Detection of Embryo Sex Chromosome by Dual Color Fluorescent In-Situ Hybridization

In order to evaluate the effects of sex chromosomal mosaicism on the accuracy of single-cell gender diagnosis, sex chromosomes of 21 normal fertilized embryos were detected by dual color fluorescent in-situ hybridization (FISH). The results showed that 4 embryos had sex chromosomal mosaicism (19%) and the remaining 17 showed uniformly XX or XY signals in all blastomeres. In conclusion, identification of sex by dual color FISH analysis of a single cell was accurate and efficient , and sex chromosomal mosaicism would not affect preimplantation gender diagnosis.

Matrix Induced Synthesis of Eu^(3+) Doped Zn_3(PO_4)_2 and LaPO_4 Phosphors by in-Situ Composing Hybrid Precursors

Using rare earth and zinc coordination polymers with aromatic carboxylic acids as the precursors, composing with the polyethylene glycol (PEG) as the dispersing media, micro crystalline phosphors Zn_3(PO_4)_2∶Eu 3+ and LaPO_4∶Eu 3+ were synthesized by in-situ co-precipitation method. X-ray diffraction and scanning electronic micrograph were used to characterize the resultant samples, whose particle size are in the range of micrometer. The emission spectra of Zn_3(PO_4)_2∶Eu 3+ (λ_ ex=245 nm) and LaPO_4∶Eu 3+ (λ_ ex=390 nm) shows that the emission for Eu 3+ in Zn_3(PO_4)_2 is dominated by the 5D_0→7F_1 (592 nm) magnetic-dipole transition,While the dominant emission for Eu 3+ in LaPO_4 is the typical hypersensitive transition 5D_0→7F_2 (618 nm).

Green Approach for <i>In-Situ</i>Growth of CdS Nanorods in Low Band Gap Polymer Network for Hybrid Solar Cell Applications

In-situ growth of CdS nanorods (NRs) has been demonstrated via solvothermal, in a low band gap polymer, poly [[4,8-bis[(2-ethylhexyl)oxy] benzo [1,2-b:4,5-b’] dithiophene-2,6-diyl] [3-fluoro-2-[(2-ethylhexyl) carbonyl] thieno [3,4-b] thiophenediyl]] (PTB7). It is a high yielding, green approach as it removes use of volatile and hazardous chemicals such as pyridine as ligand which are conventionally used to synthesize precursors of CdS (NRs). Moreover the solvothermal process is a zero emission process being a close vessel synthesis and hence no material leaching into the atmosphere during the synthesis. The PTB7:CdS nanocomposite has been characterized by SEM, XRD, FTIR, UV-visible spectroscopy techniques. The photoluminescence (PL) spectroscopy study of PTB7 with CdS NRs has shown significant PL quenching by the incorporation of CdS NRs in PTB7;this shows that CdS NRs are efficient electron acceptors with the PTB7. The PTB7:CdS is used as active layer in the fabrication of hybrid solar cells (HSC) as donor-acceptor combination in the bulk heterojunction (BHJ) geometry. The HSCs fabricated using this active layer without any additional supporting fullerene based electron acceptor has given power conversion efficiency of above 1%.

Performance-control-orientated hybrid metal additive manufacturing technologies:state of the art,challenges,and future trends

