This paper presents a feasible method for rapid detection of the interphase nuclei of uncultured amniocytes for chromosomes 18 by using our modified in situ labeling (PRINS) technique.A total of 262 independent,uncult...This paper presents a feasible method for rapid detection of the interphase nuclei of uncultured amniocytes for chromosomes 18 by using our modified in situ labeling (PRINS) technique.A total of 262 independent,uncultured amniotic fluid samples were analysed in a blind fashion before the karyotype was available.In addition,62 samples were examined by fluorescence in situ hybridization (FISH) for comparison.In more than 95% of the samples PRINS reactions with primer 18cen were successfully induced.Two samples were properly identified and correctly scored as trisomic 18.PRINS reaction could be performed automatically in less than one hour with a propgrammable thernocycler.Our studies showed that the PRINS technique is simple.rapid and cost-effective.It is as sensitive and specific as FISH;can enhance the eccuracy of standard cytogenetic analysis;and allows identification of chromosomes 18 aneuploidies in uncultrued amniocytes in significantly less time.展开更多
Subtelomeric rearrangements contribute to idiopathic mental retardation （MR）, but most children with idiopathic MR do not show any chromosome abnormalities with standard cytogenetic analysis. The primed in situ labe...Subtelomeric rearrangements contribute to idiopathic mental retardation （MR）, but most children with idiopathic MR do not show any chromosome abnormalities with standard cytogenetic analysis. The primed in situ labeling （PRINS） technique, using an oligonucleotide primer complementary to the telemetric repeat sequences （TTAGGG）, can identify chromosome telomeric abnormality （deletion） in idiopathic MR children. In this study, seventy children with idiopathic MR were enrolled and subjected to PR1NS. The results showed normal karyotype in all the children, subtelomeric rearrangements （lq del and 4q del） in 2 cases, which was confirmed by fluorescence in situ hybridization （FISH）. It was concluded that PRINS is effective for the detection of subtelomeric rearrangements and may become a routine technique for cytogenetical abnormality screening.展开更多
Objective To study the relationship between apoptosis and hypertrophy of the left ventricie (LV) before and after its regression with antihypertensive treatment. Methods Forty 12- week old spontaneously. hypertensive ...Objective To study the relationship between apoptosis and hypertrophy of the left ventricie (LV) before and after its regression with antihypertensive treatment. Methods Forty 12- week old spontaneously. hypertensive rats(SHR were randomly divided into 2 groups, one given enalapril (SHR - d grouP) and the other without treatment (SHR group). Twenty normotensive Wistar Kyoto rats of the same age served as controls (WKY group). The changes of body weight and blood pressure were observed for 3 months. The rats were killed at the 24th week of age. The weight of LV, the body weight (B W), and the ratio of LV/BW were compared between the groups. Then by in situ specific labelling of nuclear DNA fragmentation, the apoptosis in the wall of the left ventricle was compared between them. RcsuIts The index of apoptosis was SHR - d>SHR> WKY The dofference was significant. ConcIusion The hypertrophy of the LV and its regression is very’likely related with the imbalance between the proloferation and death of cells of the muscle in the LV.展开更多
The receptor for autocrine motility factor (AMFR) is known to be involved in the process of AMF-mediated cell migration and metastasis. This paper describes the procedures of non-radioactive in situ hybridization (ISH...The receptor for autocrine motility factor (AMFR) is known to be involved in the process of AMF-mediated cell migration and metastasis. This paper describes the procedures of non-radioactive in situ hybridization (ISH) detection of AMFR mRNA in both paraffin-embedded surgical sections and cultured cells using either biotinylated oligonucleotide probes or digoxigenin-labeled RNA probes. The results showed that the AMFR mRNA was expressed at an enhanced level in hyperplaJstic and malignant tissues of breast and prostate cancer patient surgical specimens, indicating that the elevated AMFR expression was associated with the tissue malignancy Moreover, AMFR mRNA was detected in both normal and earcinoma cells when cultured at a subconfluent density. However, AMFR expression was inhibited in confluent normal (3T3-A31 murine fibroblast and FHs738BL human bladder) cells while it continued to express in carcinoma (J82 human bladder)and metastatic (3T3-M murine fibroblast) cells irrespective of cell density This suggested a cell-cell contact downregulation of AMFR mRNA expression in normal but not in cancer cells. The ISH data obtained in this study are closely consistent with the AMFR protein expression pattern previously reported, implying that the differential expression of AMFR gene may be regulated and controlled at the transcriptional level.展开更多
