仙鹤草多糖的分离纯化及理化性质研究

目的从中药仙鹤草中提取仙鹤草多糖,并对其理化性质进行研究。方法利用水提醇沉法得粗多糖,并依次经纤维素DE-52柱和Superdex-200凝胶柱色谱进行纯化。采用光谱法和色谱法对其总糖含量、单糖组成、相对分子质量等理化性质进行研究。结果分离纯化的仙鹤草多糖HAPⅢ总糖含量、蛋白含量、糖醛酸含量分别为81%,3.4%,45%,由半乳糖、葡萄糖、鼠李糖、阿拉伯糖、甘露糖、岩藻糖、木糖组成,摩尔比为5.65∶2.67∶2.61∶1.86∶1.12∶1.09∶1。其平均相对分子质量为1.90×105。结论仙鹤草多糖HAPⅢ是一水溶性酸性杂多糖。     

虫草多糖的分离纯化及结构鉴定

摘 要 目的： 分离纯化冬虫夏草菌发酵液多糖,并对虫草多糖的结构进行分析。方法: 采用液体发酵法培养冬虫夏草菌,水提醇沉法提取虫草发酵液多糖EPS和菌丝体多糖IPS,联合使用葡聚糖凝胶柱色谱对虫草多糖进行分离纯化,采用凝胶过滤法测定其纯度和相对分子质量,气相色谱法分析其单糖组成,再通过硫酸咔唑法分析糖醛酸含量。结果:经分离纯化得到多糖EPS的相对分子质量（Mr）为78 kDa,多糖含量为94.8%,单糖组成为甘露糖、葡萄糖和半乳糖,摩尔比为4.5∶8.0∶1.0;糖醛酸含量为6.0%。多糖IPS 的Mr为42 kDa,多糖含量为92.5%,单糖组成为甘露糖、葡萄糖、半乳糖,摩尔比为2.8∶3.0∶;糖醛酸含量为4.5%。结论: 虫草多糖EPS和IPS均是杂多糖。     

枇杷叶多糖的组分及含量分析

目的 对枇杷叶多糖的纽分及含量进行研究.方法 枇把叶经提取分离,分为水提多糖和碱提多糖两个部分,各多糖经过脱蛋白、透析后,进行色谱分析及中性己糖、糖醛酸、蛋白质等含量测定.结果 水提多糖(FEW)的中性单糖主要由果糖、鼠李糖、甘露糖、半乳糖等组成,摩尔比为1.00:0.13:0.03:0.05;碱提多糖(FEB)的中性单糖则由果糖、鼠李糖、阿拉伯糖、甘露糖、葡萄糖、半乳糖等组成,摩尔比为1.00:0.45:0.94:0.31:0.16:0.55.各多糖中中性已糖、糖醛酸、蛋白质的含量,FEW分别为65.2%、15.8%、5.2%,FEB分别为65.9%、14.3%、14.7%.结论 枇耙叶多糖属于含蛋白质的复合杂多糖.     

桑叶多糖PMP11的分离纯化及结构初探

目的:从桑叶中分离纯化获得均一多糖PMP11,并研究其组成及初步结构。方法:桑叶经热水提取乙醇分级沉淀、脱蛋白、脱色、DEAE-52纤维素和Sephadex G-100柱层析,得到一个均一多糖组分PMP11;采用GC、HPLC、IR、NMR、Smith降解和糖醛酸的还原等方法分析其组成和初步结构。结果:PMP11主要由鼠李糖、半乳糖醛酸和葡糖醛酸组成,其摩尔比为鼠李糖∶半乳糖醛酸∶葡糖醛酸=2.27∶1.59∶1;PMP11的主链主要是以β-1→2糖苷键及β-1→3糖苷键连接的鼠李糖,侧链主要是以β-1→2糖苷键及β-1→4糖苷键连接的葡糖醛酸和半乳糖醛酸。结论:本试验首次分析桑叶多糖PMP11的初步结构,可为桑叶多糖的进一步研究提供依据。     

