Comparison of the stress distribution in base materials and thicknesses in composite resin restorations.

Resin-based composite materials are commonly used for restorations, but their dimensional changes during the polymerization could cause various clinical problems. This study evaluated the influence of a base of different materials and thicknesses on the stress magnitude and distribution in a second maxillary premolar with an MOD resin composite restoration using three-dimensional finite element analysis. A sound tooth without cavity was considered as the control group (ST), and another group was restored with composite resin without applying a base material in a MOD cavity (CR). The other three groups were restored with composite resin along with the following base materials: glass ionomer cement, low-viscosity resin, and tricalcium silicate, respectively (CR-GIC, CR-LR, and CR-TS). These three groups were further divided into two subgroups according to the thickness of the base layer: thin (0.5 mm) and thick (1.0 mm). The stress distribution was compared using the maximum principal stress after polymerization shrinkage and vertical loading with 600 N on the occlusal surface. Group ST showed the lowest stress value, and its stress propagation was confined to outer enamel surfaces only. Group CR demonstrated the highest stress distribution in the tooth-restoration interface with increased failure risk on marginal areas. The thin and thick subgroups of the three groups with a base layer had lower stress levels than Group CR. The base materials reduced the marginal stress caused by polymerization shrinkage of composite resin in MOD cavities. Different base materials and thicknesses did not affect the stress distribution.

Cytotoxicity and genotoxicity of various types of endodontic sealers in Chinese hamster ovary (CHO-K1) cells.

This study aimed to evaluate the cytotoxicity and genotoxicity of five endodontic sealers (AH Plus, MTA Fillapex, Endoseal MTA, Sealapex, and Zinc oxide eugenol) in Chinese hamster ovary cells. Cytotoxicity was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to check cell viability at 1, 3, and 7 days. Genotoxicity was assessed by cytokinesis-block micronucleus, single-cell gel electrophoresis, and γH2AX immunofluorescence assays. Cell viability of all endodontic sealers, except Endoseal MTA, on day 1 was less than 100%. Endoseal MTA showed the highest cell viability on day 7. AH Plus and Endoseal MTA showed less DNA damage than other sealers. After complete setting, AH Plus and Endoseal MTA showed low genotoxicity, which could reduce DNA damage in periapical cells, making them suitable as endodontic sealers.

Intratubular crystal formation in the exposed dentin from nano-sized calcium silicate for dentin hypersensitivity treatment.

The aim of this study is to evaluate intratubular crystal formation from the experimental material consisting of dicalcium silicate (C2S) and tricalcium silicate (C3S) with nano-scaled particle size. A total of twenty-four specimens were made by isolating 8 mm of the cervical part centered at the cementoenamel junction of extracted premolars. Twelve specimens were not treated and considered as control. The experimental material was applied to the other twelve specimens by brushing for 10,000 strokes. Each group was randomly divided into four subgroups according to the period of immersion in phosphate buffer saline (PBS) for 1, 30, 60, and 90 days each. The specimens were sectioned longitudinally and examined with scanning electron microscopy and energy dispersion X-ray spectroscopy. The intratubular crystal were formed in PBS and densely filled the dentinal tubules over time. The crystal formation occurred at a depth of more than 50 µm from the dentin surface. The Ca/P ratio of formed intratubular crystals was 1.68 after 3 months. The experimental material consisting of C2S and C3S with a nanoscale particle size can form hydroxyapatite-like crystals in dentinal tubules in PBS, and there is a possibility of reducing dentin hypersensitivity by blocking the dentinal fluid flow.

Slit-Robo expression in the leech nervous system: insights into eyespot evolution.

