In vivo fluorescence flow cytometry reveals that the nanoparticle tumor vaccine OVA@HA-PEI effectively clears circulating tumor cells

Tumor vaccine therapy offers significant advantages over conventional treatments,including reduced toxic side effects.However,it currently functions primarily as an adjuvant treatment modality in clinical oncology due to limitations in tumor antigen selection and delivery methods.Tumor vaccines often fail to elicit a sufficiently robust immune response against progressive tumors,thereby limiting their clinical efficacy.In this study,we developed a nanoparticle-based tumor vaccine,OVA@HA-PEI,utilizing ovalbumin(OVA)as the presenting antigen and hyaluronic acid(HA)and polyethyleneimine(PEI)as adjuvants and carriers.This formulation significantly enhanced the proliferation of immune cells and cytokines,such as CD3,CD8,interferon-,and tumor necrosis factor-,in vivo,effectively activating an immune response against B16–F10 tumors.In vivofluorescenceflow cytometry(IVFC)has already become an effective method for monitoring circulating tumor cells(CTCs)due to its direct,noninvasive,and long-term detection capabilities.Our study utilized a laboratory-constructed IVFC system to monitor the immune processes induced by the OVA@HA-PEI tumor vaccine and an anti-programmed death-1(PD-1)antibody.The results demonstrated that the combined treatment of OVA@HA-PEI and anti-PD-1 antibody significantly improved the survival time of mice compared to anti-PD-1 antibody treatment alone.Additionally,this combination therapy substantially reduced the number of CTCs in vivo,increased the clearance rate of CTCs by the immune system,and slowed tumor progression.Thesefindings greatly enhance the clinical application prospects of IVFC and tumor vaccines.

In Vivo Studies and Flow Cytometric Investigation on Anticancer Potential of Selenium Nanoparticles Synthesized via Aqueous Extract of Clerodendron phlomidis

Nowadays,doctors and nutritionists recommend individuals incorporate selenium-rich foods such as nuts,cereals,and mushrooms into their regular diet to maintain fitness and overall health.Selenium nanoparticles(SeNPs)exhibit strong chemopreventive capabilities.The anticipations for SeNPs with enhanced and tunable bioactive activities have led to a keen interest in phytofabrication.In this study,the aqueous extract of Clerodendron phlomidis plant leaves was utilized for the synthesis of SeNPs.In traditional Indian medicine,this plant extract is recognized as a significant anti-diabetic agent.The flavonoids tetrahydroxylflavone,7-hydroxyflavanone,and 6,4’-dimethyl-7-acetoxy-scutellarein present in this plant leaf extract demonstrate excellent anticancer activity.These secondary metabolites exhibit the ability to reduce sodium selenite into SeNPs.At a concentration of 13μg/mL,the synthesized SeNPs effectively inhibited the proliferation of the HepG2 cell line.The results suggest that the SeNPs possess promising anti-cancer potential against liver cancer and can be considered as a therapeutic agent for liver cancer treatment.Additionally,the cell cycle arrest induced by SeNPs was further confirmed by the fluorescence-activated cell sorting(FACS)method,indicating that SeNPs could efficiently differentiate cancer cells from normal cells.Notably,it showed a significant improvement in diethylnitrosamine(DEN)-induced Swiss Wistar rat groups.This scientific investigation highlights the high anti-cancer potential of SeNPs,positioning them as a promising therapeutic agent for liver cancer treatment.

Detection of cells by flow cytometry:Counting,imaging,and cell classification

The study of circulating cells in the blood stream is critical,as it covers many felds of biomed-icine,including immunology,cell biology,oncology,and reproductive medicine.In-viuo flowcytometry(IVFC)is a new tool to monitor and count cells in real time for long durations in theirnative biological environment.This review describes two main categories of IVFC,ie.,labeledand label-free IVFC.It focuses on label-free IVFC and introduces its technological developmentand related biological applications.Because cell recognition is the basis of flow cytometrycounting,this review also describes various methods for the classification of unlabeled cells,including the latest machine learning-based technologies.

