ABSTRACT
Viral load and the duration of viral shedding of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are important determinants of the transmission of coronavirus disease 2019.In this study, we examined the effects of viral doses on the lung and spleen of K18-hACE2 transgenic mice by temporal histological and transcriptional analyses. Approximately, 1×105 plaque-forming units (PFU) of SARS-CoV-2 induced strong host responses in the lungs from 2 days post inoculation (dpi) which did not recover until the mice died, whereas responses to the virus were obvious at 5 days, recovering to the basal state by 14 dpi at 1×102 PFU. Further, flow cytometry showed that number of CD8+ T cells continuously increased in 1×102 PFU-virusinfected lungs from 2 dpi, but not in 1×105 PFU-virus-infected lungs. In spleens, responses to the virus were prominent from 2 dpi, and number of B cells was significantly decreased at 1×105PFU; however, 1×102 PFU of virus induced very weak responses from 2 dpi which recovered by 10 dpi. Although the defense responses returned to normal and the mice survived, lung histology showed evidence of fibrosis, suggesting sequelae of SARS-CoV-2 infection. Our findings indicate that specific effectors of the immune response in the lung and spleen were either increased or depleted in response to doses of SARS-CoV-2. This study demonstrated that the response of local and systemic immune effectors to a viral infection varies with viral dose, which either exacerbates the severity of the infection or accelerates its elimination.
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This case report presents the successful endoscopic submucosal dissection (ESD) of a well-differentiated esophageal liposarcoma in a 51-year-old male with persistent dysphagia. The cause was initially diagnosed as a 10 cm pedunculated lesion extending from the upper esophageal sphincter to the mid-esophagus. An ESD was chosen over traditional surgery because it is less invasive. The procedure involved a precise submucosal injection and excision with special techniques to manage bleeding from a central vessel.Despite the extraction challenges owing to the size of the lesion, it was successfully removed orally. A histopathological examination of the 8.3×4.2×2.3 cm specimen revealed the characteristic features of a well-differentiated liposarcoma, including MDM2 and CDK4 positivity. The follow-up revealed no recurrence, and active surveillance has been performed since. This report highlights the versatility of ESD in treating significant esophageal tumors and provides evidence for its efficacy as a minimally invasive alternative.
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Background@#To date, various genotypes of infectious bronchitis virus (IBV) have cocirculated and in Korea, GI-15 and GI-19 lineages were prevailing. The spike protein, particularly S1 subunit, is responsible for receptor binding, contains hypervariable regions and is also responsible for the emerging of novel variants. @*Objective@#This study aims to investigate the putative major amino acid substitutions for the variants in GI-19. @*Methods@#The S1 sequence data of IBV isolated from 1986 to 2021 in Korea (n = 188) were analyzed. Sequence alignments were carried out using Multiple alignment using Fast Fourier Transform of Geneious prime. The phylogenetic tree was generated using MEGA-11 (ver.11.0.10) and Bayesian analysis was performed by BEAST v1.10.4. Selective pressure was analyzed via online server Datamonkey. Highlights and visualization of putative critical amino acid were conducted by using PyMol software (version 2.3). @*Results@#Most (93.5%) belonged to the GI-19 lineage in Korea, and the GI-19 lineage was further divided into seven subgroups: KM91-like (Clade A and B), K40/09-like, QX-like (I-IV).Positive selection was identified at nine and six residues in S1 for KM91-like and QX-like IBVs, respectively. In addition, several positive selection sites of S1-NTD were indicated to have mutations at common locations even when new clades were generated. They were all located on the lateral surface of the quaternary structure of the S1 subunits in close proximity to the receptor-binding motif (RBM), putative RBM motif and neutralizing antigenic sites in S1. @*Conclusions@#Our results suggest RBM surrounding sites in the S1 subunit of IBV are highly susceptible to mutation by selective pressure during evolution.
