High Body Mass Has No Effect on Cellular and Innate Immunity in Siberian Hamsters (<i>Phodopus sungorus</i>)

Body mass is considered to be related with immune function in animals. Our aim was to test the hypothesis that cellular and innate immunity would be suppressed in high body mass of Siberian hamsters (Phodopus sungorus). Six heavier (high body mass, HBM) and six lighter (low body mass, LBM) hamsters were selected from 28 male hamsters. Body mass, body fat mass, wet spleen mass and blood glucose levels were significantly higher in the HBM group than in the LBM group. However, phytohaemagglutinin response, serum bacteria killing capacity and white blood cells did not differ between the two groups, suggesting cellular and innate immunity was?not impaired in high body mass of hamsters. There was no correlation between cellular, innate immunity and body mass, body fat mass and glucose levels, suggesting cellular and innate immunity was?not suppressed by higher body mass, body fat mass and glucose levels. In summary, cellular and innate immunity was not impaired in the HBM hamsters compared with the LBM hamsters.

Identification of optimal reference genes in golden Syrian hamster with ethanol-and palmitoleic acid-induced acute pancreatitis using quantitative real-time polymerase chain reaction

Background:Acute pancreatitis(AP)is a severe disorder that leads to high morbidity and mortality.Appropriate reference genes are important for gene analysis in AP.This study sought to study the expression stability of several reference genes in the golden Syrian hamster,a model of AP.Methods:AP was induced in golden Syrian hamster by intraperitoneal injection of ethanol(1.35 g/kg)and palmitoleic acid(2 mg/kg).The expression of candidate genes,including Actb,Gapdh,Eef2,Ywhaz,Rps18,Hprt1,Tubb,Rpl13a,Nono,and B2m,in hamster pancreas at different time points(1,3,6,9,and 24 h)posttreatment was analyzed using quantitative polymerase chain reaction.The expression stability of these genes was calculated using Best Keeper,Comprehensive Delta CT,Norm Finder,and ge Norm algorithms and Ref Finder software.Results:Our results show that the expression of these reference genes fluctuated during AP,of which Ywhaz and Gapdh were the most stable genes,whereas Tubb,Eef2,and Actb were the least stable genes.Furthermore,these genes were used to normalize the expression of TNF-αmessenger ribonucleic acid in inflamed pancreas.Conclusions:In conclusion,Ywhaz and Gapdh were suitable reference genes for gene expression analysis in AP induced in Syrian hamster.

Characterization of SHARPIN knockout Syrian hamsters developed using CRISPR/Cas9 system

Background : SHARPIN (SHANK- associated RH domain interactor) is a component ofthe linear ubiquitination complex that regulates the NF- κB signaling pathway. To betterunderstand the function of SHARPIN, we sought to establish a novel geneticallyengineered Syrian hamster with SHARPIN disruption using the CRISPR/Cas9 system.Methods : A single- guide ribonucleic acid targeting exon 1 of SHARPIN gene was designedand constructed. The zygotes generated by cytoplasmic injection of the Cas9/gRNA ribonucleoprotein were transferred into pseudopregnant hamsters. Neonatalmutants were identified by genotyping. SHARPIN protein expression was detectedusing Western blotting assay. Splenic, mesenteric lymph nodes (MLNs), and thymicweights were measured, and organ coefficients were calculated. Histopathologicalexamination of the spleen, liver, lung, small intestine, and esophagus was performedindependently by a pathologist. The expression of lymphocytic markers and cytokineswas evaluated using reverse transcriptase- quantitative polymerase chain reaction.Results : All the offspring harbored germline- transmitted SHARPIN mutations.Compared with wild- type hamsters, SHARPIN protein was undetectable in SHARPIN −/−hamsters. Spleen enlargement and splenic coefficient elevation were spotted inSHARPIN −/− hamsters, with the descent of MLNs and thymuses. Further, eosinophilinfiltration and structural alteration in spleens, livers, lungs, small intestines, and esophagiwere obvious after the deletion of SHARPIN. Notably, the expression of CD94 and CD22 was downregulated in the spleens of knockout (KO) animals. Nonetheless,the expression of CCR3, CCL11, Il4 , and Il13 was upregulated in the esophagi. Theexpression of NF- κB and phosphorylation of NF- κB and IκB protein significantly diminishedin SHARPIN −/− animals.Conclusions : A novel SHARPIN KO hamster was successfully established using theCRISPR/Cas9 system. Abnormal development of secondary lymphoid organs andeosinophil infiltration in multiple organs reveal its potential in delineating SHARPINfunction and chronic inflammation.

