Highly Sensitive Detection of PIK3CA Mutations by Looping-Out Probes-Based Melting Curve Analysis.

PIK3CA mutations have important therapeutic and prognostic implications in various cancer types. However, highly sensitive detection of PIK3CA hotspot mutations in heterogeneous tumor samples remains a challenge in clinical settings. To establish a rapid PCR assay for highly sensitive detection of multiple PIK3CA hotspot mutations. We described a novel melting curve analysis-based assay using looping-out probes that can enrich target mutations in the background of excess wild-type and concurrently reveal the presence of mutations. The analytical and clinical performance of the assay were evaluated. The developed assay could detect 10 PIK3CA hotspot mutations at a mutant allele fraction of 0.05-0.5% within 2 h in a single step. Analysis of 82 breast cancer tissue samples revealed 43 samples with PIK3CA mutations, 28 of which were confirmed by Sanger sequencing. Further testing of 175 colorectal cancer tissue samples showed that 24 samples contained PIK3CA mutations and 19 samples were confirmed by Sanger sequencing. Droplet digital PCR supported that all mutation-containing samples undetected by sequencing contained mutations with a low allele fraction. The rapidity, ease of use, high sensitivity and accuracy make the new assay a potential screening tool for PIK3CA mutations in clinical laboratories.

Accurate Detection of Multiple Tumor Mutations in Formalin-Fixed Paraffin-Embedded Tissues by Coupling Sequence Artifacts Elimination and Mutation Enrichment With MeltArray.

Formalin-fixed paraffin-embedded (FFPE) tissues are the primary source of DNA for companion diagnostics (CDx) of cancers. Degradation of FFPE tissue DNA and inherent tumor heterogeneity constitute serious challenges in current CDx assays. To address these limitations, we introduced sequence artifact elimination and mutation enrichment to MeltArray, a highly multiplexed PCR approach, to establish an integrated protocol that provides accuracy, ease of use, and rapidness. Using PIK3CA mutations as a model, we established a MeltArray protocol that could eliminate sequence artifacts completely and enrich mutations from 23.5- to 59.4-fold via a single-reaction pretreatment step comprising uracil-DNA-glycosylase excision and PCR clamping. The entire protocol could identify 13 PIK3CA hotspot mutations of 0.05% to 0.5% mutant allele fractions within 5 hours. Evaluation of 106 breast cancer and 40 matched normal FFPE tissue samples showed that all 47 PIK3CA mutant samples were from the cancer tissue, and no false-positive results were detected in the normal samples. Further evaluation of 105 colorectal and 40 matched normal FFPE tissue samples revealed that 11 PIK3CA mutants were solely from the cancer sample. The detection results of our protocol were consistent with those of the droplet digital PCR assays that underwent sequence artifact elimination. Of the 60 colorectal samples with next-generation sequencing results, the MeltArray protocol detected 2 additional mutant samples with low mutant allele fractions. We conclude that the new protocol provides an improved alternative to current CDx assays for detecting tumor mutations in FFPE tissue DNA.

Landscape Pattern and Ecological Risk Assessment in Guilin Based on Land Use Change.

The land use and ecological risk patterns in Guilin, which is the only innovation demonstration zone under the National Sustainable Development Agenda in China with a focus on the sustainable use of natural resources, have changed significantly as a result of the combined impact of climate change and human activities, thus presenting challenges to the sustainable development of the local area. This research employs an ecological risk assessment model and spatial analysis techniques in order to analyze the spatial correlation between land use and ecological risk, and to evaluate the spatial and temporal evolution characteristics of ecological risk at the overall and county scales in Guilin. The results reveal the following: (1) A total of 1848.6 km2 land types in Guilin have changed from 2000 to 2020, and construction land has gradually expanded from the central urban area to the suburbs with increasing internal stability each year. (2) The ecological risk level in Guilin showed a decreasing trend at the city scale, but some regions still showed an increasing trend at the county distribution scale. (3) The ecological risk value in Guilin has significant spatial correlation, and the spatial distribution showed a clustering effect, which was consistent with the spatial distribution of ecological risk class areas. The research results can provide a reference for ecological risk control and sustainable development of landscape resource cities.