BACKGROUND: Slit and Robo are evolutionarily conserved ligand and receptor proteins, respectively, but the number of slit and robo gene paralogs varies across recent bilaterian genomes. Previous studies indicate that this ligand-receptor complex is involved in axon guidance. Given the lack of data regarding Slit/Robo in the Lophotrochozoa compared to Ecdysozoa and Deuterostomia, the present study aims to identify and characterize the expression of Slit/Robo orthologs in leech development. RESULTS: We identified one slit (Hau-slit), and two robo genes (Hau-robo1 and Hau-robo2), and characterized their expression spatiotemporally during the development of the glossiphoniid leech Helobdella austinensis. Throughout segmentation and organogenesis, Hau-slit and Hau-robo1 are broadly expressed in complex and roughly complementary patterns in the ventral and dorsal midline, nerve ganglia, foregut, visceral mesoderm and/or endoderm of the crop, rectum and reproductive organs. Before yolk exhaustion, Hau-robo1 is also expressed where the pigmented eye spots will later develop, and Hau-slit is expressed in the area between these future eye spots. In contrast, Hau-robo2 expression is extremely limited, appearing first in the developing pigmented eye spots, and later in the three additional pairs of cryptic eye spots in head region that never develop pigment. Comparing the expression of robo orthologs between H. austinensis and another glossiphoniid leech, Alboglossiphonia lata allows to that robo1 and robo2 operate combinatorially to differentially specify pigmented and cryptic eyespots within the glossiphoniid leeches. CONCLUSIONS: Our results support a conserved role in neurogenesis, midline formation and eye spot development for Slit/Robo in the Lophotrochozoa, and provide relevant data for evo-devo studies related to nervous system evolution.

Antiphotoaging and Skin-Protective Activities of Ardisia silvestris Ethanol Extract in Human Keratinocytes.

Ardisia silvestris is a traditional medicinal herb used in Vietnam and several other countries. However, the skin-protective properties of A. silvestris ethanol extract (As-EE) have not been evaluated. Human keratinocytes form the outermost barrier of the skin and are the main target of ultraviolet (UV) radiation. UV exposure causes skin photoaging via the production of reactive oxygen species. Protection from photoaging is thus a key component of dermatological and cosmetic products. In this research, we found that As-EE can prevent UV-induced skin aging and cell death as well as enhance the barrier effect of the skin. First, the radical-scavenging ability of As-EE was checked using DPPH, ABTS, TPC, CUPRAC, and FRAP assays, and a 3-(4-5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay was used to examine cytotoxicity. Reporter gene assays were used to determine the doses that affect skin-barrier-related genes. A luciferase assay was used to identify possible transcription factors. The anti-photoaging mechanism of As-EE was investigated by determining correlated signaling pathways using immunoblotting analyses. As-EE had no harmful effects on HaCaT cells, according to our findings, and As-EE revealed moderate radical-scavenging ability. With high-performance liquid chromatography (HPLC) analysis, rutin was found to be one of the major components. In addition, As-EE enhanced the expression levels of hyaluronic acid synthase-1 and occludin in HaCaT cells. Moreover, As-EE dose-dependently up-regulated the production of occludin and transglutaminase-1 after suppression caused by UVB blocking the activator protein-1 signaling pathway, in particular, the extracellular response kinase and c-Jun N-terminal kinase. Our findings suggest that As-EE may have anti-photoaging effects by regulating mitogen-activated protein kinase, which is good news for the cosmetics and dermatology sectors.

Complete mitochondrial genome of Aleochara (Aleochara) curtula (Goeze, 1777) (Coleoptera: Staphylinidae).

The mitochondrial genome (mitogenome) of Aleochara (Aleochara) curtula (Goeze, 1777) (Coleoptera: Staphylinidae) is reported. This mitogenome (GenBank accession no. OL675411) is 16,600 bp in size and consists of 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), and two ribosomal RNA genes (rRNA). Most PCGs use typical mitochondrial stop codon (TAR) except for cox3, which uses a single T residue. The A, G, T, and C nucleotide base composition of the mitogenome is 40.61%, 7.66%, 40.34%, and 11.39%, respectively. The phylogenetic analyses recovered the monophyly of Aleocharinae.