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Background@#The Omicron variant has become the most prevalent SARS-CoV-2 variant. Omicron is known to induce milder lesions compared to the original Wuhan strain. Fatal infection of the Wuhan strain into the brain has been well documented in COVID-19 mouse models and human COVID-19 cases, but apparent infections into the brain by Omicron have not been reported in human adult cases or animal models. In this study, we investigated whether Omicron could spread to the brain using K18-hACE2 mice susceptible to SARS-CoV-2 infection. @*Results@#K18-hACE2 mice were intranasally infected with 1 × 105 PFU of the original Wuhan strain and the Omicron variant of SARS-CoV-2. A follow-up was conducted 7 days post infection. All Wuhan-infected mice showed > 20% body weight loss, defined as the lethal condition, whereas two out of five Omicron-infected mice (40%) lost > 20% body weight. Histopathological analysis based on H&E staining revealed inflammatory responses in the brains of these two Omicron-infected mice. Immunostaining analysis of viral nucleocapsid protein revealed severe infection of neuron cells in the brains of these two Omicron-infected mice. Lymphoid depletion and apoptosis were observed in the spleen of Omicron-infected mice with brain infection. @*Conclusion@#Lethal conditions, such as severe body weight loss and encephalopathy, can occur in Omicron-infected K18-hACE2 mice. Our study reports, for the first time, that Omicron can induce brain infection with lymphoid depletion in the mouse COVID-19 model.
ABSTRACT
Foreign body ingestion is commonly seen in children. However, occasionally it may also be seen among adults and is often associated with intellectual disability, psychiatric disorders, and alcoholism. Ingestion of a magnetic foreign body may cause complications such as gastrointestinal tract perforation, wherein emergency endoscopic removal of the foreign body is generally required. Here, we report a rare case of a 59-year-old male with an intellectual disability and psychiatric disorder in whom metallic objects in the stomach cavity were accidentally discovered during abdominal CT. Esophagogastroduodenoscopy revealed several metallic objects attached to two magnets, which had been ingested several years before and had remained in the stomach cavity. The magnets and metallic objects were safely removed endoscopically using rat-tooth forceps without complications.
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Background/Aims@#The optimal treatment for acute malignant obstruction of the proximal colon (MOPC, proximal to the splenic flexure) remains challenging. Emergency resection, the traditional modality for MOPC, has shown significantly high mortality and morbidity rates, according to recent studies. This study aimed to investigate the clinical outcomes of stent vs stoma as a bridge to curative surgery for MOPC. @*Methods@#This retrospective cohort study included 72 patients who underwent endoscopic placement of a self-expanding metallic stent (SEMS) or loop ileostomy for MOPC at six referral centers between January 2011 and July 2021. Clinical and pathological characteristics, procedure-related complications, and long-term mortality rates after curative surgery were analyzed. @*Results@#During a mean follow-up period of 32 months, 30 patients (41.7%) underwent ileostomy preferentially for more proximal cancer, complete obstruction, and advanced tumor stage compared to the SEMS group. No difference was found in procedure-related complications, but five deaths were observed after ileostomy. Survival analysis for 5-year mortality after curative surgery showed no significant difference between the bridge modalities (log-rank p = 0.253). @*Conclusions@#In this study, SEMS as a bridge to surgery showed relatively safe results in terms of post-procedural mortality. However, these results should be considered when performing ileostomy in patients with more advanced malignant obstruction.
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Recently, research on rectal neuroendocrine tumors (NETs) has increased during the last few decades. Rectal NETs measuring 10 mm, suctioning is not possible due to fibrosis in the lesion, or when the snaring for modified endoscopic mucosal resection does not work well.
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Background@#As the number of large-scale studies involving multiple organizations producing data has steadily increased, an integrated system for a common interoperable format is needed. In response to the coronavirus disease 2019 (COVID-19) pandemic, a number of global efforts are underway to develop vaccines and therapeutics. We are therefore observing an explosion in the proliferation of COVID-19 data, and interoperability is highly requested in multiple institutions participating simultaneously in COVID-19 pandemic research. @*Results@#In this study, a laboratory information management system (LIMS) approach has been adopted to systemically manage various COVID-19 non-clinical trial data, including mortality, clinical signs, body weight, body temperature, organ weights, viral titer (viral replication and viral RNA), and multiorgan histopathology, from multiple institutions based on a web interface. The main aim of the implemented system is to integrate, standardize, and organize data collected from laboratories in multiple institutes for COVID-19 non-clinical efficacy testings. Six animal biosafety level 3 institutions proved the feasibility of our system. Substantial benefits were shown by maximizing collaborative high-quality non-clinical research. @*Conclusions@#This LIMS platform can be used for future outbreaks, leading to accelerated medical product development through the systematic management of extensive data from non-clinical animal studies.