Modeling cell contractility responses to acoustic tweezing cytometry

Acoustic tweezing cytometry(ATC)is a recently developed method for cell mechanics regulation.Tar-geted microbubbles,which are attached to integrins and subsequently the actin cytoskeleton,anchor,amplify and transmit the mechanical energy in an acoustic field inside the cells,eliciting prominent cy-toskeleton contractile force increases in various cell types.We propose that a mechanochemical con-version mechanism is critical for the high efficiency of ATC to activate cell contractility responses.Our models predict key experimental observations.Moreover,we study the influences of ATC parameters(ul-trasound center frequency,pulse repetition frequency,duty cycle,and acoustic pressure),cell areas,the number of ATC stimuli,and extracellular matrix rigidity on cell contractility responses to ATC.The simu-lation results suggest that it is large molecules,rather than small ions,that facilitate global responses to the local ATC stimulation,and the incorporation of visible stress fiber bundles improves the accuracy of modeling.

基于流式细胞仪鉴定番石榴倍性方法的建立及应用

【目的】利用流式细胞仪鉴定番石榴的染色体倍性,为番石榴倍性育种和杂交育种奠定基础。【方法】以番石榴为材料,比较了样品部位、离心处理和保存方式对倍性检测结果的影响。【结果】番石榴花瓣作为倍性检测材料时,收集到的细胞核DNA数目最多,CV值为1.96%,峰形效果最佳;在制备细胞核悬浮液时,过滤后不离心,直接染色后上机的测定效果最好;新鲜番石榴花瓣得到的细胞核DNA含量明显高于冷藏和其他冷冻处理组,且背景碎片少,主峰明显;-80℃冷冻处理对样本的破坏性最小,检测效果仅次于新鲜的番石榴花瓣。本研究建立的番石榴流式细胞术倍性分析的方法为:取番石榴花瓣0.50-1.00 cm^(2),加入1 mL的mGb裂解液中混合切碎,过滤后加入30μL PI染色1 min即可上机检测。【结论】利用建立的流式细胞术对33份番石榴种质资源进行倍性鉴定,共检测出二倍体32份,六倍体1份。该方法执行简便、高效准确,为番石榴种质资源倍性鉴定提供了有效方法。

Performance Evaluation of Combined Detection of Serum CEA,CYFRA21-1,CA125,and NSE in Patients with Lung Cancer by Fluorescence Flow Cytometry

Objective:To investigate the effect of combined detection of serum carcinoembryonic antigen(CEA),cytokeratin 19 fragment(CYFRA21-1),cancer antigen 125(CA125),and neuron-specific enolase(NSE)in patients with lung cancer by fluorescence flow cytometry.Methods:From August 2019 to July 2022,200 patients with lung cancer diagnosed by pathology in our hospital were retrospectively analyzed.2 mL venous blood was collected in a fasting state and centrifuged to separate the serum(containing human chorionic gonadotropin antibody[anti-hCG antibody],hepatitis B surface antibody[anti-HBs antibody],and CEA).Results:The sensitivities of CEA and CYFRA21-1 detected via enzyme-linked immunosorbent assay(ELISA)were 100%,and the detection limits were 0.5 ng/mL and 0.1 ng/mL,respectively;the sensitivities of CA125 and NSE detected via flow cytometry were 100%,and the detection limits were 10 U/mL and 2 ng/mL,respectively.Compared with ELISA,the sensitivities of CA125 and NSE detected via flow cytometry were higher.When the concentration of CEA was 10-40 ng/mL,the sensitivities of the three markers CYFRA21-1,CA125,and NSE showed no significant changes(P>0.05);when the concentration of CEA was 40-80 ng/mL,the sensitivity of CEA significantly decreased(P<0.01),but the sensitivities of the three markers CYFRA21-1,CA125,and NSE showed no significant changes(P>0.05);when the concentration of CEA was 80-200 ng/mL,the sensitivities of all four markers showed no significant changes(P>0.05).Conclusion:Compared with the double-antibody sandwich ELISA,fluorescence flow cytometry has certain advantages,including high sensitivity,good precision,short detection time,low sample usage,and low medical cost;thus,it is worthy of clinical promotion.