Evaluation of the clinical efficacy of quantitative light-induced fluorescence technology in diagnosing cracked teeth.

BACKGROUND: This retrospective study evaluated the clinical efficacy of quantitative light-induced fluorescence (QLF) technology for crack detection and the diagnosis of cracked teeth and assessed the possibility of a quantitative evaluation of cracks using QLF technology. METHODS: Patients who were clinically diagnosed with cracked teeth over a 1-year period were included. The QLF images of the corresponding symptomatic cracked teeth and asymptomatic contralateral teeth with crack lines were taken with Qraypen C (AIOBIO, Seoul, Korea). Fluorescence loss (ΔF), maximum fluorescence loss (ΔFmax), red fluorescence (ΔR), and maximum red fluorescence (ΔRmax) of the crack line were analyzed. The correlation between these parameters and sex, age, tooth position (1st premolar, 2nd premolar, 1st molar, 2nd molar), spontaneous pain (+/-), percussion test (+/-), cold test (++/+/-), and bite test (+/-) were statistically analyzed. RESULTS: A total of 66 patients were included. Twenty-four patients had asymptomatic contralateral teeth with apparent crack lines; thus, 90 teeth were analyzed. The crack lines in 84 teeth observed as red fluorescent lines on the QLF images showed ΔR values higher than the cut-off value set by the analysis program used. The patient's age and the ∣ΔF∣ and ΔR values were positively correlated. However, there was no statistically significant difference in the QLF parameters between the same patient's symptomatic tooth and the contralateral tooth. CONCLUSIONS: QLF technology is a useful assistive diagnostic device for diagnosing cracked teeth.

Development of markers using microsatellite loci of two rove beetle species, Paederus fuscipes Curtis and Aleochara (Aleochara) curtula Goeze (Coleoptera: Staphylinidae), followed by analyses of genetic diversity and population structure.

BACKGROUND: The family Staphylinidae is the most speciose beetle group in the world. The outbreaks of two staphylinid species, Paederus fuscipes and Aleochara (Aleochara) curtula, were recently reported in South Korea. None of research about molecular markers and genetic diversity have been conducted in these two species. OBJECTIVE: To develop microsatellite markers and analyze the genetic diversity and population structures of two rove beetle species. METHODS: NGS was used to sequence whole genomes of two species, Paederus fuscipes and Aleochara (Aleochara) curtula. Microsatellite loci were selected with flanking primer sequences. Specimens of P. fuscipes and A. curtula were collected from three localities, respectively. Genetic diversity and population structure were analyzed using the newly developed microsatellite markers. RESULTS: The number of alleles ranged 5.727-6.636 (average 6.242) and 2.182-5.364 (average 4.091), expected heterozygosity ranged 0.560-0.582 (average 0.570) and 0.368-0.564 (average 0.498), observed heterozygosity ranged 0.458-0.497 (average 0.472) and 0.418-0.644 (average 0.537) in P. fuscipes and A. curtula, respectively. Population structure indicates that individuals of A. curtula are clustered to groups where they were collected, but those of P. fuscipes are not. CONCLUSION: Population structures of P. fuscipes were shallow. In A. curtula, however, it was apparent that the genetic compositions of the populations are different significantly depending on collection localities.

Callerya atropurpurea suppresses inflammation in vitro and ameliorates gastric injury as well as septic shock in vivo via TLR4/MyD88-dependent cascade.