Metal additive manufacturing(AM)technologies have made significant progress in the basic theoretical field since their invention in the 1970s.However,performance instability during continuous processing,such as thermal history,residual stress accumulation,and columnar grain epitaxial growth,consistently hinders their broad application in standardized industrial production.To overcome these challenges,performance-control-oriented hybrid AM(HAM)technologies have been introduced.These technologies,by leveraging external auxiliary processes,aim to regulate microstructural evolution and mechanical properties during metal AM.This paper provides a systematic and detailed review of performance-control-oriented HAM technology,which is categorized into two main groups:energy field-assisted AM(EFed AM,e.g.ultrasonic,electromagnetic,and heat)technologies and interlayer plastic deformation-assisted AM(IPDed AM,e.g.laser shock peening,rolling,ultrasonic peening,and friction stir process)technologies.This review covers the influence of external energy fields on the melting,flow,and solidification behavior of materials,and the regulatory effects of interlayer plastic deformation on grain refinement,nucleation,and recrystallization.Furthermore,the role of performance-control-oriented HAM technologies in managing residual stress conversion,metallurgical defect closure,mechanical property improvement,and anisotropy regulation is thoroughly reviewed and discussed.The review concludes with an analysis of future development trends in EFed AM and IPDed AM technologies.

应用组织芯片和ISH技术检测人体多种肿瘤中hTER mRNA和hTERT mRNA的表达

目的 探讨组织芯片技术在原位杂交中的应用及hTER和hTERT在人体多种常见肿瘤中的表达。方法 利用自行研制的专用器具制作组织芯片 ,采用原位杂交技术 (ISH)检测 2 3 0例人体肿瘤和正常组织中hTER和hTERT基因的表达。结果 ①组织芯片中组织样本的组织学结构完整 ,满足病理学实验要求 ;采用组织芯片技术消耗的实验材料、试剂为普通方法的1 /2 3。②在人体正常组织和良性肿瘤、恶性肿瘤组织中 ,hTER和hTERT的表达与肿瘤的性质明显相关 (P <0 .0 0 5 )。结论 组织芯片技术提供了一种快速、有效的组织学方法 ,可广泛应用于原位杂交实验 ,并有可能在肿瘤生物学研究中发挥重要的作用。同时 ,hTER和hTERT表达上调有可能作为人体肿瘤诊断与鉴别诊断的重要组织学指标。

荧光原位杂交技术(FISH)在鱼类遗传学研究中的应用及前景

荧光原位杂交技术（ｆｌｕｏｒｅｓｃｅｎｃｅｉｎｓｉｔｕｈｙｂｒｉｄｉｚａｔｉｏｎ，ＦＩＳＨ）是一种在分子水平上进行细胞遗传学研究的得力工具。本文综述了近年来ＦＩＳＨ技术在鱼类的基因定位、性别鉴定、染色体变异及种间杂交等研究中的应用；并将此技术在鱼类的基因定位、早期性别鉴定、染色体重排与进化、染色体鉴别与分类及杂交鉴定等方面的应用前景作一展望。

In-situ magnetic field enhanced performances in ferromagnetic FeCo_(2)O_(4) nanofibers-based rechargeable Zinc-air batteries

Field-assisted electrocatalytic reactions are demonstrated to be sufficient strategies in enhancing the electrocatalyst activities for oxygen evolution reaction(OER).Here,we report the in-situ magnetic field enhanced electrocatalytic activity in ferromagnetic FeCo_(2)O_(4)nanofibers.Our results demonstrate that the overpotential of FeCo_(2)O_(4)nanofibers at 10 mA cm^(-2)shows a left-shift of 40 mV for the OER by applying an external magnetic field,and no obvious change has been observed in the non-ferromagnetic-order Co3O4nanofibers.Calculation results indicate that there are more overlaps between the density of states for Co3d and O 2p by applying an external magnetic field.Accordingly,the spin hybridization of 3d-2p and the kinetics of spin charge transfer are optimized in ferromagnetic FeCo_(2)O_(4),which can promote the adsorption of oxygen-intermediates and electron transfer,significantly improving its electrocatalytic efficiency.What’s more,the maximum power density of the FeCo_(2)O_(4)nanofibers based Zn-air battery(ZAB)increases from 97.3 mW cm^(-2)to 108.2 mW cm^(-2)by applying an external magnetic field,providing a new idea for the application of magnetic cathode electrocatalysts in ZABs.

