我国鬼臼类植物资源的研究

本文较为系统地整理了我国鬼臼类的原植物,它们是八角莲属(Dysosma)、桃儿七属(Podophyllum)和山荷叶属(Diphylleia)等3属共10种。并对鬼臼的原植物进行本草考证,指出鬼臼应系八角莲属植物。同时对上述3个属的8个种的样品进行了成分定性,证明这些植物均含有鬼臼脂素及其他木脂素成分。另外还利用鬼臼脂素的分子中含有次甲二氧基的特点,用比色法测定了样品中的鬼臼脂素的含量:八角莲属6个种鬼臼脂素的含量不超过1%,桃儿七为2.7～4.2%,南方山荷叶为4.9%。     

质谱导向的肾指海绵Reniochalina sp.中环肽类成分研究

目的 以质谱为导向对肾指海绵Reniochalina sp.中的环肽类成分进行研究。方法 采用质谱引导的程序性分离手段定向追踪并分离纯化海绵中的环肽类成分;通过理化常数测定、波谱数据比对确定化合物结构;利用CCK-8法对化合物进行初步细胞毒活性评价。结果 从肾指海绵Reniochalina sp.中分离获得3个环肽类化合物,分别鉴定为stylopeptide 1 (1)、hymenamide D (2)、axinastatin 2 (3)。化合物1对6种人肿瘤细胞株具有细胞毒性,IC₅₀值范围为6.09 ~ 17.26 μmol/L。结论 化合物1 ~ 3首次分离自Reniochalina属海绵,化合物1是细胞毒性环七肽。     

木兰科药用植物的研究——Ⅱ.厚朴的原植物和资源利用

本文对厚朴进行了本草考证。研究了木兰科厚朴原植物来源,包括厚朴类,姜朴类、枝子皮类和土厚朴类等多种植物。同时用红外、紫外光谱和薄层层桁等方法检查了各类厚朴中厚朴酚(magnolol)(Ⅰ)与和厚朴酚(honokiol)(Ⅱ)以及β-桉油醇(β-cudcsmol)(Ⅲ)的存在。实验证明上述成分厚朴类都含,其余各类厚朴或含少量或不含。但从植物系统位置和植物化学分类观点来看,则Ⅰ,Ⅱ,Ⅲ的分布有一定规律,即属于木兰亚属(Subgen.Magnolia)皱皮木兰组(Seot.Rytidospermum)是Ⅰ,Ⅱ,Ⅲ的分布最集中的植物群。玉兰亚属(Subgen.Pleurochasma)各组除个别种外,均不含Ⅰ,Ⅱ,Ⅲ。未研究过的土厚朴类来源于木莲属(Manglietia),如四川木莲等也含Ⅰ或Ⅱ成分。     

基于超声辅助酶解与串联质谱联用的鹿角特征肽检测

目的 检测鹿角特征肽类成分，为鹿角、鹿角胶及其相关产品的质量研究提供实验依据。方法 通过超声辅助酶解的方法，快速酶解鹿角、鹿角胶样品。采用高效液相色谱-三重四极杆质谱对酶解液进行检测，在电喷雾离子化的正离子模式下，采用多反应监测，以鹿角特征肽m/z 765.5→554.4、m/z 850.4→515.4、m/z 864.0→535.4、m/z 858.7→527.4、m/z 612.2→464.2、m/z 845.0→507.3为离子对，检测鹿角、鹿角胶及掺伪样品的鹿角特征肽。结果 该方法可在1 h内完成鹿角相关样品特征肽类成分的检测，可用于鹿角与鹿皮样品的区分以及掺伪鹿角样品的检测。结论 采用超声辅助酶解与质谱联用的方法快速、准确、专属性强，可用于鹿角相关样品的快速精准检测。     

