Liquid biopsy for non-invasive assessment of liver injury in hepatitis B patients

BACKGROUND Hepatitis B is a major public health problem in China. Accurate liver injury assessment is essential for clinical evidence-based treatment. Liver biopsy is considered the gold standard method to stage liver disease, but it is not widely used in resource-limited settings. Therefore, non-invasive liquid biopsy tests are needed. AIM To assess liver injury in hepatitis B patients using quantified cell free DNA combined with other serum biomarker as a liquid biopsy-based method. METHODS A cohort of 663 subjects including 313 hepatitis B patients and 350 healthy controls were enrolled. Ultrasound-guided liver biopsies followed by histopathological assessments were performed for the 263 chronic hepatitis B patients to determine the degree of liver injury. Cell-free DNA was quantified using a novel duplex real-time polymerase chain reaction assay. RESULTS Compared with healthy controls, patients with hepatitis B virus (HBV) infection had significantly higher plasma DNA, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), bilirubin, and HBV DNA levels (P<0.01). Serum ALT, AST, bilirubin, and plasma DNA levels of patients with markedsevere inflammation were significantly higher than those with mild-moderate inflammation (P<0.01). There was a statistically significant correlation between hepatocyte inflammation severity and serum bilirubin (R^2=0.673, P<0.01) or plasma DNA (R^2=0.597, P<0.01) levels. The areas under the curves of serum ALT, bilirubin, plasma DNA, and their combination to distinguish between patients with mild–moderate and marked-severe inflammation were 0.8059, 0.7910, 0.7921, and 0.9564, respectively. CONCLUSION The combination of plasma DNA, serum ALT, and bilirubin could be a candidate liquid biopsy for non-invasive assessment of liver injury in hepatitis B patients.

乙烯利对液体发酵瓦尼桑黄中Hispidin和Hypholomine B等酚类化合物的影响及体外活性评价

该研究旨在促进瓦尼桑黄合成Hispidin和Hypholomine B等酚类化合物,以提高瓦尼桑黄产品的药用价值、保健功能和市场竞争力。以向液体培养基中添加乙烯利的方式,对目前市面上销售和应用范围最广的瓦尼桑黄进行发酵研究。研究发现,添加乙烯利可以促进液体发酵瓦尼桑黄中Hispidin和Hypholomine B等酚类化合物的合成。乙烯利在培养基中的终浓度为2 mmol/L,添加时间点为0 d,并用1 mol/L的Na_(2)CO_(3)将pH调节至添加前水平。以Hispidin对照品计算Hispidin的平均含量,乙烯利发酵组中Hispidin的平均含量最大为液体发酵组的3.17倍。Hypholomine B的平均相对含量由HPLC图谱中的峰面积表示,前者最大为后者的2.55倍。此外,乙烯利的添加还提高了其他酚类化合物的生成累积量,乙烯利发酵组、液体发酵组和子实体组的提取物中总酚含量分别为:(3.76±0.11)%、(1.96±0.12)%和(0.77±0.04)%。体外活性评价以IC_(50)为依据,乙烯利发酵组的提取物对DPPH自由基清除率、ABTS阳离子自由基清除率和α-葡萄糖苷酶抑制率的IC_(50)分别为:150.00、106.46、16.08μg/mL,均低于液体发酵组的提取物(170.61、115.36、35.48μg/mL)和子实体组的提取物(218.10、130.51、56.83μg/mL)。

硫酸多黏菌素B在两种溶媒中的含量稳定性

目的考察硫酸多黏菌素B在成品输液及高浓度储备液中的含量稳定性,为临床给药提供依据。方法建立测定硫酸多黏菌素B含量的高效液相色谱方法,将硫酸多黏菌素B分别溶于0.9%氯化钠注射液及5%葡萄糖注射液,配成0.5、1 mg·mL^(-1)成品输液及5 mg·mL^(-1)储备液,并在4和25℃储存条件下于0、2、4、6、8、12、16、20及24 h测定多黏菌素B含量。结果4和25℃储存条件下,0.5、1 mg·mL^(-1)硫酸多黏菌素B成品输液和5 mg·mL^(-1)硫酸多黏菌素B储备液均稳定,多黏菌素B1和多黏菌素B2含量和0 h(以100%计)比较,分别在97.23%~102.63%和97.30%~102.63%范围内。结论硫酸多黏菌素B溶于0.9%氯化钠注射液及5%葡萄糖注射液,0.5、1和5 mg·mL^(-1)溶液在4和25℃储存,24 h内稳定性良好。

