PURPOSE: To establish the immortalized human corneal endothelial cell line (IHCEn) by transducing human papilloma virus (HPV) 16 E6/E7 oncogenes, and to identify their characteristics when cultivated on a lyophilized human amniotic membrane (LAM). METHODS: Primary human corneal endothelial cells (PHCEn) were infected using a retroviral vector with HPV 16 E6/E7, and transformed cells were clonally selected by G418. Growth properties and characteristics of IHCEn were compared with PHCEn by cell counting and RT-PCR of VDAC3, SLC4A4, CLCN3, FGF-1, Col IV, and Na+/K+ ATPase. IHCEn were cultured on LAM. Messenger RNA expressions of VDAC3, CLCN3, and Na+/K+ ATPase, and protein expressions of Na+/K+ ATPase and Col IV in IHCEn cultivated on LAM were investigated by RT-PCR, immunofluorescence, and immunohistochemical staining, respectively. RESULTS: Successful immortalization was confirmed by stable expression of HPV 16 E6/E7 mRNA by RT-PCR, and IHCEn exhibited typical corneal endothelial morphology. Doubling time of IHCEn was 30.15+/-10.96 hrs. Both IHCEn and PHCEn expressed VDAC3, CLCN3, SLC4A4, FGF-1, Col IV, and Na+/K+ ATPase. IHCEn cultivated on LAM showed stronger expression of VDAC3, CLCN4, and Na+/K+ ATPase mRNA than on plastic culture dish. Immunohistochemical staining and immunofluorescence revealed the positive expression of Na+/K+ ATPase and Col IV. CONCLUSIONS: IHCEn were successfully established, and LAM is a good substrate for the culture of human corneal endothelial cells.
Transfection ; Reverse Transcriptase Polymerase Chain Reaction ; Repressor Proteins/genetics/*pharmacology ; RNA, Messenger/genetics ; Protein-Tyrosine Kinases ; Oncogene Proteins, Viral/genetics/*pharmacology ; Immunohistochemistry ; Humans ; Gene Expression Regulation, Viral ; Freeze Drying ; Endothelium, Corneal/*cytology/drug effects/metabolism ; Cell Line, Transformed ; Cell Count ; Amnion

OBJECTIVES: The molecular mechanisms underlying attention-deficit hyperactivity disorder (ADHD) remain unclear. Therefore, this study aimed to identify the genetic susceptibility loci for ADHD in Korean children with ADHD. We performed a case-control and a family-based genome-wide association study (GWAS), as well as genome-wide quantitative trait locus (QTL) analyses, for two symptom traits. METHODS: A total of 135 subjects (71 cases and 64 controls), for the case-control analysis, and 54 subjects (27 probands and 27 unaffected siblings), for the family-based analysis, were included. RESULTS: The genome-wide QTL analysis identified four single nucleotide polymorphisms (SNPs) (rs7684645 near APELA, rs12538843 near YAE1D1 and POU6F2, rs11074258 near MCTP2, and rs34396552 near CIDEA) that were significantly associated with the number of inattention symptoms in ADHD. These SNPs showed possible association with ADHD in the family-based GWAS, and with hyperactivity-impulsivity in genome-wide QTL analyses. Moreover, association signals in the family-based QTL analysis for the number of inattention symptoms were clustered near genes IL10, IL19, SCL5A9, and SKINTL. CONCLUSION: We have identified four QTLs with genome-wide significance and several promising candidates that could potentially be associated with ADHD (CXCR4, UPF1, SETD5, NALCN-AS1, ERC1, SOX2-OT, FGFR2, ANO4, and TBL1XR1). Further replication studies with larger sample sizes are needed.
Adolescent* ; Case-Control Studies ; Child ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans ; Interleukin-10 ; Polymorphism, Single Nucleotide ; Quantitative Trait Loci ; Sample Size