畲药树参茎化学成分分析及抗炎活性部位高效液相指纹图谱的建立

目的 对畲药树参茎所含化学成分进行确证分析，并建立抗炎活性部位的高效液相色谱指纹图谱。方法 采用液相色谱-质谱联用、核磁共振波谱仪等分析仪器分离鉴定树参茎中的化学成分；采用高效液相色谱法建立抗炎活性部位指纹图谱：色谱柱为Waters Sunfire^TM C₁₈(4.6 mm×250 mm，5 μm)，流动相为乙腈-混合水溶液(含甲酸0.2%和四氢呋喃0.2%)，梯度洗脱。检测波长为256 nm，柱温30 ℃，体积流量为1.0 mL·mim^–1，分析时间为80 min，进样量为5 μL。结果 从树参茎中共分离鉴定出10个化合物，分别是绿原酸(1)、3,4-O-二咖啡酰基奎宁酸(2)、3,5-O-二咖啡酰基奎宁酸(3)、4,5-O-二咖啡酰基奎宁酸(4)、saikolignanoside A(5)、芦丁(6)、山柰酚-3-O-芸香糖苷(7)、紫丁香苷(8)、松柏醇(9)、芥子醛葡萄糖苷(10)；12批样品抗炎活性部位指纹图谱相似度为0.901~0.992，均符合要求。结论 树参茎中主要含有酚酸类、黄酮类、苯丙素类化合物；建立的指纹图谱不仅能体现树参茎抗炎的化学物质基础，而且鉴别方法简便、重复性好，可为树参茎的品质评价及主要抗炎活性成分的确证提供科学依据。     

西洋参茎叶总皂苷碱降解成分西洋参茎叶总皂苷碱降解成分

目的研究西洋参茎叶总皂苷碱降解成分。方法采用硅胶柱色谱并结合HPLC进行分离纯化，通过波谱分析鉴定化合物的结构。结果从西洋参茎叶总皂苷碱降解产物中分离得到9种成分，分别鉴定为：20(S)-原人参二醇(I)，20(S)-达玛-25(26)-烯-3β,12β,20-三醇(II)，24(R)-ocotillol (III)，20(S)-原人参三醇(IV)，20(S)-达玛-25(26)-烯-3β,6α,12β,20-四醇(V)，达玛-20(21),24-二烯-3β,12β-二醇(VI)，达玛-20(21),24-二烯-3β,6α,12β-三醇(VII)，20(S),24(S)-达玛-25(26)-烯-3β,6α,12β,20,24-五醇(VIII)，20(S)-达玛-23-烯-25-过氧羟基-3β,6α,12β,20-四醇(IX)。结论碱降解20位S构型未改变。V，VII，VIII，IX为4个新化合物，并利用2D-NMR技术对新化合物的氢和碳的化学位移进行了归属。其中I对HCT-8人结肠癌细胞具有较强的细胞毒活性。     

药物生物利用度遗传神经网络预测研究

目的对药物生物利用度进行遗传神经网络预测。方法将人工神经网络与遗传算法应用于药物生物利用度预测研究，提出了采用遗传算法对人工神经网络进行优化的网络模型建立方法，利用遗传算法对神经网络模型中的权重进行优化,同时运用遗传算法强大的搜寻功能,得到特定条件下模型的最优解。并以药物分子体积(V)、分子折射率(R)、脂水分配系数(lgP_C)、水合能(H)、分子极化度(P)、前线轨道能量E_HOMO和E_HOMO为网络输入参数，以药物的平均生物利用度为网络输出参数，建立了药物生物利用度遗传神经网络预测模型。结果经遗传算法优化的GA-BP神经网络模型对生物利用度的预测精度为95.9%。结论该模型可以用于药物生物利用度预测研究。     

肉苁蓉的主要化学成分及生物活性研究进展

肉苁蓉为列当科多年生草本寄生植物荒漠肉苁蓉Cistanche desertico或管花肉苁蓉Cistanche tubulosa的干燥带鳞叶的肉质茎,主要化学成分有苯乙醇苷类、环烯醚萜及其苷类、木脂素及其苷类、多糖等,具有抗衰老、保护肝脏、缓解体力疲劳、抗骨质疏松、润肠通便等多种生物活性。就肉苁蓉的主要化学成分及生物活性等方面的研究进展进行综述,为新药研发提供参考。     

