Real-time toxicity prediction of Aconitum stewing system using extractive electrospray ionization mass spectrometry

Due to numerous obstacles such as complex matrices, real-time monitoring of complex reaction systems (e.g., medicinal herb stewing system) has always been a challenge though great values for safe and rational use of drugs. Herein, facilitated by the potential ability on the tolerance of complex matrices of extractive electrospray ionization mass spectrometry, a device was established to realize continuous sampling and real-time quantitative analysis of herb stewing system for the first time. A complete analytical strategy, including data acquisition, data mining, and data evaluation was proposed and implemented with overcoming the usual difficulties in real-time mass spectrometry quantification. The complex Fuzi (the lateral root of Aconitum)–meat stewing systems were real-timely monitored in 150 min by qualitative and quantitative analysis of the nine key alkaloids accurately. The results showed that the strategy worked perfectly and the toxicity of the systems were evaluated and predicated accordingly. Stewing with trotters effectively accelerated the detoxification of Fuzi soup and reduced the overall toxicity to 68%, which was recommended to be used practically for treating rheumatic arthritis and enhancing immunity. The established strategy was versatile, simple, and accurate, which would have a wide application prospect in real-time analysis and evaluation of various complex reaction systems.     

Cycling probe technology to quantify and discriminate between wild-type varicella-zoster virus and Oka vaccine strains

Rapid differentiation between wild-type varicella zoster virus (VZV) and Oka-vaccine (vOka) strains is important for monitoring side reactions of varicella vaccination. To develop a high-throughput molecular diagnostic method for the differentiation of wild-type VZV and vOka strains based on cycling probe technology. The primers were designed to amplify common sequences spanning a single nucleotide polymorphism (SNP) in gene 62 of VZV. DNA–RNA chimeric probes (cycling probes) were designed to detect the SNP at nucleotide 105705. The cycling probe real-time PCR assays for VZV wild-type and vOka strains specifically amplified plasmids containing target sequences that ranged between 10 and 1 × 10⁶ copies per reaction. The inter- and intra-assay coefficients of variation were less than 5%. After initial validation studies, the clinical reliability of this method was evaluated using 38 swab samples that were collected from patients suspected of being zoster. Compared to the loop mediated isothermal amplification method, which is defined as the gold standard, cycling probe real-time PCR was highly sensitive and specific. The cycling probe real-time PCR technology is a reliable tool for differentiating between wild-type VZV and vOka strains in clinical samples.     

SWE联合灰阶超声评估不同分度子宫脱垂初产妇会阴体变化

目的利用实时剪切波弹性成像技术(SWE)联合灰阶超声探讨产后不同分度子宫脱垂初产妇会阴体弹性和形态的变化。方法选取2019年1月~2019年12月200例产后6~8周,患有子宫脱垂且经阴道单胎足月顺产的初产妇为观察对象。通过经阴道超声检查,结合POP分度标准,将患有Ⅰ度子宫脱垂的65例初产妇纳入B组,Ⅳ度子宫脱垂的46例初产妇纳入C组。收集同期健康已婚未育女性50例作为对照,纳入A组。分别在静息及缩肛状态下,测量三组观察对象会阴体的高度及深度,采集各组会阴体的杨氏模量值。结果三组观察对象的会阴体收缩幅度差值(高度、深度)在缩肛、静息两种状态下,差异有统计学意义(P均<0.05);静息及缩肛两种状态下健康对照组会阴体的平均杨氏模量值较Ⅰ度子宫脱垂组、Ⅳ度子宫脱垂组高,且Ⅰ度子宫脱垂组高于Ⅳ度子宫脱垂组,各组间差异均有统计学意义(P<0.05)。结论产后子宫脱垂初产妇会阴体弹性明显降低,收缩功能减弱,且会阴体收缩功能减低程度与子宫脱垂程度有关。     

人参皂苷Re对重症急性胰腺炎小鼠小肠黏膜的影响及调控机制

目的 探讨人参皂苷Re通过调控 Janus激酶2/信号转导子与转录激活子3（JAK2/STAT3）通路对重症急性胰腺炎（SAP）小鼠小肠黏膜的影响及作用机制。 方法 48只小鼠分为对照组、SAP组、SAP+人参皂苷Re组和SAP+人参皂苷Re+LY2784544组（n=12）。通过腹腔注射雨蛙肽溶液（禁食12 h后,100 μg/kg × 6次,注射间隔60 min）的方法建立SAP模型。腹腔注射人参皂苷Re（4 mg/kg,每日1次,连续7 d）。通过腹腔注射 12.7 mg/kg LY2784544来抑制JAK2/STAT3通路。检测各组胰腺指数、血清淀粉酶和炎性因子水平;检测D-乳酸和FITC-D水平分析小肠黏膜屏障功能;HE染色观察胰腺组织和小肠黏膜组织损伤情况;Western blotting检测小肠黏膜组织中凋亡蛋白Bax和Bcl-2的水平;Real-time PCR和Western blotting检测胰腺组织和小肠黏膜组织JAK2/STAT3通路水平。 结果 SAP组小鼠胰腺指数、血清淀粉酶、白细胞介素6（IL-6）和肿瘤坏死因子α（TNF-α）、FITC-D、D-乳酸、Bax蛋白水平、胰腺组织和小肠组织中JAK2和STAT3 mRNA和蛋白水平均显著高于对照组,而Bcl-2蛋白显著低于对照组（P<0.05）。SAP+人参皂苷Re组小鼠Bcl-2蛋白显著高于SAP组,其余指标显著低于SAP组（P<0.05）。SAP+人参皂苷Re+LY2784544组的Bcl-2蛋白显著高于SAP+人参皂苷Re组,其余指标显著低于SAP+人参皂苷Re组（P<0.05）。 结论 人参皂苷Re可能通过抑制JAK/STAT通路缓解炎性反应减轻SAP模型小鼠的胰腺损伤, 并通过抑制细胞的凋亡保护小肠黏膜屏障。     

