HPLC同时测定断节参中告达亭和开德苷元的含量

目的建立断节参中开德苷元、告达亭两个C21甾体类成份的分析方法。方法采用waters SunFire C18(4.6mm×250mm,5μm)色谱柱,乙腈-0.1%磷酸(43∶57)为流动相;流速:1.0mL·min^(-1);检测波长:224nm;柱温为30℃。结果开德苷元、告达亭分别在0.0109~0.544mg·mL^(-1)、0.0079~0.3955mg·mL^(-1)范围内与峰面积线性关系良好(r≥0.9996);平均加样回收率分别为99.82%、99.80%,RSD分别为1.01%、1.04%(n=6);精密度、稳定性、重复性实验结果RSD均小于1.0%。结论所建立的方法简单、准确、稳定,可用于评价断节参药材的质量。

断节参抑制α-葡萄糖苷酶活性成分研究

目的:寻找断节参中具有抑制α-葡萄糖苷酶活性的成分。方法:采用体外抑制α-葡萄糖苷酶活性模型进行追踪,用各种色谱法分离,根据理化性质和谱学数据鉴定结构,筛选出活性较强的单体化合物并进行酶活性抑制动力学研究。结果:断节参乙醇提取物的乙酸乙酯溶性部位具有显著的抑制α-葡萄糖苷酶活性,从中分离出3个化合物,其中断节参苷H和青阳参苷B两个皂苷类化合物具有较强抑制α-葡萄糖苷酶活性,IC50分别为21.90、35.32 mg·L-1,明显高于阳性对照药阿卡波糖(IC50=1 017.41 mg·L-1)。酶活性抑制动力学反应结果表明,两个皂苷对α-葡萄糖苷酶的抑制类型均为非竞争性抑制剂。结论:断节参苷H和青阳参苷B为首次报道对α-葡萄糖苷酶有抑制活性。

断节参提取物抗炎镇痛及体外抗氧化活性研究

目的:探讨断节参提取物不同极性部位的抗炎、镇痛、抗氧化作用。方法:采用角叉菜胶致小鼠足肿胀,连续给药后,测定足肿胀度、炎症组织蛋白渗出量、PGE2含量,综合评价抗炎效果;对小鼠连续灌胃给药后,观察热板痛阈值评价镇痛效果;通过清除1,1-二苯基-2-三硝基苯肼(DPPH·)自由基和还原Fe3+能力(FRAP)评价断节参提取物的体外抗氧化效果。结果:断节参提取物不同极性部位均表现出不同程度的抗炎、镇痛、抗氧化作用,尤其以乙酸乙酯部位综合效果最为明显,能抑制角叉菜胶致小鼠足肿胀,显著降低炎症组织中蛋白质渗出及PGE2生成,并具有明显的镇痛及抗氧化作用。结论:断节参提取物乙酸乙酯和正丁醇部位具有明显的抗炎、镇痛、抗氧化作用,为断节参的主要活性部位。

竹节参皂苷Ⅳa对高脂饮食小鼠小肠干细胞及其微环境Wnt/β-catenin信号通路的影响

目的:探讨竹节参皂苷Ⅳa对高脂饮食小鼠小肠干细胞及其微环境Wnt/β-catenin信号通路的影响。方法:C57BL小鼠雌雄各半随机分成4组,每组10只,即正常对照组、模型组及竹节参皂苷Ⅳa低、高剂量组。HE染色观察小肠形态学变化,免疫组化检测各组小肠干细胞数量、PCNA表达及其微环境Wnt/β-catenin信号通路的变化。结果:与正常对照组比较,模型组小肠绒毛长度、V/C值、PCNA表达量及杯状细胞数量、Bmi1阳性表达的小肠干细胞数量显著升高,小肠干细胞微环境Wnt/β-catenin信号显著上调(P<0.05或P<0.01)。与模型组比较,竹节参皂苷Ⅳa组绒毛长度、V/C值、PCNA表达量、杯状细胞数量、Bmi1阳性表达的小肠干细胞数量显著降低,小肠干细胞微环境Wnt/β-catenin信号显著下调(P<0.05或P<0.01)。结论:竹节参皂苷Ⅳa可通过Wnt/β-catenin信号通路调控小肠干细胞微环境,改善其过度增殖分化状态和抑制小肠干细胞增多,调节小肠干细胞功能。