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Patients with inflammatory bowel disease (IBD) are at risk for extraintestinal manifestations (EIM) over the course of their disease. As EIMs can involve nearly every organ, and strongly influence the quality of life, early recognition and adequate treatment are necessary to prevent severe morbidity and mortality in affected patients. Pyoderma gangrenosum is a highly severe and debilitating skin condition that occurs in 1% to 10% of ulcerative colitis (UC) patients. Thromboembolic events are also serious EIMs and usually present as deep vein thromboses in the legs or as pulmonary embolisms. A 19-year-old woman presented with bloody diarrhea lasting for 3 months and deep ulceration on the right foot. She was diagnosed with UC. The patient's skin lesions did not improve with intravenous corticosteroids and oral mesalazine. After she was started on infliximab, we observed rapid resolution of the skin lesions. She continued to complain of mild dyspnea while in the hospital. Computed tomography performed using the thromboembolism protocol revealed pulmonary thromboembolism and deep venous thrombosis. The patient underwent anticoagulant therapy with low-molecular-weight heparin, and her dyspnea gradually improved. Anticoagulation was continued with warfarin. It is rare for IBD patients to have multiple EIMs; however, this case demonstrates that multiple EIMs are a possible presentation in UC and underscores the importance of a meticulous clinical examination and adequate evaluation in the management of IBD patients presenting with EIMs.
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BACKGROUND: Immune cells express the vitamin (vit) D receptor, and vit D is a potent immune-modulator. A negative correlation between serum vit D levels and rheumatoid arthritis (RA) disease activity has been reported. Therefore, we aimed to investigate if the sufficient serum vit D level is helpful to control disease activity in RA patients treated with interleukin (IL)-6 receptor antibody tocilizumab.METHODS: RA patients taking tocilizumab were enrolled, and data were collected retrospectively. Disease activity scores (DAS) 28, serum vit D levels, modified Sharp scores of hand X-ray at the time of tocilizumab initiation, and follow-up data were analysed. Peripheral blood mononuclear cells were differentiated into T-helper (Th) 17 or osteoclasts in the presence of various concentrations of tocilizumab and/or 1,25(OH)₂D. Th17 proportions were analysed by fluorescence-activated cell sorting. Supernatant cytokine levels were determined by enzyme-linked immunosorbent assay.RESULTS: Among 98 RA patients taking tocilizumab, 34 (34.7%) had sufficient serum 25(OH)D levels (≥ 30 ng/mL) when tocilizumab was initiated. At 24 weeks, vit D sufficient patients had greater DAS28 reduction (64.6% ± 15.5% vs. 52.7% ± 20.7%, P = 0.004), and lower disease activity (91.2% vs. 70.3%, P = 0.018) or remission (82.4% vs. 57.8%, P = 0.014). These differences in DAS28 reduction and the proportion of patients with remission persisted at 48 weeks. However, there was no significant difference in hand and wrist erosion progression. In vitro, tocilizumab and 1,25(OH)₂D treatment synergistically suppressed IL-17 production and osteoclastogenesis.CONCLUSION: RA patients treated with IL-6 antibody show a better response when they have sufficient serum vit D. Tocilizumab and 1,25(OH)₂D synergistically suppress IL-17 production and osteoclast differentiation in RA patients.
Subject(s)
Humans , Arthritis, Rheumatoid , Cholecalciferol , Enzyme-Linked Immunosorbent Assay , Flow Cytometry , Follow-Up Studies , Hand , In Vitro Techniques , Interleukin-17 , Interleukin-6 , Interleukins , Osteoclasts , Retrospective Studies , Vitamin D , Vitamins , WristABSTRACT
Sjögren's syndrome (SS) is a chronic and systemic autoimmune disease characterized by lymphocytic infiltration in the exocrine glands. In SS, type I IFN has a pathogenic role, and recently, inflammasome activation has been observed in both immune and non-immune cells. However, the relationship between type I IFN and inflammasome-associated pyroptosis in SS has not been studied. We measured IL-18, caspase-1, and IFN-stimulated gene 15 (ISG15) in saliva and serum, and compared whether the expression levels of inflammasome and pyroptosis components, including absent in melanoma 2 (AIM2), NLR family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein (ASC), caspase-1, gasdermin D (GSDMD), and gasdermin E (GSDME), in minor salivary gland (MSG) are related to the expression levels of type I IFN signature genes. Expression of type I IFN signature genes was correlated with mRNA levels of caspase-1 and GSDMD in MSG. In confocal analysis, the expression of caspase-1 and GSDMD was higher in salivary gland epithelial cells (SGECs) from SS patients. In the type I IFN-treated human salivary gland epithelial cell line, the expression of caspase-1 and GSDMD was increased, and pyroptosis was accelerated in a caspase-dependent manner upon inflammasome activation. In conclusion, we demonstrate that type I IFN may contribute to inflammasome-associated pyroptosis of the SGECs of SS patients, suggesting another pathogenic role of type I IFN in SS in terms of target tissue -SGECs destruction.
