羊红膻根中羊红膻素C及羊红膻素D的分离和鉴定

从草药羊红膻Pimpinellathellungiana根中分得2个化合物，经光谱分析（IR，MS，1HNMR，13CNMR，DEPT，1H-1HCOSY，1H-13CCOSY），鉴定为：erythreo-5-n-pentyl-4-hydroxytetrahydrofuran-2-one（Ⅰ）和threo-5-n-pentyl-4-hydroxytetrahydrofuran-2-one（Ⅱ），分别命名为羊红膻素C及羊红膻素D，为2个新的天然产物。     

2，3-二氢-1-氨基-2-羟基-4-硝基茚的合成研究

最近几年,国外开发了不少以茚为母体的系列药物和兽药。如：以系列抗HIV药物L-689,502;L-685,927;L-735,524等、系列平滑肌迟缓药SARs及系列抗精神分裂药物。此外,作为农药应用,有日本开发的除草剂一茚草酮及美国杜邦公司于1992年开发,并于2001年登记上市的氨基甲酸酯类杀虫剂——茚虫威等。     

�����Բ�ͻ�����������˵��о���չ

??Previous studies demonstrate that diabetes mellitus induces cognitive impairment,leading to diabetic encephalopathy(DE), which is closely related with hippocampal synaptic plasticity impairment,including synaptic structural and functional damage. Structural damage mainly embodied in the synapse degeneration.Functional damage mainly reflects in the LTP damage, including the composition variation and functional lesions of N-methyl-D-aspartate receptors(NMDARs), ??-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors (AMPARs) and patassium channels.Abnormal synaptic plasticity may be critical in the pathogenesis of diabetic encephalopathy. In this review, we summarized the relationship between DE and synaptic plasticity impairment.     

GC法测定高良姜醇提物中两个二苯基庚烷类化合物

目的建立高良姜醇提物中2个二苯基庚烷类化合物的测定方法。方法采用GC法,色谱柱为Cp Sil 5CB(30 m×0.25 mm,0.25μm)石英毛细管柱;检测器:FID;柱温:初始温度80℃,保持2 min,以20℃/min升温到300℃,保持45 min。结果 1-苯基-7-(3’-甲氧基-4’-羟基)苯基-5-醇-3-庚酮在0.2～1.0 mg/mL(r=0.998 8)、1,7-二苯基-5-醇-3-庚酮在0.2～1.0 mg/mL(r=0.997 8)有较好的线性关系;平均回收率分别为99.6%、100.6%。结论本方法可靠、专属性强,可作为高良姜醇提物中2个二苯基庚烷类化合物的测定方法。     

Ԥ֪�ӻ�ѧ�ɷ��о�

??OBJECTIVE To study the chemical constituents of the fruits of Akebiae quinata. METHODS Various column chromatography techniques including silica gel, Sephadex LH-20, and macroporous adsorption resin column chromatography were used for fractionization and purification. The structures were identified on the basis of their physicochemical and spectroscopic evidence. RESULTS Fifteen compounds were obtained, and their structures were identified as geniposidic acid(1), 10-O-acetylgeniposidic acid(2), vomifoliol(3), p-hydroxybenzoic acid(4), protocatechuic acid(5), caffeic acid(6), tyrosol(7), palmitic acid(8), 15-nonacosanol(9), stigmasterol(10), stigmasterol-3-O-??-D-glycopyranoside(11), ??-sitosterol(12), ??-daucosterol(13), ursolic acid(14), and oleanolic acid(15). CONCLUSION Compounds 1-7, 9 and 14 were isolated from the fruits of Akebiae quinata for the first time.     

灰毛浆果楝茎的化学成分

目的研究灰毛浆果楝Cipadessa cinerascens茎的化学成分。方法采用正、反相硅胶柱色谱与Sephadex LH-20凝胶柱色谱进行分离纯化,并运用各种波谱方法对分离的化合物进行结构鉴定。结果从灰毛浆果楝茎中分离得到17个化合物,分别鉴定为羊毛甾-7-烯-3-酮-22,25-环氧-23,24-丙酮乙缩醛(1)、chisopanin M(2)、3β-羟基-5-烯-豆甾(3)、7α-羟基-4-烯-3-酮-豆甾(4)、3β-羟基-5-烯-7-酮-豆甾(5)、7α-羟基-谷甾醇(6)、22E-7α-甲氧基-5α,6α-环氧-8(14),22-二烯-3β-羟基麦角甾(7)、7β-羟基-4-烯-3-酮-胆甾(8)、3β-乙酰氧基-2β,4β-二羟基-16-酮-孕甾(9)、17α,20R-二羟基-3,16-二酮-孕甾(10)、2β,3β-二羟基-16-酮-孕甾(11)、3β,7α-二羟基-20-酮-孕甾(12)、2α,3β-二羟基-5-烯-20-酮-孕甾(13)、1,4-二烯-3,16-二酮-2-羟基雄甾(14)、5-烯-17-酮-3β,16β-二羟基雄甾(15)、芹菜素(16)和annuionone D(17)。结论除化合物14和17外,其余化合物均为首次从灰毛浆果楝中分离得到。     

