重组人源性抗HBsAg Fab抗体的肽质量图谱分析

重组人源性抗HBsAg Fab抗体具有较好的特异性和抗原结合活性,为了更好的阐明毕赤 酵母表达的重组人源性抗HBsAg Fab抗体的性质,用基质辅助激光解析飞行时间质谱(MALDI- TOF-MS)对重组Fab抗体的分子质量和肽质量图谱进行了分析。结果显示,毕赤酵母表达的重组 人源性抗HBsAg Fab抗体的分子质量为50678．49Da,与根据其一级结构计算的理论分子质量相 比多2763．84 Da,显示酵母表达的重组Fab抗体为糖蛋白。用胰蛋白酶酶解重组Fab抗体后进行 MALDI-TOF-MS分析显示,大部分的酶解肽段均能检测出来。结果表明毕赤酵母表达的重组Fab 抗体与预期的结构一致。

基于知识图谱的复杂薄壁零件机械加工工艺知识建模研究

智能工艺设计是数字孪生环境下工艺设计的核心,零件工艺知识建模是实现基于数字孪生的智能工艺设计的前提。为此,针对航空航天领域复杂薄壁零件机械加工工艺数据结构化程度低、难以重用的问题,提出了典型复杂薄壁零件机械加工工艺知识图谱的构建和质量评估方法。首先,对机械加工工艺知识组成和结构进行分析。其次,通过本体建模、知识抽取、知识储存等相关技术实现了工艺知识的可视化表征,并基于Neo4j图数据库实现机械加工工艺知识检索。最后,利用层次分析法对构建完成的知识图谱进行评估,并以框段类零件的机械加工工艺知识为验证对象,得到子图谱的综合准确度为92.28%。试验结果表明,基于知识图谱的工艺知识建模方法切实可行,有助于实现工艺知识的有效组织和重用,为数字孪生的智能工艺设计奠定基础。

融合多算法评估中英文知识图谱外延简洁性新方法

针对目前人们只是从理论上提出了针对知识图谱外延简洁性评估指标,并没有给出针对该指标规范的评估方法及流程问题,对知识图谱外延简洁性评估方法进行了研究,提出了支持中英文混合知识图谱外延简洁性评估的新方法。该方法定义了从总体层面进行分组以及分别对头实体、关系和尾实体进行评估的公式,同时为保障评估的准确性,定义了句子层面的评估公式。最后,将4种评估公式联合,得到了对知识图谱外延简洁性指标进行评估的算法。为验证所提出算法的准确性和性能,利用开放数据集OPEN KG(Knowledge Graph),对提出的算法和相关的算法进行了评估与比对,结果验证了本算法对中英文混合知识图谱简洁性评估方面的准确性、时间效率都具有一定的保障,综合性能高于相关的算法。

提高电泳图谱质量的几项技术措施

电泳技术是生命科学本科生必需掌握的一项分离技术。本文以醋酸纤维薄膜电泳为例,提出八项技术措施和细节,提高电泳图谱质量。

基于卷积神经网络的智能制造过程质量异常诊断

针对现有方法在智能制造过程中诊断能力有限和识别精度不高的问题,提出了一种与智能制造过程相适应的基于卷积神经网络的质量异常诊断模型。首先建立基于实时数据的过程质量图谱,以精准表达制造过程运行状态。其次,构建用于识别质量图谱的卷积神经网络诊断模型。最后,利用滑动窗口取值的方式对当前过程运行状态进行动态诊断,并通过某球磨过程验证了所提方法的有效性与实用性。结果表明,所提方法优于传统浅层模型,能够有效的对过程异常状态进行识别与诊断。

福建省生态环境综合信息图谱探讨

基于地学信息图谱的思想和理论,提出构建福建省生态环境综合信息图谱数据库和全省生态环境类型分类体系,实现生态环境信息数字化,增强福建省生态环境信息的可视化,为福建省生态环境的研究提供一种新的思路。

竹节参药材化学成分指纹图谱研究

建立竹节参药材化学成分的指纹图谱,为竹节参药材的质量分析及其临床应用提供依据。采用HPLC法,C18色谱柱,以0.05%磷酸水溶液和乙腈为流动相,梯度洗脱,检测波长为203 nm,建立了竹节参化学成分指纹图谱。竹节参指纹图谱共形成19个共有峰,通过对照品比对和HPLC-MS/MS分析,指认了10个共有峰,分别是人参皂苷Rg1、Re、Rb1、Rg2、Ro、Rg3、Rh2、竹节参皂苷Ⅳ、Ⅰb、Ⅳa。主成分分析和聚类分析将14批药材分为3类。主成分分析与聚类分析结果一致。所建立的竹节参化学成分指纹图谱准确、可靠、可行,对于竹节参药材的生产和质量控制提供依据。