Customizing non-diffracting structured beams.

We demonstrate a universal approach to designing and generating non-diffracting structured light beams with arbitrary shapes. Such light beams can be tailored by predefining suitable spectral phases that match the corresponding beam shapes in the transverse plane. We develop a practical spectral superposition algorithm to discuss the non-diffracting properties and experimentally confirm our numerical results. Our proposed approach differs from that of classical non-diffracting beams, which are always constructed from wave equation solutions. The various non-diffracting structured beams could help manipulate particles following arbitrary transverse shapes and are likely to give rise to new applications in optical micromachining.

Auxin and CmAP1 regulate the reproductive development of axillary buds in Chinese chestnut (Castanea mollissima).

KEY MESSAGE: Auxin accumulation upregulates the expression of APETALA1 (CmAP1) and subsequently activates inflorescence primordium development in axillary buds of chestnut. The architecture of fruiting branches is a key determinant of chestnut yield. Normally, axillary buds at the top of mother fruiting branches develop into flowering shoots and bear fruits, and the lower axillary buds develop into vegetative shoots. Decapitation of the upper axillary buds induces the lower buds to develop into flowering shoots. How decapitation modulates the tradeoff between vegetative and reproductive development is unclear. We detected inflorescence primordia within both upper and lower axillary buds on mother fruiting branches. The level of the phytohormones 3-indoleacetic acid (IAA) and trans-zeatin (tZ) increased in the lower axillary buds in response to decapitation. Exogenous application of the synthetic analogues 1-naphthylacetic acid (NAA) or 6-benzyladenine (6-BA) blocked or promoted, respectively, the development of the inflorescence primordia in axillary buds. The transcript levels of the floral identity gene CmAP1 increased in axillary buds following decapitation. An auxin response element TGA-box is present in the CmAP1 promoter and influenced the CmAP1 promoter-driven expression of ß-glucuronidase (GUS) in floral organs in Arabidopsis, suggesting that CmAP1 is induced by auxin. We propose that decapitation releases axillary bud outgrowth from inhibition caused by apical dominance. During this process, decapitation-induced accumulation of auxin induces CmAP1 expression, subsequently promoting the reproductive development of axillary buds.

Pan-genome analysis of three main Chinese chestnut varieties.

Chinese chestnut (Castanea mollissima Blume) is one of the earliest domesticated plants of high nutritional and ecological value, yet mechanisms of C. mollissima underlying its growth and development are poorly understood. Although individual chestnut species differ greatly, the molecular basis of the formation of their characteristic traits remains unknown. Though the draft genomes of chestnut have been previously released, the pan-genome of different variety needs to be studied. We report the genome sequence of three cultivated varieties of chestnut herein, namely Hei-Shan-Zhai-7 (H7, drought-resistant variety), Yan-Hong (YH, easy-pruning variety), and Yan-Shan-Zao-Sheng (ZS, early-maturing variety), to expedite convenience and efficiency in its genetics-based breeding. We obtained three chromosome-level chestnut genome assemblies through a combination of Oxford Nanopore technology, Illumina HiSeq X, and Hi-C mapping. The final genome assemblies are 671.99 Mb (YH), 790.99 Mb (ZS), and 678.90 Mb (H7), across 12 chromosomes, with scaffold N50 sizes of 50.50 Mb (YH), 65.05 Mb (ZS), and 52.16 Mb (H7). Through the identification of homologous genes and the cluster analysis of gene families, we found that H7, YH and ZS had 159, 131, and 91 unique gene families, respectively, and there were 13,248 single-copy direct homologous genes in the three chestnut varieties. For the convenience of research, the chestnut genome database was constructed. Based on the results of gene family identification, the presence/absence variations (PAVs) information of the three sample genes was calculated, and a total of 2,364, 2,232, and 1,475 unique genes were identified in H7, YH and ZS, respectively. Our results suggest that the GBSS II-b gene family underwent expansion in chestnut (relative to nearest source species). Overall, we developed high-quality and well-annotated genome sequences of three C. mollissima varieties, which will facilitate clarifying the molecular mechanisms underlying important traits, and shortening the breeding process.