超声法与闪式提取法提取刺五加中刺五加苷B的工艺对比研究

本试验采用正交试验优化超声法和闪式提取法提取刺五加中刺五加苷B的提取工艺,并对两种提取方法的最佳工艺进行对比研究。以刺五加苷B的提取率为评价指标,设计L_(9)(3^(4))超声法和闪式提取法的正交试验,以高效液相色谱法测定刺五加苷B的含量。结果显示:超声法提取率高于闪式提取法,最佳提取工艺为提取溶剂30%乙醇,超声功率200 W,提取时间0.75 h,料液比1:5(g/mL)。本研究可为刺五加在饲料领域的开发和利用提供参考。

HPLC-DAD/FLD测定婴幼儿配方奶粉中B族维生素及不确定度分析

建立了高效液相色谱法同时测定婴幼儿配方奶粉中维生素B_(2)、烟酸、烟酰胺、吡哆醇、吡哆醛的方法。样品经1%甲醇的0.1%磷酸水溶液提取并沉降蛋白后,采用Thermo Hypersil GOLD^(TM)aQ(5μm,4.6 mm×250 mm)分离,以二极管阵列检测器(DAD)和荧光检测器(FLD)串联模式检测。同时构建了不确定度评定模型,考察了不确定度的来源。结果表明,维生素B_(2)、吡哆醇、吡哆醛在0.025~4.000μg/mL,烟酸、烟酰胺在0.125~20.00μg/mL范围内,标准曲线呈现良好的线性(R^(2)>0.999),加标回收率在93.20%~108.00%(n=6),相对标准偏差(RSD)<8%。当婴幼儿配方奶粉中烟酸、烟酰胺含量为4 mg/100 g时,其扩展不确定度分别为0.300 mg/100 g(k=2)、0.182 mg/100 g(k=2);当吡哆醇、吡哆醛、VB_(2)含量为800μg/100 g时,其扩展不确定度为100μg/100 g(k=2)、71.5μg/100 g(k=2)、41.9μg/100 g(k=2),其不确定度主要来源于标准溶液配制及标准曲线拟合。该方法简便、快速、灵敏、准确,可满足婴幼儿配方奶粉中B族维生素的检测要求。

超高效液相色谱-串联质谱法测定禽用饲料中苏丹红、罗丹明B和酸性橙Ⅱ等6种染色剂含量

本研究基于超高效液相色谱-串联质谱(UPLC-MS/MS)结合QuEChERS快速净化方法,旨在建立禽用饲料中苏丹红等6种化工染料的定量分析方法。样品用乙腈提取,C_(18)+PSA净化材料净化,净化液经Hypersil GOLD C_(18)反向色谱柱分离,在电喷雾离子源(ESI)正、负离子双扫描模式下多反应监测(MRM),以标准曲线外标法定量。结果表明:6种化合物线性关系良好;相关系数(R^(2))>0.999,4种苏丹红和酸性橙Ⅱ检出限(LOD)为0.5μg/kg,定量限(LOQ)为1.0μg/kg,罗丹明B LOD为0.05μg/kg,LOQ为0.1μg/kg。添加回收率为90%~110%,相对标准偏差<10%。本研究方法具有快速、灵敏等优势,可实现禽用饲料中6种化工染料的准确定量分析,为禽用饲料的质量安全提供技术参考。

固液比与Al-Ti-B添加量对8011铝合金组织和性能的影响

8011铝合金是广泛使用的一种铸轧铝合金,其熔炼过程中的固液比和Al-Ti-B添加量等都是影响其性能的关键因素。利用光学显微镜(OM)、透射电子显微镜(TEM)和力学性能测试对比研究了固液比(25%、30%、35%、40%)和不同Al-Ti-B中间合金添加量(0.18%、0.20%、0.22%)对8011铝合金显微组织和力学性能的影响。结果表明:固液比相同时,Al-Ti-B中间合金添加量越高,8011铝合金的晶粒越细小,且分布越均匀;Al-Ti-B中间合金添加量相同时,固液比越高,8011铝合金的晶粒越细小,但也容易产生晶粒团聚的现象;当固液比为40%,Al-Ti-B中间合金添加量为0.22%时,8011铝合金的晶粒细化效果最好,力学性能最优,其抗拉强度可达到107 MPa,延伸率可达到12.4%,硬度为36.73 HV。

