ABSTRACT
METHODS@#The efficiency of electroporation-based delivery of AsCpf1/mRNA and AsCpf1/RNP to target exon 3 of leukemia inhibitory factor (Lif) into mouse zygotes was evaluated. Embryos that developed to the two-cell stage after zygote electroporation were transferred into the oviducts of surrogate mothers to produce AsCpf1-mediated LIF KO mice. The genome editing efficiency of blastocysts and pups was tested using the T7E1 assay and/or DNA sequencing. Congenital abnormalities and reproductive phenotypes in LIF KO mice produced by electroporation with AsCpf1/RNP were examined. @*RESULTS@#Survival and two-cell development of electroporated zygotes were comparable between the AsCpf1/mRNA and AsCpf1/RNP groups, whereas genome editing efficiency was relatively higher in the AsCpf1/RNP group (13.3% vs 18.1% at blastocyst and 33.3% vs 45.5% at offspring), respectively. Two mouse lines with a frameshift mutation in exon 3 of the Lif gene were established from the AsCpf1/RNP group. All congenital abnormalities of LIF KO mice produced by AsCpf1/RNP electroporation were observed. AsCpf1-mediated LIF KO mice showed postnatal growth retardation and implantation failure, both of which are major phenotypes of LIF KO mice generated by conventional gene targeting. @*CONCLUSION@#Electroporation of AsCpf1/RNP at the zygote stage is an efficient genome editing method to produce KO mice.
ABSTRACT
METHODS@#The efficiency of electroporation-based delivery of AsCpf1/mRNA and AsCpf1/RNP to target exon 3 of leukemia inhibitory factor (Lif) into mouse zygotes was evaluated. Embryos that developed to the two-cell stage after zygote electroporation were transferred into the oviducts of surrogate mothers to produce AsCpf1-mediated LIF KO mice. The genome editing efficiency of blastocysts and pups was tested using the T7E1 assay and/or DNA sequencing. Congenital abnormalities and reproductive phenotypes in LIF KO mice produced by electroporation with AsCpf1/RNP were examined. @*RESULTS@#Survival and two-cell development of electroporated zygotes were comparable between the AsCpf1/mRNA and AsCpf1/RNP groups, whereas genome editing efficiency was relatively higher in the AsCpf1/RNP group (13.3% vs 18.1% at blastocyst and 33.3% vs 45.5% at offspring), respectively. Two mouse lines with a frameshift mutation in exon 3 of the Lif gene were established from the AsCpf1/RNP group. All congenital abnormalities of LIF KO mice produced by AsCpf1/RNP electroporation were observed. AsCpf1-mediated LIF KO mice showed postnatal growth retardation and implantation failure, both of which are major phenotypes of LIF KO mice generated by conventional gene targeting. @*CONCLUSION@#Electroporation of AsCpf1/RNP at the zygote stage is an efficient genome editing method to produce KO mice.
ABSTRACT
OBJECTIVES: Early detection and treatment of an oral squamous cell carcinoma (OSCC) is critical because of its rapid growth, frequent lymph-node metastasis, and poor prognosis. However, no clinically-valuable methods of early diagnosis exist, and genetic analysis of OSCCs has yielded no biomarkers. Study DESIGN: We investigated the expression of genes associated with inflammation in OSCCs via a quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis of microarray data. Tumor and normal tissues from five patients with an OSCC were used for microarray analysis. Differentially-expressed genes, identified using permutation, local pooled error (LPE), t-tests, and significance analysis of microarrays (SAM), were selected as candidate genetic markers. RESULTS: Two groups corresponding to tissue identity were evident, implying that their differentially-expressed genes represented biological differences between tissues. Fifteen genes were identified using the Student's pairedt-test (p < 0.05) and the SAM, with a false discovery rate of less than 0.02. Based on gene expression, these 15genes can be used to classify an OSCC. A genetic analysis of functional networks and ontologies, validated by using a qRT-PCR analysis of the tissue samples, identified four genes, ADAM15, CDC7, IL12RB2 and TNFRSF8,that demonstrated excellent concordance with the microarray data. CONCLUSIONS: Our study demonstrated that four genes (ADAM15, CDC7, IL12RB2 and TNFRSF8) had potential as novel biomarkers for the diagnosis and the treatment of an OSCC
Subject(s)
Humans , Carcinoma, Squamous Cell/pathology , Odontogenic Tumor, Squamous/pathology , Mouth Neoplasms/pathology , Biomarkers, Tumor/analysis , Reverse Transcriptase Polymerase Chain Reaction/methods , Gene ExpressionABSTRACT
OBJECTIVES: Early detection and treatment of an oral squamous cell carcinoma (OSCC) is critical because of its rapid growth, frequent lymph-node metastasis, and poor prognosis. However, no clinically-valuable methods of early diagnosis exist, and genetic analysis of OSCCs has yielded no biomarkers. STUDY DESIGN: We investigated the expression of genes associated with inflammation in OSCCs via a quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis of microarray data. Tumor and normal tissues from five patients with an OSCC were used for microarray analysis. Differentially-expressed genes, identified using permutation, local pooled error (LPE), t-tests, and significance analysis of microarrays (SAM), were selected as candidate genetic markers. RESULTS: Two groups corresponding to tissue identity were evident, implying that their differentially-expressed genes represented biological differences between tissues. Fifteen genes were identified using the Student's paired t-test (p<0.05) and the SAM, with a false discovery rate of less than 0.02. Based on gene expression, these 15 genes can be used to classify an OSCC. A genetic analysis of functional networks and ontologies, validated by using a qRT-PCR analysis of the tissue samples, identified four genes, ADAM15, CDC7, IL12RB2 and TNFRSF8, that demonstrated excellent concordance with the microarray data. CONCLUSIONS: Our study demonstrated that four genes (ADAM15, CDC7, IL12RB2 and TNFRSF8) had potential as novel biomarkers for the diagnosis and the treatment of an OSCC.
Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Squamous Cell/genetics , Early Detection of Cancer/methods , Mouth Neoplasms/genetics , Biomarkers, Tumor/analysis , Carcinoma, Squamous Cell/chemistry , DNA, Neoplasm/analysis , Female , Gene Expression Profiling , Gene Expression Regulation, Neoplastic , Humans , Male , Middle Aged , Mouth Neoplasms/chemistryABSTRACT
PURPOSE: The aim of this study is to evaluate the relationship between abdominal subcutaneous fat thickness measured by ultrasonography (US) and serum lipid profile and liver transaminases in obese children. METHODS: One hundred and sixty-six children diagnosed with obesity from May 2001 to December 2013 were included in this study. Data on serum lipid profile and liver transaminases were collected from clinical records. Abdominal subcutaneous fat thickness and grade of hepatic steatosis were evaluated by US. RESULTS: Of the 166 children, 107 were diagnosed with hepatic steatosis by US, 46 with grade I, 56 with grade II, and five children with grade III. According to the grade of hepatic steasosis, the average values of midline abdominal subcutaneous fat thickness and right flank abdominal subcutaneous fat thickness measured 2.9+/-0.8 cm and 1.9+/-0.7 cm in the normal group, 3.3+/-0.8 cm and 2.0+/-0.7 cm in grade I, 3.8+/-0.8 cm and 2.3+/-0.8 cm in grade II, and 4.1+/-0.8 cm and 2.8+/-1.4 cm in grade III, respectively. Abdominal subcutaneous fat thickness correlated with grade of hepatic steatosis (p<0.01). In addition, abdominal subcutaneous fat thickness correlated with concentration of serum lipids and liver transaminases in the age group of 12-14 years (p<0.01). CONCLUSION: Abdominal subcutaneous fat thickness measured by US can be used as a reliable predictor of possible hyperlipidemia and steatohepatitis in children, especially during the adolescent stage.
Subject(s)
Adolescent , Child , Humans , Fatty Liver , Hyperlipidemias , Liver , Obesity , Subcutaneous Fat, Abdominal , Transaminases , UltrasonographyABSTRACT
During 2011~2013, a total of 729 samples for 19 types of medicinal plant were collected from Seoulyekryungsi in Seoul, Korea, and investigated for the presence of aflatoxins. The samples were analyzed using immunoaffinity column cleanup and high-performance liquid chromatography coupled to a fluorescence detector after post-column derivatization. Aflatoxins were found in 124 out of the 729 analyzed samples: 65 containing aflatoxin B1 (AFB1), 24 with aflatoxin B2 (AFB2), 15 with aflatoxin G1 (AFG1), and 20 samples with aflatoxin G2 (AFG2). The ranges for positive samples were 0.1~404.7 microg/kg for AFB1, 0.1~10.0 microg/kg for AFB2, 0.1~635.3 microg/kg for AFG1, 0.1~182.5 microg/kg for AFG2, and 0.1~1,043.9 microg/kg for total aflatoxins. Most of the medicinal plant samples (721, 98.9%) were below legal limits, but 8 samples exceeded the legal limits of 10 and 15 microg/kg established by the Korean standard for AFB1 and total aflatoxins (the sum of AFB1, AFB2, AFG1 and AFG2), respectively.
Subject(s)
Aflatoxin B1 , Aflatoxins , Chromatography, Liquid , Fluorescence , Incidence , Korea , Plants, Medicinal , SeoulABSTRACT
PURPOSE: The purposes of this study are to determine the incidence of postoperative ileus after colorectal surgery, to analyze its clinical features, and to identify the risk factors for its development. METHODS: We reviewed the cases of 263 patients with mechanical ileus among 3,237 patients who underwent colorectal surgery in our clinic between June 1989 and December 2000. RESULTS: A total of 263 (8.1%) patients of postoperative ileus were documented, 193 (73.4%) cases occurred during the 1st. year. Postoperative ileus is influenced by the initial site of surgery; the rectum has more impact than the colon (P=0.028). The causes of postoperative ileus were adhesion, recurrence of cancer, and parastomal hernia. Adhesion (81.1%) was the most common cause of ileus, and cancer recurrence (18.0%) was the second. However, in postoperative ileus requiring surgery, cancer recurrence increased with time (2 year: 58.1%). The cases receiving postoperative adjuvant radiation therapy presented a significant increase in the incidence of postoperative ileus (10.3% vs 6.7% P=0.01) and in the requirement for surgical treatment (4.6% vs 2.7%, P=0.04). Patients with a temporary stoma presented a significant increase in the incidence of postoperative ileus than patients with a permanent stoma (P=0.001). The frequency of prior episodes of ileus was the strongest predictor of recurrence. CONCLUSIONS: There is a high risk of adhesion-related problems after colorectal surgery. The risk factors are associated with rectal surgery, postoperative radiation therapy, and a temporary stoma.
Subject(s)
Humans , Colon , Colorectal Surgery , Hernia , Ileus , Incidence , Intestinal Obstruction , Postoperative Complications , Rectum , Recurrence , Risk FactorsABSTRACT
No abstract available.