AHPN������ĺϳɼ�����RAR�õĽ�ϻ����о�

??OBJECTIVE To study the synthesis and activities of AHPN derivatives. METHODS Starting from p-bromophenol and 1-adamantanol, a series of AHPN derivatives were synthesized by substitution reaction, condensation reaction, oxidation reaction and reduction reaction. These new compounds were characterized by ¹H-NMR, ¹³CNMR and HR-MS. Biacore technique was used to test the derivatives?? combining activities with RAR??. RESULTS Four compounds, 7c, 6c, 6e, and 6h, exhibited significant combining activities with RAR?? compared with AHPN. The introduction of phosphoric acid groups and nitrogen heterocyclic ring increased the activities of these compounds. CONCLUSION Compounds 7c, 6c, 6e, and 6h show significant combining activities with RAR??, which are worthy of further study.     

高良姜根茎中一个新的二芳基庚烷

目的：对高良姜（Alpinia officinarum Hance）根茎化学成分进行研究。方法：运用多种层析方法进行分离纯化,通过波谱解析进行结构鉴定。结果：分离鉴定了5个二芳基庚烷类化合物,分别为：5-乙氧基-7-（4-羟基-3-甲氧基苯基）-1-苯基-3-庚酮（1）,5-羟基-1,7-二苯基-3-庚酮（2）,5-羟基-7-（4-羟基-3-甲氧基苯基）-1-苯基-3-庚酮（3）,5-甲氧基-7-（4-羟基-3-甲氧基苯基）-1-苯基-3-庚酮（4）,（E）-7-（4-羟基-3-甲氧基苯基）-1-苯基-4-烯-3-庚酮（5）。结论：化合物1为新的二芳基庚烷类化合物。     

螺内酯杂质的分离与结构鉴定

 目的研究药物螺内酯中所含的杂质。方法应用色谱方法进行分离和纯化,NMR和MS等波谱解析化学结构。结果从螺内酯母液中分离得到2个化合物,分别鉴定为7α-乙酰硫基-17β-甲氧基-孕甾-4-烯-3-酮(2)和17β-羟基-3-氧代孕烷-4-烯-21-羧酸-γ-内酯(3)。结论化合物2为新化合物。     

八角茴香油精馏釜底残液中1个新的苯甲醛类化合物

目的研究八角茴香油精馏釜底残液的化学成分。方法运用多种色谱方法进行分离纯化,根据理化性质和波谱数据鉴定化合物的结构。结果从八角茴香油精馏釜底残液中得到2个化合物,分别鉴定为(E)-1,4-二(4-甲氧基苯基)-3-丁烯-2-酮(1)和4-羟基-3-(3-甲基-3-丁烯-2-基)苯甲醛(2)。结论化合物1的核磁数据为首次报道,化合物2为新化合物。     

�˲�����Rg1�ԡ�Ѫ΢͸��̽��������������о�

??OBJECTIVE To establish a measuring method for microdialysis probe recovery of ginsenoside Rg₁ and investigate the effects of flow rate, concentration and using times of probe on the recovery in vivo and in vitro. METHODS Dialysis method and retrodialysis method were used for the study. The concentration of ginsenoside Rg₁ in brain and blood dialysate was determined by LC-MS/MS and the probe recovery was calculated. RESULTS The recoveries of brain and blood microdialysis probes showed good stability within 10 h, with average values of 17.0% and 34.4% respectively for ginsenoside Rg₁ at 1.5 ??L??min^-1. Concentrations (50,200,500,1 000 ng??mL^-1) had no obvious effect on recovery. At the same concentration, the recovery of brain and blood probes for ginsenoside Rg₁ decreased with the increase of flow rate (0.5, 1.0, 1.5, 2.0, 3.0 ??L??min^-1) in vitro and in vivo. The dialysis recoveries of brain and blood probes in vitro were (40.6??4.3)%, (23.5??2.3)%, (17.7??0.8)%, (12.2??1.1)%, (8.8??0.6)% and (70.6??3.6)%, (46.0??2.1)%, (32.9??1.6)%, (25.6??0.7)%, (18.2??1.3)%, respectively. The recoveries of dialysis and retrodialysis in vitro were approximately equal, and the recovery detected by retrodialysis in vivo was similar with the in vitro results. Probe used for no more than 3 times still kept high transmittance by flushing with 2% heparin sodium and ultrapure water successively. CONCLUSION Retrodialysis method can be used to study brain and blood probe recovery in vivo, and microdialysis can be used for simutaneous pharmacokinetic studies of ginsenoside Rg₁ in intercelluar fluid and blood.     

