RESUMEN
BACKGROUND: The role of CXCL10 in progression and prognosis of colorectal cancer (CRC) has been studied for years, yet results remain controversial. AIM: This study aims to explore the relationship between CXCL10 and CRC progression and prognosis. METHODS: We evaluated plasma CXCL10 in CRC patients using ELISA. We also performed a meta-analysis of the associations between CXCL10 and overall survival (OS), disease-free survival (DFS), disease-specific survival (DSS), relapse-free survival (RFS), and clinicopathological features. Finally, correlations between CXCL10 and methylation or immune infiltration were performed using TCGA data. RESULTS: ELISA analysis showed that CXCL10 was associated with age, red blood cells, blood platelets, and blood urea nitrogen. A separate analysis of 3,763 patients from 24 studies revealed that there were significant associations between low CXCL10 expression and OS (HR 1.25, 95% CI 1.01-1.53), DFS (HR 1.65, 95% CI 1.17-2.34), and RFS (HR 1.43, 95% CI 1.20-1.71) in CRC. Additionally, downregulated CXCL10 expression was significantly correlated with age [odds ratio (OR) 1.31, 95% CI 1.13-1.52], metastasis (OR 1.34, 95% CI 1.11-1.63), recurrence (OR 1.46, 95% CI 1.16-1.83), tumor location (OR 1.88, 95% CI 1.58-2.24), differentiation (OR 0.57, 95% CI 0.35-0.93), microsatellite instability (OR 0.23, 95% CI 0.15-0.35), BRAF mutation (OR 1.62, 95% CI 1.25-2.08), p53 mutation (OR 0.28, 95% CI 0.16-0.47), and CIMP (OR 0.27, 95% CI 0.17-0.43). Furthermore, significant associations were observed between CXCL10 and methylation and immune infiltration. CONCLUSIONS: The study suggests that CXCL10 might be a potential target for the treatment of CRC. TRIAL REGISTRATION: NCT03189992. Registered 4 June 2017, https://www.clinicaltrials.gov/ct2/show/study/NCT03189992?term=NCT03189992&rank=1 .
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Quimiocina CXCL10/sangre , Neoplasias Colorrectales/sangre , Recurrencia Local de Neoplasia/sangre , Biomarcadores de Tumor/sangre , Biomarcadores de Tumor/genética , Quimiocina CXCL10/genética , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/cirugía , Progresión de la Enfermedad , Femenino , Humanos , Masculino , Persona de Mediana Edad , Mutación , Recurrencia Local de Neoplasia/patología , Recurrencia Local de Neoplasia/cirugía , Valor Predictivo de las Pruebas , Tasa de SupervivenciaRESUMEN
Burkitt lymphoma (BL) is a highly malignant non-Hodgkin's lymphoma that is closely related to the abnormal expression of genes. Familial acute myelogenous leukemia related factor (FAMLF; GenBank accession No. EF413001.1) is a novel gene that was cloned by our research group, and miR-181b is located in the intron of the FAMLF gene. To verify the role of miR-181b and FAMLF in BL, RNAhybrid software was used to predict target site of miR-181b on FAMLF and real-time quantitative PCR (RQ-PCR) was used to detect expression of miR-181b and FAMLF in BL patients, Raji cells and unaffected individuals. miR-181b was then transfected into Raji and CA46 cell lines and FAMLF expression was examined by RQ-PCR and western blotting. Further, Raji cells viability and proliferation were detected by MTT and clone formation, and Raji cell cycle and apoptosis were detected by flow cytometry. The results showed that miR-181b can bind to bases 21-42 of the FAMLF 5' untranslated region (UTR), FAMLF was highly expressed and miR-181b was lowly expressed in BL patients compared with unaffected individuals. FAMLF expression was significantly and inversely correlated to miR-181b expression, and miR-181b negatively regulated FAMLF at posttranscriptional and translational levels. A dual-luciferase reporter gene assay identified that the 5' UTR of FAMLF mRNA contained putative binding sites for miR-181b. Down-regulation of FAMLF by miR-181b arrested cell cycle, inhibited cell viability and proliferation in a BL cell line model. Our findings explain a new mechanism of BL pathogenesis and may also have implications in the therapy of FAMLF-overexpressing BL.
