Strategies for the treatment of invasive ductal carcinoma of the pancreas and how to achieve zero mortality for pancreaticoduodenectomy

Although various therapeutic modalities are available for carcinoma of the pancreas, “curative resection” is the most important. Thus, the aim of surgery for carcinoma of the pancreas is local complete resection of the carcinoma. Carcinoma of the head of the pancreas invades through the pancreatic parenchyma, following the arteries, veins, and especially nerves between the parenchyma and fusion fascia, and then spreads horizontally toward the superior mesenteric artery or celiac axis. We suggest techniques for resection of the extrapancreatic nerve plexus in the head of the pancreas during a Whipple procedure for carcinoma of the pancreas, from the perspective of surgical anatomy and pathology, to achieve “curative resection”. We suggest that: (1) en-bloc resection of the right side of the superior nerve plexus and the first and second nerve of the pancreatic head should be performed. With this technique, it is possible to avoid cutting these nerves. It is easy to perform this procedure, as follows. First, the superior mesenteric artery and vein are encircled with tape. Next, the superior mesenteric artery should be moved to the right side of the superior mesenteric vein under this vein. In addition, (2) the entire cut end of the nerve plexus should be investigated during the operation, using frozen specimens, and confirmed to be negative for cancer. If the cut end is positive for cancer, additional resection of the nerve plexus should be performed to achieve curative resection. It is impossible to completely determine whether the cut end of the nerve plexus is positive or negative for carcinoma after surgery, because the cut end is long and some specimens are deformed by formalin fixation; thus, it is difficult to identify the true surgical cut end. With regard to reconstruction, we perform a modified Child method with pancreaticojejunostomy (end-to-side), choledochoduodenostomy (also end-to-side), and gastrojejunostomy with Braun’s anastomosis. The greater omentum is set around the pancreaticojejunostomy to prevent pancreatic juice from spreading in the abdomen. Careful management of the intraabdominal drainage tubes after the operation is crucial. With the operative procedure and postoperative controls described above, operative mortality was zero in 114 consecutive patients in our series who underwent pancreaticoduodenectomy.     

ELISA检测低硒小鼠组织GSH—Px的含量——兼论GSH—Px含量与活性的关系

昆明小鼠30只分别饲以低硒(14.7ng/g)及补硒(500ng/g)等饲料27天,用 ELISA 检测心、肝、肾、肝细胞线粒及胞质 GSH—px 含量,结果表明肝>肾>心,低硒动物组织及肝亚细胞成分酶含量明显下降,补硒则可维持酶含量和活性,本方法灵敏性高,方法简便。低硒时动物组织内 GSH—px 含量下降幅度不及全血酶活性大,揭示低硒早期,动物组织内可能存在无活性蛋白多肽,作者对其发生机制进行了探讨。     

UPLC-MS/MS测定大鼠血浆多柔比星浓度方法的建立及毒代动力学应用

目的 建立一种简便、快速、灵敏的测定大鼠多柔比星血药浓度的超高效液相-质谱联用（UPLC-MS/MS）法,并将其应用于注射用盐酸多柔比星大鼠体内毒代动力学实验。方法 采用ACQUITY UPLC^® BEH C₁₈（50 mm×2.1 mm,1.7 μm）色谱柱,流动相为0.1%甲酸（含2 mmol/L甲酸铵）水溶液-乙腈,梯度洗脱。体积流量为0.4 mL/min,进样量为10 μL。采用电喷雾离子源（ESI）,多反应监测（MRM）方式扫描,以正离子方式进行检测,蛋白沉淀法提取样品。用于定量分析的离子对分别为多柔比星m/z 544.43→m/z 397.08,内标地西泮m/z 285.02→154.40。SD大鼠30只,按体质量随机分为3组,分别单次iv 52.2、61.4、72.3 mg/m²盐酸多柔比星后测定血药浓度,并用DAS 3.1.4软件计算毒代参数。结果 血浆中内源性物质不干扰待测物和内标的测定,多柔比星在0.5～100 ng/mL范围内线性关系良好,定量下限为0.5 ng/mL。多柔比星在0.5、1、20、80 ng/mL 4个浓度的批内批间精密度RSD值为3.21%～12.79%。多柔比星在1、80 ng/mL的提取回收率和基质效应分别为102.00%～103.75%和79.27%～89.34%。SD大鼠分别单次iv给予注射用盐酸多柔比星52.2、61.4、72.3 mg/m²后,多柔比星在大鼠体内的AUC_0-t分别为（2 318.78±282.65）、（3 203.11±829.41）和（3 326.96±546.04） ng·h/mL,C_0.083h分别为（1 720.50±851.19）、（3 363.00±1 458.84）和（2 156.50±919.90）ng/mL。结论 建立的UPLC-MS/MS分析方法灵敏度高、样品处理方法简单、样品分析时间短,可以应用于大鼠多柔比星毒代动力学试验中。     