家蚕幼虫关联基因在胚胎期的表达(英文)

用蛋白质表达谱分析技术研究家蚕胚胎发育时期的基因表达.用蛋白质双向电泳技术(2-DE)分离蛋白质,用I mage Master Elite图谱分析软件比较分析12个家蚕胚胎发育特异时期的2-DE图谱,发现一些蛋白斑点丰度高,并且一旦出现就在后续特异时期一直表达,推测这些蛋白可能在胚胎发育和代谢方面起重要作用,从中选择5个点分析它们的序列和功能.用基质辅助激光解吸电离质谱(MALDI-TOF-MS)和数据库检索鉴定蛋白质,发现5个幼虫期功能蛋白分别在4个胚胎发育特异时期表达:保幼激素结合蛋白(J HBP)基因在缩短期表达,表皮生长因子受体在头胸分化期表达,家蚕幼虫表皮蛋白和副腺特异肽在毛瘤发生期表达,淀粉酶在转青期表达.这些幼虫功能蛋白都在胚胎特定的生理期功能需要下出现,表明幼虫关联基因在胚胎期有序而及时地表达.

质谱技术在微生物检测和鉴定中的应用

随着医学技术的蓬勃发展,质谱技术在医学中的应用越加广泛,质谱技术能够通过样品中离子产生的质量图谱对于其分子构成进行分析,广泛用于临床中细菌的快速鉴定中。在生物学中常用MALDI-TOF质谱仪对于革兰氏阴性菌种与阳性菌种进行鉴定,通过质谱法对于微生物进行鉴定是基于不同物种不同的特征光谱来进行区别的。本文通过对近年来相关文献的查阅,介绍了临床基于该技术的应用与样品的制备,简述了微生物检测分类,分析了该技术进行微生物检测的局限性,阐述了在细菌鉴定方面所可能遇见的挑战与应用情况,并就其日后的发展进行展望。

Study and Application of Fingerprints of Ginkgo biloba Leaves Preparations

Aim To establish a method for determination of Ginkgo biloba L, its extractand preparations with HPLC fingerprints, so as to control the quality of the preparations. MethodsHPLC-DAD method was used to determine the constituents in preparations. Diamonsil? C_(18) (200mm X 4.6 mm, 5 μm) was used as analytical column, and acetonitrile/KH_2PO_4 was used as mobilephase with gradient elu-tion. The column temperature was at 24 ℃. The HPLC profile of chemicalconstituents of control sample and preparations were analyzed using similarity software. Results Thefingerprints of different preparations from different companies were slightly different because ofthe different preparing procedures. Mean while, the fingerprints of different batches of the samepreparation from the same company were similar to each other and the technology of each preparationwas stable. Conclusion This method is accurate, reproducible , simple, and can be used as ananalytical method for the routine quality control of Ginkgo biloba preparations.

Diagnostic model of saliva protein finger print analysis of patients with gastric cancer

AIM:To explore the method for early diagnosis of gastric cancer by screening the expression spectrum of saliva protein in gastric cancer patients using mass spectrometry for proteomics.METHODS:Proportional peptide mass fingerprints were obtained by analysis based on proteomics matrix-assisted laser desorption ionization time-of-flight/mass spectrometry.A diagnosis model was established using weak cation exchange magnetic beads to test saliva specimens from gastric cancer patients and healthy subjects.RESULTS:Significant differences were observed in the mass to charge ratio(m/z) peaks of four proteins(1472.78 Da,2936.49 Da,6556.81 Da and 7081.17 Da) between gastric cancer patients and healthy subjects.CONCLUSION:The finger print mass spectrum of saliva protein in patients with gastric cancer can be established using gastric cancer proteomics.A diagnostic model for distinguishing protein expression mass spectra of gastric cancer from non-gastric-cancer saliva can be established according to the different expression of proteins 1472.78 Da,2936.49 Da,6556.81 Da and 7081.17 Da.The method for early diagnosis of gastric cancer is of certain value for screening special biological markers.