The Clinical Value of Detecting the Level of Exfoliated Cells in Pleural Effusion by Flow Cytometry in the Differential Diagnosis of Non-

Objective:To explore the value of flow cytometry(FCM)in detecting the level of exfoliated cells in pleural effusion in the differential diagnosis of non-small cell lung cancer and benign lung diseases.Methods:Clinical data of patients with non-small cell lung cancer who were hospitalized in Hebei hospital from June 2019 to March 2022 were collected.A total of 98 patients were included,and 63 patients with alveolar lung disease were screened during the same period,and the two groups of patients were analyzed.Results:Compared with alveolar lung disease group,FCM detection and analysis showed that the level of exfoliated cells in the pleural effusion of non-small cell lung cancer(NSCLC)patients was 99(3-969)/100,000,and patients with alveolar lung disease was 4(0~19)/100,000.Additionally,compared with the alveolar lung disease group,the level of exfoliated cells in the pleural effusion of patients with non-small cell lung cancer(NSCLC)was significantly increased(P<0.001).The diagnostic efficacy of FCM for detecting pleural fluid exfoliated cells in non-small cell lung cancer was assessed using ROC curves and using 95%CI(-11.1,-13.2)with a sensitivity of 0.75 and specificity of 0.94,and the diagnostic efficacy of FCM for detecting pleural fluid exfoliated cells in alveolar lung disease was assessed using 95%CI(-11.1,-13.2)with a sensitivity of 0.71 and specificity of 0.87.Conclusion:Flow cytometry has a wider range of clinical applications,simple operation,low cost,and high sensitivity,which makes it of great significance in disease diagnosis.

基于流式细胞术对不同倍性草莓基因组大小的测定

以不同倍性草莓资源的嫩叶为材料,根据待测样品倍性预估基因组大小后分别选择水稻“日本晴”(基因组大小为430 Mb)、番茄(基因组大小为900 Mb)、玉米“B73”(基因组大小为2.3 Gb)作为内参,利用流式细胞仪测定样品的基因组大小。结果表明,二倍体资源,森林草莓基因组大小为(233±0.78)Mb,绿色草莓为(234±0.82)Mb;四倍体资源,西南草莓基因组大小为(492±2.12)Mb,伞房草莓(雄株)为(488±7.07)Mb,伞房草莓(雌株)为(491±4.95)Mb,伞房草莓雄株和雌株基因组大小均为首次测定;六倍体资源,麝香草莓基因组大小为(644±1.73)Mb;八倍体资源,栽培品种“章姬”基因组大小为(793±7.78)Mb,“蒙特瑞”为(761±0.00)Mb;十倍体资源,野生的择捉草莓基因组大小为(938±6.36)Mb,栽培品种“桃熏”为(1007±6.36)Mb。

流式细胞术检测细胞周期影响因素及不同免疫细胞亚群周期分析

细胞周期检测对于了解细胞增殖状态、细胞功能研究、药物筛选与评估等领域具有重要意义。流式细胞仪分析细胞周期是目前较为经典且广泛应用的检测手段之一。碘化丙啶(propidium iodide,PI)单染是最常用的基于流式的检测细胞周期方法,然而,该方法在使用过程有诸多处理因素会对检测结果造成影响。此外,不同免疫细胞亚群在不同细胞周期阶段的分布差异也有助于研究免疫反应,了解疾病状态。本文以B16-F10细胞系为例,采用PI单染法,从固定条件、上机条件和软件分析3个角度,评价多种处理因素对细胞周期检测结果的影响。根据结果分析,在进行细胞周期检测时,建议取3×10^(6)个细胞,用300μL预冷PBS重悬细胞后逐滴滴入700μL预冷的无水乙醇,放置于4℃或-20℃过夜固定,低速收样,收样速率为每秒400~600颗粒数,去黏连后需收集至少3000个细胞。另外,通过EdU、PI双指标染色可以精确划分细胞周期,而细胞表面标志物染色联合Ki-67和PI染色法,可在不进行细胞分选的情况下进行不同免疫细胞亚群周期分析。本文较为系统的探讨了PI单染法检测细胞周期的影响因素,提供了标准的实验操作流程。建立了EdU联合PI检测细胞周期和针对不同免疫细胞亚群周期分析方案,拓宽了细胞周期检测方式。