紫茎泽兰中的萜类成分研究

目的 对紫茎泽兰中的萜类成分及其药理活性进行研究。方法 采用D101、HP20大孔树脂、硅胶柱色谱、葡聚糖凝胶色谱、高效液相色谱等多种分离分析手段，MS、¹H-NMR和¹³C-NMR等多种波谱技术以及ECD计算鉴定化合物结构，并采用MTT法对分离得到的单体化合物进行细胞毒活性的筛选。结果 从紫茎泽兰95%乙醇提取物的石油醚和乙酸乙酯萃取部位中分离得到10个化合物，分别鉴定为泽兰酮D (1)、thymoquinol 5-O-β-glucopyranoside (2)、thymoquinol2-O-β-glucopyranoside (3)、2R^*,3S^*-toxol-7-O-β-D-glucopyranoside (4)、6-甲氧基-山柰酚-7-O-β-D-葡萄糖苷(5)、万寿菊素-4’-甲氧基-7-O-β-D-葡萄糖苷(6)、6-hydroxykaempferol-7-O-β-D-glucopyranoside (7)、eupatonriochromene (8)、demethoxyencecalin (9)、encecalin (10)。化合物10对食管癌细胞的IC₅₀值为178.4 μmol·L^-1。结论 其中化合物1为新化合物，化合物6为首次从泽兰属中分离得到，化合物2，3，7为首次从紫茎泽兰中分离得到。化合物10对食管癌细胞Eca-109具有一定的细胞毒活性。     

Isolation and characterization of surfactin produced by <Emphasis Type="Italic">Bacillus polyfermenticus</Emphasis> KJS-2

Bacillus polyfermenticus KJS-2 (BP-KJS-2) was used to produce a lipopeptide-type surfactin. To accomplish this, a surfactin-producing BP-KJS-2 was fermented by soybeans. The surfactin was then purified by a procedure including ethanol treatment and preparative chromatography. Next, the biochemical structure of the purified surfactin was analyzed by electrospray ionization mass spectrometry (ESI-MS) and high-resolution ESI Q-Tof mass spectrometry (Q-Tof MS). In addition, the masses of the four peaks were determined to be 1007, 1021, 1035, and 1049 m/z revealing that the compound was mixture with quasi-molecular ions. Taken together, these findings indicated that the lipopeptide had a cyclic structure and amino acid composition of Gln-Leu-Leu-Leu-Val-Asp-Leu-Leu, and that the major lipopeptide product of BP-KJS-2 is the surfactin isoform. In addition, this lipopeptide showed strong antimicrobial activity against bacteria at the level of 0.05 mg/mL.     

Enhanced Immunostimulant Activity and Protective Effect of a Synthetic Lipopeptide after Liposomization Against Plasmodium berghei Infection in Mice

The immunostimulant activity of non-pyrogenic, sugar-free immunomodulator lipopeptide, Ala-D-Glu(Gly-Lys-CO. C₁₁H₂₃)-NH₂ (comp. no 84/201), and its iiposomized formulation has been studied. Liposomization of this lipopeptide significantly enhanced its antigen specific as well as nonspecific immune responses, as compared to the free lipopeptide. The liposomized formulation of lipopeptide significantly stimulated both the antibody and delayed-type hypersensitivity responses in Balb/c mice, and also enhanced nonspecifically the macrophage migration index, phagocytic activity and incorporation of ¹⁴C glucosamine in peritoneal macrophages of the mice that received pretreatment with this preparation. Further, the mice that received pretreatment with the liposomized preparation strongly resisted lethal P. berghei infection and consequently survived for longer period of times. These results indicate that liposomization of the compound no 84/201 significantly improves its ability to enhance not only antigen-specific immune response but also the nonspesific host's resistance against infections.     