小毛足鼠(Phodopus roborovski)巢区和活动距离的初步研究

１９９５～１９９７年在鄂尔多斯沙地采用标志重捕法对小毛足鼠（Ｐｈｏｄｏｐｕｓｒｏｂｏｒｏｖｓｋｉ）的巢区和活动距离进行了调查．结果表明，该鼠的巢区和活动距离雄性分别为３５３６．１±４４３．３ｍ２和１１２．７±８．０ｍ、雌性分别为３３７５．０±３７９．８ｍ２和１１０．１±７．２ｍ，二者均不存在显著的性别差异，也不存在显著的年龄差异和季节差异．巢区面积和活动距离呈极显著的线性相关（ｒ＝０．８０７＞ｒ０．０１＝０．２８３，ｄｆ＝８７）．

小毛足鼠(Phodopus roborovskii)食性和食量的研究

小毛足鼠饲喂试验结果以及颊囊食物统计结果表明，该鼠在饲喂条件下食物资源利用谱相对较宽，但在野外条件下食物资源的利用主要以植物种子为主；并结合具体情况探讨了造成野外条件下该鼠食物资源利用范围趋于特化的原因．估算了小毛足鼠的采食效率为０．０７６％～０．１４％．在以上研究基础上。

无镁诱导仓鼠原代皮质神经元电生理学特性

目的无镁细胞外液建立癫痫放电模型,利用全细胞膜片钳技术,检测仓鼠原代皮质神经元的电生理学特性。方法采用生后1~2 d新生叙利亚仓鼠,分离大脑皮质培养原代神经元培养至第12天,分别予有镁细胞外液(有镁组)和无镁细胞外液(无镁组)孵育3 h,3 h后均更换为正常孵育液继续培养24 h。利用全细胞膜片钳技术,电压钳模式下记录神经元兴奋性突触后电流(excitatory postsynaptic currents,EPSC),电流钳模式下记录神经元兴奋性突触后电位(excitatory postsynaptic potentials,EPSP)。结果与有镁组相比,无镁组仓鼠原代皮质神经元EPSC[(124.38±75.15)Hz vs.(33.93±22.32)Hz,P<0.001]及EPSP[(37.05±38.37)Hz vs.(5.63±9.52)Hz,P<0.01]的频率升高,且差异有统计学意义;而两组之间EPSC及EPSP的振幅、曲线下面积和半宽度的差异无统计学意义(P>0.05)。结论无镁处理后仓鼠原代皮质神经元兴奋性升高,仓鼠原代皮质神经元可用于构建癫痫细胞模型。

基于肠道菌群探讨粗壮女贞总苷对高脂血症金黄仓鼠的降脂作用机制

目的评价粗壮女贞总苷(total phenylpropanoid glycosides extracted from Ligustrum robustum(Roxb.)Blume,LRTPG)对高脂血症金黄仓鼠的降脂作用并探讨其对肠道菌群的调节作用。方法60只仓鼠随机分为对照组、模型组、阳性药组、LRTPG-L组、LRTPG-M组、LRTPG-H组。高脂饲料诱导造模成功后连续给药4周,进行血脂和肝脂含量检测,并采集对照组、模型组和LRTPG-H组仓鼠结直肠部位成形粪便进行16S rDNA测序。结果LRTPG对高脂血症仓鼠具有降低血清TG、TC、LDL-C水平和肝脏TG、TC含量的作用。肠道菌群测序结果显示,与模型组比较,LRTPG在门水平降低厚壁菌门相对丰度,增加拟杆菌门和疣微菌门相对丰度(P<0.01);科水平上明显升高克里斯滕森菌科、消化球菌科、疣微菌科相对丰度(P<0.05或P<0.01);属水平上明显升高颤螺菌属、颤杆菌克属、解黄酮菌属和阿克曼氏菌属的相对丰度(P<0.05或P<0.01)。这些菌群的变化均有益于LRTPG降脂作用的发挥。结论LRTPG可能通过改善高脂饮食导致的金黄仓鼠肠道菌群紊乱发挥降脂功效。

