Fragmentation stability and retention time-shift obtained by LC-MS/MS to distinguish sialylated N-glycan linkage isomers in therapeutic glycoproteins

Sialylated N-glycan isomers withα2-3 orα2-6 linkage(s)have distinctive roles in glycoproteins,but are difficult to distinguish.Wild-type(WT)and glycoengineered(mutant)therapeutic glycoproteins,cytotoxic T lymphocyte-associated antigen-4-immunoglobulin(CTLA4-Ig),were produced in Chinese hamster ovary cell lines;however,their linkage isomers have not been reported.In this study,N-glycans of CTLA4-Igs were released,labeled with procainamide,and analyzed by liquid chromatography-tandem mass spectrometry(MS/MS)to identify and quantify sialylated N-glycan linkage isomers.The linkage isomers were distinguished by comparison of 1)intensity of the N-acetylglucosamine ion to the sialic acid ion(Ln/Nn)using different fragmentation stability in MS/MS spectra and 2)retention time-shift for a selective m/z value in the extracted ion chromatogram.Each isomer was distinctively identified,and each quantity(>0.1%)was obtained relative to the total N-glycans(100%)for all observed ionization states.Twenty sialylated N-glycan isomers with onlyα2-3 linkage(s)in WT were identified,and each isomer's sum of quantities was 50.4%.Furthermore,39 sialylated N-glycan isomers(58.8%)in mono-(3 N-glycans;0.9%),bi-(18;48.3%),tri-(14;8.9%),and tetra-(4;0.7%)antennary structures of mutant were obtained,which comprised mono-(15 N-glycans;25.4%),di-(15;28.4%),tri-(8;4.8%),and tetra-(1;0.2%)sialylation,respectively,with onlyα2-3(10 N-glycans;4.8%),bothα2-3 andα2-6(14;18.4%),and onlyα2-6(15;35.6%)linkage(s).These results are consistent with those forα2-3 neuraminidase-treated N-glycans.This study generated a novel plot of Ln/Nn versus retention time to distinguish sialylated N-glycan linkage isomers in glycoprotein.

Design,delivery and efficacy testing of therapeutic nucleic acids used to inhibit hepatitis C virus gene expression in vitro and in vivo

Despite major achievements in the treatment ofchronic hepatitis C with the combination ofinterferons and the nucleoside analog ribavirin themajority of patients with chronic hepatitis C virus(HCV) infection cannot be treated effectively.Toimprove this response rate we used antisensetechnologies to inhibit HCV translation as possibleadditional option for experimental treatment.Antisense oligodeoxynucleotides(ODN) are

Therapeutic and Medicinal Uses of <i>Aloe vera</i>: A Review

The plant Aloe vera is used in Ayurvedic, Homoeopathic and Allopathic streams of medicine, and not only tribal community but also most of the people for food and medicine. The plant leaves contains numerous vitamins, minerals, enzymes, amino acids, natural sugars and other bioactive compounds with emollient, purgative, anti-microbial, anti inflammatory, antioxidant, aphrodisiac, anti-helmenthic, antifungal, antiseptic and cosmetic values for health care. This plant has potential to cure sunburns, burns and minor cuts, and even skin cancer. The external use in cosmetic primarily acts as skin healer and prevents injury of epithelial tissues, cures acne and gives a youthful glow to skin, also acts as extremely powerful laxative.

Continuous Wear of Hydrogel Contact Lenses for Therapeutic Use

Purpose: To investigate the efficacy and complications of continuous wear of etafilcon A for therapeutic use. Materials and Methods: The subjects were 228 eyes of 219 outpatients prescribed contact lens (CL) for one week of continuous therapeutic wear during 10 years. The reason for prescription of CLs, the primary disease, the duration of CL wear and the complications were assessed retrospectively. Results: The predominant reason for prescription of CLs was relief of pain or a foreign-body sensation (62.3%) and protection of the corneal epithelium (20.6%). The primary disease was post-penetrating keratoplasty (36.8%), followed by corneal epithelial erosion (14.5%), post-lamellar keratoplasty (14.0%) and bullous keratopathy (12.2%). The average duration of wearing single lens was 6.5 ± 3.2 days. The average duration of wearing CLs in total was 9.2 ± 10.7 months. The most frequent problem associated with continuous wear of CLs was their dropping out of CLs (12.3%). The complications associated with CLs included conjunctivitis with papillary hyperplasia, corneal erosion and superficial punctate keratitis, but corneal ulcer and corneal infiltrates were not found. Conclusion: Serious complications were not shown changing the lenses every week to keep to the prescribed time limit for continuous therapeutic wear, even if corneal epithelial barrier function is impaired.