基于流式细胞仪提高乳制品商业无菌检验时效性的研究

目的基于流式细胞仪探究提高乳制品商业无菌检验项目时效性。方法以地衣芽孢杆菌CMCC(B)63552和蜡样芽孢杆菌CMCC(B)63303菌株制备定量参考品。以4种灭菌乳为基质,制备染菌浓度分别为800、8000、80000 CFU/mL的菌悬液,测试流式细胞仪法灵敏度。制备染菌浓度为8 CFU/样品和80 CFU/样品的灭菌乳和调制乳样品,经(36±1)℃保温(24±2)h后,按照GB 4789.26法和流式细胞仪法进行检测,并分析比较两种方法检测结果。结果在灭菌乳基质中,染菌浓度为800 CFU/mL时,流式细胞仪法对人工污染地衣芽孢杆菌样品的阳性检出率为75%,而对人工污染蜡样芽孢杆菌的样品均未检出阳性;染菌浓度为8000 CFU/mL和80000 CFU/mL时,流式细胞仪法对地衣芽孢杆菌和蜡样芽孢杆菌污染样品的阳性检出率均为100%。对人工污染8 CFU和80 CFU地衣芽孢杆菌和蜡样芽孢杆菌的灭菌乳和调制乳样品,经保温处理后,流式细胞仪法检出率为100%,GB 4789.26法中未见样品膨胀,但是镜检和培养结果检出率为100%。结论当样品中染菌浓度≥8000 CFU/mL时,流式细胞仪法可稳定检出。人工污染地衣芽孢杆菌和蜡样芽孢杆菌的灭菌乳和调制乳样品经保温处理(24±2)h后,流式细胞仪法和GB 4789.26法检出率均为100%。

基于流式细胞术的江西10个油茶农家品种基因组大小测定

油茶农家品种具有较高经济价值和地方特色,是油茶育种的重要资源,然而目前有关油茶的遗传基础研究较少。为了探明江西省油茶农家品种的基因组大小,为油茶农家品种的遗传评价与合理利用提供基础,文章采用流式细胞术对江西省10个油茶农家品种的基因组大小进行测定,结果表明,江西油茶农家品种白皮中子、观音桃、茅岗大果、珍珠子、二水桃、霜降红皮、赣萍茶、石市红皮、遂中子、夏讲的基因组大小分别为:5 480.06、8 531.31、8 751.11、8 545.69、8 422.12、8 756.84、8 477.28、8 591.20、8 763.49、8 684.92 Mb,白皮中子的基因组大小接近其他农家品种的2/3,赣石84-8的基因组大小与除白皮中子外的其他油茶农家品种的基因组大小接近。

基于虚拟实验数据库的流式细胞术线上教学应用研究

目的探讨线上教学在流式细胞术及其应用课程实验课教学中的应用效果。方法研究时间为2022年3月至2023年12月。选取首都医科大学2019级和2021级选修流式细胞术及其应用课程的37名硕士研究生为对照组,2022级和2023级选修流式细胞术及其应用课程的31名硕士研究生为试验组。对照组接受实验课线下教学,试验组接受实验课线上和线下混合教学模式教学。对试验组实验课线上教学模式满意度进行调查和评价,比较两组流式数据分析成绩和实验操作成绩。结果调查显示,试验组线上教学模式提升流式数据分析能力和线上教学模式满意度最高(96.7%),线上教学模式提升科研能力满意度最低(83.9%)。试验组流式数据分析成绩和实验操作成绩均高于对照组(P<0.05)。结论基于虚拟实验数据库的流式细胞术线上教学可提高研究生流式细胞术数据分析及实验操作能力。