RP-HPLC法检测甜梦口服液(甜梦合剂)中淫羊藿苷、紫丁香苷和23-乙酰泽泻醇B的含量

目的建立同时检测甜梦口服液(甜梦合剂)中的淫羊藿苷、紫丁香苷和23-乙酰泽泻醇B含量的RP-HPLC法。方法利用Shimadzu CLC ODS C 18(4.6 mm×250 mm,5μm)分离甜梦口服液中3种成分(淫羊藿苷、紫丁香苷和23-乙酰泽泻醇B)。测定波长:淫羊藿苷为271 nm、紫丁香苷为265 nm和23-乙酰泽泻醇B为207 nm;柱温30℃,进样体积10μl。流动相:0.3%磷酸溶液(流动相A)-乙腈(流动相B),洗脱程序:0~4 min(95%A)、4~7 min(95%~78%A)、7~10 min(78%~38%A)、10~16 min(38%~26%A)、16~23 min(26%~19%A)、23~28 min(19%A)、28~28.5 min(19%~95%A)。外标法检测3种成分的含量。结果紫丁香苷、23-乙酰泽泻醇B和淫羊藿苷分别在0.1962~19.62μg/ml,0.7544~75.44μg/ml和0.1966~19.66μg/ml质量浓度范围内存在良好的线性关系,线性系数R 2均大于0.995;平均加标回收率分别为99.47%,98.68%和100.26%;仪器精密度和方法重复性RSD(n=6)均在5.0%以内。结论建立的RP-HPLC法具有线性范围广、检测结果准确等优点,可以用于甜梦口服液(甜梦合剂)供试品的质量控制。

A novel synthesis of pyrazolo[3,4-b]pyridine derivatives through multi-component reaction in ionic liquid

An efficient and novel procedure for the preparation of pyrazolo[3,4-b]pyridine derivatives through multi-component reaction of aldehyde, 5-amino-3-methyl-1-phenylpyrazole and malononitrile or cyanoacetate in [bmim][BF4] is described in this paper. Advantages of the method presented here include mild conditions, high yields together with a green nature and ease of recovery and reuse of the reaction medium.

One-pot three-component synthesis of tetrahydrobenzo[b]pyrans catalyzed by cost-effective ionic liquid in aqueous medium

A simple and ef ficient method is proposed for the synthesis of tetrahydrobenzo[b]pyrans with aromatic aldehydes, active methylene compounds, and dimedone using basic ionic liquid catalyst in water. The procedure offers several advantages including short reaction time, good yield, easy procedure, and good recyclability of catalysts, which may be a practical alternative to conventional processes for preparation of 4-hpyrans.

Characteristics of gas-liquid diaphragm discharge and its application on decolorization of brilliant red B in aqueous solution

A simple gas-liquid diaphragm discharge reactor was designed and characteristics of the discharge and its application on decolorization of brilliant red B in an aqueous solution were investigated. The results showed that strong oxidizing agents such as ·OH and ·O radicals were generated. Average electron temperature of the discharge was 0.72 eV, 1.15 eV and 0.83 eV with air, oxygen and argon as the discharge gas, respectively. Solution p H and conductivity changed little when oxygen or argon was used as the discharge gas; however, these two parameters changed significantly when the discharge was performed in air. During the discharge treatment,the characteristic absorption peaks of brilliant red B gradually decreased where the decolorization followed the first-order kinetics. With 10 min of discharge, the decolorization of brilliant red B（30 mg L^-1） can reach 96%, 81% and 62% in the cases of oxygen, argon and air,respectively. The analysis of by-products showed that the brilliant red B molecule can be effectively destroyed in this discharge mode.