断节参的化学成分研究

目的研究云南产断节参的化学成分。方法采用反复硅胶柱层析和葡聚糖凝胶柱层析的色谱方法对断节参乙酸乙酯提取部位进行分离纯化,通过理化性质和光谱(1H-NMR、13C-NMR)数据进行结构鉴定。结果从断节参根乙酸乙酯提取部位中分离得到7个化合物,分别鉴定为:白首乌二苯酮(1)、2,5-二羟基苯乙酮(2)、2,5-二羟基苯甲酸乙酯(3)、4-羟基苯乙酮(4)、2,4-二羟基苯乙酮(5)、4-羟基苯甲酸(6)、β-谷甾醇(7)。结论所分离化合物均为首次从断节参植物中分离得到。

HPLC法同时测定断节参中5种苯乙酮

目的建立断节参中5种苯乙酮类化合物的HPLC定量测定方法。方法断节参甲醇提取液分析采用HanbonC18色谱柱（4.6 mm×250 mm,5μm）,以甲醇（A）-0.1%乙酸水溶液（B）为流动相,柱温30℃,体积流量1.0m L/min,检测波长265 nm。结果 2,5-二羟基苯甲酸甲酯、4-羟基苯乙酮、白首乌二苯酮、2,5-二羟基苯乙酮、2,4-二羟基苯乙酮分别在2.66～53.2μg/m L、4.56～91.2μg/m L、4.10～82.0μg/m L、2.18～43.6μg/m L,1.53～30.6μg/m L范围内与峰面积呈良好线性关系（r〉0.999 0）,它们的平均加样回收率为98.5%～101.4%,RSD为1.4%～2.8%。结论 5种苯乙酮成分在22 min内就能达到完全分离,方法简便易行且稳定可靠。

断节参的HPLC指纹图谱研究

目的建立断节参药材的高效液相色谱法(high performance liquid chromatography,HPLC)指纹图谱,为断节参质量控制提供依据。方法采用Hanbon-C18(4.6 mm×250 mm,5μm)色谱柱,以甲醇(A)-0.5%乙酸水溶液(B)为流动相进行梯度洗脱,体积流量1.0 mL·min^-1,柱温30℃,检测波长265 nm。对所收集的9个不同产地批次的断节参药材建立HPLC指纹图谱,采用国家药典委员会"中药色谱指纹图谱相似度评价系统(2012版)"对结果进行分析处理,并对主要成分峰进行指认。结果9批不同产地断节参药材的HPLC指纹图谱中,确定了16个共有峰,指认出的3个峰分别为:2,5-二羟基苯甲酸甲酯、白首乌二苯酮、2,4-二羟基苯乙酮。各批次样品相似度在0.91以上。结论本试验所建立的指纹图谱重复性好,简便可行,可作为断节参的质量控制和评价的依据。

响应面法优化断节参C21甾体总苷醋酸水解条件

目的:通过Box-Behnken响应面法优化断节参C21甾体总苷的醋酸水解条件。方法:以告达亭苷元含量为指标,考察醋酸浓度(A)、水解时间(B)、水解温度(C)3个因素对断节参C21甾体总苷醋酸水解的影响,并进一步优化其水解工艺。结果:C21甾体总苷的醋酸水解最佳工艺为醋酸浓度5.6%、水解时间6.5 h、水解温度91℃,此条件下告达亭含量为6.79 mg/g。结论:该条件下优化的断节参总皂苷的水解工艺稳定、可行,可为断节参的水解工艺操作提供参考。

竹节参总皂苷对大鼠心肌梗死的保护作用

目的:观察竹节参总皂苷对大鼠心肌梗死的保护作用,并初步探讨其发挥作用的机制。方法:通过结扎左冠状动脉前降支建立大鼠心肌梗死模型,模型成功后随机分为模型组、竹节参总皂苷组(50 mg/kg、100 mg/kg),连续灌胃给药4周,假手术组和模型组给予等体积的生理盐水。4周后进行心功能检测;取心脏,称重后计算心脏重量指数;HE、Masson、苦味酸-天狼星红染色,观察心肌组织形态并计算胶原容积分数。取血清,检测SOD、CAT、GSH-Px的活性和MDA的含量;ELISA法检测TNF-α、TGFβ炎症因子的含量。结果:模型组大鼠心功能明显受损,心脏重量指数升高,并发生纤维化;血清中抗氧化物酶的活性降低,脂质过氧化物含量升高,且炎症因子含量升高,与假手术组比较有显著性差异。竹节参总皂苷(50 mg/kg、100 mg/kg)能使以上症状得到显著改善。结论:竹节参总皂苷对心肌梗死大鼠具有保护作用,其作用机制可能与降低氧化应激反应和炎症损伤相关。