AfCHIL, a Type IV Chalcone Isomerase, Enhances the Biosynthesis of Naringenin in Metabolic Engineering.

Naringenin is an essential precursor for all flavonoids, and effectively promoting naringenin production is crucial in metabolic engineering. The interaction between plant metabolic enzymes ensures metabolic flux. The effect can effectively improve the natural product synthesis of engineering microbial systems. In this study, chalcone isomerase genes in Allium fistulosum have been identified. The expression of AfCHIL is closely related to the accumulation of anthocyanins, and the expression of AfCHIL and AfCHS was highly synchronized. Yeast two-hybrid and firefly luciferase complementation imaging assay further confirmed AfCHIL physically interacted with AfCHS/AfCHI. The bioconversion experiment confirmed that AfCHIL reduced the derailment produced by AfCHS and increased the yield of naringenin. In addition, a system of biosynthesis naringenin involved in AfCHS was constructed, and these results suggested that the potential function between CHS with CHIL advanced naringenin production effectively. In conclusion, this study illustrated the function of AfCHIs in Allium fistulosum and provided new insight into improving the synthesis efficiency of naringenin.

Clinical outcomes of partial sensory root rhizotomy on patients with recurrence of multiple sclerosing trigeminal neuralgia after percutaneous balloon compression.

OBJECTIVE: To investigate the clinical outcomes of partial sensory root rhizotomy (PSR) on patients with recurrence of multiple sclerosing trigeminal neuralgia(TN-MS) after percutaneous balloon compression (PBC). METHODS: 21 patients with recurrence of TN-MS after PBC were treated with PSR between January 2012 and July 2018. The visual analogue score (VAS) of participants before and after PBC/PSR were observed, and the postoperative recurrence rate of PBC/PSR were recorded, and the postoperative complications were also followed up. RESULTS: The VAS score reducing to 0-3 points after treatment was defined as effective and ≥4 points as invalid or recurrence. The effective rates of PSR at 1 day, 6 months, 12 months and 18 months after operation were 100%, 100%, 95% and 81%, respectively. The VAS scores of participants after PBC/PSR were significantly lower than those before PBC/PSR (all P<0.05), and the VAS scores at 1 day, 6 months, 12 months and 18 months after PSR were lower than those after PBC (all P<0.05). The postoperative recurrence rates at 6 months, 12 months and 18 months after PSR were significantly lower than those after PBC (all P<0.05). After PSR, all 21 patients had facial sensation loss, and one patient had intracranial infection, and none occurred decrease in masticatory muscle strength, weakened corneal reflex, intracranial hemorrhage, facial paralysis and cerebrospinal fluid leakage. CONCLUSION: PSR had a lower pain recurrence rate and a more significant reduction in VAS score compared with PBC, and it could be recommended to treat patients with recurrence of TN-MS after PBC.

Landscape Pattern and Ecological Risk Assessment in Guangxi Based on Land Use Change.