PURPOSE: This study was performed to evaluate the effectiveness of conventional chest radiography and abdominal CT for early detection of pulmonary metastases after curative surgery for colorectal cancer. METHODS: We retrospectively reviewed 138 cases of pulmonary metastases from a group of colorectal-cancer patients, who were recruited from 1994 to 2002 at Samsung Medical Center, Sungkyunkwan University School of Medicine, and who had been surgically treated with a curative resection. RESULTS: The detection rates for pulmonary metastases were 34.1% by conventional chest radiography, 50.0% by abdominal CT, and 15.9% by other means. For stage I and II tumors, conventional chest radiography was superior to abdominal CT (45.7% vs. 34.3%, P<0.05) for detecting pulmonary metastases. On the contrary, for stage III tumors, abdominal CT was superior to conventional chest radiography (55.3% vs. 30.1%, P<0.05). Compared with stage I and II, pulmonary metastases in stage III had a tendency to be more numerous, bilateral, and extra-pulmonary. They also had a low detection rate by conventional chest radiography and a higher detection rate by abdominal CT, and they were associated with poor survival. CONCLUSIONS: Conventional chest radiography is no more useful in detecting early pulmonary metastases after curative colorectal surgery than abdominal CT, especially for stage III tumors. We propose the use of routine chest CT or extended abdominal CT for screening of occult lung metastases in stage III colorectal cancer patients.
Colorectal Neoplasms* ; Colorectal Surgery ; Humans ; Lung ; Mass Screening ; Neoplasm Metastasis* ; Radiography ; Retrospective Studies ; Thorax* ; Tomography, X-Ray Computed

PURPOSE: The aim of this study was to compare the clinical characteristics between hepatic resection and radiofrequency ablation (RFA) in hepatic metastases of colorectal cancer. METHODS: Among 183 patients who were diagnosed as having colorectal cancer with hepatic metastases from May 1999 to Dec. 2002, excluding 56 patients who did not undergo a hepatic resection or RFA due to multiple hepatic metastases or other distant metastases, 127 patients who were treated with a pure hepatic resection (N=68), pure RFA (N=35), or a hepatic resection with RFA (N=24) synchronous or metachronous were reviewed in this study. The study included metastatic hepatic tumor size, number, distribution, disease-free survival rate, and overall survival rate. RESULTS: The mean hepatic tumor sizes in the resection group, the RFA group, and the resection with RFA group were 3.3 cm, 3.0 cm, and 2.5 cm, respectively, but the differences in the sizes had no statistical significance (P>0.1). In the view of the number of hepatic metastases, single metastases were the most prevalent kind in the resection group and the RFA group (64.7% and 60.0%) while multiple metastases were the most prevalent kind in the resection with RFA (20/24, 83.3%). In the resection and the RFA groups, a unilobar distribution was the most common (88.2% and 68.6%), but a bilobar distribution was the most common (87.5%) in the resection with RFA group. The disease-free survival rates were 42.2% (resection group), 30.7% (RFA group), and 22.2% (resection with RFA group) in the third year (P=0.65). The overall survival rates were 70.9% (resection group), 68.4% (RFA group), and 62.9% (resection with RFA group) in the third year (P=0.19). CONCLUSIONS: There were no significant statistical differences in the disease-free survival and the overall survival rates between the three groups. Radiofrequency ablation (RFA) is considered as not only a complementary but also an alternative treatment tool to hepatic resection in the treatment of hepatic metastases of colorectal cancer and has a similar survival rate.
Catheter Ablation* ; Colorectal Neoplasms* ; Disease-Free Survival ; Humans ; Neoplasm Metastasis* ; Survival Rate