柱前衍生-高效毛细管电泳法测定刺糖多糖中单糖的组成

目的:分析测定刺糖多糖中单糖的组成。方法:将刺糖多糖水解成单糖,1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生化,采用高效毛细管电泳法在245 nm紫外检测分离测定各单糖。结果:刺糖多糖中阿拉伯糖、葡萄糖、鼠李糖、半乳糖、甘露糖、葡萄糖醛酸、半乳糖醛酸的摩尔比为2.80∶38.61∶5.28∶3.76∶1.91∶3.82∶2.45,且刺糖多糖中葡萄糖的含量为35.65%。结论:本法测定刺糖多糖的单糖组分操作简便,灵敏度高,结果准确可靠,可用于刺糖多糖单糖组成测定和质量控制。     

天葵子多糖的提取与鉴定

目的提取天葵子多糖,并对其进行定性定量分析。方法采用水提醇沉法提取天葵子多糖,用苯酚-硫酸比色法测定多糖粗提物中的总糖含量,咔唑-硫酸法测定酸性多糖含量,双缩脲法测定蛋白质含量,凝胶过滤法测定相对分子质量(Mr),气相色谱法确定天葵子多糖的单糖组成及摩尔比。并对其进行红外光谱分析。结果总糖含量为80.4%,酸性糖含量为6.8%,蛋白质含量为16.6%,Mr约为6.3×104,天葵子多糖含葡萄糖、半乳糖、甘露糖、阿拉伯糖、岩藻糖,摩尔比例为6.23∶2.04∶3.15∶1.03∶1。多糖结构含非O-型呋喃糖苷键。结论天葵子多糖为5种单糖组成的混合物。     

鹰嘴豆多糖的分离纯化及其抗氧化作用研究

目的对鹰嘴豆果实多糖进行分离纯化得到均一多糖,对分离得到的均一多糖的抗氧化活性进行研究。方法利用热水对鹰嘴豆多糖进行提取,乙醇沉淀得到粗多糖;粗多糖经DEAE-Cellulose 52和Sephacryl S-300柱层析纯化,得到均一多糖CA-1b;高效凝胶渗透色谱法(HPGPC)验证CA-1b纯度和相对分子质量;酸水解、GC-MS对CA-1b进行糖组分分析;DPPH·清除评价均一多糖的体外抗氧化活性。结果鹰嘴豆果实中分离得到的多糖CA-1b为均一多糖,相对分子质量为1.33×106 Da,由阿拉伯糖(Ara)、甘露糖(Man)、葡萄糖(Glc)和半乳糖(Gal)组成,其摩尔比为2.2∶3.1∶6.2∶1.0。DPPH·清除实验表明,CA-1b具有良好的抗氧化活性,且呈现一定的量效关系,半数清除质量浓度IC50为18.2μg·mL-1。结论研究结果可为鹰嘴豆的深加工和进一步研究开发奠定理论基础。     

扁玉螺多糖的提取、分离和结构分析

目的从扁玉螺(Neverita didyma)中提取和分离纯化多糖,并对其基本理化性质和结构组成进行分析。方法依次采用水提和碱提方法从扁玉螺中提取粗多糖,采用DEAE Sepharose FF阴离子交换和Sephacryl S-300凝胶柱层析对粗多糖进行分离纯化,并对其总糖、蛋白、氨基糖、糖醛酸和硫酸根含量,相对分子质量和单糖组成进行分析。对多糖纯化组分采用甲基化、气质联用(GC-MS)、红外光谱(IR)、电喷雾质谱(ESI-MS)和核磁共振波谱(NMR)对其化学结构进行分析。结果扁玉螺水提粗多糖(BYL-S)中不含有糖醛酸和硫酸基,其单糖组成只含Glc,进一步分离得到了4种水溶性多糖组分。碱提粗多糖(BYL-J)单糖组成相对复杂,除含有Glc外,还含有Man、GlcN、GalN、Gal和Fuc,其摩尔比为Glc∶Man∶GlcN∶GalN∶Gal∶Fuc=78.9∶1.7∶3.4∶2.2∶5.2∶5.6,进一步分离得到了3种多糖组分。对水提多糖纯化组分BYL-S2的结构分析表明其是以α-(1→4)为主链,含有少量β-(1→3,4)和β-(1→3)分支的D-吡喃型葡聚糖。结论从扁玉螺中提取分离得到了7种多糖组分,并确定了一种水溶性葡聚糖的结构,为扁玉螺多糖结构和活性的深入研究提供了基础。     