Comparison of long-lasting therapeutic effects between succimer and penicillamine on hepatolenticular degeneration

ＩＮＴＲＯＤＵＣＴＩＯＮＨｅｐａｔｏｌｅｎｔｉｃｕｌａｒｄｅｇｅｎｅｒａｔｉｏｎ（ＨＬＤ）ｉｓａｎａｕｔｏｓｏｍａｌｒｅｃｅｓｉｖｅｄｉｓｏｒｄｅｒｔｈａｔｃａｕｓｅｓｃｈａｎｇｅｓｉｎｔｈｅｂａｓａｌｇａｎｇｌｉａａｎｄｌｉｖｅｒｔｈａｔｒｅｓｐｅ...

Functional dyspepsia of ulcer-dysmotility type:clinical incidence and therapeutic strategy

Ｆｕｎｃｔｉｏｎａｌｄｙｓｐｅｐｓｉａｏｆｕｌｃｅｒｄｙｓｍｏｔｉｌｉｔｙｔｙｐｅ：ｃｌｉｎｉｃａｌｉｎｃｉｄｅｎｃｅａｎｄｔｈｅｒａｐｅｕｔｉｃｓｔｒａｔｅｇｙＷＡＮＧＸｉａｏＺｈｏｎｇａｎｄＬＩＮＧｕＺｈｅｎＳｕｂｊｅｃｔｈｅａａｄｉｎｇｓ...

Therapeutic role of template-based lymphadenectomy in urothelial carcinoma of the upper urinary tract

Lymphadenectomy for urothelial carcinoma of the upper urinary tract has attracted the attention of physicians. The mapping study of lymphatic spread has shown that a relatively wide area should comprise the regional nodes for tumors of the right renal pelvis or the right upper two-thirds of the ureter. A prospective study showed that an anatomical templatebased lymphadenectomy significantly improved patient survival in tumors of the renal pelvis. This benefit was more evident for patients with p T2 stage tumors or higher. The risk of regional node recurrence is significant reduced by template-based lymphadenectomy,which is likely to be associated with improved patient survival. The removal of lymph node micrometastases is assumed to be the reason for therapeutic benefit following lymphadenectomy. The number of resected lymph nodes can be used to assess the quality of lymphadenectomy,but not to determine the extent of lymphadenectomy. The guidelines currently recommend lymphadenectomy for patients with muscle-invasive disease,even though the current recommendation grades are still low. The present limitation of lymphadenectomy is the lack of standardization of the extent of lymphadenectomy and the randomized trials. Further studies are warranted to collect the evidence to support lymphadenectomy.

Controversies regarding transplantation of mesenchymal stem cells

Mesenchymal stem cells(MSCs)have tantalized regenerative medicine with their therapeutic potential,yet a cloud of controversies looms over their clinical tran-splantation.This comprehensive review navigates the intricate landscape of MSC controversies,drawing upon 15 years of clinical experience and research.We delve into the fundamental properties of MSCs,exploring their unique immuno-modulatory capabilities and surface markers.The heart of our inquiry lies in the controversial applications of MSC transplantation,including the perennial debate between autologous and allogeneic sources,concerns about efficacy,and lingering safety apprehensions.Moreover,we unravel the enigmatic mechanisms surro-unding MSC transplantation,such as homing,integration,and the delicate balance between differentiation and paracrine effects.We also assess the current status of clinical trials and the ever-evolving regulatory landscape.As we peer into the future,we examine emerging trends,envisioning personalized medicine and innovative delivery methods.Our review provides a balanced and informed perspective on the controversies,offering readers a clear understanding of the complexities,challenges,and potential solutions in MSC transplantation.