复方石榴皮药膜对金黄地鼠皮脂腺斑形态学及雄激素受体表达的影响

目的:观察复方石榴皮药膜对金黄地鼠皮脂腺斑大小、组织形态及雄激素受体的影响,探讨其在痤疮治疗中的作用机制。方法:将15只天生具有雄激素依赖性的雄性金黄地鼠随机分为空白组、阳性对照组、药膜组,每组5只。分别于两侧皮脂腺斑处涂抹蒸馏水、0.025%维A酸乳膏、复方石榴皮药膜,连续给药28 d,于第0、14、28天测量金黄地鼠双侧皮脂腺斑面积大小并取部分皮脂腺斑行组织病理检测,免疫组织化学检查雄激素受体(AR)表达水平。结果:药膜组给药第14、28天分别与给药前比较,皮脂腺斑面积减小,差异均有统计学意义(P<0.05)。阳性对照组给药第28天与给药第14天比较,皮脂腺斑面积减小,差异有统计学意义(P<0.05)。给药第28天,药膜组与阳性对照组皮脂腺斑面积大小比较,差异有统计学意义(P<0.05)。给药第28天,与空白组比较,药膜组、阳性对照组皮脂腺斑厚度减小,差异均有统计学意义(P<0.05);药膜组、阳性对照组皮脂腺斑AR表达减少,差异均有统计学意义(P<0.05)。结论:复方石榴皮药膜可减小金黄地鼠痤疮模型皮脂腺斑面积、减轻皮脂腺斑厚度,其作用机理可能与抑制雄激素受体表达有关。

不同固定液对光镜下金黄地鼠海马体神经元的影响

目的比较不同固定液对光镜下金黄地鼠海马体神经元的影响,为金黄地鼠海马体神经元固定液的选择提供参考。方法将21只11周龄雄性SPF级金黄地鼠随机分为7组,分别使用4%多聚甲醛、10%中性甲醛、Bouin’s、Carnoy、Davidson’s、Zenker、Helly溶液进行心脏灌注,剥离脑壳取出完整大脑,在对应的固定液中固定48 h,并制作成石蜡切片,经HE染色后在光镜下观察海马体神经元的形态特征。通过匈牙利3DHISTECH公司数字切片扫描系统测量尺测量神经元细胞直径,比较固定液对细胞收缩的影响。结果4%多聚甲醛、10%中性甲醛、Bouin’s溶液展现了良好的固定效果,固定后的大脑硬度适中,HE染色后的切片着色适中、色泽鲜艳、神经元各种细胞形态对比清晰,4%多聚甲醛因固定后切片无龟裂而更为理想。不同固定液固定后的海马体神经元存在细胞收缩方面的极显著差异(P<0.01),使用Carnoy、Davidson’s固定后的神经元各种细胞收缩最严重。结论4%多聚甲醛、10%中性甲醛、Bouin’s溶液是金黄地鼠海马体神经元较为理想的固定液,4%多聚甲醛溶液建议作为首选。

啮齿类动物Playfighting的行为学研究进展

Playfighting是一种常见的社会性互动形式,具有互惠性和合作性等特点。其贯穿于动物的整个生命周期,并且会通过影响动物的社交、认知和运动等进一步影响大脑的执行功能。啮齿类动物的playfighting行为因具有易识别、易量化等特点,而被广泛研究。本文主要以大鼠和金仓鼠为例,整理概述了近年来啮齿类动物playfighting行为的研究现状,包括行为模式及其相关影响因素,希望为今后进一步研究playfighting行为提供参考。

哺乳动物细胞表达系统研究进展

哺乳动物细胞表达系统具有强大的翻译后修饰功能,表达的重组蛋白更接近人源构象,因此被广泛用于治疗性重组蛋白的生产,其中应用最广泛的是中华仓鼠卵巢细胞。基因组学、转录组学、蛋白质组学以及代谢组学等研究的不断深入促进了哺乳动物细胞表达系统的不断完善。本文立足于哺乳动物细胞表达系统的研究现状,对表达载体的优化、宿主细胞和位点特异性重组等方面的最新进展进行综述。