保健食品中B族维生素的快速分析方法研究

目的:建立同时分析烟酰胺、维生素B1、维生素B6、维生素B2且前处理简单的快速检测方法。方法:样品用0.1%三氟乙酸溶液超声提取,C18色谱柱分离,以0.1%三氟乙酸-乙腈为流动相梯度洗脱,紫外检测器检测。结果:方法对4种B族维生素的分离效果明显,分离度全部超过1.5,标准曲线呈直线回归,相关系数大于0.999 5,检测结果与国家标准方法一致。结论:该方法操作更便捷,分析速度快,灵敏度和准确度高,适用于保健食品中4种B族维生素含量的检测。

Ultrasonic study on organic liquid and binary organic liquid mixtures by using Schaaffs＇ collision factor theory

Based on Schaaff＇s collision factor theory （CFT） in liquids, the equations for nonlinear ultrasonic parameters in both organic liquid and binary organic liquid mixtures are deduced. The nonlinear ultrasonic parameters, including pressure coefficient, temperature coefficients of ultrasonic velocity, and nonlinear acoustic parameter B/A in both organic liquid and binary organic liquid mixtures, are evaluated for comparison with the measured results and data from other sources. The equations show that the coefficient of ultrasonic velocity and nonlinear acoustic parameter B/A are closely related to molecular interactions. These nonlinear ultrasonic parameters reflect some information of internal structure and outside status of the medium or mixtures. From the exponent of repulsive forces of the molecules, several thermodynamic parameters, pressure and temperature of the medium, the nonlinear ultrasonic parameters and ultrasonic nature of the medium can be evaluated. When evaluating and studying nonlinear acoustic parameter B/A of binary organic liquid mixtures, there is no need to know the nonlinear acoustic parameter B/A of the components. Obviously, the equation reveals the connection between the nonlinear ultrasonic nature and internal structure and outside status of the mixtures more directly and distinctly than traditional mixture law for B/A, e.g. Apfel＇s and Sehgal＇s laws for liquid binary mixtures.

Co-rotational Oldroyd Fluid B Model for Spinning Flow of Liquid Crystalline Polymer

The relationship between the extensional viscosity and material parameters was studied through the analytical formulas of stress and extensional viscosity. The differential equations were solved to obtain the relationship between extensional viscosity and strain rates. The results obtained qualitatively agree with the experimental results. The study makes it practicable to simulate the rheologic behaviors of spinning flow of liquid crystalline polymer using co-rotational Oldroyd fluid B model.

超高效液相色谱-串联质谱法测定水果及谷物中多抗霉素B残留量

建立了一种快速有效的超高效液相色谱串联质谱法检测分析水果及谷物中多抗霉素B残留量的方法。色谱柱为Waters CORTECS®HILIC(150 mm×2.1 mm,2.7μm),流动相为0.1%甲酸(含5 mmol·L^(-1)甲酸铵)水溶液和乙腈,梯度洗脱,流速为0.40 mL·min-1。采用多反应监测模式(Multiple Reaction Monitoring,MRM),采用ESI正电离源,定量离子对508.1/305。样品经0.2%甲酸水溶液超声提取,离心取上清液,经由PLS柱(亲水亲脂平衡固相萃取柱)净化,过膜后直接测定,外标法定量。结果表明,多抗霉素B在2~50μg·L^(-1)线性关系良好,相关系数r2>0.999。在3个浓度加标水平条件下,即多抗霉素B浓度为0.10 mg·kg^(-1)、0.20 mg·kg^(-1)和1.00 mg·kg^(-1)时,回收率为91.1%~112.3%,相对标准偏差为6.7%~10.1%。该方法操作简便、回收率高,可用于批量水果及谷物中多抗霉素B残留的检测。