Identification of embryonic chromosomal abnormality using FISH-based preimplantaion genetic diagnosis

Objective: Embryonic chromosomal abnormality is one of the main reasons for in vitro fertilization (IVF) failure. This study aimed at evaluating the value of Fluorescence in-situ Hybridization (FISH)-based Preimplantation Genetic Diagnosis (PGD) in screening for embryonic chromosomal abnormality to increase the successful rate of IVF. Method: Ten couples, four with high risk of chromosomal abnormality and six infertile couples, underwent FISH-based PGD during IVF procedure. At day 3, one or two blastomeres were aspirated from each embryo. Biopsied blastomeres were examined using FISH analysis to screen out embryos with chromosomal abnormalities. At day 4, embryos without detectable chromosomal abnormality were transferred to the mother bodies as in regular IVF. Results: Among 54 embryos screened using FISH-based PGD, 30 embryos were detected to have chromosomal abnormalities. The 24 healthy embryos were implanted, resulting in four clinical pregnancies, two of which led to successful normal birth of two healthy babies; one to ongoing pregnancy during the writing of this article; and one to ectopic pregnancy. Conclusion: FISH-based PGD is an effective method for detecting embryonic chromosomal abnormality, which is one of the common causes of spontaneous miscarriages and chromosomally unbalanced offsprings.

Characterization of DYRK2(dual-specificity tyrosine-phosphorylation-regulated kinase 2) from Zebrafish(Dario rerio)

Proteins of the DYRK(dual-specificity tyrosine-phosphorylation-regulated kinase) family are characterized by the presence of a conserved kinase domain and N-terminal DH box.DYRK2 is involved in regulating key developmental and cellular processes,such as neurogenesis,cell proliferation,cytokinesis,and cellular differentiation.Herein,we report that the ortholog of DYRK2 found in zebrafish shares about 70% identity with that of human,mouse,and chick.RT-PCR showed that DYRK2 is expressed maternally and zygotically.In-situ hybridization results show that DYRK2 is expressed in somite cells that will develop into muscles.Our results provide preliminary evidence for investigating the in-vivo function of DYRK2 in zebrafish muscle development.

Characterization of progenies of Triticum aestivum- Psathyrostachys juncea derivatives by using genomic in-situ hybridization

Using genomic in-situ hybridization (GISH) technique, 7 translocation-addition lines, 6 transloca-tion and translocation-addition lines, 2 ditelosomic addition lines and 1 translocation line were identified from Triticum aestivum L.-Psathyrostachys juncea (Fisch. ) Nevski intergeneric hybrids, of which translocation-addition and translocation and translocation-addition lines were not found in other reports. No substitutions and disomic additions were detected in the hybrids and breakages occurred in all P. juncea chromosomes studied. Results have shown that the improved GISH technique is a rapid and economical method for use in this field.

Manipulating the ionic nanophase of Nafion by in-situ precise hybridization with polymer quantum dot towards highly enhanced fuel cell performances

Hierarchical-structure materials hold great promise for numerous applied domains such as fuel cell,sensor,and optic.However,the developments are significantly impeded by the lack of efficient strategy permitting precise and efficient decoration of specific confined space.Here,an in-situ precise hybridization strategy is proposed to efficiently manipulate the nanostructure of membrane nanochannels.Typically,Nafion ionic nanochannels are impregnated with precursors via heat swelling,followed by microwave-assisted condensation to form polymer quantum dot network.The formation of polymer quantum dot network significantly improves the stability and functionality of ionic nanophase(i.e.,ionic nanochannel).This helps hybrid membrane achieving enhanced proton conduction and methanol barrier properties,resulting in over ten times increase in proton/methanol selectivity.These then impart prominent device performances for both hydrogen and methanol fuel cells with the elevation of~100%.Importantly,such function manipulation of ionic nanochannels is achieved with fully maintaining function of backbone nanophase.Besides,the regulation of physical topology and chemical environment of ionic nanochannel also brings optimization of gas and ion separation properties.This facile and versatile strategy may open up a new avenue for decorating confined space of many hierarchical-structure materials.