BACKGROUND: Callerya atropurpurea is a traditional plant in a tropical zone discovered to have anti-inflammatory functions. PURPOSE: we want to investigate the mechanism related to anti-inflammation of C. atropurpurea ethanol extract (Ca-EE) both in vitro and in vivo. STUDY DESIGN: Murine macrophage cells and mouse models for gastritis and septic shock were conducted to evaluate the abilities of Ca-EE in anti-inflammation. METHODS: Ca-EE was tested by HPLC and LC-MS/MS. NO outcome was checked by Griess reagent test. Cell viabilities were evaluated using MTT assay. Inflammatory cytokines were determined via RT-PCR and ELISA. The mechanism of Ca-EE in anti-inflammation was investigated by luciferase reporter gene assay and immunoblot in transcription level and protein level respectively. Gastric injury and septic shock administrated with Ca-EE were studied by H&E, PCR, and immunoblot. RESULTS: Ca-EE significantly decreased LPS-induced NO production, but hardly stimulated the expression of NO itself. It not only showed no cytotoxicity, but also protected cells from LPS damage. Moreover, Ca-EE decreased TLR4 expression, altered MyD88 recruitment and TRAF6, and suppressed the phospho-Src/PI3K/AKT. Ca-EE inhibited downstream signaling P38, JNK and NF-κB. Finally, Ca-EE alleviated HCl/EtOH-induced gastritis and LPS/poly (I:C)-induced septic shock through the previously mentioned signaling cascades. CONCLUSION: Ca-EE exhibited an integrated and promising mechanism against TLR4-related inflammation, which shows potential for treating gastritis, septic shock, and other inflammatory diseases.

The complete mitochondrial genome and phylogenetic analysis of Euurobracon yokahamae (Hymenoptera: Braconidae).

Euurobracon yokahamae is a parasitoid wasp found solely in Asia, and is endangered in some countries. The complete mitochondrial DNA sequence of E. yokahamae was sequenced using next-generation sequencing (NGS). The mitogenome of this species is 14,974bp long and encodes for 13 protein-coding genes (PCGs), 22 transfer RNAs, and 2 ribosomal RNAs. Maximum likelihood phylogenetic analysis of the mitochondrial genome of braconid species was performed. Tree topology showed that E. yokahamae was closely related to another species of the same genus.

Cocculus hirsutus ameliorates gastric and lung injuries by suppressing Src/Syk.

BACKGROUND: Cocculus hirsutus (L.) W. Thedo., a traditionally well-known plant, has confirmed antitumor properties as well as acute and chronic diuretic effects. However, little is known about its inflammatory activities and the potential effect on inflammatory disease treatment. PURPOSE: Our aim in this study was to explore additional beneficial properties of C. hirsutus ethanol extract (Ch-EE) such as anti-inflammatory activity in vitro and in vivo as well as its underlying mechanisms and to provide a theoretical basis for its role as a candidate natural drug in clinical gastritis and lung disease therapy. STUDY DESIGN: RAW264.7 cells, HEK293T cells, peritoneal macrophages, and mouse models of acute gastritis and acute lung injury were used to assess the anti-inflammatory activity of Ch-EE. METHODS: Decreases in LPS-induced nitric oxide (NO) production and cytokine expression by RAW264.7 cells after Ch-EE treatment were evaluated by Griess assays and PCR, respectively. Transcription factor activity was assessed through luciferase reporter gene assay, and protein expression was determined by Western blotting analysis. Overexpression assays and cellular thermal shift assays were executed in HEK293T cells. Our two in vivo models were an HCl/EtOH-induced gastritis model and an LPS-induced lung injury model. Changes in stomach lesions, lung edema, and lung histology were examined upon treatment with Ch-EE. Components of Ch-EE were determined by liquid chromatography-mass spectrometry. RESULTS: LPS-induced nitric oxide production and Pam3CSK4- and L-NAME-induced NO production were inhibited by Ch-EE treatment of RAW264.7 cells. Furthermore, LPS-induced increases in transcript levels of iNOS, COX2, CCL12, and IL-1ß were reduced by Ch-EE treatment. Ch-EE decreased both MyD88- and TRIF-induced NF-κB promotor activity. Proteins upstream of NF-κB, namely p-p50, p-p65, p-IκBα, p-AKT1, p-Src, and p-Syk, were all downregulated by Ch-EE. Moreover, Src and Syk were targets of Ch-EE. Ch-EE treatment reduced the size of inflammatory stomach lesions induced by HCl/EtOH, lung edema, and accumulation of activated neutrophils caused by LPS. CONCLUSIONS: These results strongly suggest that Cocculus hirsutus can be developed as a promising anti-inflammatory remedy with Src- and Syk-inhibitory functions targeting diseases related to gastritis and lung injury.