超快速液相色谱法同时测定刺五加中刺五加苷B和刺五加苷E的含量

目的建立一种同时测定刺五加中刺五加苷B和刺五加苷E含量的超快速液相色谱法。方法采用Shim-Pack XR-ODS柱(75 mm×3.0 mm,2.2μm);H_(2)O(A)-CH3OH(B)为流动相,梯度洗脱(0~1 min,10%B→35%B;1~5 min,35%B→50%B;5~8 min,50%B→60%B);流速:0.4 mL·min^(-1);柱温:30°C;进样量:5μL;平衡时间:5 min;检测波长:215 nm。结果刺五加苷B和刺五加苷E分别在10~400μg·mL^(-1)(r=0.9990)和5~200μg·mL^(-1)(r=0.9991)质量浓度范围内与峰面积呈良好的线性关系;平均回收率(n=9)分别为98.9%和98.2%;6批样品中刺五加苷B与刺五加苷E的含量范围分别为0.130%~0.145%和0.0708%~0.0776%。结论与《中国药典》方法比较,该方法稳定性和专属性好,分析时间短,可用于刺五加药材的质量控制研究。

ctDNA在B细胞淋巴瘤中的研究进展

循环肿瘤DNA(ctDNA)是肿瘤患者体液循环中一种广泛存在的游离DNA片段,相较于肿瘤实体病灶,ctDNA具有取材方便、安全无创、信息全面等优势。ctDNA检测可应用于B细胞淋巴瘤的治疗前、治疗中和治疗后等各个阶段,在肿瘤辅助诊断、预后分层、靶向治疗、疗效评估、微小残留病灶(MRD)监测及预测复发等方面具有重要指导意义。本文主要就ctDNA在B细胞淋巴瘤中的相关检测手段、应用进展和前景展望等作一概述。

多功能净化柱-光化学衍生高效液相色谱法测定成品植物油和花生原油中黄曲霉毒素B1

为了快速、准确测定成品植物油和花生原油中黄曲霉毒素B1(AFB1)的含量,建立了采用多功能净化柱对油样进行前处理,再结合光化学衍生高效液相色谱法测定植物油中AFB1含量的方法,对前处理提取剂、高效液相色谱法的分析条件(色谱柱、进样量)进行优化,再通过与国标中免疫亲和柱法进行比较,对所建立的方法进行评价。结果表明:优化的条件为以乙腈-水溶液(84+16)为提取剂,采用Zorbax Eclipse XDB-C18色谱柱(4.6 mm×150 mm,5μm),进样量10μL;建立的方法加标回收率和精密度良好,定量限为0.2μg/kg,低于GB 2761—2017规定的最低限量值要求,可满足企业生产的监测需求;将建立的方法用于测定浅色植物油中AFB1时,检测结果与免疫亲和柱法相比相对误差为0~18.0%,符合国标要求,但对于深色植物油测定的相对误差超出国标要求。综上,建立的方法可用于快速、准确检测花生油(成品油和原油)及成品玉米油、亚麻籽油、葵花籽油、浅黄色菜籽油、黄色芝麻油中AFB1的含量。

B酸离子液体催化棉籽油酯交换制备生物柴油

制备了5种对水稳定性好、带—SO3H官能团的磺酸类B酸离子液体,并以它们作催化剂进行了棉籽油酯交换反应制备生物柴油的研究。实验结果表明,这些磺酸类B酸离子液体的催化活性高,它们的催化活性与含氮官能团的种类和碳链长度有关,其中,吡啶丁烷磺酸硫酸氢盐离子液体的催化活性最高。考察了吡啶丁烷磺酸硫酸氢盐离子液体的用量、醇与油摩尔比、反应温度对酯交换反应的影响及离子液体的稳定性,实验结果表明,在反应温度170℃、n(甲醇)∶n(棉籽油)∶n(离子液体)=12∶1∶0.057、反应时间5h的条件下,产物中脂肪酸甲酯的质量分数可达92.0%,且离子液体的稳定性好,可循环使用。

固相萃取-高效液相色谱检测葡萄酒中罗丹明B

建立固相萃取富集净化,高效液相色谱紫外检测葡萄酒中禁用色素罗丹明B的方法。葡萄酒样品中的罗丹明B经C18固相萃取,40%乙醇溶液清洗净化,80%的甲醇溶液洗脱后,用高效液相色谱紫外检测器进行测定。结果表明:罗丹明B在1.0～10.0μg/mL范围呈现良好的线性关系,相关系数0.9924;在1.5～3.5μg/mL添加水平时,平均回收率为72.06%,相对标准偏差为2.87%(n=5);定量测出限为0.25μg/mL。表明该方法灵敏度高,重复性良好,操作简便快速,适用于葡萄酒中罗丹明B的检测。