Due to ecological environmental fragility and soil erosion in Guangxi, studies of landscape patterns and associated ecological risks are needed to guide sustainable land development and ecologically sensitive land management. This study assesses dynamic spatial and temporal change patterns in land use and ecological risks based on 30 m land-use data, analyzes spatial correlations with ecological risks, and explores natural and socio-economic factor impacts on ecological risks. The results reveal: (1) A rapid and sizeable construction land increase in Guangxi from 2000 to 2018 associated mainly with loss of woodland and grassland. (2) Guangxi had the highest number of arable land patches from 2000 to 2018, and the distribution tended to be fragmented; moreover, the construction land gradually expanded outward from concentrated areas to form larger aggregates with increasing internal stability each year. (3) Guangxi ecological risk levels were low, low-medium, and medium, with significantly different spatial distributions observed for areas possessing different ecological risk levels. Regional ecological risk gradually decreased from the middle Guangxi regions to the surrounding areas and was positively correlated with spatial distribution. (4) Socio-economic factor impacts on ecological risk exceeded natural factor impacts. These results provide guidance toward achieving ecologically sensitive regional land-use management and ecological risk reduction and control, it can also provide a reference for ecological risk research in other similar regions in the world.

Periostin promotes the proliferation and metastasis of osteosarcoma by increasing cell survival and activates the PI3K/Akt pathway.

BACKGROUND: Silencing of the periostin gene (POSTN) can inhibit the biological process of several different cancers, and this inhibition may be related to down-regulation of PI3K/AKT signaling. However, the effect of POSTN on the progression, proliferation, and invasion of osteosarcoma (OS) remain unclear. METHODS: We used the Gene Expression Omnibus (GEO) database to screen datasets on in situ OS and lung metastases to identify core genes and potential pathways. We used additional bioinformatics tools to identify protein-protein interactions (PPIs) and gene networks, and selected the top seven genes whose expression had the strongest correlations with other genes. RESULTS: The results indicated that POSTN was a major hub gene. Subsequent analysis of gene expression profiles showed that POSTN was highly expressed in 262 cases with sarcoma and expression was closely related to poor prognosis. We also performed enrichment analysis to identify differentially expressed genes and used real-time PCR, western blotting, and immunohistochemistry analyses to measure POSTN expression in cells and tissues. Transfection of a POSTN-shRNA plasmid into cultured OS cells (Saos-2) effectively inhibited the proliferation, invasion, and migration of these cells. Taken together, our results suggest that POSTN may play a role in promoting the proliferation and metastasis of OS by activation of the PI3K/Akt signaling pathway. CONCLUSIONS: Our results provide a preliminary characterization of the mechanism by which POSTN may regulate the migration and invasion of OS cells and also provide a theoretical basis for identifying biomarkers that have potential use for the diagnosis and treatment of OS.

Swallowtail-type diffraction catastrophe beams.

We demonstrate a universal approach for generating high-order diffraction catastrophe beams, specifically for Swallowtail-type beams (abbreviated as Swallowtail beams), using diffraction catastrophe theory that was defined by potential functions depending on the control and state parameters. The three-dimensional curved caustic surfaces of these Swallowtail catastrophe beams are derived by the potential functions. Such beams are generated by mapping the cross sections of the high-order control parameter space to the corresponding transverse plane. Owing to the flexibility of the high-order diffraction catastrophe, these Swallowtail beams can be tuned to a diverse range of optical light structures. Owing to the similarity in their frequency spectra, we found that the Swallowtail beams change into low-order Pearcey beams under given conditions during propagation. Our experimental results are in close agreement with our simulated results. Such fantastic catastrophe beams that can propagate along curved trajectories are likely to give rise to new applications in micromachining and optical manipulation, furthermore, these diverse caustic beams will pave the way for the tailoring of arbitrarily accelerating caustic beams.

Abruptly autofocusing circular swallowtail beams.

In this Letter, to the best of our knowledge, we report the first experimental demonstration of a new family of autofocusing beams, circular swallowtail beams (CSBs), based on the high-order swallowtail catastrophe, which were determined by potential functions depending on the state and control parameters. The dynamics of the CSBs is discussed here. These types of CSBs tend to automatically focus without external components. Numerical results showed the focal intensity increased significantly, and it was as much as 110 times in the initial plane when the radius of the main ring was 40. Additionally, in contrast to previous circular Pearcey and Airy beams, these CSBs appeared to have more diversity and tunability due to having more propagation trajectories and intensity distribution structures due to high-order diffraction catastrophe. The numerical simulations were verified by our experimental results. These diverse CSBs could have new applications in flexible optical manipulation. These various CSBs could be beneficial for potential applications in optical trapping, medical treatment, or micromachining.