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BACKGROUND@#Immune cells express the vitamin (vit) D receptor, and vit D is a potent immune-modulator. A negative correlation between serum vit D levels and rheumatoid arthritis (RA) disease activity has been reported. Therefore, we aimed to investigate if the sufficient serum vit D level is helpful to control disease activity in RA patients treated with interleukin (IL)-6 receptor antibody tocilizumab.@*METHODS@#RA patients taking tocilizumab were enrolled, and data were collected retrospectively. Disease activity scores (DAS) 28, serum vit D levels, modified Sharp scores of hand X-ray at the time of tocilizumab initiation, and follow-up data were analysed. Peripheral blood mononuclear cells were differentiated into T-helper (Th) 17 or osteoclasts in the presence of various concentrations of tocilizumab and/or 1,25(OH)₂D. Th17 proportions were analysed by fluorescence-activated cell sorting. Supernatant cytokine levels were determined by enzyme-linked immunosorbent assay.@*RESULTS@#Among 98 RA patients taking tocilizumab, 34 (34.7%) had sufficient serum 25(OH)D levels (≥ 30 ng/mL) when tocilizumab was initiated. At 24 weeks, vit D sufficient patients had greater DAS28 reduction (64.6% ± 15.5% vs. 52.7% ± 20.7%, P = 0.004), and lower disease activity (91.2% vs. 70.3%, P = 0.018) or remission (82.4% vs. 57.8%, P = 0.014). These differences in DAS28 reduction and the proportion of patients with remission persisted at 48 weeks. However, there was no significant difference in hand and wrist erosion progression. In vitro, tocilizumab and 1,25(OH)₂D treatment synergistically suppressed IL-17 production and osteoclastogenesis.@*CONCLUSION@#RA patients treated with IL-6 antibody show a better response when they have sufficient serum vit D. Tocilizumab and 1,25(OH)₂D synergistically suppress IL-17 production and osteoclast differentiation in RA patients.
ABSTRACT
Sjögren's syndrome (SS) is a chronic and systemic autoimmune disease characterized by lymphocytic infiltration in the exocrine glands. In SS, type I IFN has a pathogenic role, and recently, inflammasome activation has been observed in both immune and non-immune cells. However, the relationship between type I IFN and inflammasome-associated pyroptosis in SS has not been studied. We measured IL-18, caspase-1, and IFN-stimulated gene 15 (ISG15) in saliva and serum, and compared whether the expression levels of inflammasome and pyroptosis components, including absent in melanoma 2 (AIM2), NLR family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein (ASC), caspase-1, gasdermin D (GSDMD), and gasdermin E (GSDME), in minor salivary gland (MSG) are related to the expression levels of type I IFN signature genes. Expression of type I IFN signature genes was correlated with mRNA levels of caspase-1 and GSDMD in MSG. In confocal analysis, the expression of caspase-1 and GSDMD was higher in salivary gland epithelial cells (SGECs) from SS patients. In the type I IFN-treated human salivary gland epithelial cell line, the expression of caspase-1 and GSDMD was increased, and pyroptosis was accelerated in a caspase-dependent manner upon inflammasome activation. In conclusion, we demonstrate that type I IFN may contribute to inflammasome-associated pyroptosis of the SGECs of SS patients, suggesting another pathogenic role of type I IFN in SS in terms of target tissue -SGECs destruction.
ABSTRACT
BACKGROUND@#Immune cells express the vitamin (vit) D receptor, and vit D is a potent immune-modulator. A negative correlation between serum vit D levels and rheumatoid arthritis (RA) disease activity has been reported. Therefore, we aimed to investigate if the sufficient serum vit D level is helpful to control disease activity in RA patients treated with interleukin (IL)-6 receptor antibody tocilizumab.@*METHODS@#RA patients taking tocilizumab were enrolled, and data were collected retrospectively. Disease activity scores (DAS) 28, serum vit D levels, modified Sharp scores of hand X-ray at the time of tocilizumab initiation, and follow-up data were analysed. Peripheral blood mononuclear cells were differentiated into T-helper (Th) 17 or osteoclasts in the presence of various concentrations of tocilizumab and/or 1,25(OH)₂D. Th17 proportions were analysed by fluorescence-activated cell sorting. Supernatant cytokine levels were determined by enzyme-linked immunosorbent assay.@*RESULTS@#Among 98 RA patients taking tocilizumab, 34 (34.7%) had sufficient serum 25(OH)D levels (≥ 30 ng/mL) when tocilizumab was initiated. At 24 weeks, vit D sufficient patients had greater DAS28 reduction (64.6% ± 15.5% vs. 52.7% ± 20.7%, P = 0.004), and lower disease activity (91.2% vs. 70.3%, P = 0.018) or remission (82.4% vs. 57.8%, P = 0.014). These differences in DAS28 reduction and the proportion of patients with remission persisted at 48 weeks. However, there was no significant difference in hand and wrist erosion progression. In vitro, tocilizumab and 1,25(OH)₂D treatment synergistically suppressed IL-17 production and osteoclastogenesis.@*CONCLUSION@#RA patients treated with IL-6 antibody show a better response when they have sufficient serum vit D. Tocilizumab and 1,25(OH)₂D synergistically suppress IL-17 production and osteoclast differentiation in RA patients.