蒸汽凝结水两种回收方式的技术经济比较

本文以某油气加工企业为例从技术和经济两个方面对蒸汽凝结水的两种加收方式进行了比较，并着重说明了密闭式回收方式进行了比较，并着重说明了密闭式回收方式的工艺原理，为同类企业的供热提供了一个节能降耗的范例。

Analysis of the Constituents in the Chinese Drug Notoginseng by Liquid Chromatography-Electrospray Mass Spectrometry

To develop a HPIX-UV-MS method for identifying the constituents in theChinese drug Notoginseng (the root of Panax notoginseng). Methods A Phenomenex Luna C_(18) column(250 mm x 4.6 mm ID, 5 μm) was utilized. Water containing 0.005% formic acid (A) and acetonitrilecontaining 0.005% formic acid (B) were used as gradient eluents. UV spectra were recorded in range195 - 400 nm. Both positive and negative ion ESI modes were used. Results The constituents inNotoginseng were well separated and detected. Fourteen compounds were identified by comparing theirretention time and ESI-MS data with those obtained from the reference compounds. Forty-one compoundswere deduced by data analysis of MS and literature; among them, yesanchinosides-H and -E,chikusetsusaponin-L_5, malonyl-ginsenoside-R_(g_1), the isomers of notoginsenosides-J, -A, -R_1, -G,-R_2, and ginsenoside-Rh_3 were discovered in Notoginseng for the first time. Conclusion Thismethod gives high sensitivity and good separation, and is suitable for identifying the constituentsin Notoginseng. This result is helpful for further phytochemical research on Notoginseng. Based onthis result, further quality control can be studied.

Rock skeleton models and seismic porosity inversion

By substituting rock skeleton modulus expressions into Gassmann approximate fluid equation, we obtain a seismic porosity inversion equation. However, conventional rock skeleton models and their expressions are quite different from each other, resuling in different seismic porosity inversion equations, potentially leading to difficulties in correctly applying them and evaluating their results. In response to this, a uniform relation with two adjusting parameters suitable for all rock skeleton models is established from an analysis and comparison of various conventional rock skeleton models and their expressions including the Eshelby-Walsh, Pride, Geertsma, Nur, Keys-Xu, and Krief models. By giving the two adjusting parameters specific values, different rock skeleton models with specific physical characteristics can be generated. This allows us to select the most appropriate rock skeleton model based on geological and geophysical conditions, and to develop more wise seismic porosity inversion. As an example of using this method for hydrocarbon prediction and fluid identification, we apply this improved porosity inversion, associated with rock physical data and well log data, to the ZJ basin. Research shows that the existence of an abundant hydrocarbon reservoir is dependent on a moderate porosity range, which means we can use the results of seismic porosity inversion to identify oil reservoirs and dry or water-saturated reservoirs. The seismic inversion results are closely correspond to well log porosity curves in the ZJ area, indicating that the uniform relations and inversion methods proposed in this paper are reliable and effective.

Research Advances on Panax japonicus and its Approximation Varieties in Tujia Nationality

Panax japonicus and its approximation varieties,such as Rhizoma Panacis Majoris and Panax japonicus C. A. Mey. var.major （Burk.） C.Y. Wu et K.M. Feng belong to Panax,which are less commonly used traditional Chinese medicine. Because of similar traits and effectiveness,they were always used as one type of medicine for a long time. Aiming at this phenomenon,the chemical composition and contents of P. japonicus and its approximation varieties from different area were compared in order to provide a chemical basis for clarifying the classification of the genus.