Hyperplasia suppressor gene inhibits the proliferation and metastasis of glioma cells by targeting rho family proteins.

AIM: To investigate the effect of the hyperplasia suppressor gene (HSG) on human glioma cell invasion and its possible mechanism. METHODS: Human glioma U251 cells were infected with recombinant viral vectors carrying the HSG gene sequence (HSG overexpression group) and HSG interference sequence (HSG suppression group). The negative control group with no-load virus transcription and a blank control group with only PBS treatment were set up. CCK-8 assay, cell scratch healing test, transwell migration, and invasion test were used to detect the effect of HSG expression on proliferation, migration and invasion of U251 glioma cells. Cell immunofluorescence and cell adhesion test were used to analyze the effect of HSG expression on cytoskeleton formation and adhesion ability of U251 cells. Gene chip technology was employed to preliminarily explore the effect of HSG expression change on the inherent gene expression in U251 cells. The expression of Rho family key molecule mRNA and protein was detected by light quantitative PCR and western blot. RESULTS: After 24 h of transcription with the recombinant virus vector, the cells showed a green color under an inverted fluorescence microscope. HSG expression increased in the HSG overexpression group (P < 0.01), and decreased in the HSG inhibition group (P < 0.01). Compared with the two control groups, the proliferation, scratch healing rate, migrating cell number, invasive cell number and adhesion cell number in the HSG overexpression group were markedly lower. After HSG overexpression, the morphology of U251 cells changed; filamentous pseudopods shortened and partially flaked. However, after HSG inhibition, the pseudopods grew toward both ends and were arranged axially. The overexpression of HSG inhibited the expression of rho family proteins (RhoA, Rock1, Rock2, Rac1, and Cdc42). CONCLUSION: The overexpression of HSG inhibits the progression of glioma U251 cells by regulating the expression of rho family proteins.

Chloroplast genomes of seven species of Coryloideae (Betulaceae): structures and comparative analysis.

Coryloideae is a subfamily in the family Betulaceae consisting of four extant genera: Carpinus, Corylus, Ostrya, and Ostryopsis. We sequenced the plastomes of six species of Corylus and one species of Ostryopsis for comparative and phylogenetic analyses. The plastomes are 159-160 kb long and possess typical quadripartite cp architecture. The plastomes show moderate divergence and conserved arrangement. Five mutational hotspots were identified by comparing the plastomes of seven species of Coryloideae: trnG-atpA, trnF-ndhJ, accD-psaI, ndhF-ccsA, and ycf1. We assembled the most complete phylogenomic tree for the family Betulaceae using 68 plastomes. Our cp genomic sequence phylogenetic analyses placed Carpinus, Ostrya, and Ostryopsis in a clade together and left Corylus in a separate clade. Within the genus Corylus, these analyses indicate the existence of five subclades reflecting the phylogeographical relationships among the species. The data offer significant genetic information for the identification of species of the Coryloideae, taxonomic and phylogenetic studies, and molecular breeding.

Heterologous Expression of SvMBD5 from Salix viminalis L. Promotes Flowering in Arabidopsis thaliana L.