CD19^(+)CD27^(+)IgD^(+)B细胞与甲状腺相关性眼病患者疾病活动度相关性研究

目的:探讨甲状腺相关性眼病(thyroid-associated ophthalmopathy,TAO)患者外周血CD19^(+)CD27^(+)IgD^(+)B细胞比例变化与病情活动度的关系。方法:采用流式细胞术检测TAO患者和健康对照组CD19^(+)CD27^(+)IgD^(+)B细胞比例,分析TAO患者CD19^(+)CD27^(+)IgD^(+)B细胞比例与健康对照组的差异,并分析CD19^(+)CD27^(+)IgD^(+)B细胞比例与TAO疾病活动度之间的相关性。结果:TAO患者CD19^(+)CD27^(+)IgD^(+)B细胞比例明显低于健康对照(P<0.05);活动性TAO患者CD19^(+)CD27^(+)IgD^(+)B细胞比例细胞显著低于非活动性TAO患者;CD19^(+)CD27^(+)IgD^(+)B细胞比例与TAO患者CAS评分呈显著负相关(r=-0.7128,P<0.001)。结论:TAO患者CD19^(+)CD27^(+)IgD^(+)B细胞比例降低,与疾病活动度呈负相关。

B超监测梅花鹿性控精液腹腔镜输精试验

梅花鹿常规冻精与性控冻精解冻后进行品质鉴定,结合B超监测卵泡有选择的对60只同期发情处理的梅花鹿进行腹腔镜精准输精。数据分析显示,同期发情处理后梅花鹿卵巢B超监测结果与腹腔镜输精受胎率存在明显的正相关;X和Y型冻精产仔率为83.33%,而常规冻精产仔率为65.31%,X、Y型冻精与常规冻精组间差异显著(P<0.05);常规冻精组、X冻精组与Y冻精组所产后代性别比(公/母)分别为9:8、0:10和16:1,X、Y型冻精与常规冻精组间差异显著(P<0.05),X、Y型冻精与常规冻精所产后代出生及60 d时的体重差异不显著(P <0.05)。结果表明,结合B超监测有选择的进行梅花鹿性控精液腹腔镜人工输精,可显著提高受胎率,降低成本,技术可行。

免疫表型在鉴别急性早幼粒细胞白血病与HLA-DR阴性急性髓系白血病中的应用

目的探讨免疫表型在急性早幼粒细胞白血病(APL)和HLA-DR阴性的急性髓系白血病(AML)之间的鉴别诊断中的价值。方法回顾性研究2014年至2024年间收治的42例APL患者和28例初治及复发的HLA-DR阴性AML患者。通过流式细胞免疫表型分析,比较两组患者CD64、MPO、CD7、CD11c、CD9、CD123等抗原的阳性表达率。利用χ2检验进行统计学分析,并应用ROC曲线评估CD9和CD123的表达模式,以及CD64^(+)MPO^(+)CD7^(-)CD11c^(-)对APL的诊断价值。结果AML组患者的CD64、CD9、MPO阳性率显著低于APL组,而CD11c、CD7阳性率显著高于APL组,差异具有统计学意义(P<0.05)。CD64^(+)MPO^(+)CD7^(-)CD11c^(-)的综合抗原积分表达模式在鉴别诊断APL方面的ROC曲线下面积(AUC^(ROC))为0.859,敏感性为93.8%,特异性为75.0%,CD9/CD123的鉴别诊断APL的AUC^(ROC)为0.919,敏感性为83.3%,特异性为84.0%;而联合应用CD9/CD123和综合抗原积分的诊断模式,AUC^(ROC)为0.955,敏感性为83.3%,特异性为100%。结论CD9/CD123表达模式及CD64^(+)MPO^(+)CD7^(-)CD11c^(-)联合应用可作为鉴别诊断APL与HLA-DR阴性的AML的重要依据.