Cissus subtetragona Planch. Ameliorates Inflammatory Responses in LPS-induced Macrophages, HCl/EtOH-induced Gastritis, and LPS-induced Lung Injury via Attenuation of Src and TAK1.

Several Cissus species have been used and reported to possess medicinal benefits. However, the anti-inflammatory mechanisms of Cissus subtetragona have not been described. In this study, we examined the potential anti-inflammatory effects of C. subtetragona ethanol extract (Cs-EE) in vitro and in vivo, and investigated its molecular mechanism as well as its flavonoid content. Lipopolysaccharide (LPS)-induced macrophage-like RAW264.7 cells and primary macrophages as well as LPS-induced acute lung injury (ALI) and HCl/EtOH-induced acute gastritis mouse models were utilized. Luciferase assays, immunoblotting analyses, overexpression strategies, and cellular thermal shift assay (CETSA) were performed to identify the molecular mechanisms and targets of Cs-EE. Cs-EE concentration-dependently reduced the secretion of NO and PGE2, inhibited the expression of inflammation-related cytokines in LPS-induced RAW264.7 cells, and decreased NF-κB- and AP-1-luciferase activity. Subsequently, we determined that Cs-EE decreased the phosphorylation events of NF-κB and AP-1 pathways. Cs-EE treatment also significantly ameliorated the inflammatory symptoms of HCl/EtOH-induced acute gastritis and LPS-induced ALI mouse models. Overexpression of HA-Src and HA-TAK1 along with CETSA experiments validated that inhibited inflammatory responses are the outcome of attenuation of Src and TAK1 activation. Taken together, these findings suggest that Cs-EE could be utilized as an anti-inflammatory remedy especially targeting against gastritis and acute lung injury by attenuating the activities of Src and TAK1.

Folate-receptor-targeted NIR-sensitive polydopamine nanoparticles for chemo-photothermal cancer therapy.

We propose the use of folate-receptor-targeted, near-infrared-sensitive polydopamine nanoparticles (NPs) for chemo-photothermal cancer therapy as an enhanced type of drug-delivery system which can be synthesized by in situ polymerization and conjugation with folic acid. The NPs consist of a Fe3O4/Au core, coated polydopamine, conjugated folic acid, and loaded anti-cancer drug (doxorubicin). The proposed multifunctional NPs show many advantages for therapeutic applications such as good biocompatibility and easy bioconjugation. The polydopamine coating of the NPs show a higher photothermal effect and thus more effective cancer killing compared to Fe3O4/Au nanoparticles at the same intensity as near-infrared laser irradiation. In addition, the conjugation of folic acid was shown to enhance cancer cellular uptake efficiency via the folate receptor and thus improve chemotherapeutic efficiency. Through in vitro cancer cell treatment testing, the proposed multifunctional NPs showed advanced photothermal and chemotherapeutic performance. Based on these enhanced anti-cancer properties, we expect that the proposed multifunctional NPs can be used as a drug-delivery system in cancer therapy.

A Magnetically Actuated Microscaffold Containing Mesenchymal Stem Cells for Articular Cartilage Repair.