当归酸性多糖的分析

目的:建立当归酸性多糖的分离方法,对当归酸性多糖的组成进行分析。方法:采用化学分析方法结合高效尺寸排阻色谱、气相色谱等仪器分析方法,对当归酸性多糖的分离、多糖纯度和重均相对分子质量、糖残基组成进行研究。结果:得到重均相对分子质量分别为7.4×10~5,1.1×10~5,5.7×10~5,2.3×10~5,1.4×10~4的5种当归酸性多糖 APS-b1、APS-b2、APS-c1、APS-c2和 APS-c3,糖醛酸含量分别为13.2%,15.2%,33.4%,30.3%,35.7%。结论:当归酸性多糖组分复杂,各组分单糖组成主要为葡萄糖、半乳糖、阿拉伯糖、鼠李糖、甘露糖、木糖中的4～6种以不同比例构成。     

当归多糖X—C—3—Ⅱ的分离纯化与组成研究

目的对中药岷当归[Angelicasinensis(Oliv)Diels]的水溶性成分进行研究,分离出有活性的单一的多糖组分.方法采用热水提取、乙醇沉淀、DEAE-SephadexA-25柱层析进行分离,凝胶色谱法测定相对分子质量,利用气相色谱法鉴定多糖组分中所含单糖的种类和它们之间的摩尔比.结果分离得到一个多糖组分X-C3-Ⅱ,相对分子质量为1.0×105,其中所含单糖的种类和它们之间的摩尔比为葡萄糖半乳糖阿拉伯糖鼠李糖半乳糖醛酸=56.022.118.91.91.1.结论x-C-3-Ⅱ为首次从该植物中分离得到,并且为均一性多糖.     

In vitro enzymatic evaluation of some pyrazolo[1,5-a]pyrimidine derivatives: Design,synthesis, antioxidant,anti-diabetic,anti-Alzheimer,and anti-arthritic activities with molecular modeling simulation

The strategy of utilizing nitrogen compounds in various biological applications has recently emerged as a powerful approach to exploring novel classes of therapeutics to face the challenge of diseases. A series of pyrazolo[1,5-a]pyrimidine-based compounds 3a–l and 5a–f were prepared by the direct cyclo-condensation reaction of 5-amino-1H-pyrazoles 1a, b with 2-(arylidene)malononitriles and 3-(dimethylamino)-1-aryl-prop-2-en-1-ones, respectively. The structures of the new pyrazolo[1,5-a]pyrimidine compounds were confirmed via spectroscopic techniques. The in vitro biological activities of all pyrazolo[1,5-a]pyrimidines 3a–l and 5a–f were evaluated by assaying total antioxidant capacity, iron-reducing power, the scavenging activity against 1-diphenyl-2-picryl-hydrazyl (DPPH) and 2, 2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals, anti-diabetic, anti-Alzheimer, and anti-arthritic biological activities. All compounds displayed good to potent bioactivity, and three compounds 3g, 3h , and 3l displayed the most active derivatives. Among these derivatives, compound 3l exhibited the highest antioxidant (total antioxidant capacity [TAC] = 83.09 mg gallic acid/g; iron-reducing power [IRP] = 47.93 µg/ml) and free radicals scavenging activities with (DPPH = 18.77 µg/ml; ABTS = 40.44%) compared with ascorbic acid (DPPH = 4.28 µg/ml; ABTS = 38.84%). Furthermore, compound 3l demonstrated the strongest inhibition of α-amylase with a percent inhibition of 72.91 ± 0.14 compared to acarbose = 67.92 ± 0.09%. Similarly, it displayed acetylcholinesterase inhibition of 62.80 ± 0.06%. However, compound 3i showed a significantly higher inhibition percentage for protein denaturation and proteinase at 20.66 ± 0.00 and 26.42 ± 0.06%, respectively. Additionally, some in silico ADMET properties were predicted and studied. Finally, molecular docking simulation was performed inside the active site of α-amylase and acetylcholinesterase to study their interactions.     