GZMB在乳腺癌和乳腺良性肿物患者中的表达及与T淋巴细胞水平的关系研究

目的:探讨颗粒酶B(granzyme B,GZMB)在乳腺癌和乳腺良性肿物患者中的表达及与T淋巴细胞水平的关系。方法:选取2022年1月至2023年8月南阳市第一人民医院收治的乳腺癌患者63例为观察组,根据肿瘤分期分为T_(1)N_(0)M_(0)组、T_(2)N_(0)M_(0)组、T_(2)N_(1)M_(0)组、T_(3)N_(0)M_(0)组及T_(3)N_(1)M_(0)组;选择同期治疗的乳腺良性肿物患者9例为对照组;采用流式细胞仪测定各组GZMB及T淋巴细胞水平;对乳腺癌患者GZMB与T淋巴细胞参数完成相关性分析。结果:观察组乳腺癌患者GZMB水平高于对照组(P<0.05);CD_(3)^(+)、CD_(4)^(+)及CD_(4)^(+)/CD_(8)^(+)水平低于对照组(P<0.05);乳腺癌不同分期下CD_(8)^(+)水平无统计差异(P>0.05);乳腺癌患者中T_(3)N_(1)M_(0)组GZMB水平高于其余4组(P<0.05),CD_(3)^(+)、CD_(4)^(+)及CD_(4)^(+)/CD_(8)^(+)水平低于其余4组(P<0.05);乳腺癌患者分期越高,GZMB水平越高,CD_(3)^(+)、CD_(4)^(+)及CD_(4)^(+)/CD_(8)^(+)水平越低;Pearson相关性结果表明,乳腺癌患者GZMB水平与CD_(3)^(+)、CD_(4)^(+)及CD_(4)^(+)/CD_(8)^(+)水平呈负相关(P<0.05)。结论:GZMB在乳腺癌患者中呈高表达,其表达水平在不同分期中存在差异,且与T淋巴细胞水平存在相关性,可指导临床诊疗。

次氯酸钠与酶联合对不锈钢表面细菌生物被膜的清除作用

定植于设备不同位置的细菌生物被膜是食品工业中应用清洁和消毒程序面临的最大挑战。该研究目的是通过人工培养生物被膜进行不同菌株生物被膜形成能力筛选,采用酶、次氯酸钠以及二者联合的方式对不锈钢试材上的成熟膜进行清除,通过平板计数、流式细胞仪等手段去探究不同处理方式的差异。结果表明,酶的抗生物被膜功效因生物被膜形成阶段的不同而体现出差异,在生物被膜形成期间,酶处理往往比对成熟生物被膜更有效,部分酶的参与会导致细菌产生更多的生物被膜基质。细菌计数显示,3种处理方式分别导致枯草芽孢杆菌(Bacillus subtilis)CICC 10900、金黄色葡萄球菌(Staphylococcus aureus)ATCC 6538细胞群减少0.64~3.08 lg CFU/cm^(2)、0.81~2.19 lg CFU/cm^(2);流式细胞仪结果表明,经处理后7.09%~12.04%、22.43%~23.45%、31.33%~37.78%的细胞受到损伤或死亡。使用酶制剂可以帮助次氯酸钠等二级制剂更有效地渗透生物被膜,增强对细菌细胞的清除功效。该研究结果可对食品工业在实施生物被膜清除程序时提供数据支撑和理论参考。