This study proposes a magnetically actuated microscaffold with the capability of targeted mesenchymal stem cell (MSC) delivery for articular cartilage regeneration. The microscaffold, as a 3D porous microbead, is divided into body and surface portions according to its materials and fabrication methods. The microscaffold body, which consists of poly(lactic-co-glycolic acid) (PLGA), is formed through water-in-oil-in-water emulsion templating, and its surface is coated with amine functionalized magnetic nanoparticles (MNPs) via amino bond formation. The porous PLGA structure of the microscaffold can assist in cell adhesion and migration, and the MNPs on the microscaffold can make it possible to steer using an electromagnetic actuation system that provides external magnetic fields for the 3D locomotion of the microscaffold. As a fundamental test of the magnetic response of the microscaffold, it is characterized in terms of the magnetization curve, velocity, and 3D locomotion of a single microscaffold. In addition, its function with a cargo of MSCs for cartilage regeneration is demonstrated from the proliferation, viability, and chondrogenic differentiation of D1 mouse MSCs that are cultured on the microscaffold. For the feasibility tests for cartilage repair, 2D/3D targeting of multiple microscaffolds with the MSCs is performed to demonstrate targeted stem cell delivery using the microscaffolds and their swarm motion.

Korean species of the genus Neoperla Needham, 1905 (Plecoptera: Perlidae).

Two new species of the perlid genus Neoperla, N. adamantea Murányi & Li, sp. n. and N. goguryeo Murányi & Li, sp. n. are described from both sexes, collected in the Kumgang and Myohyang Mountains of North Korea by personnel of the Hungarian Natural History Museum during the 1970's. Two additional species, N. coreensis Ra, Kim, Kang & Ham, 1994 and N. ussurica Sivec & Zhiltzova, 1996 are redescribed from North Korean specimens. Neoperla quadrata Wu & Claassen, 1934 is redescribed on the basis of the holotype from Sichuan Province, China, and the Korean occurrence of this species requires confirmation. Korean and global distributions of the five Neoperla species reported from the peninsula are depicted on maps.

Korean species of the genus Perlomyia Banks, 1906 (Plecoptera: Leuctridae).

Eight species of the genus Perlomyia (Plecoptera: Leuctridae) are reported from Korea, six are new records for the Korean Peninsula. Two species, known only as unassociated females are described under informal unnamed designations. Previous Korean records are discussed, taxonomic characters of the Korean specimens and Korean distribution are presented for these species. Zoogeographic notes on the Asian Perlomyia are also given.

Phylogeny, biogeography and the stepwise evolutionary colonization of intertidal habitat in the Liparocephalini based on morphological and molecular characters (Coleoptera: Staphylinidae: Aleocharinae).

A phylogenetic analysis of the tribe Liparocephalini Fenyes is presented based on morphological and molecular characters. The data set comprised 50 adult morphological characters, partial COI (907 bp), COII (366 bp) and 12S rDNA (325-355 bp), and nearly complete sequences of 18S rDNA (1768-1902 bp) for 21 species. Eighteen species of liparocephaline beetles from all eight genera and three outgroups, are included. The sequences were analysed separately and simultaneously with morphological characters by direct optimization in the program POY4 and by partitioned Bayesian analysis for the combined data. The direct optimization (DO) tree for the combined data under equal weighting, which also shows a minimum incongruence length difference value, resulted in a monophyletic Liparocephalini with the following patterns of phylogenetic relationships (outgroup ((Baeostethus, Ianmoorea) (Paramblopusa ((Amblopusa, Halorhadinus) (Liparocephalus, Diaulota))))). A sensitivity analysis using 16 different parameter sets for the combined data shows the monophyly of the liparocephalines and all its genera under all parameter sets. Bayesian analysis resulted in topological differences in comparison with the DO tree under equal weighting only in the position of the genus Paramblopusa and clade (Amblopusa + Halorhadinus), which were reversed. Historical biogeography and the stepwise evolutionary colonization of intertidal habitat in the Liparocephalini are discussed. Based on the biogeographical analyses, we hypothesize that the ancestor of the Liparocephalini occurred along the Panthallassan Ocean, the direct antecedent of the Pacific Ocean, followed by repeated dispersals to the Nearctic from the Palearctic. We also hypothesize that ancestors of the Liparocephalini appear to have arisen in the littoral zone of beaches and then colonized rocky reef areas in the low tidal zone later through high- to mid-tide zones. © The Willi Hennig Society 2009.