ABSTRACT
Duodenal leiomyosarcoma is a rare condition with a poor prognosis. Early diagnosis of duodenal leiomyosarcoma is challenging because it presents with nonspecific symptoms and endoscopic biopsies usually do not enable a definitive diagnosis. Duodenal leiomyosarcomas are diagnosed on the basis of the histopathological identification of a mesenchymal lesion composed of malignant tumor cells that on immunohistochemical examination is positive for smooth muscle actin and desmin. We report the case of a 38-year-old man who presented with gastrointestinal bleeding and obstruction who was diagnosed with duodenal leiomyosarcoma after surgical resection.
Subject(s)
Adult , Humans , Actins , Biopsy , Desmin , Diagnosis , Duodenal Obstruction , Early Diagnosis , Gastrointestinal Hemorrhage , Hemorrhage , Leiomyosarcoma , Muscle, Smooth , PrognosisABSTRACT
Since primary Sjögren's syndrome (pSS) is an autoummune disease of B cell hyperactivity and pathologic autoantibody response, follicular helper T (Tfh) cells and follicular regulatory T (Tfr) cells are suggested to be key players in pSS. We examined subsets of Tfh and Tfr cells from the blood in pSS patients, and whether these subsets represent disease activity, glandular inflammation, or autoantibody responses in pSS. Circulating Tfh and Tfr cells, along with their specific subsets, were identified from the peripheral blood of 18 pSS patients and 14 age- and sex-matched healthy controls (HCs) using flow cytometry analysis. Blood Tfr and Tfh cell ratios were increased in pSS patients compared with HCs. The CCR7(lo)PD-1(hi) subset of circulating Tfh cells was increased in pSS patients with high degree of focal lymphocytic sialadenitis; whereas circulating Tfh cells did not differ between pSS patients and HCs. The frequency of CCR7(lo)PD-1(hi) Tfh cells was significantly correlated with disease activity scores and differentiated B cells. PD-1 expression on blood Tfh and Tfr cells showed positive correlations with IL-21 in pSS. Increasing trend of blood Tfr cells was observed in pSS patients, and blood Tfr cells (particularly Th1 and Th17 subsets) represented hypergammaglobulinemia in pSS. In summary, circulating CCR7(lo)PD-1(hi) Tfh cells indicated disease activity and glandular inflammation in pSS. Circulating Tfr cells, shifted toward Th1 and Th17 subsets, indicated ongoing IgG production in pSS. Subsets of circulating Tfh or Tfr cells could be biomarkers for disease monitoring and patient stratification in pSS.
Subject(s)
Humans , Autoantibodies , B-Lymphocytes , Biomarkers , Flow Cytometry , Hypergammaglobulinemia , Immunoglobulin G , Inflammation , Sialadenitis , T-Lymphocyte Subsets , T-Lymphocytes , T-Lymphocytes, Helper-Inducer , T-Lymphocytes, RegulatoryABSTRACT
The circulation and infection of avian influenza virus (AIV) in zoos and backyard flocks has not been systematically investigated. In the present study, we surveyed the birds including those in live bird markets (LBMs) and evaluated co-circulation of AIVs among them. Overall, 26 H9N2 AIVs and one H6N2 AIV were isolated from backyard flocks and LBMs, but no AIVs were isolated from zoo birds. Genetic analysis of the HA and NA genes indicated that most of the H9N2 AIVs showed higher similarities to AIVs circulating in domestic poultry than to those in wild birds, while the H6N2 AIV isolate from an LBM did to AIVs circulating in migratory wild birds. In serological tests, 15% (391/2619) of the collected sera tested positive for AIVs by competitive-ELISA. Among them, 34% (131/391) of the sera tested positive for AIV H9 antigen by HI test, but only one zoo sample was H9 positive. Although AIVs were not isolated from zoo birds, the serological results indicated that infection of AIVs might occur in zoos. It was also confirmed that H9N2 AIVs continue to circulate and evolve between backyard flocks and LBMs. Therefore, continuous surveillance and monitoring of these flocks should be conducted to control further epidemics.