HPV16/18型表达和细胞病理学检查与宫颈癌及癌前病变的相关性研究

目的探讨细胞病理学和原位杂交技术(ISH)检测宫颈组织高危型HPV(16/18型)感染与宫颈癌及癌前病变发生的相关性。方法运用细胞病理学和原位杂交技术检测218例宫颈组织HPV16/18型的表达水平及病变情况,判定HPV16/18型感染对宫颈癌及癌前病变的筛查价值。结果218例低度鳞状上皮内瘤变(LSIL)以上患者HPV16/18检出率分别为:鳞状上皮轻度非典型增生(CIN)Ⅰ16.67%(15/90例)、鳞状上皮中度非典型增生(CIN)Ⅱ48.43%(31/64例)、鳞状上皮重度非典型增生(CIN)Ⅲ91.11%(41/45例),鳞状细胞癌(SCC)84.21%(16/19例)。CINⅢ和SCC组与CINⅠ、Ⅱ组间比较有非常显著性差异(P<0.01);CINⅢ与SCC组间比较差异无显著性(P>0.01)。结论原位杂交技术检测HPV16/18型对诊断宫颈癌及癌前病变具有敏感度、特异度高的特点;若原位杂交技术与细胞病理学诊断协同检测,可更早发现宫颈癌和癌前病变,并对宫颈病变干预和治疗具有重要的指导意义。

小鼠Y染色体RNA探针的标记和应用

目的 标记小鼠Y染色体RNA探针 ,检验该探针在检测外周血涂片和脑组织切片Y染色体阳性细胞的效率。 方法 以小鼠Y染色体重复序列pY35 3/BcDNA为模板标记RNA探针。以雌性小鼠为对照 ,用原位杂交法检测小鼠外周血涂片Y染色体阳性细胞 ;用荧光原位杂交法检测雄性小鼠脑组织切片Y染色体阳性细胞。 结果 Y染色体原位杂交法在外周血涂片Y染色体阳性的检测灵敏度和特异度均为 1 0 0 %,在脑切片检测Y染色体阳性细胞的灵敏度和特异度分别为 85 6 7%和 1 0 0 %。

H9N2亚型禽流感病毒原位杂交检测方法的建立

根据已知禽流感病毒(Avianinfluenzavirus,AIV)核蛋白(NP)基因序列,选取其中保守区域设计出1对引物,以RT-PCR方法扩增出543bp的NP基因片段,采用地高辛标记制备探针。MDCK细胞人工感染H9N2亚型禽流感病毒后,不同时间取样制备细胞涂片,进行原位杂交,研究了禽流感病毒对MDCK细胞的感染,探讨了原位杂交的条件及其应用于检测AIV感染的可行性。结果显示,感染24h后,原位杂交可检测出细胞内的病毒RNA,并且杂交信号的强度与感染时间的长短有关。研究表明,原位杂交检测禽流感病毒,不但能很好地表现病毒和细胞的位置关系,而且具有较好的特异性和灵敏性,为禽流感病毒的组织原位杂交检测奠定了实验基础。

原位杂交技术检测与中药干预治疗宫颈高危型HPV感染的对比研究

目的:探讨原位杂交技术(ISH)检测宫颈高危型HPV(16/18型)感染与宫颈癌及癌前病变发生的相关性,分析中药干预和治疗HPV16/18型感染的临床价值。方法:运用原位杂交技术检测218例宫颈组织HPV16/18型的表达水平,判定HPV16/18型感染对宫颈癌及癌前病变的诊断价值;应用中药清热解毒扶正方干预和治疗HPV感染患者,并观察用药前后变化。结果:218例患者HPV16/18检出率分别为:CINⅠ16.67%(15/90例)、CINⅡ48.43%(31/64例)、CINⅢ91.11%、(41/45例)、SCC84.21%(16/19例);实验组52例,对HPV感染者用清热解毒、活血化瘀方剂局部熏蒸外洗和益气温阳、清热扶正方剂水煎口服,显效率34.62%(18/52例);对照组51例,显效率11.54%(6/51例),两组间比较有显著性差异(P<0.01)。结论:原位杂交技术检测HPV16/18型对诊断宫颈癌及癌前病变具有早期、敏感、特异的特点;中药清热解毒扶正方治疗HPV感染者疗效显著,若二者协同使用对宫颈癌的早期筛查和干预治疗,具有较高的临床使用价值和推广前景。