Clinacanthus nutans:a review of the medicinal uses,pharmacology and phytochemistry

Clinacanthus nutans Lindau is known as snake grass belonging to the Acanlhaceae family.This plant has diverse and potential medicinal uses in traditional herbal medicine for treating skin rashes,insects and snake bites,lesions caused by herpes simplex virus,diabetes,and gout in Malaysia.Indonesia.Thailand and China.Phylochemieal investigations documented the varied contents of bioaclive compounds from litis plant namely flavonoids,glycosides,glycoglyeerolipids.cerebrosides and monoacylmonogalatosylglycerol.The pharmacological experiment proved that various types of extracts and pure compounds from this species exhibited a broad range of biological properties such as anti-inflammatory,antiviral,antioxidant,and anti-diabetic activities.The lindings of toxicity study showed that extracts from this plant did not show any toxicity thus it can be used as strong therapeutic agents for specific diseased conditions.However,further experiments on chemical components and their mode of action showing biological activities are required to elucidate the complete phytochemical profile and assess to confirm their suitability tor future drugs.This review summarizes the medicinal uses,phytochemistry and pharmacology of this plant in order to explore its therapeutic potential and gaps necessitating for prospected research work.

High-intensity focused ultrasound extracorporeal ablation of liver tissuesin rabbits

ＨｉｇｈｉｎｔｅｎｓｉｔｙｆｏｃｕｓｅｄｕｌｔｒａｓｏｕｎｄｅｘｔｒａｃｏｒｐｏｒｅａｌａｂｌａｔｉｏｎｏｆｌｉｖｅｒｔｉｓｓｕｅｓｉｎｒａｂｂｉｔｓＣＨＥＮＧＳｈｕＱｕｎ１，ＺＨＯＵＸｉｎＤａ１，ＴＡＮＧＺｈａｏＹｏｕ１，ＹＵＹａｏ１，Ｂ...

血小板膜糖蛋白特异性抗体在ITP患儿中的表达及其与疗效、出血程度的关系分析

目的分析血小板膜糖蛋白(GP)特异性抗体在免疫性血小板减少症(ITP)患儿中的表达情况及其与疗效、出血程度的关系。方法选取2018年2月至2020年12月于唐山市妇幼保健院儿科就诊的102例ITP患儿为研究对象,并将其纳入病例组,另选取同期同院进行体检的110例健康儿童纳入健康组。病例组患儿均接受糖皮质激素冲击疗法,以4周为1个疗程,共治疗5个疗程,并随访3个月。检测并比较两组GP特异性抗体表达情况,统计ITP患儿出血部位并分析不同GP特异性抗体出血程度、疗效及预后。结果病例组抗GPⅠb/Ⅸ抗体、抗GPⅡb/Ⅲa抗体表达水平高于健康组,差异有统计学意义(t值分别为16.521、51.718,P<0.05);102例ITP患儿抗GPⅠb/Ⅸ抗体阳性、抗GPⅡb/Ⅲa抗体阳性、双抗体阳性、双抗体阴性检出率分别为15.69%、4.90%、30.39%、49.02%,0级、1级、2级出血率分别为16.67%、50.00%、33.33%;且0级出血以双抗体阳性占比(0)最低,1级出血以抗GPⅡb/Ⅲa抗体阳性占比(80.00%)最高,2级出血以抗GPⅠb/Ⅸ抗体阳性、双抗体阳性占比(62.50%、48.39%)较高,差异有统计学意义(H=38.237,P<0.05);102例ITP患儿治疗5个疗程后有效率为78.43%,随访3个月后完全缓解率为68.63%;双抗体阴性ITP患儿有效率、完全缓解率均高于抗GPⅠb/Ⅸ抗体阳性、抗GPⅡb/Ⅲa抗体阳性、双抗体阳性ITP患儿,差异有统计学意义(χ^(2)值分别为19.866、27.709,P<0.05)。结论ITP患儿抗GPⅠb/Ⅸ抗体、抗GPⅡb/Ⅲa抗体表达水平较高,且其不同表达水平与患儿出血程度、疗效及预后均有密切关系,临床可通过监测GP特异性抗体来对ITP患儿进行早期诊断,并对其出血程度、疗效、预后等进行预测。