The anti-arthritic effect of ursolic acid on zymosan-induced acute inflammation and adjuvant-induced chronic arthritis models.

Ursolic acid (UA) is pentacyclic triterpenoic acid that naturally occurs in many medicinal herbs and plants. In this study, we examined the possible suppressive effect of UA extracted from Oldenlandia diffusa on zymosan-induced acute inflammation in mice and complete Freund's adjuvant (CFA)-induced arthritis in rats. UA treatment (per oral) dose-dependently (25-200 mg kg(-1)) suppressed zymosan-induced leucocyte migration and prostaglandin E2 (PGE(2)) production in the air pouch exudates. Since the maximal effective dose of UA was 50 mg kg(-1) in the zymosan experiment, we used this dose of UA in a subsequent study using an adjuvant-induced rheumatoid arthritis model. UA treatment (50 mg kg(-1), per oral, once a day for 10 days) was started from day 12 after adjuvant injection. UA dramatically inhibited paw swelling, plasma PGE(2) production and radiological changes in the joint caused by CFA injection. Moreover, UA significantly suppressed the arthritis-induced mechanical and thermal hyperalgesia as well as the spinal Fos expression, as determined by immunohistochemistry, which was increased by CFA injection. In addition, overall anti-arthritic potency of UA was comparable with ibuprofen (100 mg kg(-1), oral) while UA did not induce significant gastric lesions as compared with the ibuprofen treatment group. These findings strongly suggest that UA is a useful suppressive compound for rheumatoid arthritis treatment with low risk of gastric problems.

[Resolution of ABO Discrepancies by ABO Genotyping.].

BACKGROUND: Before a blood transfusion, both red cell and serum typing need to be matched for ABO tests on the donor and patient (recipient). When a mismatch exists in the tests, additional ABO genotyping and serological tests are required for the resolution of the discrepancy. We performed ABO genotyping on a series of blood donors and patients with ABO discrepancies to assist in resolving their blood groups. METHODS: We examined 46 samples with ABO discrepancies from a random pool of donors recruited at Gwangju-Chonnam Red Cross Blood Center and from patients at Chonnam National University Hospital between May 2004 and July 2005. ABO genotyping was performed on all samples with an allele specific polymerase chain reaction for differentiation of A, B,O, cis-AB, A(var) (784 G>A), and B(var) (547 G>A) alleles; routine serologic tests were also performed. Exon 6 and 7 of ABO gene from five samples were sequenced. RESULTS: The genotypes of 18 donors/patients with weakened A or B antigen expressions consisted of 4 cases of cis-AB/O (3 A(2)B(3), 1 A(2)B); 5 cases of cis-AB/A (5 A(1)B(x or el)); 2 cases of A/O (1 O, 1 A(m or x)); 1 case of B/O (1 B(m or x)); 4 cases of A/B (1 A(2)B , 1 A(1)B(x or el), 2 A(1)B(3)); and 2 cases of A(var)/B (2 A(w)B). On the other hand, the genotypes of 28 samples with unexpected serum reactions included 18 cases of A/O (16 A(1), 2 A(int)); 7 cases of A/A (5 A(1), 1 A(1)B(x or el), 1 A(1)B(w)); and 3 cases of O/O (1 O, 2 B(w)). CONCLUSIONS: ABO genotyping is useful for differentiating the ABO discrepancies that were difficult to resolve by serological tests. The most frequent unusual red cell reactions were weak A and B antigen expressions, which were resulted from the ABO subgroup alleles including cis-AB allele, whereas the most frequent unusual serum reactions were caused by decreased anti-B titers.