Cell-derived microvesicles(MVs) are secreted from almost all kinds of mammalian cells into the extracellular space, and play crucial roles in intercellular communication and transporting biomolecules between cells. Ho...Cell-derived microvesicles(MVs) are secreted from almost all kinds of mammalian cells into the extracellular space, and play crucial roles in intercellular communication and transporting biomolecules between cells. However, there is a great challenge for visualizing and monitoring of MVs’ bio-behaviors due to the limitations of existing labeling methods. Herein, we report the first paradigm of designer cell-self-implemented labeling of MVs secreted from living mammalian MCF-7 cells in situ using the intracellular-synthesized fluorescent quantum dots(QDs) during the formation of MVs. By elaborately coupling intracellular biochemical reactions and metabolism pathways, the MCF-7 cells can be illuminated brightly by intracellular-biosynthesized fluorescent CdSe QDs. Simultaneously, intracellular-synthesized QDs can be in situ encapsulated by the secreted MVs budding from the plasma membrane of the fluorescing cells to label the MVs with an efficiency of up to 89.9%. The whole labeling process skillfully combines designer precise cell-tuned intricate synthesis of CdSe QDs with mild in-situ labeling via cell-selfimplementation just after feeding the cell with suitable chemicals, which is structure-or function-nondestructive and much more straightforward and milder than those by chemical conjugation or indirect encapsulation with conventional fluorogenic labels.展开更多
Analyzing undiluted whole human blood is a challenge due to its complex composition of hematopoietic cellular populations,nucleic acids,metabolites,and proteins.We present a novel multi-functional microfluidic acousti...Analyzing undiluted whole human blood is a challenge due to its complex composition of hematopoietic cellular populations,nucleic acids,metabolites,and proteins.We present a novel multi-functional microfluidic acoustic streaming platform that enables sorting,enrichment and in situ identification of cellular subsets from whole blood.This single device platform,based on lateral cavity acoustic transducers(LCAT),enables(1)the sorting of undiluted donor whole blood into its cellular subsets(platelets,RBCs,and WBCs),(2)the enrichment and retrieval of breast cancer cells(MCF-7)spiked in donor whole blood at rare cell relevant concentrations(10 mL^(−1)),and(3)on-chip immunofluorescent labeling for the detection of specific target cellular populations by their known marker expression patterns.Our approach thus demonstrates a compact system that integrates upstream sample processing with downstream separation/enrichment,to carry out multi-parametric cell analysis for blood-based diagnosis and liquid biopsy blood sampling.展开更多
文摘This paper presents a feasible method for rapid detection of the interphase nuclei of uncultured amniocytes for chromosomes 18 by using our modified in situ labeling (PRINS) technique.A total of 262 independent,uncultured amniotic fluid samples were analysed in a blind fashion before the karyotype was available.In addition,62 samples were examined by fluorescence in situ hybridization (FISH) for comparison.In more than 95% of the samples PRINS reactions with primer 18cen were successfully induced.Two samples were properly identified and correctly scored as trisomic 18.PRINS reaction could be performed automatically in less than one hour with a propgrammable thernocycler.Our studies showed that the PRINS technique is simple.rapid and cost-effective.It is as sensitive and specific as FISH;can enhance the eccuracy of standard cytogenetic analysis;and allows identification of chromosomes 18 aneuploidies in uncultrued amniocytes in significantly less time.
文摘Subtelomeric rearrangements contribute to idiopathic mental retardation （MR）, but most children with idiopathic MR do not show any chromosome abnormalities with standard cytogenetic analysis. The primed in situ labeling （PRINS） technique, using an oligonucleotide primer complementary to the telemetric repeat sequences （TTAGGG）, can identify chromosome telomeric abnormality （deletion） in idiopathic MR children. In this study, seventy children with idiopathic MR were enrolled and subjected to PR1NS. The results showed normal karyotype in all the children, subtelomeric rearrangements （lq del and 4q del） in 2 cases, which was confirmed by fluorescence in situ hybridization （FISH）. It was concluded that PRINS is effective for the detection of subtelomeric rearrangements and may become a routine technique for cytogenetical abnormality screening.