高原低氧对大鼠左乙拉西坦药代动力学特征及脑血分布的影响

目的:高原低氧暴露易引起药物药动学参数及脑血分布的改变,其中包括多种P糖蛋白(P-glycoprotein,P-gp)的底物。抗癫痫药物左乙拉西坦是P-gp的底物,但高原低氧暴露是否改变其药代动力学特征及脑血分布尚不明确。本研究旨在探讨高原低氧对大鼠左乙拉西坦药代动力学特征及脑血分布的影响。方法:将Wistar大鼠分为低海拔对照组、高海拔组、溶剂对照组和P-gp诱导组,其中高海拔组大鼠急性暴露于4010 m高原环境24 h后口服或静脉注射左乙拉西坦,口服给药后分别于第0.083、0.25、0.5、0.83、1.25、2、4、6、8、10、12、24 h采集血浆,静脉注射给药后分别于第5、45、60、120、240 min采集血浆及脑组织;P-gp诱导组大鼠连续3 d腹腔注射地塞米松后,静脉注射左乙拉西坦,于第120 min采集血浆及脑组织。采用高效液相色谱串联质谱(high performance liquid chromatography-tandem mass spectrometry,HPLC-MS/MS)测定血浆和脑内药物浓度,采用蛋白质印迹法检测大鼠血脑屏障P-gp的表达。结果:与低海拔对照组相比,高海拔组大鼠左乙拉西坦药时曲线下面积(area under the curve,AUC)下降14.69%(P<0.01),平均驻留时间(mean residence time,MRT)下降15.42%(P<0.01),血浆清除率(clearance,CL)升高16.67%(P<0.01),静脉注射给药后第5、45、120、240 min的脑/血药物浓度比分别降低22.82%(P<0.05)、12.42%(P<0.05)、17.40%(P<0.01)和13.22%(P<0.01),血脑屏障P-gp表达升高86.3%(P<0.05);与溶剂对照组相比,P-gp诱导组大鼠血脑屏障P-gp表达升高56.3%(P<0.05),脑/血药物浓度比下降19.3%(P<0.05)。结论:急性高原低氧暴露改变了左乙拉西坦在大鼠体内的药代动力学特征,并降低了其在大鼠体内的脑血分布。左乙拉西坦脑血分布的下降可能与血脑屏障P-gp的表达上调有关。

The role of drug utilization evaluation in medical sciences

Background:Drug utilization evaluation(DUE)is defined by the World Health Organization(WHO)and focuses on the medical,social,and economic consequences of pharmaceutical marketing,distribution,prescribing,and usage in society.The WHO recommends a physician to every 1000 people.According to the recent data from the Health Ministry in 2019,in which 1.16 million doctors are of active population with just 80%,or 0.9 million,practicing.As a result,a ratio of 0.68 doctors for every 1000 people,which is much below as per the WHO reports.This article describes history,types,WHO guidelines,need and purpose of DUE.Objective:The main aim of this paper is to provide information about the rational use of medication in outpa­tient and inpatient department with special emphasis of DUEs.It also provides awareness directly to healthcare professionals,researchers,academicians,pharmacist and nurses to reduce the irrationality of medicines.Methods:The method used to compile this review information gathered from websites,Google scholar,PubMed,Research gate,and studies published on DUE from July 20 to Oct 22 were included as source of information.Results:We studied more than 35 published study on DUE,that reveals most of the physicians prescribed branded drugs not generic drugs,but WHO prescribing indicator allows to prescribe generic drugs in the hospital pharmacy to maintain better inventory control.It may also help to prevent pharmacist misunderstanding during dispensing.Conclusion:The use of generic prescription names avoids the possibility of medication product duplication and lowers patient costs.It is important to remember that incorrect medication prescriptions have impact on both patients and their family members.WHO indicators identify irrational prescribing behaviours to make therapy more rational and cost-effective.