����������׻����������⿹�Ҹβ���ʵ���о�

??OBJECTIVE To evaluate the anti-hepatitis B virus (HBV) activity of herpetrione nanosuspension (PEDX-NS) both in vitro and in vivo. METHODS HepG2 2.2.15 cells and duck hepatitis B virus (DHBV) infected ducks as in vitro and in vivo models were used to compare the anti-HBV activity of PEDX-NS and PEDX coarse suspension (PEDX-CS). RESULTS In the HepG2 2.2.15 cell, PEDX-NS effectively suppressed the secretion of the HBV antigens (HBsAg and HBeAg) in a dose-dependent manner with significant difference from PEDX-CS (P<0.05 or P<0.01). In the in vivo evaluation, PEDX-NS with high dose (100 mg??kg^-1) and middle dose (60 mg??kg^-1) significantly reduced the serum HBV DNA level (P<0.05 or P<0.01) and the effect was better than that of PEDX-CS (P<0.05 or P<0.01). CONCLUSION The result revealed that PEDX-NS exhibits anti-HBV activity both in vitro and in vivo and its effect was superior to that of PEDX-CS. The mechanism is probably that the small particle size of PEDX-NS provides a large specific surface area that resulted in better absorption in vivo, thus enhancing its anti-HBV activity.
     

�׹��ٺ����������յ��˹����� HeLaϸ���������ü����ӻ���

??OBJECTIVE To investigage the effects of celastrol-triggered HeLa cells autophagy and the molecular mechanisms in vitro and in vivo. METHODS The antiproliferative effect of celastrol was detected using MTT assay. Apoptotic rate and cell cycle were evaluated using flow cytometric analysis. Autophagy was detected using fluorescence microscope. Protein expression was evaluated using Western blotting. Tumor growth was evaluated by subcutaneous xenograft model in vivo. RESULTS Celastrol inhibited HeLa cells proliferation and induced HeLa cells autophagy and cell cycle arrest at G₀/G₁ phase, but not induced HeLa cell apoptosis in vitro. The protein expression of Beclin 1 was up-regulated and the conversion from LC3 ?? to LC3 ?? was increase in HeLa cells in vitro after treatment with celastrol. Moreover, celastrol promoted the protein expression of PTEN??p-ERK1/2??p-MEK1/2 and inhibited the phosphorylated of Akt, p70S6K and mTOR in HeLa cells. After pretreatment with 3-methyladenine (5 mmol??L^-1), the antiproliferative and induced-autophagy effects of celastrol were reversed. Furthermore, celastrol inhibited tumor growth and the protein expression of p-Akt and p-mTOR, but up-regulated the protein expression of LC3 ?? and Beclin 1 in vivo. CONCLUSION Antitumor effect of celastrol dependent on cells autophagy in HeLa cells via inhibition of Akt/mTOR signaling pathway in vitro and in vivo.     

��ʯ����Ī�ᶨ������΢͸�������ʵ��о�

??OBJECTIVE To evaluate the effects of drug concentration and perfusion rate on the recoveries of self-made linear microdialysis probes for further ocular pharmacokinetic study. METHODS Brimonidine tartrate was selected as the model drug. The in vitro recovery was determined using positive dialysis and retrodialysis at different perfusion rates and drug concentrations. And the in vivo recovery was determined using retrodialysis method. RESULTS The microdialysis recoveries of brimonidine tartrate were inversely proportional to perfusion rate,while independent of drug concentration. The positive dialysis and retrodialysis recoveries in vitro were different at 1.0 ??L??min^-1, but no significant difference at 2.0 and 3.0 ??L??min^-1. The in vitro recoveries were greater than those in vivo. CONCLUSION The self-made microdialysis probe has stable recovery and can be used in ocular pharmacokinetic study of brimonidine tartrate.     