文摘Objective To study the relationship between apoptosis and hypertrophy of the left ventricie (LV) before and after its regression with antihypertensive treatment. Methods Forty 12- week old spontaneously. hypertensive rats(SHR were randomly divided into 2 groups, one given enalapril (SHR - d grouP) and the other without treatment (SHR group). Twenty normotensive Wistar Kyoto rats of the same age served as controls (WKY group). The changes of body weight and blood pressure were observed for 3 months. The rats were killed at the 24th week of age. The weight of LV, the body weight (B W), and the ratio of LV/BW were compared between the groups. Then by in situ specific labelling of nuclear DNA fragmentation, the apoptosis in the wall of the left ventricle was compared between them. RcsuIts The index of apoptosis was SHR - d>SHR> WKY The dofference was significant. ConcIusion The hypertrophy of the LV and its regression is very’likely related with the imbalance between the proloferation and death of cells of the muscle in the LV.
文摘The receptor for autocrine motility factor (AMFR) is known to be involved in the process of AMF-mediated cell migration and metastasis. This paper describes the procedures of non-radioactive in situ hybridization (ISH) detection of AMFR mRNA in both paraffin-embedded surgical sections and cultured cells using either biotinylated oligonucleotide probes or digoxigenin-labeled RNA probes. The results showed that the AMFR mRNA was expressed at an enhanced level in hyperplaJstic and malignant tissues of breast and prostate cancer patient surgical specimens, indicating that the elevated AMFR expression was associated with the tissue malignancy Moreover, AMFR mRNA was detected in both normal and earcinoma cells when cultured at a subconfluent density. However, AMFR expression was inhibited in confluent normal (3T3-A31 murine fibroblast and FHs738BL human bladder) cells while it continued to express in carcinoma (J82 human bladder)and metastatic (3T3-M murine fibroblast) cells irrespective of cell density This suggested a cell-cell contact downregulation of AMFR mRNA expression in normal but not in cancer cells. The ISH data obtained in this study are closely consistent with the AMFR protein expression pattern previously reported, implying that the differential expression of AMFR gene may be regulated and controlled at the transcriptional level.
基金the National Natural Science Foundation of China(21535005,91859123,21705111).
文摘Cell-derived microvesicles(MVs) are secreted from almost all kinds of mammalian cells into the extracellular space, and play crucial roles in intercellular communication and transporting biomolecules between cells. However, there is a great challenge for visualizing and monitoring of MVs’ bio-behaviors due to the limitations of existing labeling methods. Herein, we report the first paradigm of designer cell-self-implemented labeling of MVs secreted from living mammalian MCF-7 cells in situ using the intracellular-synthesized fluorescent quantum dots(QDs) during the formation of MVs. By elaborately coupling intracellular biochemical reactions and metabolism pathways, the MCF-7 cells can be illuminated brightly by intracellular-biosynthesized fluorescent CdSe QDs. Simultaneously, intracellular-synthesized QDs can be in situ encapsulated by the secreted MVs budding from the plasma membrane of the fluorescing cells to label the MVs with an efficiency of up to 89.9%. The whole labeling process skillfully combines designer precise cell-tuned intricate synthesis of CdSe QDs with mild in-situ labeling via cell-selfimplementation just after feeding the cell with suitable chemicals, which is structure-or function-nondestructive and much more straightforward and milder than those by chemical conjugation or indirect encapsulation with conventional fluorogenic labels.
基金This work was supported by the NSF Center for Advanced Design and Manufacturing of Integrated Microfluidics(CADMIM)(Award Nos.IIP-1362165 and IIP-1362048)Schlumberger Faculty for the Future Award(Award No.SF-202940)the National Cancer Institute of the National Institutes of Health under award no.P30CA062203.
文摘Analyzing undiluted whole human blood is a challenge due to its complex composition of hematopoietic cellular populations,nucleic acids,metabolites,and proteins.We present a novel multi-functional microfluidic acoustic streaming platform that enables sorting,enrichment and in situ identification of cellular subsets from whole blood.This single device platform,based on lateral cavity acoustic transducers(LCAT),enables(1)the sorting of undiluted donor whole blood into its cellular subsets(platelets,RBCs,and WBCs),(2)the enrichment and retrieval of breast cancer cells(MCF-7)spiked in donor whole blood at rare cell relevant concentrations(10 mL^(−1)),and(3)on-chip immunofluorescent labeling for the detection of specific target cellular populations by their known marker expression patterns.Our approach thus demonstrates a compact system that integrates upstream sample processing with downstream separation/enrichment,to carry out multi-parametric cell analysis for blood-based diagnosis and liquid biopsy blood sampling.