HPLC波长切换法同时测定杜仲双降袋泡剂中7种成分的含量

目的建立高效液相(HPLC)波长切换法同时测定杜仲双降袋泡剂中桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁及槲皮素7种成分含量的方法。方法以80%甲醇为溶剂,加热回流提取;色谱柱:Hydrosphere C18(4.6 mm×200 mm,5μm);柱温:30℃;流动相:乙腈-0.1%磷酸溶液,梯度洗脱;检测波长:208 nm(桃叶珊瑚苷)、235 nm(京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷)、270 nm(芦丁、槲皮素);流速:0.9 ml/min。结果桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁、槲皮素的质量浓度分别在10.97~274.25μg/ml(r=0.999 7)、9.78~244.50μg/ml(r=0.999 8)、6.86~171.50μg/ml(r=0.999 6)、2.47~61.75μg/ml(r=0.999 7)、8.11~202.75μg/ml(r=0.999 1)、4.59~114.75μg/ml(r=0.999 9)、1.85~46.25μg/ml(r=0.999 5)范围内线性关系良好;平均加样回收率分别为99.85%、98.92%、97.52%、97.08%、98.51%、97.10%、96.91%,RSD分别为0.93%、0.74%、1.26%、1.37%、0.88%、1.05%、1.33%。结论所建立的HPLC波长切换法可以同时测定杜仲双降袋泡剂中桃叶珊瑚苷、京尼平苷酸、绿原酸、京尼平苷、松脂醇二葡萄糖苷、芦丁和槲皮素的含量,方法简便准确、灵敏度高,可用于杜仲双降袋泡剂的质量控制。     

基于限定日剂量和ABC-改进VEN法的2018年亳州市人民医院质子泵抑制剂应用分析

目的:了解亳州市人民医院(以下简称“我院”)质子泵抑制剂(proton pump inhibitor,PPI)的使用情况,明确需重点管控的PPI品规,向临床提供关于PPI选择的相关建议。方法:调查2018年我院PPI消耗数据,采用限定日剂量(defined daily dose,DDD)法和ABC-改进VEN法进行统计分析,并根据分析结果和管理优先级别确定Ⅰ(高)、Ⅱ(低)和Ⅲ(中)组。结果:ABC-改进VEN法基于减轻患者负担和医院成本对PPI进行分类,便于管理。涉及的PPI共18个品规,销售金额合计14051911.21元;其中Ⅰ组包含AN类和DDD分析加入的品规,共5个,销售金额为9405764.53元(占66.94%),代表药品包括注射用泮托拉唑钠(40 mg)、注射用雷贝拉唑钠(20 mg);Ⅱ组包含CV类5个品规,销售金额为886138.62元(占6.31%),代表药品为奥美拉唑肠溶片(20 mg);Ⅲ组包含10个品规,销售金额为3760008.06元(占26.76%),代表药品为奥美拉唑肠溶片(40 mg)。结论:我院PPI应用结构不甚合理,应重点监控Ⅰ组药品,减少不合理用药情况,向临床推荐Ⅲ组药品,并保障供应。     

基于一测多评法对丁蔻理中丸中6种成分的质量控制研究

目的:建立一测多评法同时测定丁蔻理中丸中白术内酯Ⅲ、白术内酯Ⅰ、丁香酚、6-姜辣素、8-姜酚、10-姜酚6种成分的含量。方法:以白术内酯Ⅰ为内标物,建立白术内酯Ⅰ与白术内酯Ⅲ、丁香酚、6-姜辣素、8-姜酚、10-姜酚之间的校正因子,计算待测成分含量,并将外标法测定值与一测多评法计算值进行对比,验证所建立一测多评法的准确性。结果:白术内酯Ⅲ、白术内酯Ⅰ、丁香酚、6-姜辣素、8-姜酚和10-姜酚分别在1.09~21.80μg·mL-1、0.71~14.20μg·mL-1、13.67~273.40μg·mL-1、6.04~120.80μg·mL-1、1.46~29.20μg·mL-1、1.78~35.60μg·mL-1范围内线性关系良好;各成分一测多评法计算值与外标法实测值无显著性差异。结论:利用校正因子对丁蔻理中丸中6种成分的含量测定是可行的,一测多评法可以用于丁蔻理中丸的质量评价研究。     

Comparison of seven different reagents of peroxidase method for small and dense low density lipoprotein cholesterol (sdLDL‐C) measurement

BackgroundWe validated the performance of seven different reagents of peroxidase method for sdLDL‐C in two automatic analyzers that are common in Chinese laboratories.MethodsSeven commercially available sdLDL‐C assays were analyzed with the Beckman AU5400 and Mindray BS2000 automatic analyzers. A total of 336 blood samples were collected and the reference interval was also validated in 298 apparently healthy individuals. Serum samples were used for method comparison of precision, recovery, lower limit of detection, comparison and concurrence analysis, as well as reference interval for the Mindray reagent.ResultsThe repeatability CV% of the seven sdLDL‐C assays were 0.81%~3.66% for Mindray BS2000 and 0.76%~3.91% for Beckman AU5400, while Total CVs for Mindray BS2000 sdLDL‐C assay were 1.34%~4.81%, and that of Beckman AU5400 were 2.25%~10.33%. The measured recovery rates of sdLDL‐C assays were within the allowable ±10% deviation range. There was no obvious difference between the reagents in the lower limit detection. There was a difference between the validation results of the reference range and the manufacturer''s.BSBE, Mindray, and Dongou had a high degree of association with DENKA SEIKEN on Mindray BS2000, while BSBE, Mindray, Dongou and Merit Choice had a high degree of association with DENKA SEIKEN on Beckman AU5400. Passing–Bablok regression showed excellent linear correlation between BSBE and Mindray and DENKA SEIKEN and on Beckman AU5400.ConclusionsOur results indicate that the basic performance can meet the testing requirements, but the comparability between them is still insufficient.     