TGF-β1与Smad2基因在肝细胞肝癌中的表达及意义

目的:通过检测TGF-β1和Smad2基因在肝细胞肝癌(HCC)中的表达,初步探讨TGF-β/Smad信号通路中激活型Smad(R-Smad)与HCC的发生、发展之间的可能关系。方法:采用免疫组化ABC法及原位杂交法(ISH)检测41例HCC组织及癌旁组织TGF-β1、Smad2蛋白、Smad2 mRNA的表达,5例外伤性肝破裂手术切除标本作正常对照,比较正常对照、癌组织、癌旁组织中上述三者表达的差异,并进行图像分析和统计学分析。结果:正常对照组TGF-β1、Smad2蛋白、Smad2 mRNA均呈阴性表达。TGF-β1在HCC组织中阳性表达率为75.6%(31/41),在癌旁组织中为95.1%(39/41),二者比较差异显著(P<0.05);Smad2蛋白在HCC组织中阳性表达率为43.9%(18/41),在癌旁组织中为56.1%(23/41),二者比较无显著差异(P>0.05);Smad2 mRNA在HCC组织中阳性表达率为61.0%(25/41),在癌旁组织中为82.9%(34/41),二者比较有显著差异(P<0.05)。HCC组织中TGF-β1、Smad2蛋白、Smad2 mRNA的表达与病理分级均无显著相关性(P>0.05)。癌旁组织中Smad2蛋白的阳性面积百分比、吸光值及Smad2 mRNA的阳性面积百分比均显著高于HCC组织(P<0.05)。结论:TGF-β1和Smad2基因过表达可能在HCC的发生、发展中发挥作用。

一氧化氮合酶mRNA和蛋白在骨巨细胞瘤中的表达及意义

背景与目的:近年的研究发现,一氧化氮在恶性肿瘤的发生和发展中起着重要作用,且在正常骨细胞中检测到一氧化氮合酶(nitric oxide synthase,NOS)的活性,但目前未见有关骨巨细胞瘤(giant cell tumors of bone,GCT)NOS的活性及其与GCT病理分级和肿瘤复发关系的报道。本研究探讨GCT组织中NOS mRNA和蛋白的表达,及其与GCT病理分级、肿瘤复发的关系。方法;用NOS的cDNA探针检测14例冻存GCT组织中原生型一氧化氮合酶(constitutive NOS,cNOS)、诱导型一氧化氮合酶(inducible NOS,iNOS)mRNA的表达,用抗NOS1、NOS2、NOS3的多克隆抗体检测42例GCT石蜡包埋组织中NOS1、NOS2、NOS3蛋白的表达。结果:(1)在14例冻存的GCT组织中多核巨细胞(multinuclear giant cell,MGC)cNOS和iNOS mRNA的阳性率分别为78.6％和57.1％,单核细胞(mononuclaer cell,MC)的阳性率均为 35.7％;(2)病理分级Ⅱ、Ⅲ级MGC的cNOS mRNA阳性率明显高于Ⅰ级MGC(P=0.008);(3)在42例GCT的石蜡包埋组织中,MGC中NOS1、NOS2、NOS3蛋白的阳性率分别为85.7％、59.5％和31.0％,MC的阳性率分别为54.8％、28.6％和14.3％;(4)病理分级Ⅱ、Ⅲ级CCT组织中MC的NOS1阳性率明显高于Ⅰ级组(P=0.006);(5)复发组GCT组织MC的NOS1阳性率明显高于无复发组(P=0.018)。