Pharmacokinetics, efficacy, and safety of LMWHs in venous thrombosis and stroke in neonates, infants and children

Since the early nineties it has been shown that low molecular weight heparin (LMWH) has significant advantages over unfractionated heparin and oral anticoagulants for both the treatment and the prevention of thrombosis, not only in adults, but also in children. The present review was based on an 'EMBASE', 'Medline' and 'PubMed' search including literature published in any language since 1980 on LMWH in neonates, infants and children. It included paediatric pharmacokinetic studies, the use of LMWH in children with venous thrombosis, LMWH administration in paediatric patients with ischaemic stroke, and its use in order to prevent symptomatic thromboembolism in children at risk. An increasing rate of off-label use of LMWH in children has been reported, showing that LMWHs offer important benefits to children with symptomatic thromboembolic events and poor venous access. Two well-conducted pharmacokinetic studies in this age group showed that neonates and younger infants require higher LMWH doses than older children to achieve the targeted anti-Xa levels, due to an increased extra vascular clearance. Recurrent symptomatic thromboses under LMWH occur in approximately 4% of children treated for venous thrombosis, and in 7% of children treated for stroke; major bleed was documented in 3% of children with therapeutic target LMWH anti-Xa levels, whereas minor bleeding was reported in approximately 23% of children receiving either therapeutic or prophylactic doses, respectively. Further randomized controlled trials are recommended to evaluate the optimum duration and application for different LMWH indications in children.     

Intestinal glutathione: determinant of mucosal peroxide transport, metabolism, and oxidative susceptibility

The intestine is a primary site of nutrient absorption and a critical defense barrier against dietary-derived mutagens, carcinogens, and oxidants. Accumulation of oxidants like peroxidized lipids in the gut lumen can contribute to impairment of mucosal metabolic pathways, enterocyte dysfunction independent of cell injury, and development of gut pathologies, such as inflammation and cancer. Despite this recognition, we know little of the pathways of intestinal transport, metabolism, and luminal disposition of dietary peroxides in vivo or of the underlying mechanisms of lipid peroxide-induced genesis of intestinal disease processes. This chapter summarizes our current understanding of the determinants of intestinal absorption and metabolism of peroxidized lipids. I will review experimental evidence from our laboratory and others (Table 1) supporting the pivotal role that glutathione (GSH) and reduced nicotinamide adenine dinucleotide phosphate (NADPH) play in mucosal transport and metabolism of lipid hydroperoxides and how reductant availability can be compromised under chronic stress such as hypoxia, and the influence of GSH on oxidative susceptibility, and redox contribution to genesis of gut disorders. The discussion is pertinent to understanding dietary lipid peroxides and GSH redox balance in intestinal physiology and pathophysiology and the significance of luminal GSH in preserving the integrity of the intestinal epithelium.     

Pharmacokinetics of sildenafil and its metabolite,N-desmethylsildenafil,in rats with liver cirrhosis and diabetes mellitus,alone and in combination

1. Pharmacokinetics of sildenafil and its metabolite, N-desmethylsildenafil, in humans and rats with liver cirrhosis (LC) and diabetes mellitus (DM), alone and in combination (LCD) did not seem to be reported.

2. Sildenafil was administered intravenously (10?mg/kg) and orally (20?mg/kg) to control, LC, DM, and LCD rats. Expression of intestinal CYP isozymes in those rats was also measured.

3. In LC, DM, and LCD rats, the areas under the curve (AUCs) of intravenous sildenafil were significantly greater (by 195%, 54.2%, and 127%, respectively) than controls. In LC and LCD rats, AUCs of oral sildenafil were significantly greater (3010% and 2030%, respectively) than controls.