基于非标记分析方法应用四极杆-静电场轨道阱-线性离子阱三合一高分辨质谱仪鉴定中国仓鼠卵巢细胞膜蛋白

目的 探讨应用四极杆-静电场轨道阱-线性离子阱(Orbitrap fusion lumos)三合一高分辨质谱仪鉴定中国仓鼠卵巢(Chinese hamster overy,CHO)细胞的全蛋白质组和膜蛋白质组的非标记(label-free)分析方法,并评价其应用效果,为研究膜蛋白相关传染病的发生机制提供方法学基础。方法 贴壁培养中国仓鼠卵巢细胞,提取、富集和酶解处于对数生长期细胞的全蛋白和膜蛋白,用高效液相色谱仪联用四极杆-静电场轨道阱-线性离子阱三合一高分辨质谱仪对酶解后的多肽进行分析,利用Maxquant软件和Uniprot数据库进行搜库及鉴定,最后对鉴定膜蛋白进行基因本体论(gene ontology,GO)功能分类。结果 全蛋白组一共检测出14 300个肽段,搜库得到2708个蛋白,其中可信蛋白2002个(FDR<1%且特异性肽段>1);膜蛋白组一共检测出13 179个肽段,搜库得到2594个蛋白,其中可信蛋白1873个。在鉴定的全蛋白组和膜蛋白组中定量结果前20的蛋白中,全蛋白组中与膜相关的蛋白有4个,占比20%;膜蛋白组中与膜相关的蛋白有10个,占比50%。在膜蛋白组中,GO功能分类表明分子功能上主要涉及结合和催化活性,在生物学进程上主要涉及细胞进程、代谢过程等。结论 膜蛋白组中与膜相关的蛋白显著高于本实验的全蛋白组,基于非标记分析方法应用Orbitrap fusion lumos的分析技术是鉴定细胞膜蛋白的有效方法。

Expression of hepatitis B virus genes in early embryonic cells originated from hamster ova and human spermatozoa transfected with the complete viral genome

Aim： To detect the expression of hepatitis B virus （HBV） genes （HB S and C genes） in early embryonic cells after introducing motile human sperm carrying HBV DNA into zona-free hamster oocytes via the in vitro fertilization （IVF） technique. Methods： Human sperm-mediated HBV genes were delivered into zona-free hamster oocytes by the IVF method. Polymerase chain reaction （PCR） was used to detect HB S and pre-Core/Core （pre-C/C） coding genes both in one- and two-cell embryos. Reverse transcription-PCR （RT-PCR） analysis was used to study the expression of the two genes. Fluorescence in situ hybridization （FISH） analysis using the full-length HBV DNA as the hybridization probe was performed to confirm the integration of viral DNA in the host embryonic genome. Results： Both HB S and pre-C/C coding genes are present and transcribed in one- and two-cell embryos originated from hamster ova IVF with human spermatozoa carrying HBV DNA sequences. Conclusion： Sperm-mediated HBV genes are able to replicate and express themselves in early embryonic cells. These results provide direct evidence that HBV DNA could transmit vertically to the next generation via the male germ line.

Modulating Effects of Chlorogenic Acid on Lipids and Glucose Metabolism and Expression of Hepatic Peroxisome Proliferator-activated Receptor-α in Golden Hamsters Fed on High Fat Diet

Objective To examine the effects of chlorogenic acid （CGA） on lipid and glucose metabolism under a high dietary fat burden and to explore the possible role of peroxisome proliferator-activated receptor-α （PPAR-α） in these effects. Methods Twenty male golden hamsters were randomly divided into CGA treatment group （n=10, given peritoneal injection of CGA solution prepared with PBS, 80 mg CGA/kg body weight daily）, and control group （n=10, given PBS i.p. at the average volume of the treatment group）. Animals in both groups were given 15% high fat diet. Eight weeks after treatment with CGA, the level of biochemical parameters in fasting serum and tissues and the expression of hepatic mRNA and protein PPAR-α were determined. Results Eight weeks after treatment with CGA, the levels of fasting serum triglyceride （TG）, free fatty acid （FFA）, total cholesterol （TC）, low density lipoprotein cholesterol （LDL-C）, high density lipoprotein cholesterol （HDL-C）, glucose （FSG）, and insulin （FSI） were significantly lower in the GGA treatment group than in the control group. CGA also led to higher activity of hepatic lipase （HL） lower contents of TG and FFA in liver, and lower activity of lipoprotein lipase （LPL） in skeletal muscle. Furthermore, CGA significantly elevated significantly elevated the expression level of mRNA and protein expression in hepatic PPAR-α. Conclusion CGA can modify lipids and glucose metabolism, which may be attributed to PPAR-α facilitated lipid clearance in liver and improved insulin sensitivity.