应用RNA原位杂交技术研究马尾松针叶中银松素合酶的基因表达

以马尾松(Pinus massonianaLamb.)银松素合酶(PS)基因为模板,体外转录合成带地高辛标记的反义RNA全长及特异探针,并用这2个探针与马尾松针叶组织石蜡切片进行原位杂交,研究该基因在马尾松针叶中的表达特性以及外界诱导因素对该基因表达的调节特性。结果表明,该基因的转录表达主要集中在针叶的韧皮部;紫外线照射和酵母提取液处理均可使该基因的转录水平明显增加。

川芎嗪对大鼠CIR顶叶皮质神经元超微结构及Caspase-9mRNA表达的影响

目的探讨川芎嗪治疗脑缺血再灌注损伤的机制。方法采用Bannisters方法复制大鼠脑缺血再灌注动物模型,利用川芎嗪治疗;在透射电镜下观察顶叶皮质神经元超微结构变化;采用核酸原位杂交技术观察治疗前后额顶叶皮质神经元半胱天冬蛋白酶基因(caspase-9mRNA)的表达变化。结果模型组顶叶皮质神经元水肿,质膜、线粒体膜破裂;溶酶体、内质网和高尔基复合体破坏;caspase-9mRNA数密度增高,着色加深(灰度值下降)。川芎嗪治疗组顶叶皮质神经元质膜、核膜、内质网、线粒体和溶酶体损伤明显减轻;caspase-9mRNA阳性神经元减少,着色变浅。结论川芎嗪下调脑缺血再灌注顶叶皮质神经元中凋亡相关基因caspase-9mRNA的表达并具有神经保护作用。

MMP-9及TIMP-1mRNA在甲状腺肿瘤中的表达及临床意义

目的:研究MMP-9及TIMP-1mRNA在不同类型甲状腺肿瘤组织及不同临床病理特征甲状腺乳头状癌(PTC)中的表达,探讨其在甲状腺癌浸润、转移中的作用。方法:采用石蜡切片、原位杂交法检测了20例甲状腺乳头状癌,20例甲状腺腺瘤,20例结节性甲状腺肿和20例癌旁正常甲状腺组织中MMP-9、TIMP-1mRNA的表达。结果:与癌旁正常甲状腺比较各类型肿瘤组织中MMP-9mRNA杂交信号均呈增强的趋势(P<0.01),与甲状腺癌比较,结节性甲状腺肿和甲状腺腺瘤组织中MMP-9mRNA杂交信号均呈减弱的趋势(P<0.05);同时,MMP-9mRNA在甲状腺癌中的表达与肿瘤的大小、临床分期、淋巴结转移、预后指数呈正性调节关系(MMP-9mRNA表达高者恶性度高、预后差,P<0.05或P<0.01)。TIMP-1mRNA在癌旁正常甲状腺组织、良性甲状腺结节及甲状腺癌中均有不同程度的表达,与癌旁正常甲状腺组织比较各类型肿瘤组织中TIMP-1mRNA表达均呈增加的趋势,但只有甲状腺癌组明显高于正常甲状腺组,差异有显著(P<0.05);与甲状腺癌比较,结节性甲状腺肿和甲状腺腺瘤组织中TIMP-1mRNA表达没有明显改变(P>0.05);同时TIMP-1mRNA在甲状腺癌中的表达与临床病理特征呈负性调节关系(TIMP-1mRNA表达低者恶性度高、预后差,P<0.05或P<0.01)。结论:MMP-9及TIMP-1mRNA的表达与甲状腺癌的浸润、转移密切相关,MMP-9及TIMP-1mRNA表达有望成为判断甲状腺乳头状癌浸润、转移和预后较可靠的指标。