Irbesartan is a new selective angiotensin II subtype 1 receptor antagonist. We evaluated the efficacy and tolerability of irbesartan in patients with mild to moderate hypertension and renal disease. On 24 hypertensive patients, oral irbesartan 150mg a day was administered. In cases whose seated diastolic blood pressure did not decrease to 85mmHg after treatment for 4 weeks, the dose of irbesartan was increased to 300mg per day. Every 4 weeks, blood pressure, heart rates, and adverse effects were monitored. And we assessed WBC counts, hemoglobin, hematocrits, platelets, creatinine, BUN, total protein, albumin, fasting blood sugar, total cholesterol, AST, ALT, alkaline phosphatase, total bilirubin, sodium, potassium, calcium, uric acid and urine protein/creatinine ratio to evaluate the change of renal and hepatic function and other adverse effects. Seated systolic blood pressure was decreased from 157.1+/-3.1mmHg to 135.5+/-3.7mmHg, and seated diastolic blood pressure was also decreased from 99.2+/-1.7mmHg to 84.3+/-2.5mmHg. Irbesartan was effective in lowering blood pressure in 20 among 24 patients, and the effective rate of this drug was 83.3%. After treatment, a non clinically significant increase of heart rates and statistically significant decrease of total cholesterol level were noted. There was no dose-related adverse effect. We conclude that irbesartan is a safe and effective angiotensin II subtype 1 receptor antagonist for lowering blood pressure in patients with mild to moderate hypertension and renal disease.
Alkaline Phosphatase ; Angiotensin II ; Bilirubin ; Blood Glucose ; Blood Pressure ; Calcium ; Cholesterol ; Creatinine ; Fasting ; Heart Rate ; Hematocrit ; Humans ; Hypertension* ; Potassium ; Sodium ; Uric Acid

BACKGROUND AND OBJECTIVES: Patients with Kawasaki disease (KD) are clinically heterogeneous because its diagnosis is based solely on clinical observation and there are no definitive biomarkers. We dissected the clinical heterogeneity of KD patients using the KD-associated genetic variants. METHODS: We performed a genetic association analysis in several KD subgroups categorized by clinical characteristics using the KD-associated variants of the B lymphoid tyrosine kinase (BLK; rs6993775) and Fc gamma receptor II a (FCGR2A; rs1801274) in a large number of case (n=1,011) and control (n=4,533) samples. RESULTS: BLK and FCGR2A were very significantly associated with KD in Korean KD patients (odds ratio [OR],1.48; p=4.63×10⁻¹¹ for BLK, and OR, 1.26; p=1.42×10⁻⁴ for FCGR2A). However, in KD subgroup analysis, we found that neither BLK nor FCGR2A were associated with either incomplete Kawasaki disease (iKD) type patients or those older than 5 years of age (p>0.2), suggesting that patients with iKD or those older than 5 years of age are a unique subgroup of KD. In genetic association analysis after excluding iKD patients and those older than 5 years old, we found that BLK was associated with all KD subgroups, whereas FCGR2A was specifically associated with male KD patients younger than 1 year of age (OR, 2.22; p=2.35×10⁻⁵). CONCLUSIONS: KD is a clinically and genetically heterogeneous disease. These findings will provide new insights into the clinical and genetic heterogeneity of KD.
Biomarkers ; Diagnosis ; Genetic Heterogeneity ; Genome-Wide Association Study ; Humans ; Male ; Mucocutaneous Lymph Node Syndrome ; Polymorphism, Single Nucleotide ; Population Characteristics ; Protein-Tyrosine Kinases

Purpose: Platelet function test (PFT) results and genotype hold unique prognostic implications in East Asian patients. The aim of the PTRG-DES (Platelet function and genoType-Related long-term proGnosis in Drug-Eluting Stent-treated Patients with coronary artery disease) consortium is to assess the clinical impact thereof on long-term clinical outcomes in Korean patients with coronary artery disease during dual antiplatelet therapy (DAPT) including clopidogrel. Materials and Methods: Searching publications on the PubMed, we reviewed clopidogrel treatment studies with PFT and/or genotype data for potential inclusion in this study. Lead investigators were invited to share PFT/genotype results, patient characteristics, and clinical outcomes to evaluate relationships among them. Results: Nine registries from 32 academic centers participated in the PTRG-DES consortium, contributing individual patient data from 13160 patients who underwent DES implantation between July 2003 and August 2018. The PTRG-PFT cohort was composed of 11714 patients with available VerifyNow assay results. Platelet reactivity levels reached 218±79 P2Y12 reaction units (PRU), and high on-clopidogrel platelet reactivity based on a consensus-recommended cutoff (PRU >208) was observed in 55.9%. The PTRGGenotype cohort consisted of 8163 patients with candidate genotypes related with clopidogrel responsiveness. Of those with cytochrome P450 (CYP) 2C19 genotype, frequencies of carrying one and two loss-of-function allele (s) (*2 or *3) were 47.9% (intermediate metabolizers) and 14.2% (poor metabolizers), respectively. Conclusion The PTRG-DES consortium highlights unique values for on-clopidogrel platelet reactivity and CYP2C19 phenotype that may be important to developing optimal antiplatelet regimens in East Asian patients.