PKA含量动力学检测方法的研究与建立

目的建立激肽释放酶原激活剂(prekallikrein activator, PKA)含量的动力学检测方法,并对该方法进行验证。方法对比不同样品稀释缓冲液的pH、离子强度,各步骤的孵育时间、孵育温度等因素,确定检测方法的最适条件。并对该方法的准确度、专属性、精密度、线性、稳定性和耐用性进行方法学验证。结果确定以0.05 mol/L Tris-HCl缓冲液(pH8.5、含0.15 mol/L NaCl)稀释样品,在37℃下与激肽释放酶原(PK)试剂混合孵育20 min后加入底物S-2302,检测反应前10 min内吸光度变化率△A405/min。方法验证结果表明PKA含量在(0.5~4.0)IU/mL范围内,△A405/min与含量呈良好的线性关系,标准曲线相关系数R2>0.99,校正标样回收率在96.9%~103.7%。专属性验证表明人血白蛋白和静注人免疫球蛋白(pH4)的辅料、低pH及蛋白质含量等因素对该方法检测PKA含量均无明显影响,各溶液的加标回收率分别为98.0%(0.9%NaCl溶液)、95.3%(0.46%辛酸钠溶液)、96.7%(10%麦芽糖溶液,pH4.0)、94.0%(20%BSA)、94.0%(5%BSA,pH4.0)。在(0.5~4.0)IU/mL内,该方法准确度和精密度均符合要求。其中质控品批内回收率在96.4%~109.5%, CV在0.2%~6.9%,批间回收率在101.5%~102.9%, CV在2.6%~5.9%。PK试剂和S-2302在室温放置6 h内,线性、准确度和精密度均能符合要求,质控品回收率在94.9%~109.9%。采用新建的动力学法和《中国药典》(ChP)2015年版终点法检测本公司生产的20批次人血白蛋白样品中PKA含量的结果对比表明2种分析方法差异无统计学意义(P>0.05),2种方法的相关系数为0.99805(n=20)。结论建立的PKA含量动力学检测方法,具有良好的线性、专属性、准确度、精密度、稳定性、耐用性,相比《中国药典》方法,新建方法更节省分析时间,更为方便、准确、快速,可实现快速测定人血白蛋白和静注人免疫球蛋白(pH4)PKA含量测定。     

Application of Pop-off Method to Bone Marrow and Peripheral Blood Specimens for Purposes of Electron Microscopy

In this paper, we describe a pop-off method applicable to the hematological field. Bone marrow or peripheral blood specimens from patients were placed on a clean glass slide and fixed immediately in 2% glutaraldehyde solution for 10 minutes. For the DAB reaction, the slide was immersed in the DAB reagent for 30 minutes, and post fixed with 1 % OsO₄ solution for 1 hour. Specimens on the slide were washed with buffer solution, dehydrated and polymerized directly on the slide. A gelatin capsule filled with Epon mixed monomer was then reversed over the specimen. After polymerization was completed, the specimen was popped off from the slide to the capsule and trimmed carefully to prepare for ultrathin sectioning. This method allows the entire sequence of tissue preparation to be carried out on the slide, from fixation to embedding, and even, especially in scanty specimens, including the DAB reaction. Electron microscopic findings in specimens prepared by this technique show excellent preservation and the absence of specific artifacts.     

Development of a loop-mediated isothermal amplification assay for the detection of porcine hokovirus

Hokoviruses have recently been detected as pathogens belonging to the family Parvoviridae, which comprises porcine hokovirus (PHoV) and bovine hokovirus (BHoV). In this study, we developed a loop-mediated isothermal amplification (LAMP) assay for the rapid, specific and sensitive detection of PHoV. A set of four primers specific for six regions within the PHoV VP1/2 genes was designed using online software. The reaction temperature and time were optimized at 65 °C and 60 min, respectively. LAMP products were detected by agarose gel electrophoresis or by visual inspection of a color change caused by a fluorescent dye. The method was highly specific for PHoV, and no cross-reaction was observed with porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine bocavirus (PBoV), pseudorabies virus (PRV), porcine reproductive and respiratory syndrome virus (PRRSV), classic swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The detection limit was approximately 10 copies per reaction, which was 10 times more sensitive than conventional PCR. Furthermore, the efficiency of detection of PHoV in clinical samples was comparable to that of PCR and sequencing. These results show that the LAMP assay is a simple, rapid, sensitive and specific method for detecting PHoV. It does not require specialized equipment and can be used to detect PHoV both in the laboratory and in the field.