4. In LC, DM, and LCD rats, significantly greater AUCs of intravenous sildenafil were due to the slower hepatic extraction of sildenafil (because of decrease in the protein expression of hepatic CYP2C11 and 3A subfamily in LC and LCD rats, and CYP2C11 in DM rats). In LC and LCD rats, greater magnitude of increase in AUCs of oral sildenafil than those after the intravenous administration could be mainly due to the decrease in the intestinal extraction of sildenafil (because of decrease in the protein expression of intestinal CYP2C11 in LC and LCD rats).

     

Comparative study of the hydrolytic metabolism of methyl-, ethyl-, propyl-, butyl-, heptyl- and dodecylparaben by microsomes of various rat and human tissues

Abstract

1.?Hydrolytic metabolism of methyl-, ethyl-, propyl-, butyl-, heptyl- and dodecylparaben by various tissue microsomes and plasma of rats, as well as human liver and small-intestinal microsomes, was investigated and the structure–metabolic activity relationship was examined.

2.?Rat liver microsomes showed the highest activity toward parabens, followed by small-intestinal and lung microsomes. Butylparaben was most effectively hydrolyzed by the liver microsomes, which showed relatively low hydrolytic activity towards parabens with shorter and longer alkyl side chains.

3.?In contrast, small-intestinal microsomes exhibited relatively higher activity toward longer-side-chain parabens, and showed the highest activity towards heptylparaben.

4.?Rat lung and skin microsomes showed liver-type substrate specificity. Kidney and pancreas microsomes and plasma of rats showed small-intestinal-type substrate specificity.

5.?Liver and small-intestinal microsomal hydrolase activity was completely inhibited by bis(4-nitrophenyl)phosphate, and could be extracted with Triton X-100. Ces1e and Ces1d isoforms were identified as carboxylesterase isozymes catalyzing paraben hydrolysis by anion exchange column chromatography of Triton X-100 extract from liver microsomes.

6.?Ces1e and Ces1d expressed in COS cells exhibited significant hydrolase activities with the same substrate specificity pattern as that of liver microsomes. Small-intestinal carboxylesterase isozymes Ces2a and Ces2c expressed in COS cells showed the same substrate specificity as small-intestinal microsomes, being more active toward longer-alkyl-side-chain parabens.

7.?Human liver microsomes showed the highest hydrolytic activity toward methylparaben, while human small-intestinal microsomes showed a broadly similar substrate specificity to rat small-intestinal microsomes. Human CES1 and CES2 isozymes showed the same substrate specificity patterns as human liver and small-intestinal microsomes, respectively.     

近海浅海海水弗朗西斯菌的分离培养和鉴定

目的　对广东地区近海浅海海水弗朗西斯菌进行分离培养,以了解弗朗西斯菌在近海浅海海水中的分布情况。方法　在广东近海浅海地区水域内,选取茂名市电白区水东湾（2019年10月4日）以及惠州市两大湾（巽寮湾、双月湾,2019年8月12日）进行多点采集水样,分别19份和29份。利用前期探索出针对分离弗朗西斯菌的有效海水前处理手段（浓缩、酸处理）以及分离培养基（BHI-2216E-GVPC和BCYEα-2216E-GVPC）进行分离培养,依据弗朗西斯菌的菌落特征,初步将可疑菌株进行分纯培养,并对纯培养后的可疑菌株进行传统形态特征、生理生化特征、16S rRNA分子特征等鉴定。结果　19份水东湾水样中有14份（阳性率为73.7%）成功分离到共60株目标菌（含邻近兼性胞内寄生菌）;29份惠州地区水样中有17份（阳性率为58.6%）成功分离到共79株目标菌（含邻近兼性胞内寄生菌）。经菌落特征、细菌形态学检查、生化反应以及测序结果比对后,水东地区60株目标菌分离出弗朗西斯菌属（含潜在新种）12株（20.0%）,潜在新科2株（2.9%）,惠州地区79株目标菌分离出弗朗西斯菌属（含潜在新种）22株（27.8%）,潜在新科18株（22.8%）。结论　广东地区近海浅海海水中弗朗西斯菌菌种资源丰富,且存在不同的该菌属内新种以及可能存在邻近新属。     

Pharmacokinetics and elucidation of the rates and routes of N-glucuronidation of PF-592379, an oral dopamine 3 agonist in rat,dog, and human

1. PF-592379 is a potent, selective agonist of the dopamine 3 receptor, for the treatment of male erectile dysfunction and female sexual dysfunction.