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Linfoma de Burkitt/genética , Linfoma de Burkitt/metabolismo , Regulación Neoplásica de la Expresión Génica , MicroARNs/metabolismo , Proteínas/metabolismo , Adolescente , Adulto , Apoptosis/genética , Línea Celular Tumoral , Proliferación Celular/genética , Supervivencia Celular/genética , Niño , Preescolar , Regulación hacia Abajo/genética , Femenino , Humanos , Lactante , Masculino , MicroARNs/genética , Proteínas/genética , Adulto JovenRESUMEN
This study aimed to study the role of rapamycin (RAPA) in modulating the interaction between gδ T cells and dendritic cells (DCs) in a lipopolysaccharide (LPS)-induced acute lung injury mouse model. Mice were injected with LPS to establish the acute lung injury model or LPS + RAPA to assess the role of RAPA in modulating cell interactions. Mice were injected with PBS or RAPA alone as controls. gδ T cells and DCs were isolated from all mice and assessed by flow cytometry and fluorescence microscopy. The isolated gδ T cells and DCs were cultured independently or co-cultured to study their interactions. Enzyme-linked immunosorbent assay was performed to assess the expression of the cytokines, namely, interferon (IFN)-γ, interleukin (IL)-4, tumor necrosis factor (TNF)-α and IL-12 in the individually cultured or co-cultured gδ T cells and DCs, and reverse transcription-polymerase chain reaction (RT-PCR) was employed to investigate the levels of relevant mRNAs. Our study found that co-culture of gδ T cells and DCs from mice treated with LPS + RAPA have reduced expression of IFN-γ and IL-4 (but not TNF-α and IL-12) compared to mice treated with LPS only. These results were confirmed by RT-PCR, where the levels of IFN-γ and IL-4 mRNA were also reduced. This study may provide useful information in understanding the interaction between gδ T cells and DCs in the LPS-induced lung injury model in mice.
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Lesión Pulmonar Aguda/inmunología , Células Dendríticas/inmunología , Inmunosupresores/farmacología , Interferón gamma/metabolismo , Interleucina-4/metabolismo , Sirolimus/farmacología , Linfocitos T/inmunología , Lesión Pulmonar Aguda/etiología , Animales , Técnicas de Cocultivo , Células Dendríticas/efectos de los fármacos , Interferón gamma/genética , Interleucina-12/genética , Interleucina-12/metabolismo , Interleucina-4/genética , Lipopolisacáridos/toxicidad , Ratones , Ratones Endogámicos C57BL , Receptores Inmunológicos/genética , Receptores Inmunológicos/metabolismo , Linfocitos T/efectos de los fármacos , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Isolation of high-quality RNA is important for assessing sperm gene expression, and semen purification methods may affect the integrity of the isolated RNA. This study evaluated the effectiveness of the sperm swim-up method for seminal RNA isolation. Frozen semen samples in straws from three bulls of proven fertility were purified by the swim-up method. RNA extraction was carried out using the E.Z.N.A.(TM) Total RNA kit II, with non-swim-up sperm as a control. Total sperm RNA was analyzed by UV spectrophotometry, reverse transcription polymerase chain reaction (RT-PCR), and agarose gel electrophoresis, and expression of the sex-determining region on the Y chromosome (SRY), leptin (LEP), and ribosomal protein subunit 23 (RPS23) genes, were determined. 18S RNA was used as a positive control. Fewer somatic cells were found in sperm swim-up samples than in the non-swim-up counterparts (0 x 10(3) vs 17.33 ± 2.52 x 10(3) sperm, P < 0.05). In addition, high-quality RNA was obtained in about 2 h, with no significant difference between groups. Interestingly, the yields of RNA fragments containing ≥200 nucleotides were significantly reduced in sperm swim-up samples (0.92 ± 0.41 x 10(7) sperm) compared with the non-swim-up samples (1.36 ± 0.33 x 10(7) sperm, P < 0.05). After RT-PCR, clear bands representing SRY, LEP, and RPS23 in sperm cDNA were observed on agarose gel electrophoresis. Finally, no bands corresponding to 18S RNA were found in RNA samples from the sperm swim-up group. Our findings suggest that small amounts of sperm RNA can be efficiently extracted from frozen straw semen samples using the swim-up technique.