胃切除术后小肠套叠的危险因素分析

目的 分析成人胃切除术后小肠套叠的危险因素。方法 选择2010年1月至2018年12月安徽医科大学第一附属医院普外科确诊为胃部疾病并行手术的患者9 825例作为研究对象。将术后发生小肠套叠的9例患者作为病例组,从其余患者中选择36例患者（1：4）作为对照组,要求性别相同、年龄相差不超过1岁。分析术后小肠套叠与性别、体质指数（BMI）、吻合方式等因素的关系。结果 病例组患者中位BMI为21.67（19.81,23.09）kg/m²,低于对照组,差异有统计学意义（P<0.05）。病例组3例（33.33%）患者发生术中吻合器上肠管套叠,对照组2例（5.56%）,两组患者术中吻合器上肠管套叠发生率的差异有统计学意义（P<0.05）。多因素logistics回归分析显示,术中吻合器上肠管套叠是胃切除术后小肠套叠的危险因素（OR=7.508）,而BMI则是术后小肠套叠的保护因素（OR=0.609）,差异均有统计学意义（P<0.05）。结论 术前BMI高的患者术后发生小肠套叠的风险更低,而术中吻合器上肠管套叠的患者更容易出现术后小肠套叠。     

Synthetic Glycopeptide-Based Delivery Systems for Systemic Gene Targeting to Hepatocytes

Purpose. To design, synthesize, and test synthetic glycopeptide-baseddelivery systems for gene targeting to hepatocytes by systemicadministration.Methods. All peptides were synthesized by the solid phase methoddeveloped using Fmoc chemistry on a peptide synthesizer. The bindingof galactosylated peptides to HepG2 cells and accessibility of thegalactose residues on particle surface was demonstrated by acompetition assay using ¹²⁵I-labeleld asialoorosomucoid and RCA lectinagglutination assay, respectively. DNA plasmid encoding chloramphenicolacetyl transferase (CAT) gene was complexed with a tri-galactosylatedpeptide (GM245.3) or tri-galactosylated lipopeptide (GM246.3) in thepresence of an endosomolytic peptide (GM225.1) or endosomolyticlipopeptide (GM227.3) to obtain DNA particles of 100–150 nm insize. The plasmid/peptide complexes were added to HepG2 cell culturesor intravenously administered by tail vein injection into normal miceor rats. Plasmid uptake and expression was quantified by qPCR andELISA, respectively.Results. Multiple antennary glycopeptides that have the ability tocondense and deliver DNA plasmid to hepatocytes were synthesized andcomplexed with DNA plasmid to obtain colloidally stable DNA/peptidecomplexes. Addition of DNA/GM245.3/GM225.1 peptide complexes(1:3:1 (–/+/–)) to HepG2 cell cultures yielded CAT expression intransfected cells. The transfection efficiency was significantly reducedin the absence of galactose ligand or removal of endosomolytic peptide.Intravenous administration of DNA/GM245.3 peptide complexes (1:0.5(–/+)) into the tail vein of normal rats yielded DNA uptake in theliver. Substitution of GM245.3 by galactosylated lipopeptide GM246.3resulted in more stable DNA particles, and a 10-fold enhancement inliver plasmid uptake. CAT expression was detectable in liver followingintravenous administration of DNA/GM246.3 complexes. Addition ofendosomolytic lipopeptide GM227.3 into the complexes(DNA/GM246.3/GM227.3 (1:0.5:1 (–/+/–))) yielded a 5-fold increase inCAT expression. Liver expression was 8-fold and 40-fold higher thanlung and spleen, respectively, and localized in the hepatocytes only.The transfection efficiency in liver was enhanced by increasing DNAdose and injection volume. The plasmid uptake and expression in liverusing DNA/GM246.3/GM227.3 complexes was 100-200-fold higherthan DNA formulated in glucose. Tissue examination and serumbiochemistry did not show any adverse effect of the DNA/GM246.3/GM227.3 (1:0.5:1 (–/+/–)) complexes after intravenous delivery.Conclusions. Gene targeting to hepatocytes was achieved by systemicadministration of a well-tolerated synthetic glycopeptide-baseddelivery system. The transfection efficiency of this glycopeptide deliverysystem was dependent on peptide structure, endosomolytic activity,colloidal particle stability, and injection volume.     