Role of the Chinese Herbal Medicine Xianhuayin on the Reversal of Premalignant Mucosal Lesions in the Golden Hamster Buccal Pouch

Aim To investigate the role of the Chinese herbal medicine Xianhuayin on the reversal of 7,12-dimethylbenz[a]anthracene （DMBA）-induced premalignant mucosal lesions in the oral buccal pouch of golden hamsters. Methodology The animals were randomly divided into a non-diseased control group （n=5） and an experimental group including 50 animals in which the buccal mucosa had been painted with DMBA （0.5% in acetone） to generate an oral mucosa premalignant lesion. Animals in the experi- mental group were further divided into Xianhuayin-treated group （n=30）, untreated prem＇alignant lesion group （n=10） and normal saline （NS）-treated group （n=10）. The cheek （buccal） pouch mucosa of the golden hamsters in each group was observed with light and electron microscopy eight weeks after intragastric administration with NS or Xianhuayin. Results In the non-diseased control group, the buccal mucosa was keratinized and stratified squamous epithelium under a light microscope. In the untreated premalignant lesion group, variable degrees of epithelial dysplasia was observed. The irregular epithelial mucosa gradually became distinct in the Xianhuayin-treated group. Scanning electronic microscopic （SEM） analysis showed that surface of the cells exhibited honeycomb structures in the hamster of untreated- group. The cells were morphologically irregular, overlapped and loosened in the untreated premalignant lesion group. Most of the cell surface exhibited honeycomb structure in the Xianhuayin-treated group. Transmission electronic micro- scopic （TEM） analysis showed that buccal mucosal epithelial cells were morphologically regular in the non-diseased control group. Desmosomes and tonofibrils were reduced and the nucleus was morphologically irregular in the untreated premalignant lesion group. In the Xianhuayin-treated group, the widening intercellular gap was gradually reduced, desmosomes and the cells becoming morphologically regular. No significant difference was observed between the hamsters in NS-treated group and those in the untreated premalignant lesion group. Significant therapeutic efficacy was observed in the group receiving Xianhuayin. Conclusion Xianhuayin is effective in the reversal of DMBA-induced premalignant lesions in the buccal pouch of golden hamsters.

Pravastatin activates PPARα/PPARγexpression in the liver and gallbladder epithelium of hamsters

BACKGROUND:Our earlier study with cultured gallbladder epithelial cells demonstrated that statins(HMG-CoA reductase inhibitors)activate the expression of PPARαand PPARγ, consequently blocking the production of pro-inflmmatory cytokines.The present study used hamsters to investigate the effects of pavastatin on PPARα/PPARγexpression in the liver and gallbladder epithelium,and to determine whether pravastatin suppresses cholesterol crystal formation in the gallbladder. METHODS:A total of 40 Golden Syrian male hamsters(4 weeks old)were randomly assigned to four groups(basal diet control; basal diet+pavastatin;high cholesterol diet;high cholesterol diet+pravastatin).All hamsters were 11 weeks old at the end of the experiment.The liver,gallbladder and bile were harvested. Immunohistochemical staining and Western blotting for PPARαand PPARγwere performed in the liver and gallbladder. A drop of fresh bile was examined for cholesterol crystals under a microscope. RESULTS:In the gallbladder and liver of the hamsters, pravastatin activated the PPARαand PPARγexpression of gallbladder epithelial cells and hepatocytes,and particularly the response of PPARγwas much stronger than that of PPARα. Pravastatin suppressed the formation of cholesterol gallstones or crystals in the gallbladder. CONCLUSION:Pravastatin is an effective medication to activate PPARs(especially PPARγ)in the liver and the gallbladder epithelium of hamsters,and contributes to the prevention of gallstone formation.

Effects of 13 T Static Magnetic Fields (SMF) in the Cell Cycle Distribution and Cell Viability in Immortalized Hamster Cells and Human Primary Fibroblasts Cells

Magnetic resonance image （MRI） systems with a much higher magnetic flux density were developed and applied for potential use in medical diagnostic. Recently, much attention has been paid to the biological effects of static, strong magnetic fields （SMF）. With the 13 T SMF facility in the Institute of Plasma Physics, Chinese Academy of Sciences, the present study focused on the cellular effects of the SMF with 13 T on the cell viability and the cell cycle distribution in immortalized hamster cells, such as human-hamster hybrid （AL） cells, Chinese hamster ovary （CHO） cells, DNA double-strand break repair deficient mutant （XRS-5） cells, and human primary skin fibroblasts （AG1522） cells. It was found that the exposure of 13 T SMF had less effect on the colony formation in either nonsynchronized or synchronized AL cells. Moreover, as compared to non-exposed groups, there were slight differences in the cell cycle distribution no matter in either synchronized or nonsynchronized immortalized hamster ceils after exposure to 13 T SMF. However, it should be noted that the percentage of exposed AG1522 cells at G0/G1 phase was decreased by 10% as compared to the controls. Our data indicated that although 13 T SMF had minimal effects in immortalized hamster cells, the cell cycle distribution was slightly modified by SMF in human primary fibroblasts.