Regulation of Bcl-2 Family in TIP30-Induced Apoptosis in Hepatoblastoma Cells

Objective: To investigate the role of TIP30 in the apoptotic signal pathwayin HepG2, and Hep3B and Hu-7 hepatoblastoma cell lines. Methods: In order to confirm whether TIP30conducted Bcl-2 family was involved in apoptosis signal pathway, MTT assay, in situ 3' end labellingof DNA assay and Western blot were carried out to detect the diverse apoptotic function of TIP30and the regulation of Bcl-2 family. Results: TIP30 induced apoptosis as evidenced by morphologicalchanges in hepatoblastoma cells, which was accompanied by up-regulating Bax and Bad proteins andstimulating them from cytoplasm to mitochondria, and down-regulating Bcl-xl, while it had no effecton the level of Bak protein. Conclusion: TIP30 induced apoptosis partly by modulating the proteinlevels of members of Bcl-2 family in hepatoblastoma cells. Elucidating the mechanism by which TIP30induces cell death might establish it as an anticancer factor.

Effects of photoperiod on daily activity rhythm of juvenile sea cucumber, Apostichopus japonicus (Selenka)

In nature, Apostichopus japonicus exhibits a behavioral characteristic of emerging at night and sheltering during the day. Hence, it was commonly believed that longer and darker light conditions are better for a population of A. japonicus. In aquaculture situations therefore, animals have commonly been cultured without lighting, especially during juvenile-hatching culture. However, how the length of darkness each day affects .4.japonicus and what the mechanism is of behavioral response of A. japonicus to different photoperiods remain unclear, and are specifically addressed in this study. We applied nine photoperiod treatments （light：dark （LD） cycles at 0：24, 3：21, 6：18, 9：15, 12：12, 15：9, 18：6, 21：3, and 24：0 under 500Ix） to 60 individuals （29.73±0.23 g） per treatment （with 3 replicates）. The enviroramental conditions were set as： water temperature = 16.9±0.3℃, dissolved oxygen〉6.0 rag/L, ammonia〈0.3 mg/L, pH=7.8-8.2, and salinity=30-32. Under experimental conditions of continuous darkness as well as continuous light, cyclic nocturnal activity patterns of A. japonicus （viz. the animals emerged and fed at night and sheltered during the day） were observed. However, they spent more and less time, respectively, moving and feeding under continuous darkness and continuous light, than those under a natural light cycle. Under photoperiods with 6-12 h of light, the animals showed one sheltering behavior transition and two emerging behavior transitions. These behavior transitions appeared to be governed by some internal physiological factors and induced by daily light variation. The behavior of the animals was significantly affected by different photoperiods, and the distribution rate （DR） of the animals emerging at ＂daytime＂ increased with lengthening ＂light time＂ （except for the LD 3：21 cycle）. We also found that there was no significant difference in mean DR per day under LD cycles of 6：18, 9：15, 15：9, and 12：12. Mean DRs per day were significantly lower for these photoperiods, than for those under LD cycles of 0：24 and 3：21, and significantly higher than for those under LD cycles of 18：6, 21：3 and 24：0. These results imply that 6-15 h light per day were the similar photoperiods to natural sunlight. From the viewpoint of activity and feeding behaviors, these might be the optimal photoperiods for rearing A. japonicus.

Screening and functional evaluation of the glucose-lowering active compounds of total saponins of Baibiandou(Lablab Semen Album)

Objective To screen forα-glucosidase inhibitor active compounds in the total saponins of Baibiandou(Lablab Semen Album)based on UHPLC-Q-Exactive Orbitrap MS technology and to evaluate its hypoglycemic activity in vivo.Methods Acarbose was used as the positive control,and the median inhibitory concentration(IC50)was used as the evaluation index ofα-glucosidase inhibitory activity to establish an in vitroα-glucosidase inhibition model.Further,UHPLC-Q-Exactive Orbitrap MS technology was used to screen and identify the active compounds ofα-glucosidase inhibitors in the total saponins of Baibiandou(Lablab Semen Album)in order to further verify the activity of the main active monomer and to perform homologous modeling and molecular docking of yeast-derivedα-glucosidase and human-derivedα-glucosidase,while the hypoglycemic activity was evaluated in diabetic mice.Results This study successfully identified 15 compounds with potentialα-glucosidase inhibitory activity,including Chikusetsusaponin IVa,from the total saponins of Baibiandou(Lablab Semen Album).Simultaneously,we verified the activity of the main active monomer Chikusetsusaponin IVa,and showed that it has strongα-glucosidase inhibitory activity.Theα-glucosidase inhibitory concentration IC50 was(565.2±1.026)μg/m L,and the IC50 of acarbose,which was lower than the positive control,was(706.6±1.058)μg/m L.The docking energies of Chikusetsusaponin IVa were–6.1 and–7.7 kcal/mol with yeast-derivedα-glucosidase and human-derivedα-glucosidase molecules,respectively.Both showed strong binding activity,and the levels of alanine aminotransaminase(ALT),aspartate aminotransaminase(AST),UREA,Creatinine(CREA),and cholesterol(CHO)were significantly decreased by Chikusetsusaponin IVa(P<0.05).In addition,it could repair damaged liver and pancreas cells of diabetic mice to some extent.Conclusion This study provides a basis for screeningα-glucosidase inhibitors and structural modifications of the total saponins of Baibiandou(Lablab Semen Album).