禽致病性大肠杆菌HlyE蛋白的免疫原性研究

为评估禽致病性大肠杆菌(APEC)溶血素HlyE蛋白的免疫原性,本研究对APEC溶血素HlyE蛋白进行原核表达和纯化,并对表达的重组蛋白进行SDS-PAGE和western blot分析,将纯化蛋白利用透析袋在4℃透析后,利用血琼脂平板对其溶血活性进行检测。SDS-PAGE和western blot结果显示,表达并纯化到分子量约为36 ku的重组HlyE蛋白(rHlyE),经ND-2000超微量核酸蛋白测定仪测定纯化后蛋白浓度为0.65 mg/mL;溶血活性检测结果显示,rHlyE具有溶血活性。以透析后的rHlyE作为抗原,按照50μg/只的剂量免疫小鼠,对照组于相同时间点注射等量PBS,共免疫3次间隔14 d,并于首免后不同时间采血,采用间接ELISA方法检测两组小鼠血清特异性抗体水平,并于三免后18 d以2 LD_(50)的APEC菌液攻毒小鼠,观察7 d内小鼠的死亡情况;于首免后28 d剖杀各组小鼠取其脾脏制备脾淋巴细胞,采用流式细胞术分别检测CD3^(+)、CD4^(+)、CD8^(+)T淋巴细胞亚型比率,对rHlyE的免疫原性进行评估。间接ELISA检测结果显示,该蛋白能够诱导机体产生体液免疫应答,分泌高表达量的IgG抗体,抗体水平于三免后15 d达到最高水平。rHlyE免疫攻毒保护试验结果显示,免疫组小鼠基本无明显临床症状,7 d内存活率达80%;而对照组小鼠表现明显临床症状,于攻毒后3 d内全部死亡。流式细胞术结果显示,与对照组相比,免疫组小鼠的CD3^(+)、CD4^(+)和CD8^(+)T淋巴细胞亚型比率均升高。上述结果表明,rHlyE在大肠杆菌BL21中为部分可溶性表达,将其免疫小鼠后可诱导小鼠产生较高水平的体液免疫应答,并且可对小鼠产生较好的免疫保护效果。本研究明确了APEC HlyE蛋白的免疫原性,为APEC免疫保护蛋白的筛选以及疫苗研发提供了借鉴与参考。

蝴蝶兰种质资源倍性鉴定及育性相关性研究

为探究蝴蝶兰种质资源倍性与育性的关系,本研究采用染色体计数法结合流式细胞术鉴定蝴蝶兰属33个品种(种或变种)的倍性水平,并通过完全双列杂交试验探究不同倍性蝴蝶兰种质资源的育性。染色体计数结果表明,二倍体和三倍体蝴蝶兰均有5个,均占15.15%;四倍体蝴蝶兰有10个,占30.30%;非整倍体蝴蝶兰有13个,占39.39%。蝴蝶兰不同品种之间染色体大小构成存在差异。流式细胞术检测结果表明,蝴蝶兰属25个品种(种或变种)的流式细胞术倍性估计结果与染色体计数结果相符。供试材料的流式直方图均呈现出3~4个峰,且循环值均大于0.1。杂交育性结果表明,作为亲本杂交的四倍体蝴蝶兰具有最高的坐果率和萌发率;在三倍体和非整倍体蝴蝶兰中,不同品种之间的育性差异较大,同一品种作为父本或母本杂交和自交后,其育性差异也较大。上述结果表明,大多数蝴蝶兰杂交品种的倍性为多倍体或非整倍体,蝴蝶兰的育性与其倍性水平和染色体大小构成相关;流式细胞术的倍性估计适用于蝴蝶兰种内以及含有小而均匀染色体的杂交品种;供试蝴蝶兰叶片均存在内多倍体化且不同倍性蝴蝶兰叶片的内多倍体化模式存在显著差异。本研究为蝴蝶兰种质资源鉴定、亲本选配、遗传改良以及多倍体育种提供了参考依据。

从Genomics,Proteomics到Cytomics,还是从Cytometry到Cytomics

Cytomics是一个特殊名词,她将cyto-与-omics连在一起,代表着一个崭新的领域。Cyto-主要来自分析细胞学(analyticalcytology或cytometry),而-omics主要来自蛋白质组学(proteomics)及其决定者基因组学(genomics)。经过三十余年的发展,分析细胞学可以通过分子探针将异质性的或混杂的细胞群体分为不同的细胞组份(cytomes),而基因组学和蛋白质组学的技术已能鉴定出某一物种的整套基因组和蛋白质组。因此,我们认为,细胞组学的任务是分离细胞组,并在基因组和蛋白质组层面确定其赖以存在和转变的分子基础。细胞组学的实施需要两大系列技术,一是细胞组分离技术,二是分离后再分析技术。这些技术的发展将使我们最终理解基因组并利用之,也必将拓展出更新的领域。