Rho相关激酶抑制剂Y27632对小鼠腔前卵泡体外发育的作用

目的 探讨Rho相关激酶（ROCK）抑制剂Y27632在小鼠腔前卵泡体外发育中的作用。 方法 取出生12.5 d的小鼠卵巢,机械法收集单枚腔前卵泡,采用超低吸附96孔板模拟3D环境对其进行培养。设置对照组和含有Y27632的实验组,通过形态学观察、卵泡直径的变化、成熟卵泡的数量、成熟卵母细胞纺锤体的变化等来评估卵泡的整体发育情况。采用Real-time PCR技术检测颗粒细胞中卵泡刺激素受体（FSHR）、卵母细胞中骨形态发生蛋白15（BMP15）、生长分化因子9（GDF9）等以及凋亡相关基因（Bad、Bax和Caspase-3）的表达情况,同时,用细胞存活染料检测卵泡发育过程中颗粒细胞的凋亡情况。 结果 ROCK抑制剂Y27632对卵泡直径增长和基本发育指标（存活数、成腔率和成熟率）无明显影响（P>0.05）,但对照组卵母细胞成熟后纺锤体组装出现异常。培养8 d后,与对照组相比,实验组颗粒细胞特异性基因FSHR上调;卵母细胞特异性基因BMP15和GDF9上调,而叉头框O3（FoxO3)下调;凋亡基因Bax、Bad和Caspase3表达下调,实验组卵泡内的颗粒细胞凋亡明显少于对照组。 结论 小鼠卵泡体外发育过程中,ROCK抑制剂Y27632能够抑制颗粒细胞的凋亡,防止卵母细胞纺锤体组装异常,进而提高卵泡体外发育质量。     

超声介入医师训练实时影像虚拟导航系统辅助肝癌消融的学习曲线分析

目的分析超声介入医师行实时影像虚拟导航系统（RVS）辅助肝癌消融的学习曲线。 方法回顾性分析2018年10月至2019年9月就诊于浙江中医药大学附属杭州市西溪医院的,由同一名具有5年以上超声引导下肝癌消融经验的医师,行RVS引导下辅助肝癌消融的第1~60例肝癌患者资料。按照治疗顺序分为A、B、C组,每组各20例。采用方差分析比较各组间RVS术前融合时间和肿瘤消融时间的差异,组间两两比较采用LSD-t检验;采用Fisher确切概率法分析患者组间肿瘤1次性消融成功率和术后1个月肿瘤的完全灭活率的差异。绘制RVS辅助肝癌消融的术前融合时间曲线图。 结果A组的RVS术前融合时间、肿瘤消融时间明显大于B组及C组［（20.9±6.7）min vs （9.7±1.2）min vs （9.6±2.7）min;（23.1±7.9）min vs （19.6±5.0）min vs （19.2±3.7）min］,差异具有统计学意义（t=7.4,P＜0.001;t=1.9,P=0.035）,B组与C组之间比较,差异均无统计学意义（P均＞0.05）;A组的肿瘤1次性消融成功率［74.1%（20/27）］小于B组［96.4%（27/28）］和C组［96.4%（27/28）］,差异具有统计学意义（P均=0.025）,B组与C组之间比较,差异无统计学意义（P＞0.05）。3组RVS辅助肝癌消融术后1个月肿瘤完全灭活率比较［92.6%（25/27） vs 92.9%（26/28） vs 92.9%（26/28）］,差异均无统计学意义（P均＞0.05）。操作者积累约20例患者后,术前融合时间趋于稳定,学习进入平台期。 结论对于期望熟练掌握RVS辅助肝癌消融的介入医师,操作者积累约20例患者RVS辅助肝癌消融可以明显缩短术前融合时间和肿瘤消融时间,提高融合精准度、团队契合度及肿瘤1次性消融成功率,之后学习曲线进入平台期。     

Assessment of Diffuse Thyroid Disease by Strain Ratio in Ultrasound Elastography

The goal of this study was to explore the value of strain ratio from real-time elastography in the semi-quantitative assessment of diffuse thyroid disease. Fifty-one patients with primary hyperthyroidism, 70 with Hashimoto's thyroiditis, 8 with subacute thyroiditis and 43 with normal healthy thyroids were recruited to measure the strain ratio (SR) of thyroid tissue and sternocleidomastoid muscle (on the same side of the thyroid). SR values of all groups were subjected to statistical analysis. The SRs (mean ± standard deviation) of patients with hyperthyroidism, Hashimoto's thyroiditis and subacute thyroiditis were 2.30 ± 1.08, 7.04 ± 7.74 and 24.09 ± 13.56, respectively. The SR of the control group was 1.76 ± 0.54. SR values ranked in ascending order were control group < hyperthyroidism group < Hashimoto's thyroiditis group < subacute thyroiditis group. There were statistically significant (p < 0.05) differences in thyroid hardness between groups with different diffuse thyroid diseases. SR values of the hyperthyroidism and control groups did not statistically differ (p > 0.05). It is feasible to assess diffuse thyroid disease with strain ratios obtained with ultrasound elastography.     

两种方法检测LGC盲样及效果探讨

目的：检测LGC盲样中的小肠结肠炎耶尔森氏菌并对过程和结果进行探讨。方法①分离培养法。②实时荧光PCR法。结果两者均检出小肠结肠炎耶尔森氏菌。结论两种方法的有效结合对目标菌检出有更重要的意义。