BACKGROUND AND OBJECTIVES: Patients with Kawasaki disease (KD) are clinically heterogeneous because its diagnosis is based solely on clinical observation and there are no definitive biomarkers. We dissected the clinical heterogeneity of KD patients using the KD-associated genetic variants. METHODS: We performed a genetic association analysis in several KD subgroups categorized by clinical characteristics using the KD-associated variants of the B lymphoid tyrosine kinase (BLK; rs6993775) and Fc gamma receptor II a (FCGR2A; rs1801274) in a large number of case (n=1,011) and control (n=4,533) samples. RESULTS: BLK and FCGR2A were very significantly associated with KD in Korean KD patients (odds ratio [OR],1.48; p=4.63×10⁻¹¹ for BLK, and OR, 1.26; p=1.42×10⁻⁴ for FCGR2A). However, in KD subgroup analysis, we found that neither BLK nor FCGR2A were associated with either incomplete Kawasaki disease (iKD) type patients or those older than 5 years of age (p>0.2), suggesting that patients with iKD or those older than 5 years of age are a unique subgroup of KD. In genetic association analysis after excluding iKD patients and those older than 5 years old, we found that BLK was associated with all KD subgroups, whereas FCGR2A was specifically associated with male KD patients younger than 1 year of age (OR, 2.22; p=2.35×10⁻⁵). CONCLUSIONS KD is a clinically and genetically heterogeneous disease. These findings will provide new insights into the clinical and genetic heterogeneity of KD.

Background: Coronary artery disease patients undergoing percutaneous coronary intervention (PCI) often exhibit reduced left ventricular ejection fraction (LVEF). However, the impact of LV dysfunction status in conjunction with platelet reactivity on clinical outcomes has not been previously investigated. Methods: From the multicenter PTRG-DES (Platelet function and genoType-Related long-term prognosis in DES-treated patients) consortium, the patients were classified as preserved-EF (PEF: LVEF ≥ 50%) and reduced-EF (REF: LVEF< 5 0%) group by echocardiography. Platelet reactivity was measured using VerifyNow P2Y 12 assay and high platelet reactivity (HPR) was defined as PRU ≥ 252. The major adverse cardiac and cerebrovascular events (MACCEs) were a composite of death, myocardial infarction, stent thrombosis and stroke at 5 years after PCI. Major bleeding was defined as Bleeding Academic Research Consortium bleeding types 3–5. Results: A total of 13,160 patients from PTRG-DES, 9,319 (79.6%) patients with the results of both PRU and LVEF were analyzed. The incidence of MACCE and major bleeding was higher in REF group as compared with PEF group (MACCEs: hazard ratio [HR] 2.17, P < 0.001, 95% confidence interval [CI] 1.85–2.55; major bleeding: HR 1.78, P < 0.001, 95% CI 1.39–2.78).The highest rate of MACCEs was found in patients with REF and HPR, and the difference between the groups was statistically significant (HR 3.14 in REF(+)/HPR(+) vs. PEF(+)/HPR(-) group,P <0.01, 95% CI 2.51–3.91). The frequency of major bleeding was not associated with the HPR in either group. Conclusion LV dysfunction was associated with an increased incidence of MACCEs and major bleeding in patients who underwent PCI. The HPR status further exhibited significant increase of MACCEs in patients with LV dysfunction in a large, real-world registry.Trial Registration: ClinicalTrials.gov Identifier: NCT04734028