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ARN/química , Análisis de Semen/veterinaria , Semen/metabolismo , Animales , Bovinos , Leptina/genética , Leptina/metabolismo , Masculino , ARN/genética , ARN Ribosómico 18S/genética , Proteínas Ribosómicas/genética , Proteínas Ribosómicas/metabolismo , Semen/citología , Análisis de Semen/métodos , Análisis de Semen/normas , Cromosoma Y/genéticaRESUMEN
Chitinase is an important pathogenesis-related protein in plants, and it can accumulate when induced by salicylic acid (SA) or other elicitors. Here, we found that chitinase mRNA levels were 4.5-times greater when peanut seedlings were sprayed with 1.5 mM SA, as compared to water. The upstream promoter sequence of the chitinase gene was cloned by TAIL-PCR and the potential cis-regulatory elements in this promoter were predicted by the cis-element databases PLACE and plantCARE. Elements in the promoter related to SA induction and disease resistance response included AS-1, GT1-motif, GRWAAW, TGTCA, W-box, and WB-box. The full-length promoter (P) and a series of 5'-deleted promoters (P1-P5) were cloned and then substituted for the 35S promoter of pCAMBIA1301-xylA, which carries the xylose isomerase gene as the selectable marker and GUS as the reporter gene. Six plant expression vectors (pCAMBIA1301-xylA-P-pCAMBIA1301-xylA-P5) were obtained. The six expression vectors were then transferred into onion epidermal cells and peanut plants by Agrobacterium-mediated transformation. Both the full-length and deleted promoters resulted in GUS staining of the onion epidermis cells when induced by SA. In onion epidermis cells, GUS enzyme activity was greater after SA induction. In transgenic peanut plants, GUS mRNA levels were greater after SA induction. Consideration of the cis-regulatory elements predicted by PLACE and plantCARE suggested that AS-1, GRWAAW, and W-box are positive regulatory elements in P2 and P3 and that GT1-motif and TGTCA are negative regulatory elements between P and P2.
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Arachis/enzimología , Arachis/genética , Quitinasas/genética , Regiones Promotoras Genéticas/genética , Regulación de la Expresión Génica de las Plantas/genética , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/enzimología , Plantas Modificadas Genéticamente/genéticaRESUMEN
Osteoporosis is the most common bone disease, affecting millions of people worldwide and leading to significant morbidity and high costs. Monacolin K, an extract of red yeast rice (RYR, Hongqu), plays important roles in the management of dyslipidemia, coronary heart disease, and diabetes. Our study aimed to investigate the protective effect of monacolin K on ovariectomy-induced bone loss in rats. Fifty female Sprague-Dawley rats were randomly divided into a sham-operated and five ovariectomized (OVX) groups: OVX with vehicle, OVX with fluvastatin, and OVX with RYR extract of three graded doses. Bone mineral density (BMD), biochemical markers, and cell viability were analyzed by dual energy X-ray absorptiometry, enzyme-linked immunosorbent assay, and 3(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays. Gene expression was evaluated by real-time polymerase chain reaction amplification and western blot. Our results showed that administration of RYR extract markedly increased the bone mineral density in OVX rats. Moreover, RYR extract decreased the levels of bone turnover markers, including osteocalcin and tartrate resistant acid phosphatase activity. The MMT assay revealed that RYR extract treatment significantly improved the osteoblast viabilities in a dose-dependent manner (P < 0.05). At the molecular level, we further demonstrated that RYR extract enhanced the expression of Bmp2 and Bmp4 both at the mRNA and protein levels. Collectively, these data suggested RYR extract could protect against osteoporosis in ovariectomized rats, most likely through activation of BMP2/4 expression.