USE OF CELL-BASED BIOASSAY FOR THE QUALITY CONTROL OF BOTANICAL THERAPEUTICS

The single most important factor in modernization efforts of traditional oriental medicines is the limited understanding of active compounds responsible for claimed therapeutic effects.Indeed,the unknown identity of biologically active molecules generates major difficulties associated with botanical therapeutics in general.This includes management of raw materials,poor understanding of

冠状动脉内注射替罗非班改善直接PCI患者近期预后临床研究

目的:探讨盐酸替罗非班不同给药途径对ST段回落率、主要心脏不良事件(MACE)以及TIMI血流分级的影响。方法:选择符合直接PCI适应证的急性ST抬高性心肌梗死(STE-MI)患者88例,随机分为静脉应用组43例和冠脉应用组45例,观察心电图ST段回落情况和30d内不良事件的发生情况,比较术前及术后TIMI血流分级。结果:冠脉组术后1hST段完全回落率高于静脉组(P<0.05),冠脉组MACE发生率低于静脉组(P<0.05),TIMI血流分级变化无显著性差异(P>0.05)。结论:直接PCI时,冠脉内注射盐酸替罗非班安全有效,可以显著改善术后1hST段回落率,减少术后30d心脏不良事件的发生。

化疗栓塞后肝细胞癌细胞凋亡和耐药基因表达的改变

目的：研究肝细胞癌患者经动脉化疗栓塞（ＴＡＣＥ）治疗后肿瘤的细胞凋亡与耐药基因表达的情况及两者的关 系。方法：经病理证实的肝细胞癌９８例，单纯手术５７例，４种介入治疗后Ⅱ期手术切除４１例，用流式细胞仪检测细胞凋 亡，用链霉亲合素－生物素酶复合物免疫组化检测各标本中Ｐｇｐ。结果：肿瘤细胞凋亡率和Ｐｇｐ阳性百分数在单纯手术组和 介入Ａ、Ｂ、Ｃ、Ｄ各组分别为１．７７％、６．１％、８．７％、１３．３％、１５．７％（经秩和检验，除Ｃ、Ｄ组间比较Ｐ＞０．０５，其余两两比较，Ｐ 值均＜０．０５＝和５２．６％、１００．０％、６２．５％、４６．７％、１８．２％（各介入组与单纯手术组相比，Ｘ２值分别为３．９６、０．０２、０．１７、４．３９，Ｐ 值分别＜０．０５、＞０．０５、＞０．０５、＜０．０５）。两指标改变相关系数 ｒ＝－０．９６５５，ｔ＝５．２４３，Ｐ＜０．０２５。结论：碘油、乙醇、明胶 海绵多材料栓塞细胞凋亡率增加，而耐药基因表达减少，两者呈负相关，多材料联合栓塞优于单材料栓塞和单纯化疗。

糖蛋白药物的多糖结构解析进展

细胞表达的治疗性单克隆抗体多数是糖蛋白。附着在蛋白质上的多糖直接影响蛋白质药物的稳定性、生物活性及免疫原性。因此,应对糖蛋白产品上的多糖进行充分分析,以控制产品质量。然而,糖蛋白上多糖的复杂性对检测带了艰巨挑战。介绍了糖蛋白上多糖分析的最新进展,以及利用凝集素芯片技术的高通量多糖分析法,重点介绍了用于检测糖基化位点、糖链结构和含量的测定分析方法,以期为糖蛋白药物的研发提供参考。

乌司他丁对创伤性休克患者术后呼吸功能及炎性因子的影响

目的:探讨术后注射乌司他丁治疗创伤性休克患者的临床疗效。方法:选择创伤性休克手术患者53例,根据术后治疗方案分为治疗组和对照组,对照组27例采用常规糖皮质激素治疗,治疗组26例在对照组基础上给予乌司他丁静注。分析和比较两组术后第1、2、3、5天的呼吸功能,白细胞计数和血清C反应蛋白水平及住院时间。结果:两组术后第1、2、3、5天PaO_2/FiO_2比值、FIO_2及白细胞计数比较均无统计学差异(P>0.05),但治疗组术后第5天动脉血氧饱和度显著高于对照组(P<0.01),术后第1天血清C反应蛋白水平为(5.7±0.6)μg/ml,术后第2天血清C反应蛋白水平为(14.3±1.1)μg/ml,均显著低于对照组,差异具有统计学意义(P<0.01)。两组术后机械辅助呼吸时间、ICU滞留时间、住院时间比较,治疗组显著少于对照组(P<0.05)。结论:创伤性休克患者术后早期注射乌司他丁有益于抑制术后全身炎症反应,改善患者呼吸功能。