����������-VEGF165���弰��תȾ֬����ϸ��������о�

??OBJECTIVE To clone target gene by RT-PCR method, construct VEGF₁₆₅ lentivirus vectors, transfect adipose tissue derived stem cells (ADSCs) and verify the expression of VEGF₁₆₅ in vitro and in vivo. METHODS RT-PCR technology was employed to clone VEGF₁₆₅ gene, and this gene was cloned to lentivirus vector pLVX-EF1??-IRES2-AcGFP1 to construct a lentiviral vector pLVX-EF1??-VEGF₁₆₅-IRES2-AcGFP1. Bacterial colonies PCR and sequencing analysis were used for identification. Then, Lenti-X 293T cells were transfected with main vector pLVX-EF1??-VEGF₁₆₅-IRES2-AcGFP1, packaging plasmid gag-pro, vpr-pol, Tet-Off^TM, tat-IRES-rev and coating plasmid env(VSV-G). Lentiviral vectors were packaged and the titer was determined. ADSCs were isolated by collagenase digestion method, then cultured, and identificated by morphology, immunofluorescence and multi-directional differentiation. ADSCs was transfected with packaged VEGF₁₆₅ lentivirus. Immunofluorescence and ELISA were used to detect the expression of VEGF₁₆₅ in vitro. ADSCs transfected with VEGF₁₆₅ lentivirus were injected into nude mice. ELISA was used to detect the expression of VEGF₁₆₅. RESULTS The VEGF₁₆₅ gene fragment was cloned successfully, and the lentiviral vector plasmid pLVX-EF1??-HVEGF₁₆₅-IRES2-AcGFP1 was confirmed to contain VEGF₁₆₅ gene fragment as shown by bacterial colonies PCR. DNA sequencing analysis confirmed that VEGF₁₆₅ gene sequencing was exactly the same with that reported by Genbank. After transfection, a large number of Lenti-X 293T cells with green fluorescence were observed by fluorescent microscopy. The concentration of the virus titer was 1??10⁸ TU??mL^-1. ADSCs were identified by morphology, immunofluorescence and multi-directional differentiation methods, in line with the literature reported ADSCs characteristics. There were about 90% of ADSCs which could express VEGF₁₆₅ after being transfected with the viruses by immunofluorescence detection, also, VEGF₁₆₅ protein was proved by ELISA to express stably and efficiently in vitro and in vivo. CONCLUSION Lentiviral vectors expressing VEGF₁₆₅ are successfully constructed by cloning target gene with RT-PCR method. After transfection, ADSCs expressing VEGF165 protein stably in vitro and in vivo can be obtained.
     

��������ģ���о���Ƭ����ԭ�����յ����ü������������

??OBJECTIVE To investigate the enhancing effect of borneol on the absorption of chlorogenic acid (CGA) with two different biological models and to evaluate the correlation of these two models. METHODS The in situ rat perfusion model was used to investigate the enhancing effect of borneol on the absorption of CGA. The in vivo pharmacokinetics study was carried out to evaluate the correlation of the two-compartment open model.RESULTS With the in situ rat perfusion model, the absorption rate constant (Ka) of CGA was 0.083 4 h^-1,which was up to 1.58-fold higher by borneol. The results of in vivo pharmacokinetics study in rats suggested that 0.04% borneol could increase the relative bioavailability by 116.02% and maximum plasma concentration increased slightly. The consistent result of these two different absorption models was provided (r??0.941). CONCLUSION Borneol could promote the oral absorption of CGA.
     

��ҩ���˶�NDM-1����Ʋ����˾����־��������������о�

??OBJECTIVE To explore the bacteriostasis and plasmid elimination activities of different extracted parts of traditional Chinese medicine Radix Scutellariae Baicalensis on NDM-1 Acinetobacter calcoaceticus.METHODS The in vitro antibacterial effect of the extracts from Radix Scutellariae Baicalensis was studied. Inhibition zone and minimum inhibitory concentration(MIC) of the alcohol extract and water decoction were examined by using MH agar plates and microdilution susceptibility testing. The growth curve of the NDM-1 Acinetobacter calcoaceticus was tested after being incubated with alcohol extract and water decoction at sub-MIC. At three time points after incubation with different extracts at sub-MIC, photocopy dish method was used to screen plasmid-cured strains of NDM-1 Acinetobacter calcoaceticus. The plasmid-elimination rates and phenotypic changes were compared.RESULTS Both the alcohol extract and water decoction of Radix Scutellariae Baicalensis inhibited the growth of NDM-1 Acinetobacter calcoaceticus. The MICs were 1.56 mg??mL^-1 for the alcohol extract and 6.25 mg??mL^-1 for the water decoction. The growth curve showed that the antibacterial effect of the alcohol extract was more obvious. Both the alcohol extract and water decoction of Radix Scutellariae Baicalensis had some degrees of plasmid elimination effect. The plasmid-elimination rates in the alcohol extract group were higher than those in the water decoction group. The plasmid-elimination rates were 61.27% for the alcohol extract and 49.78% for the water decoction, respectively.CONCLUSION Radix Scutellariae Baicalensis can inhibit the growth of NDM-1 Acinetobacter calcoaceticus and eliminate the drug-resistant plasmid effectively and has the potential to be used to control the spread of pan-drug resistant Acinetobacter strains or be an adjuvant treatment method for clinical infections. Its alcohol extract has better effect.     