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Productos Biológicos/uso terapéutico , Resorción Ósea/tratamiento farmacológico , Resorción Ósea/etiología , Osteoporosis/tratamiento farmacológico , Ovariectomía , Extractos Vegetales/uso terapéutico , Animales , Productos Biológicos/farmacología , Biomarcadores/metabolismo , Peso Corporal/efectos de los fármacos , Densidad Ósea/efectos de los fármacos , Proteínas Morfogenéticas Óseas/metabolismo , Remodelación Ósea/efectos de los fármacos , Resorción Ósea/complicaciones , Proliferación Celular/efectos de los fármacos , Femenino , Tamaño de los Órganos/efectos de los fármacos , Osteoporosis/complicaciones , Ratas Sprague-Dawley , Transducción de Señal/efectos de los fármacos , Útero/efectos de los fármacos , Útero/patologíaRESUMEN
We performed a 1-year cluster-randomized field trial to assess the effect of standardized management of chronic obstructive pulmonary disease (COPD) on lung function and quality of life (QOL) measures in patients in China. We used the Global Initiative for Chronic Obstructive Lung Disease (GOLD) treatment guidelines and assessed indexes including pulmonary function, QOL, quality-adjusted life years (QALY), Medical Research Council (MRC) dyspnea scale, 6-min walk distance (6-MWD), number of emergency visits, and frequency of hospitalization. Of a total of 711 patients with chronic cough and asthma, 132 were diagnosed as having COPD and 102 participated in this study [intervention group (N = 47); control group (N = 55)]. We found that adherence to GOLD guidelines had a perceivable impact on 6-MWD, MRC dyspnea scale score, and QOL. The average QALY increased by 1.42/person/year in the intervention group, but declined by 0.95/person/year in the control group. We conclude that standardized management improves disease severity, QOL, and QALY in COPD patients when treatment protocols adhere to GOLD guidelines.
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Enfermedad Pulmonar Obstructiva Crónica/diagnóstico , Enfermedad Pulmonar Obstructiva Crónica/terapia , Caminata/fisiología , Anciano , Estudios de Casos y Controles , China , Análisis Costo-Beneficio , Disnea/diagnóstico , Disnea/fisiopatología , Femenino , Adhesión a Directriz , Humanos , Masculino , Persona de Mediana Edad , Cooperación del Paciente , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Enfermedad Pulmonar Obstructiva Crónica/prevención & control , Calidad de Vida , Años de Vida Ajustados por Calidad de Vida , Pruebas de Función RespiratoriaRESUMEN
The Rab protein family belongs to a superfamily of ras-like GTP-binding proteins. Rab proteins regulate many steps of membrane trafficking. In this study, three Rab family members, Rab5B, Rab6A, and Rab7, designated LvRab5B, LvRab6A, and LvRab7, were cloned from Litopenaeus vannamei. The full-length cDNA sequences of LvRab5B, LvRab6A, and LvRab7 were 1383, 873, and 767 nucleotides in length and they encoded proteins of 211, 212, and 205 amino acids, respectively. Using qRT-PCR, the mRNA expression levels of the three proteins were determined in the hepatopancreas of L. vannamei at different stages after infectious hypodermal and hematopoietic necrosis virus and white spot syndrome virus challenge. The results indicated that the mRNA expression levels of LvRab5B, LvRab6A, and LvRab7 were all significantly up-regulated after virus injection, suggesting that these genes may play essential roles in the immune response to viral infection in shrimp.