乌司他丁对冠状动脉搭桥术后患者认知功能障碍的影响

目的：探讨不同剂量乌司他丁对冠状动脉搭桥术患者术后认知功能障碍的影响及其可能的作用机制。方法：选择2013年1月至2014年6月在南京医科大学附属南京医院就诊择期行冠状动脉搭桥术的老年患者127例，分为三组：大剂量乌司他丁组（1．6万U／kg）、小剂量乌司他丁组（0．8万U／kg）和对照组（等容量的生理盐水）。所有患者在手术当日和次日晨8时测血浆皮质醇浓度，并在术前、开胸、术毕、术后6h和术后24h分别检测IL-6、IL-10、TNF—α．和S100β蛋白水平。术前1d、术后1周和3个月分别应用精神神经测试组合量表评估患者认知功能的变化判定有无术后认知功能障碍，计算各组患者术后认知功能障碍的发生率，并且比较术后1周是否发生术后认知功能障碍患者S100β蛋白水平。结果：最终93例患者完成研究，三组患者一般资料比较差异均无统计学意义（均P〉0．05）。与术前相比较，对照组患者术后24h血浆皮质醇浓度升高明显（P〈0．01），且大剂量和小剂量乌司他丁组患者术后血浆皮质醇浓度均较对照组低（均P〈0．01）；三组患者术毕、术后6h和24h血浆IL-6、IL-10、TNF-α水平及S100β蛋白水平均高于术前（均P〈0．05）；大剂量和小剂量鸟司他丁组患者在术毕、术后6h和24h血浆IL-6、TNF-α水平浇以及在术后6hSlOOl3蛋白水平均比相对应时间点的对照组患者降低（均P〈0．05），但大剂量和小剂量乌司他丁组患者组问差异无统计学意义（P〉0．05）。大剂量和小剂量乌司他丁组患者术后1周认知功能障碍发生率（25．8％和23．3％）均低于对照组（50．0％），差异有统计学意义（均P〈0．05）；而大剂量和小剂量乌司他丁组术后3个月的术后认知功能障碍发生率（12．9％和16．7％）与对照组（28．1％）比较，差异均无统计学意义（P〉0．05）。术后认知功能障碍组（n=31）在术后24h血清S100β蛋白水平高于非术后认知功能障碍组（n=62），其差异具有统计学意义（P〈0．05）。结论：乌司他丁可降低冠状动脉搭桥术患者术后1周的术后认知功能障碍的发生率，其机制可能与减轻炎性反应及脑损伤有关。

斑蝥酸钠逆转K562/AO_2细胞多药耐药的机制初探

目的观察斑蝥酸钠(SCA)对多药耐药的白血病细胞(K 562/AO2)细胞是否有逆转作用,并初步探讨其逆转机制。方法应用M TT测定多柔比星(阿霉素,ADM)对K 562/AO2细胞的半数抑制浓度(IC50);应用逆转录聚合酶链式反应(RT-PCR)检测K 562、K 562/AO2细胞表达m dr1基因水平;用流式细胞术(FCM)检测其P-gp蛋白表达的水平。结果M TT结果显示,无毒剂量的SCA与ADM联合应用比单纯加ADM(同等剂量)对K 562/AO2细胞的IC50降低了1.51倍;耐药细胞(K 562/AO2)经SCA处理后m dr1基因与P-gp蛋白表达均下降(P<0.05)。结论SCA对K 562/AO2有一定的逆转作用,其机制可能与下调m dr1基因和P-gp蛋白表达有关。