2. In vivo, PF-592379 has low-moderate clearance relative to liver blood flow of 6.3 and 8.5?ml/min/kg in dog and 44.8 and 58.2?ml/min/kg in rat. It has high permeability in Caco-2 cells and was completely absorbed in rat and dog pharmacokinetic studies with an oral bioavailability of 28% in both rats and 61 and 87% in the dogs. These data are consistent with the physicochemical properties of PF-592379, which indicate complete absorption by the transcellular route.

3. Elimination of PF-592379 was predominantly metabolic in nature. In vitro routes of metabolism studies indicate that metabolism in the rat is a combination of P450 mechanisms and N-glucuronidation, whereas in dog and human, N-glucuronidation is the major route. NMR analysis indicates that N-glucuronidation is non-quaternary in nature and occurs on both the pyridyl amine and ring nitrogen. Rates of clearance via N-glucuronidation were predicted to be low in humans compared with acyl or phenolic glucuronidation. PF-592379 was predicted to have complete absorption from the gastrointestinal tract and an oral bioavailability of >60% in the clinic.

4. Clinical data verified that PF-592379 is a low clearance compound in human, with a mean oral clearance of 6.5?ml/min/kg following a 200?mg oral dose. PF-592379 has ideal pharmacokinetic properties for an oral D3 agonist, intended for on demand dosing.

     

The risk of fragility fractures following initiation of gabapentin and pregabalin—A Danish,nationwide, high-dimensional propensity score-matched cohort study

Gabapentin and pregabalin have been associated with an increased risk of fragility fractures. Due to differences in pharmacokinetics, we aimed to assess the fracture-risk difference between the two medicines. We performed a Danish nationwide new user, high-dimensional propensity score-matched cohort study to assess the 90-day risk of fragility fractures among adults, from January 1996 to December 2018. We applied a high-dimensional propensity score to match new users of gabapentin with new users of pregabalin in a 1:1 intention-to-treat approach. Hazard ratios (HRs), incidence rates (IRs) and incidence rate difference (IRD) were obtained. We identified 388 236 eligible patients of which 294 223 and 98 869 initiated gabapentin and pregabalin, respectively. We included 48 272 matched pairs for further analysis. The mean age was 56 (IQR 44–69) years, and the average follow-up was approximately 11 500 person-years (PY). The IRs of fragility fractures were 23.7 (95%CI 21.0–26.7) and 23.2 (95%CI 20.5–26.2) per 1000 PY for gabapentin and pregabalin-exposed patients, respectively. This yielded an HR of 1.02 (95%CI 0.86–1.21) when using gabapentin as the intervention drug and pregabalin as the reference drug. The IRD was estimated to 0.5 PY (95%CI −3.5–4.5). In conclusion, short-term risk of fragility fractures among gabapentin initiators was not different compared to those initiating pregabalin.     

洛铂腔内灌注治疗恶性胸腹腔积液病人的临床观察

目的 探讨洛铂腔内灌注治疗恶性胸腹腔积液的近期疗效及药物毒副作用.方法 对确诊为恶性胸腹腔积液的56例病人采用抛硬币法随机分为洛铂(LBP)组和顺铂(DDP)组,每组28例,均采用腔内灌注,比较两组间近期疗效、不良反应及生活质量改善情况.结果 LBP组总有效率为75.0%,而DDP组总有效率为46.4%,LBP组明显高于DDP组(P<0.05).进一步分析显示,在胸腔积液中LBP组有效率(85.7%)明显优于DDP组(50.0%)(P<0.05),在腹腔积液中有效率(64.3%)也高于DDP组(41.7%),但差异无统计学意义(P>0.05).LBP组白细胞减少、胃肠道反应及肾功能损害的发生率明显低于DDP组,且无Ⅲ、Ⅳ级毒副反应发生(P<0.05).LBP组与DDP组生活质量改善率分别为85.7%和60.7%,LBP组明显高于DDP组(P<0.05).