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Regulación de la Expresión Génica , Penaeidae/genética , Proteínas de Unión al GTP rab/genética , Proteínas de Unión al GTP rab5/genética , Secuencia de Aminoácidos , Animales , Western Blotting , Clonación Molecular , Densovirinae , Perfilación de la Expresión Génica , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Análisis de Secuencia de ADN , Virus del Síndrome de la Mancha Blanca 1 , Proteínas de Unión al GTP rab/química , Proteínas de Unión al GTP rab/metabolismo , Proteínas de Unión al GTP rab5/química , Proteínas de Unión al GTP rab5/metabolismo , Proteínas de Unión a GTP rab7RESUMEN
We investigated the association between polymorphisms in the adiponectin gene (APM-1) and atherosclerotic cerebral infarction (ACI) in a Chinese Han population of Hainan Province. Polymerase chain reaction-restriction fragment length polymorphism and gene sequencing were used to analyze the distribution of APM-1 +45T/G and +276G/T genotypes and their alleles in 120 ACI patients and 120 healthy controls. No statistical correlation was found in the frequency and distribution of the genotype 45T/G between the ACI group and the control group. Genotypic frequencies of GG, GT, and TT at the APM-1 +276 locus were 70.0% (84/120), 25.0% (30/120), and 5.0% (6/120), respectively, in the ACI group, while these values were 52.5% (63/120), 37.5% (45/120), and 10.0% (12/120), respectively, in the control group. The frequency of the G allele was 82.5% (198/240) in the ACI group and 71.25% (171/240) in the control group. The T allele frequency was 17.5% (42/240) in the ACI group and 28.75% (69/240) in the control group. Polymorphisms at the APM-1 -276 locus in the case-controlled groups showed significant differences in the genotype distribution and al-lele frequency between the 2 groups (P = 0.041). The occurrence of ACI in the Hainan Chinese Han population may be associated with +276G/T polymorphisms but not with +45T/G polymorphisms in the APM-1 gene.
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Adiponectina/genética , Infarto Cerebral/genética , Frecuencia de los Genes/genética , Arteriosclerosis Intracraneal/genética , Adiponectina/biosíntesis , Pueblo Asiatico/genética , Secuencia de Bases , Estudios de Casos y Controles , China , Femenino , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Polimorfismo de Nucleótido Simple/genética , Análisis de Secuencia de ADNRESUMEN
MicroRNAs (miRNAs) are known to play an important role in regulating both adaptive and innate immunity. Pacific white shrimp (Litopenaeus vannamei) is the most widely farmed crustacean species in the world. However, little is known about the role miRNAs play in shrimp immunity. To understand the impact of viral infection on miRNA expression in shrimp, we used high-throughput sequencing technology to sequence two small RNA libraries prepared from L. vannamei under normal and white spot syndrome virus (WSSV) challenged conditions. Approximately 19,312,189 and 39,763,551 raw reads corresponding to 17,414,787 and 28,633,379 high-quality mappable reads were obtained from the two libraries, respectively. Twelve conserved miRNAs and one novel miRNA that were highly expressed (>100 RPM) in L. vannamei were identified. Of the identified miRNAs, 8 were differentially expressed in response to the virus infection, of which 1 was upregulated and 7 were downregulated. The prediction of miRNA targets showed that the target genes of the differentially expressed miRNAs were related to immunity, apoptosis, and development functions. Our study provides the first characterization of L. vannamei miRNAs in response to WSSV infection, which will help to reveal the roles of miRNAs in the antiviral mechanisms of shrimp.
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Inmunidad Innata/genética , MicroARNs/genética , Penaeidae/genética , Virus del Síndrome de la Mancha Blanca 1/genética , Animales , Regulación de la Expresión Génica , MicroARNs/biosíntesis , Penaeidae/virología , Virus del Síndrome de la Mancha Blanca 1/patogenicidadRESUMEN
Blumea balsamifera DC is a member of the Compositae family and is frequently used as traditional Chinese medicine. Blumea balsamifera is rich in monoterpenes, which possess a variety of pharmacological activities, such as antioxidant, anti-bacteria, and anti-viral activities. Farnesyl diphosphate synthase (FPS) is a key enzyme in the biosynthetic pathway of terpenes, playing an important regulatory role in plant growth, such as resistance and secondary metabolism. Based on the conserved oligo amino acid residues of published FPS genes from other higher plant species, a cDNA sequence, designated BbFPS, was isolated from B. balsamifera DC using polymerase chain reaction. The clones were an average of 1.6 kb and contained an open reading frame that predicted a polypeptide of 342 amino acids with 89.07% identity to FPS from other plants. The deduced amino acid sequence was dominated by hydrophobic regions and contained 2 highly conserved DDxxD motifs that are essential for proper functioning of FPS. Phylogenetic analysis indicated that FPS grouped with other composite families. Prediction of secondary structure and subcellular localization suggested that alpha helices made up 70% of the amino acids of the sequence.