两面针碱全合成及活性研究进展

两面针碱是传统中药两面针根的提取物,属于苯并菲啶类生物碱。研究发现两面针碱具有多种生物活性,对肿瘤、疟疾等多种疾病有效。笔者对两面针碱全合成方法及近年来的活性研究进行综述。     

Lipophilization of Somatostatin Analog RC-160 Improves Its Bioactivity and Stability

Purpose. Acromegaly is a symptomatically disabling condition, resulting from a growth hormone (GH) secreting pituitary tumor. The somatostatin analog RC-160 is known to potently inhibit hypersecretion of GH, from pituitary adenomas. However, the therapeutic potential of RC-160, is limited by its short serum half life. To overcome this limitation, fatty acids with carbon chain lengths ranging from 4 to 18 were conjugated to RC-160. Methods. The GH-inhibitory activity of these lipopeptides, as well as their binding profile to somatostatin receptors, on the rat pituitary adenoma cell line GH3 was studied in vitro. The relative stability of lipophilized RC-160 towards degradation by crude papaya protease was also determined. Results. The long chain lipopeptides, like myristoyl-RC-160 (carbon chain length = 14) were found to exhibit greater receptor affinity and GH-inhibitory activity, as compared to their counterparts of lower chain lengths. However, the receptor affinity and GH-inhibitory activity of stearoyl-RC-160 (carbon chain length = 18), was found to lower than RC-160 and its lipophilized derivatives. Unlike RC-160, the myristoylated derivative was found to have significantly greater resistance to protease and serum degradation (p < 0.01). Conclusions. Lipophilization of RC-160 with long chain fatty acids improves its stability and GH-inhibitory activity. The activity of lipophilized RC-160 seems to increase with increasing hydrophobicity of the lipopeptide, and reaches a maxima at myristoyl-RC-160 for GH3. Hence, optimizing the hydrophobicity should be an important consideration governing the design and synthesis of bioactive lipopeptides.     

Solid-phase synthesis and application of double-fluorescent-labeled lipopeptides,containing a CTL-epitope from the measles fusion protein

Abstract: The mechanism which enables lipopeptides to induce cytotoxicity is not known. By preparing fluorescent-labeled lipopeptides one might unravel the mechanism of their entry into the cell and their intracellular pathway. A method of preparing double-fluorescent-labeled peptides by solid-phase chemistry is described. As model peptides we have chosen analogs of the sequence RRYPDAVYL, which occurs in the measles fusion protein (F_438–446) and is an epitope for cytotoxic T lymphocytes. The peptides Pal-K^TMRKKKRRYPDAVK^FLL ( 7 ) and Pal-K^FLKKKRRYPDAVK^TMRL ( 8 ), in which Pal is palmitoyl and K^TMR and K^FL are N^ε-carboxytetramethylrhodamine- and N^ε-carboxyfluorescein-labeled lysyl residues, respectively, were prepared and obtained in ≈ 30% yield after purification by high-performance liquid chromatography. The fluorescence of fluorescein and tetramethylrhodamine in lipopeptide Pal-K^TMRKKKRRYPDAVK^FLL ( 7 ) was quenched to 98–99% due to intramolecular interaction of the labels. On incubation with trypsin (i.e. cleavage at the KKKRR-site) the fluorescence of both labels was restored. The intracellular routing of lipopeptide Pal-K^TMRKKKRRYPDAVK^FLL was studied with human melanoma cell line, Mel/J, which was transfected with human leukocyte antigen B*2705. It appeared that the double-fluorescent-labeled lipopeptide was able to induce antigen-specific cytotoxicity. Furthermore, preliminary confocal microscopical studies indicated that this lipopeptide is observed intracellularly.     