Methyl-CpG-binding domain (MBD) proteins have diverse molecular and biological functions in plants. Most studies of MBD proteins in plants have focused on the model plant Arabidopsis thaliana L. Here we cloned SvMBD5 from the willow Salix viminalis L. by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed the structure of SvMBD5 and its evolutionary relationships with proteins in other species. The coding sequence of SvMBD5 is 645 bp long, encoding a 214 amino acid protein with a methyl-CpG-binding domain. SvMBD5 belongs to the same subfamily as AtMBD5 and AtMBD6 from Arabidopsis. Subcellular localization analysis showed that SvMBD5 is only expressed in the nucleus. We transformed Arabidopsis plants with a 35S::SvMBD5 expression construct to examine SvMBD5 function. The Arabidopsis SvMBD5-expressing line flowered earlier than the wild type. In the transgenic plants, the expression of FLOWERING LOCUS T and CONSTANS significantly increased, while the expression of FLOWERING LOCUS C greatly decreased. In addition, heterologously expressing SvMBD5 in Arabidopsis significantly inhibited the establishment and maintenance of methylation of CHROMOMETHYLASE 3 and METHYLTRANSFERASE 1, as well as their expression, and significantly increased the expression of the demethylation-related genes REPRESSOR OF SILENCING1 and DEMETER-LIKE PROTEIN3. Our findings suggest that SvMBD5 participates in the flowering process by regulating the methylation levels of flowering genes, laying the foundation for further studying the role of SvMBD5 in regulating DNA demethylation.

Generation of spirally accelerating optical beams.

We developed a generalized spectral phase superposition approach for generating accelerating optical beams along arbitrary trajectories. Such beams can be customized by predefining an appropriate superimposed phase pattern that consists of multiple sub-phases. We generated a spirally accelerating beam in a three-dimensional space and developed an algorithm to improve the uniformity of the intensity along the trajectory by introducing phase-shift factors. We also experimentally verified our numerical simulations. The proposed approach breaks the conventional convex trajectory restrictions. These various accelerating beams would pave the way for optically moving particles along a desired trajectory. The generation of such arbitrary accelerating beams is likely to give rise to new applications in flexible optical manipulation, wave front control, and optical transportation and guidance of particles.

Transcriptome Sequencing Analyses between the Cytoplasmic Male Sterile Line and Its Maintainer Line in Welsh Onion (Allium fistulosum L.).

Cytoplasmic male sterility (CMS) is important for exploiting heterosis in crop plants and also serves as a model for investigating nuclear-cytoplasmic interaction. The molecular mechanism of cytoplasmic male sterility and fertility restoration was investigated in several important economic crops but remains poorly understood in the Welsh onion. Therefore, we compared the differences between the CMS line 64-2 and its maintainer line 64-1 using transcriptome sequencing with the aim of determining critical genes and pathways associated with male sterility. This study combined two years of RNA-seq data; there were 1504 unigenes (in May 2013) and 2928 unigenes (in May 2014) that were differentially expressed between the CMS and cytoplasmic male maintainer Welsh onion varieties. Known CMS-related genes were found in the set of differentially expressed genes and checked by qPCR. These genes included F-type ATPase, NADH dehydrogenase, cytochrome c oxidase, etc. Overall, this study demonstrated that the CMS regulatory genes and pathways may be associated with the mitochondria and nucleus in the Welsh onion. We believe that this transcriptome dataset will accelerate the research on CMS gene clones and other functional genomics research on A. fistulosum L.

[Effects of GABAB receptor expression level on cognitive impairment and Arc/Arg3.1 expression in induced epileptic rats model].

OBJECTIVE: To investigate the effects of GABAB receptor on cognitive impairment by using pilocarpine induced kindled rats model and also to check early gene (Arc/Arg3.1) expression. METHODS: Pilocarpine induced kindled rats were divided into four groups (Group normal, Baclofen, CGP and Kindled) randomly, and every group included 20 rats.We checked their cognitive impairment by using passive avoidance test and water maze test.The expression of GABAB receptor (GB1, GB2) and Arc/Arg3.1 was tested by immunohistochemical staining, RT-PCR and Western blot. RESULTS: Passive avoidance test showed four Group rats shuttle times were 6.8±0.6, 1.2±0.2, 5.4±0.5, 3.6±0.3, incubation period were 26.1±3.9, 152.2±12.9, 65.8±7.0, 91.2±9.1, and water maze test had the same trend, with values in epilepsy groups significantly lower than the normal group of rats, which meant cognitive dysfunction.The above results also showed Baclofen further inhibited the learning and memory ability of the rats and CGP35348 promoted the learning and memory ability.The results of the Arc/Arg3.1 and GB1, GB2 level detection showed that epilepsy groups had significantly higher expression levels of Arc/Arg3.1 and GB1, GB2 than the normal group.Comparison among epilepsy groups showed that Baclofen group expressed lower levels of Arc/Arg3.1 and expressed higher levels of GB1, GB2, however CGP35348 group expressed higher levels of Arc/Arg3.1 and expressed lower levels of GB1, GB2. CONCLUSIONS: GABAB receptor can affect the ability of spatial learning and memory of epileptic rats by regulating Arc/Arg3.1.

