Review of Economic Evaluations

The management of patients with metastatic hormone-sensitive prostate cancer (mHSPC) has been significantly modified by the availability of innovative but expensive treatments, increasing the economic burden of prostate cancer. Here, we aimed to systematically identify and review published economic evaluations (EEs) related to the treatment of mHSPC and assess their quality. A systematic search was performed of the PubMed and Cochrane databases. Three reviewers independently selected EEs by defined inclusion and exclusion criteria. They extracted all data from each EE (general information, study population, data about the EE, interventions and comparators, and outcomes). They also assessed the quality of the selected EEs according to Drummond's checklist. Fourteen EEs published between 2016 and 2021 were eligible for the systematic review. The EEs found ADT + docetaxel to be the most cost-effective of all available treatments as a first-line strategy for mHSPC (abiraterone acetate plus prednisone, enzalutamide, and apalutamide). Five EEs showed that a simple price reduction of abiraterone acetate of 50% to 75% could change the results to render this treatment also cost-effective relative to that with docetaxel. Twelve EEs were of high quality, with a Drummond score ≥ 7. Analysis of the 14 EEs identified by our systematic review, amongst which 78.6% met high quality standards, showed that ADT + docetaxel tends to be the most cost-effective alternative for mHSPC. These results were assessed by sensitivity analysis. The data provided by this systematic review help to provide a better understanding of these treatments and the better use of healthcare resources.     

论炔雌醇环丙孕酮治疗多囊卵巢综合症疗效分析

目的：探讨炔雌醇环丙孕酮治疗多囊卵巢综合症的临床效果。方法对该院2011年6月—2014年6月收治的采取炔雌醇环丙孕酮治疗的112例PCOS患者进行分析,观察治疗前后经量、经期变化及LH、FSH、E2、T、PRL情况,观察治疗后卵巢及卵泡变化。结果治疗后经量恢复107例,周期30.6±4.3d,较治疗前差异有统计学意义（P＜0.05）;治疗后LH、FSH、E2、T、PRL水平较治疗前均有下降,以LH、FSH、T水平下降显著,E2及PRL水平无明显改变;治疗后卵巢体积及卵泡数较治疗前明显变小、变少。结论炔雌醇环丙孕酮片可有效改善PCOS患者临床症状和内分泌情况,对恢复月经周期和排卵有重要作用,是治疗PCOS的有效药物,值得临床使用。     

醋酸泼尼松联合维生素B12治疗成人慢性唇炎临床疗效观察

目的：观察醋酸泼尼松片联合维生素B12注射液治疗成人慢性唇炎的疗效。方法：将56例成人慢性唇炎患者采用随机数字表法分为两组,对照组26例采用常规治疗,试验组30例采用醋酸泼尼松片联合维生素B12注射液治疗。随访1年,以唇红部黏膜干燥、脱屑、皲裂和充血、水肿、糜烂愈合情况来判断临床疗效。结果：试验组慢性唇炎自觉症状及体征的显愈率和总有效率均明显高于对照组,差异均有统计学意义（P<0.05）。结论：醋酸泼尼松片联合维生素B12注射液治疗成人慢性唇炎具有明显临床疗效,无明显不良反应。