冬虫夏草人工繁育品、野生品及亚香棒虫草中氨基酸比较

目的：比较冬虫夏草人工繁育品、野生品与亚香棒虫草中水解和游离氨基酸的含量。方法：采用氨基酸分析仪测定冬虫夏草人工繁育品、野生品以及亚香棒虫草中17种氨基酸的含量。结果：冬虫夏草人工繁育品的水解氨基酸含量为16.776%~19.080%,野生冬虫夏草为14.857%~21.959%,亚香棒虫草为13.043%~14.933%。冬虫夏草人工繁育品的游离氨基酸含量为1.767%~2.373%,野生冬虫夏草为1.753%~2.521%,亚香棒虫草为2.856%~3.197%。结论：冬虫夏草人工繁育品和野生品中氨基酸含量基本一致,和亚香棒虫草有显著性差异。本研究为冬虫夏草的鉴别及人工繁育品的进一步开发利用提供科学依据。     

N-Acyl-(α,γ Diaminobutyric Acid)n Hydrazide as an Efficient Gene Transfer Vector in Mammalian Cells in Culture

Purpose. This study investigates the structure/activity relationship of a series of N-acyl-peptides (lipopeptides) for the transfection of mammalian cells. Methods. Lipopeptides comprising 1 to 3 basic amino-acids and a single fatty acid chain were synthesized. Transfecting complexes between lipopeptide, plasmid DNA and dioleoyl phosphatidylethanolamine were prepared and applied on cells in culture. Transfection efficiency was evaluated by measuring -galactosidase activity 48 h post-transfection. Lipopeptide-DNA binding was also investigated by physical means and molecular modelling. Results. Besides the length of the fatty acid chain, the nature of the basic amino-acid and the C-terminal group were crucial parameters for high transfection efficiency. The N-acyl-(diaminobutyric acid)n derivatives were the most potent transfecting agents among those tested and induced a -galactosidase activity 2 to 20 times higher than the N-acyl-lysine, -ornithine or -diaminopropionic acid derivatives. Furthermore, a hydrazide C-terminal modification greatly enhanced transfection efficiency for all compounds tested. The reason why , -diaminobutyric acid hydrazide-based lipopeptides were the most potent in transfection is not fully understood but could be related to their high DNA binding. Conclusions. Poly- or oligo-diaminobutyric acid containing or not a hydrazide C-terminus could advantageously be used in peptide-based gene delivery systems.     

2015—2017年北京怀柔医院呼吸科重症监护病房感染性疾病病原菌分布及耐药性分析

目的 探究2015-2017年北京怀柔医院呼吸科重症监护病房（RICU）感染性疾病病原菌分布及耐药性,为临床合理选用抗菌药物及有效控制感染提供参考。方法 回顾性分析2015年1月-2017年12月在北京怀柔医院RICU住院治疗的发生感染的860例患者的病原学检测结果及药敏结果,探讨其病原菌分布及耐药性状况。结果 共分离出3 800株病原菌,主要来源于痰液（63.34%）。其中革兰阳性菌926株,占24.37%,主要以金黄色葡萄球菌、肺炎链球菌和溶血葡萄球菌为主;革兰阴性菌2 482株,占65.31%,主要以大肠埃希菌、鲍曼不动杆菌和铜绿假单胞菌为主;真菌392株,占10.32%,主要以白假丝酵母为主。革兰阴性菌中大肠埃希菌对碳青霉烯类药物（如亚胺培南）、阿米卡星及头孢哌酮等耐药率低,对多黏菌素E高度敏感;鲍曼不动杆菌具有较强的耐药性,仅对多黏菌素E高度敏感;铜绿假单胞菌对多黏菌素E和头孢哌酮耐药率低;革兰阳性菌中金黄色葡萄球菌、肺炎链球菌以及溶血葡萄球菌对万古霉素及替考拉宁的敏感率均较高,而对大环内酯类药物（红霉素、庆大霉素等）的耐药率均较高。结论 北京怀柔医院RICU感染性疾病病原菌以革兰阴性菌为主,且耐药情况较严重。定期监测感染病原菌的分布状况及其耐药性对临床上抗菌药物的合理选择具有重要意义。