TELOMERASE ACTIVITY DURING 7,12-DIMETHYLBENZ [a] ANTHRACENE-INDUCED HAMSTER BUCCAL POUCH CARCINOGENESIS

Objective: To investigate the roles of telomerase activity (TA) in relation to hamster buccal pouch tumor progression. Methods: male hamster were treated three times weekly with 0.5% of 7, 12-dimethyl-benzanthracene (DMBA) over a IS weeks experimental period. Hamsters were sacrificed at 3, 6, 9, 12 and 15 weeks after treatment. Telomerase activity of hamster buccal pouch tissue were measured along with the analyses of the formation of DMBA-induced hamster buccal pouch tumors. Results: DMBA-induced squamous cell carcinomas were found at the 6th week after dosing. Telomerase activity elevation began at the 3rd week and was increasing to a plateau at the 12th week. Conclusion: Our results show that telomerase activity in the target tissue may be detected at the early stage of the DMBA-induced hamster buccal pouch tumor formation and suggests that telomerase activity may be used as a biomarker for an early clinical detection of buccal pouch cancer.

The role of leptin in striped hamsters subjected to food restriction and refeeding

Food restriction （FR） and refeeding （Re） have been suggested to impair body mass regulation and thereby making it easier to regain the lost weight and develop over-weight when FR ends. However, it is unclear if this is the case in small mammals showing seasonal forging behaviors. In the present study, energy budget, body fat and serum leptin level were measured in striped hamsters that were exposed to FR-Re. The effects of leptin on food intake, body fat and genes expressions of several hypothalamus neuropeptides were determined. Body mass, fat content and serum leptin level decreased during FR and then increased during Re. Leptin supplement significantly attenuated the increase in food intake during Re, decreased genes expressions of neuropepetide Y （NPY） and agouti-related protein （AgRP） of hypothalamus and leptin of white adipose tissue （WAT）. Hormone-sensitive lipase （HSL） gene expression of WAT increased in leptin-treated hamsters that were fed ad libitum, but decreased in FR-Re hamsters. This indicates that the adaptive regulation of WAT HSL gene expression may be involved in the mobilization of fat storage during Re, which partly contributes to the resistance to FR-Re-induced overweight. Leptin may be involved in the down regulations of hypothalamus orexigenic peptides gene expression and consequently plays a crucial role in controlling food intake when FR ends.

Glycoproteomics analysis of plasma proteins associated with Opisthorchis viverrini infection-induced cholangiocarcinoma in hamster model

Objective: To apply lectin affinity chromatography and glycoproteomics-based LC-MS/MS to preliminarily investigate the possible potential plasma biomarkers of Opisthorchis viverrini(OV)-associated CCA in OV/dimethylnitrosamine(DMN)-induced CCA hamster model. Methods: Nine Syrian hamsters were divided into 3 groups as follows(n=3 each): normal(healthy control group); OV group; and OV/DMN group(CCA group). Pooled plasma samples collected from animals in each group at the 6th month post-infection with OV metacercarae were subjected to glycoproteomics analysis. Glycoproteins in the pooled sample from each group were initially isolated by concanavilin A(Con A)-based affinity chromatography. The expression of glycoproteins isolated by both enrichment methods were determined using LCMS/MS. Results: Among the 24 Con A-binding glycoproteins isolated, two proteins, N-myc downstream regulated gene 1(NDRG1) and fetuin-B(FETUB) were found up-regulated only in the samples from the OV and control groups, but not in the OV/DMN(CCA) groups. On the other hand, one protein, i.e., NSFL1 cofactor p47 isoform x3(NSFL1C) was found only in the samples from OV/DMN(CCA) and control groups, but not in the OV group. The remaining 21 proteins were upregulated in the samples from all groups. Conclusions: NDRG1, FETUB and NSFL1 C glycoproteins isolated by Con A-based affinity chromatography could be potential biomarkers for CCA. Plasma samples with negative for NDRG1 and FETUB proteins but positive for NSFL1 C are likely to be OV-associated CCA. Nevertheless, this conclusion remains to be confirmed whether this battery test can discriminate OV-associated CCA from other risk factors.