[The clinical analysis of plakophilin-2 gene mutation in patients with arrhythmogenic right ventricular cardiomyopathy/dysplasia].

OBJECTIVE: The purpose of this study was to screen genetic variations in plakophilin-2 (PKP2) gene in patients with arrhythmogenic right ventricular cardiomyopathy/dysplasia (ARVC/D) and investigate the differences in clinical features between mutation and no-mutation groups. METHODS: Thirty unrelated Chinese patients clinically diagnosed with ARVC/D and 50 healthy controls were included. Genomic DNA was isolated from peripheral blood samples. PCR and direct sequencing were used to detect variations in PKP2 gene. RESULTS: Eight PKP2 mutant variants were identified in 10 ARVC/D patients (8 men, 2 women). Among the eight mutation, three (c.2194C>T, c. 1170+ 1G>A and c. 810_813delGGTC) were novel mutation. Clinical features of the PKP2 mutation group were similar to those of the non-mutation group. CONCLUSIONS: The rate of PKP2 mutation is 33.3% (10/30) in ARVC/D patients. The penetrance of PKP2 mutation for ARVC/D tends to be higher in man patients. No significant differences could be detected in phenotype characteristics between patients with and without PKP2 mutation.

Improved long-term survival associated with stent deployment during percutaneous coronary interventions: results from a registry of 3399 patients.

INTRODUCTION: The use of stents in percutaneous coronary intervention (PCI) improves procedural success and reduces restenosis. However, few studies have had a sufficient sample size or adequate follow-up to determine whether this advantage results in a positive effect on mortality. METHODS: A total of 3399 patients undergoing PCI (stented [with dual antiplatelet therapy]: n = 2456, nonstented [balloon PCI or rotational atherectomy]: n = 942) at a single institution from 1994 to 2001 were followed up prospectively (43 +/- 22 and 54 +/- 25 months, respectively) for acute and long-term clinical outcomes. RESULTS: Angiographic success (< 50% residual stenosis) (99.7% vs 97.7%, P < .001) and acute gain (3.02 +/- 0.55 vs 2.08 +/- 0.62 mm, P < .001) were both greater for stented lesions. Likewise, procedural complications of death (0.04% vs 0.4%, P = .02) and dissection (4.9% vs 8.0%, P = .001) were lower in the stent group, as were rates of 6-month clinical restenosis (10.3% vs 16.3%, P < .001). Eight-year mortality (12.0% vs 18.2%, hazard ratio = 0.78, P = .009) was lower among the stent group, as was long-term major adverse cardiac events (36.2% vs 50.6%, P < .001), but no difference in long-term myocardial infarction was found (6.5% vs 7.6%, P = .28). In multivariable Cox regression, stent use (hazard ratio = 0.76, 95% CI [0.58-0.99], P = .04) remained associated with significantly reduced mortality. CONCLUSION: This large prospective study demonstrates that, in addition to a general improvement in procedural success and a reduced need for repeat revascularization, the use of stents with dual antiplatelet therapy was associated with a significant reduction in long-term mortality. Consideration should be given for the use of stents whenever feasible during PCI.