Optimization of DsbA Purification from Recombinant Escherichia coli Broth Using Box-Behnken Design Methodolog

Disulfide bond formation protein A （DsbA） is one of the important helper proteins for folding in protein synthesis in vivo. In this study, purification of recombinant DsbA was investigated by examining four important factors with Box-Behnken design method, a statistic-based design of experiments. The optimal operation conditions were obtained by adopting the effectiveness coefficient method on the multi-objective problem, which takes the protein recovery, purification efficiency and throughput of ion-exchange chromatography into account. After the optimization, protein recovery of 96.8% and purity higher than 95% DsbA was achieved, and the productivity was （377.9±1.7） mg soluble DsbA per liter broth. The purified protein was identified by peptide mass fingerprinting matching the record of gil2624856, a mutant of DsbA. The DsbA was preliminarily applied to the refolding of denatured lysozyme in vitro.

基于全方特征图谱质量表征的芩连制剂质量评价研究

目的建立芩连制剂特征图谱质量表征方法,以全面表征芩连制剂质量,并应用于芩连制剂的质量评价。方法采用UHPLC-DAD多波长分析方法,Waters Cortecs T3(100 mm×2.1 mm,1.6μm)色谱柱,以乙腈(含0.02%甲酸)-(10 mol/L乙酸铵-0.09%甲酸缓冲液)为流动相,梯度洗脱,体积流量0.3 mL/min,柱温30℃,检测波长230、330 nm。结果建立了芩连制剂特征图谱质量表征方法,指认了38个特征峰,且检测了12个指标性成分的含量,平均加样回收率为83.58%~106.76%;测得11批芩连制剂中12个指标性成分质量分数分别为连翘酯苷A 0.21~9.08 mg/g、盐酸黄柏碱0.39~1.91 mg/g、芍药内酯苷0.13~9.28 mg/g、芍药苷3.02~20.24 mg/g、甘草苷0.19~0.51 mg/g、黄芩苷12.36~27.64 mg/g、1,2,3,4,6-五-O-倍酰-D-葡萄糖(PGG)0.22~3.63 mg/g、汉黄芩苷1.92~6.12 mg/g、连翘苷0.22~4.10 mg/g、盐酸小檗碱5.37~16.85 mg/g、盐酸巴马汀0.53~2.26mg/g、甘草酸0.45~2.06mg/g。不同厂家及其不同批次芩连制剂中成药的质量表征差异主要来源于黄芩(峰22、38、27)、赤芍[峰2、21(PGG)、7(芍药内酯苷)]、连翘[峰36(连翘酯素)、30]、黄连(峰24)、黄连-川黄柏(峰8、4)、连翘-赤芍(峰3)。结论建立的特征图谱质量表征方法可全面地表征与评价芩连制剂中成药,为该类品种的质量控制提供了科学依据,亦为中成药的质量控制提供方法学借鉴。

Murine brain tissues with human cytomegalovirus infection: a proteomic study

To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from murine brain tissues by human cytomegalovirus（HCMV） infection and paired murine brain tissues and to identify the differential expression proteins. Methods： Forty Kunming mice were randomly divided into infection group （20） injected with HCMVAD169 and control group （20） injected with saline into their brain. After 30 days, the murine brain tissues by HCMV infection and paired murine brain tissues were separated by two-dimensional gel electrophoresis（2-DE）, analyzed by Image Master 2D software, and identified by peptide mass fingerprint（PMF） and database searching, and make Western blotting analyses the differential expression of individual proteins. Results： Well resolved, reproducible 2-D maps of the above tissues were obtained. Some of the different proteins identified by mass spectrometry（MS） were matched in the SWISS-2D PAGE database, Western blotting analyses were further carried out to verify the differential expression of individual proteins. Conclusion： These data will be valuable for studying the diagnosis of disease at an early stage, mechanisms of pathogenic and the key to the development of anti-HCMV medicine.

Chemical fingerprinting of Su-He-Xiang-Wan and attribution of major characteristic peaks for its quality control by GC-MS

A simple and facile gas chromatography-mass spectrometer （GC-MS） fingerprint of Su-He-Xiang-Wan （SHXW） was developed, the similarity analysis was conducted, and attribution of the major characteristic peaks was identified for SHXW quality control. GC-MS analysis was performed on a QP2010 instrument （Shimadzu, Japan） equipped with a capillary column of RTX-5MS. The column temperature was initiated at 50℃, held for 5 min, increased at the rate of 3 ℃/min to 120 ℃, held for 2 min, and then increased at the rate of 4 ℃/min to 220℃, held for 10 min. Helium carrier gas was used at a constant flow rate of 1.3 mL/min. Mass conditions were ionization voltage, 70 eV; injector temperature, 250℃; ion source temperature, 250 ℃; splitting ratio, 30：1; full scan mode in the 40-500 Da mass ranges with rate of 0.2 s per scan. Attribution of the major characteristic peaks was identified for SHXW by comparing the chemical standards, references of Chinese herbal medicines and the negative controls of prescription samples （NC） of SHXW. With the help of the temperature-programmed retention indices （PTRIs） used together with mass spectra and chemical standards, 25 major characteristic peaks have been identified. Nine volatile medicinal materials were identified in the prescription of SHXW by attributing to the 27 major characteristic peaks. The results demonstrate that the proposed method is a powerful approach to quality control of complex herbal medicines.