��˾ƥ��/����������þ��������΢�轺�Ҽ����Ʊ�����ҩ������

??OBJECTIVE To prepare compound aspirin and esomeprazole magnesium enteric-coated pellet capsules and evaluate the drug release in vitro/in vivo. METHODS The aspirin pellet cores were prepared by using extrusion-spheronization method, and the esomeprazole magnesium-containing drug pellets were prepared with fluidized bed. By using fluidized bed coating method, the two kinds of drug-containing pellets were respectively coated with enteric layer to obtain enteric-coated pellets. After determining the loading capacity by measuring drug content, the two kinds of drug-containing pellets were filled into No.1 capsules. In vitro release was evaluated by measuring release percentage. The in vivo release behavior was evaluated by determination of pharmacokinetic parameters in rats. RESULTS The cumulative release percentage of the two drugs was less than 5% in 2 h in 0.1 mol??L^-1 hydrochloric acid solution. The cumulative release percentage of aspirin was more than 70% in 45 min in pH 6.8 PBS and it was more than 80% in 30 min for esomeprazole magnesium. Aspirin was metabolized to salicylic acid in plasma and its main pharmacokinetic parameters were as follows:t_1/2=9.47 h, MRT_0-??=14.43 h, t_max=3.00 h, ??_max=51.34 mg??L^-1, AUC _0-24=703.39 mg??h??L^-1, AUC _0-??=860.52 mg??h??L^-1. The pharmacokinetic parameters for esomeprazole magnesium were as follows:t_1/2=3.72 h, MRT_0-??=7.44 h, t_max=1.50 h, ??_max=2.71 mg??L^-1, AUC_0-24=11.89 mg??h??L^-1, AUC_0-??=13.79 mg??h??L^-1. CONCLUSION The formulation of compound enteric-coated pellet capsules is reasonable, and the preparation technology has good reproducibility. The drug release is located in the intestinal tract, thus esomeprazole magnesium can antagonize the gastrointestinal side effects of aspirin and aspirin can produce better antithrombotic effect .     

����UHPLC-Q-TOF-MS�����İ�ͷ������B7�ڴ���������Ĵ�л�������

??OBJECTIVE To identify the metabolites of Pulsatilla oleanolic acid 3-??-L-rhamnopyranosyl (1??2) -??-L-pyran arabinoside (B7) in rat plasma, bile, urine, feces, and in vitro liver microsomes incubation system. METHODS An ultra-high performance liquid chromatography combined with electrospray ionization quadrupole time of flight mass spectrometry (UHPLC-Q-TOF-MS) method was developed and successfully applied to the study on the metabolites of B7 in rat plasma, bile, urine, feces, and in vitro liver microsomes incubation system after oral administration. RESULTS A total of 23 metabolites were identified, of which 12 metabolites were present in rat liver microsomes incubation system. CONCLUSION Deglycosylation, demethylation glucuronidation, and hydroxylation are the major metabolic transformation forms of B7 in rats in vivo and in vitro.     

�����Ǽ�ˮ�����͸����������������������

??OBJECTIVE To study evodiamine complex nanoemulsion oil-in-water (EPBCN) in vitro and in vivo evaluation, about release in vitro and pharmacokinetics. METHODS Release in pH 1.2 hydrochloric acid and pH 6.8 PBS solutions and oral bioavailability in rats of EPBCN would be studied. The bioequivalence between evodiamine (ED) and EPBCN was judged. RESULTS In vitro release of EPBCN was about 2.61 times higher than that of ED. The oral bioavailability of EPBCN was 7.35 folds than that of ED. 90% confidence interval of area under concentration-time curve (AUC_{0-72 h}) and peak concentration (??_max) exceeded the standard range. CONCLUSION EPBCN can remarkably increase the release in vitro and oral bioavailability in rats, and they do not have bioequivalence.