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Asteraceae/enzimología , Asteraceae/genética , Genes de Plantas , Geraniltranstransferasa/genética , Análisis de Secuencia de ADN , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Evolución Molecular , Geraniltranstransferasa/química , Modelos Moleculares , Datos de Secuencia Molecular , Filogenia , Estructura Secundaria de Proteína , Alineación de Secuencia , Análisis de Secuencia de ProteínaRESUMEN
A total of 160 Rongchang pigs (26.76±1.78 kg) were randomly assigned to 5 dietary treatment groups until their body weight (BW) reached 90 kg. The diets were supplemented with 0, 0.5, 1.0, 1.5, and 2.0% conjugated linoleic acid (CLA). Our results showed that the 1.0 to 2.0% CLA-fed pigs had less back fat deposition when their BW reached 90 kg than the pigs that received less than 1% CLA. During the 30 to 60 kg growing period, 1.0, 1.5, and 2.0% CLA treatments improved pork quality by significantly reducing the pork pH (P<0.01) and color value (P<0.05), but they increased marble scaling (P<0.01). Similarly, the 1.5 and 2.0% CLA-fed pigs had more marble than other pigs when their BW reached 90 kg. Furthermore, CLA significantly affected the expression of muscle fiber-type genes. The 1.5% CLA-fed pigs exhibited the highest mRNA expression of MyHC1 and MyHC2a (P<0.05) at 60 kg BW. At 90 kg BW, the highest expression of MyHC1 and MyHC2a (P<0.05) was found in the 2.0% CLA group. However, MyHC2x was downregulated in the CLA-fed pigs at this time. In addition, CLA supplements did not evidently alter mRNA expression of MyHC2b at all times. These results demonstrate that CLA could affect carcass traits and improve the meat quality of growing-finishing pigs by altering the expression of genes related to muscle growth and development; 1-1.5% CLA was the most appropriate CLA dose.
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Calidad de los Alimentos , Regulación de la Expresión Génica , Ácidos Linoleicos Conjugados/metabolismo , Carne/normas , Fibras Musculares Esqueléticas/metabolismo , Carácter Cuantitativo Heredable , Alimentación Animal , Animales , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Ácidos Linoleicos Conjugados/farmacología , Fibras Musculares Esqueléticas/efectos de los fármacos , ARN Mensajero/genética , PorcinosRESUMEN
Ossification of the posterior longitudinal ligament (OPLL) of the cervical spine is a complex multifactorial disease. Patients with OPLL commonly present with symptoms in their 40s or 50s. The genetic basis of OPLL remains poorly understood. Exome capture combined with massively parallel DNA sequencing has been proposed as an efficient strategy to search for disease-causing genes of both monogenic and multigenic disorders. To identify candidate pathogenic genes associated with OPLL, we performed whole exome sequencing (WES) on two unrelated southern Chinese OPLL patients. The entire DNA coding region of the candidate genes was amplified by PCR and Sanger sequenced. The common single nucleotide polymorphisms were analyzed by association studies. WES revealed p.T265S/PTCH1, p.P1232L/PTCH1, and p.T902S/COL17A1 mutants in the two female cases with mixed OPLL. These were confirmed by Sanger sequencing. p.P1232L/PTCH1, p.N1374D/COL17A1 and p.T902S/COL17A1 were subsequently identified in three males with continuous OPLL and one female with mixed OPLL. The association studies indicated that the SNPs rs805698 and rs4918079 in COL17A1 were significantly associated with OPLL. This study suggests that WES may be a practical approach to revealing significant genetic involvement in OPLL. Variants of the PTCH1 and COL17A1 genes may contribute to the development of OPLL.