Development of the fingerprints of crude Pu-erh tea and ripened Pu-erh tea by high-performance liquid chromatography

A simple and accurate high-performance liquid chromatography（HPLC）method to generate the fingerprints of crude Pu-erh tea（CPT）and ripened Pu-erh tea（RPT）is described.A suitable chromatographic system was established using a linear gradient elution with acetonitrile and water containing 0.6% formic acid as the mobile phase and a detection wavelength of 300 nm.HPLC analysis identified 24 common peaks among RPT and 21 common peaks among CPT.This study revealed that crude Pu-erh tea and ripened Pu-erh tea contained some similar major constituents,but with distinctive peaks,which may be caused by the difference in the fermentation process.This HPLC fingerprint method could be used to evaluate and authenticate crude Pu-erh tea and ripened Pu-erh tea.

High-performance liquid chromatographic fingerprint analysis of Oxytropis falcata Bunge and Oxytropis chiliophylla Royle

A simple, sensible and reliable HPLC-DAD fingerprint analysis method for the raw materials of Oxytropisfalcata and Oxytropis chiliophylla, both of which were used as ＂Er-Da-Xia＂ in Tibetan medicines, was developed and then subsequently applied to analyze samples collected from different locations or times. 19 common fingerprint peaks for O. falcata, 24 for O. chiliophylla, and 11 for the two herbs were designated respectively, including 7 identified characteristic peaks existing in both herbs and 1 uniquely presenting in O. chiliophylla. Although there were some slight differences in the chemicals of O. falcata and O. chiliophylla, the main components of both herbs were consistent generally. The results provided scientific basis, at least from the chemical point of view, for the reasonablity of two herbs being used as the same drug in Tibetan medicines and for the necessary of further investigation on their detailed chemical and pharmacological differences.

Chemical analysis of the Hedysarum multijugum root by HPLC fingerprinting

The root of Hedysarum multijugum（RHM） is recorded as a folk herbal medicine in China and is sometimes used as a substitute for Hedysari Radix, which is a famous traditional Chinese medicine derived from the roots of Hedysarum polybotrys. In the present study, a sensible, reliable, and reproducible HPLC-DAD fingerprint analysis method for RHM was developed and then subsequently applied to analyze RHM samples from different origins. The chemical constituents of the RHM samples were generally consistent, although it was slightly affected by the local environment of the plant. In addition, the chemical constituency of RHM was shown to be significantly different from that of Hedysari Radix, suggesting that RHM is not suitable as a substitute for Hedysari Radix, at least from the chemical point of view.

Development of a fingerprint of Melilotus suaveolens Ledeb by high-performance liquid chromatography

A simple and accurate high-performance liquid chromatography method （HPLC） was described to generate the metabolite fingerprints of Melilotus. suaveolens Ledeb of different harvesting times and different parts of the plant. A suitable ehro- matographic system was established using a linear gradient elution with acetonitrile and 0.1% phosphoric acid （H3PO4） as the mobile phase and a detection wavelength of 210 nm. HPLC analysis identified 12 common peaks and 8 common fingerprint peaks among the different parts of the herb. We found that the constituents varied at different harvesting times and in different parts of the plant. The HPLC fingerprint method could be used to evaluate the quality ofM. suaveolens Ledeb and Melilotus genus herbs. The same strategy can also be applied for the development of fingerprints for the quality control of other herbal medicines.

泵送混凝土配合比设计与控制创新模式研究之二——泵送混凝土配合比理论模型及应用

本研究是前文泵送混凝土配合比设计与控制创新模式研究之一(《商品混凝土》2009年第二期)的延伸和继续。围绕着单掺粉煤灰和双掺粉煤灰、矿渣微粉两方案,泵送混凝土材料组、度质量图谱的广义泵送混凝土配合比组成理论模型开展深入理论探索,组成材料合理配制逐级演绎推算,创建品质较优的泵送混凝土配合比总体框架理论模型,为编绘可控质量图谱提供序列准(经验)混凝土配合比,做系统试配,绘制泵送混凝土配合比专用尺。