Seasonal Changes in Food Uptake by the Sea Cucumber Apostichopus japonicus in a Farm Pond: Evidence from C and N Stable Isotopes

This study investigated the seasonal changes in carbon (C) and nitrogen (N) stable isotope values of several typical food sources of Apostichopus japonicus in a farm pond, including particulate organic matter (POM), macroalgae, benthic microalgae and animals such as nematode and copepod. The stable isotope technique was used to quantify relative contributions of various sources to the food uptake by A. japonicus. The results showed that significant changes occurred in the C and N stable isotope values of sea cucumber food sources due to the seasonality of micro-or macroalgae prosperity and the fluctuation of environmental conditions. The sea cucumber A. japonicus exhibited corresponding alterations in feeding strategy in response to the changes in food conditions. Calculation with a stable isotope mixing model showed that macroalgae was the principal food source for A. japonicus throughout the 1-yr investigation, with the relative contribution averaging 28.1% - 63.2%. The relative contributions of other food sources such as copepod and nematode, POM, benthic microalgae to the total food uptake by sea cucumber averaged 22.6% - 39.1%, 6.3% - 22.2%, 2.8% - 6.5%, and 2.8% - 4.2%, respectively. Together these results indicated that the seasonal changes in food sources led to the obvious temporal differences in the relative contribution of various food sources utilized by A. japonicus. Such findings provide the basic scientific information for improving the aquaculture techniques of A. japonicus, particularly for optimizing the food environment of A. japonicus culture in farm ponds.

Multiplicity of Solutions for Nonlinear Biharmonic Equation Involving Critical Parameter and Critical Exponent

By using variational method, the multiplicity of solutions for nonlinear biharmonic equation involving critical parameter and critical exponent are established.

Run-to-run Optimization for Fed-batch Fermentation Process with Swarm Energy Conservation Particle Swarm Optimization Algorithm

An iterative optimization strategy for fed-batch fermentation process is presented by combining a run-to-run optimization with swarm energy conservation particle swarm optimization (SEC-PSO). SEC-PSO, which is designed with the concept of energy conservation, can solve the problem of premature convergence frequently appeared in standard PSO algorithm by partitioning its population into several sub-swarms according to the energy of the swarm and is used in the optimization strategy for parameter identification and operation condition optimization. The run-to-run optimization exploits the repetitive nature of fed-batch processes in order to deal with the optimal problems of fed-batch fermentation process with inaccurate process model and unsteady process state. The kinetic model parameters, used in the operation condition optimization of the next run, are adjusted by calculating time-series data obtained from real fed-batch process in the run-to-run optimization. The simulation results show that the strategy can adjust its kinetic model dynamically and overcome the instability of fed-batch process effectively. Run-to-run strategy with SEC-PSO provides an effective method for optimization of fed-batch fermentation process.

Study on dynamics of central pattern generator for a quadrupedal robot

Based on Matsuoka＇s central pattern generator （CPG） model and taking quadruped as an example, the dynamics of CPG model was investigated through the single-parameter-analysis method and the numerical simulation technique. Simulation results indicate that the CPG model exhibits complex dynamics, while each parameter has specifically definitive influence trends on the CPG output. These conclusions were applied to control a quadrupedal robot to walk in different gaits, clear obstacle, and walk up- and down-slope successfully.