Subject(s)
Animals , Birds , Influenza in Birds , Korea , Molecular Epidemiology , Poultry , Serologic Tests , VirusesABSTRACT
It is difficult to distinguish between circulating lymphoma cells and non-lymphoma cells which are lymphocytes, activated lymphocytes, monocytes, and leukemic cells. The presence of circulating lymphoma cells in the peripheral blood is infrequent but the incidence is probably significantly higher than has been reported in morphologic studies of peripheral blood smears. So the morphologic evaluations of the circulating lymphoma cells and non-lymphoma cells are needed. We experienced that circulating lymphoma cells were found in the peripheral blood smears in the 4 cases of non-Hodgkin's lymphomas. They were consisted of mantle cell lymphoma (1 case), peripheral T cell lymphoma (1 case), adult T cell leukemia/lymphoma (1 case), cutaneous T cell lymphoma (1 case) and diagnosed from their lymph node biopsies. Those circulating lymphoma cells were morphologically observed by light and electron microscopic methods. Using those morphological features, we think that detection rate of circulating lymphoma cells can be improved in the patient with lymphoma.
Subject(s)
Adult , Humans , Biopsy , Incidence , Leukemia , Lymph Nodes , Lymphocytes , Lymphoma , Lymphoma, Mantle-Cell , Lymphoma, Non-Hodgkin , Lymphoma, T-Cell, Cutaneous , Lymphoma, T-Cell, Peripheral , MonocytesABSTRACT
BACKGROUND: Because moving means can be easily shiftable according to their crude data, we made a selective expected ranges to calculate the moving means. Bull's mean (exponential factor, P=0.50) and Exponentially Adjusted Moving Mean (EAMM, P=0.66) were assessed. we studied to determine appropriately expected range and exponential factor. METHODS: We made the target values from RBC indices being measured with H-2 hematology autoanalyzer from 800 patients and the expected range from red cell indices data of additional 600 patients. Both moving means using this expected ranges were calculated. The % difference of Bull's mean and EAMM was compared and total mean of (deltaBull's mean/deltaBatch mean) and (deltaEAMM/deltaBatch mean) was compared. RESULTS: The target values were MCV: 90.6 fL, MCH: 29.8 pg. MCHC: 32.8 g/dL. The expected ranges were within +/-6% of their target values. Among the 20 batches obtained from expected range, there were no above +/-3% difference of red cell indices in both moving means. The comparison between % difference of Bull's mean and that of EAMM showed no difference. Total mean of (deltaEAMM/deltaBatch mean) was higher than that of (deltaBull's mean/deltaBatch mean). CONCLUSIONS: The % difference results of Bull's mean and EAMM were basically similiar within the expected range but EAMM method was more sensitive than Bull's mean method under the aspect of specimen effects, so EAMM was more detectable than Bull's mean on the quality control of red cell indices.
Subject(s)
Humans , Erythrocyte Indices , Hematology , Quality ControlABSTRACT
BACKGROUND: For the early diagnosis of tsutsugamushi disease, polymerase chain reaction (PCR) using skin specimen might be a precise and useful diagnostic tool. The purpose of this study is to detect and identify the serotype of the Orientia tsutsugamushi from the skin lesions of the patients with tsutsugamushi disease by nested PCR. METHODS: Nested PCR was used to diagnose tsutsugamushi disease and identify the serotype of the O. tsutsugamushi; Karp, Kato, Gilliam or Boryong/Kuroki. The primer sets were derived from serotype-specific DNA sequences encoding the 56kDa antigen of O. tsutsugamushi. The PCR products were analyzed by using direct cyclic sequencing. RESULTS: The serotype-specific DNA bands with 1% agarose gel electrophoresis of amplified DNAs by nested PCR were observed in all 11 skin biopsy specimens from 8 patients with tsutsugamushi disease. Among 8 patients, 7 were proved to be infected with Boryong/Kuroki strains and one with Karp. One Karp strain showed one base mutation with amino acid sequence variation, but all the Boryong/ Kuroki strains showed no mutation. CONCLUSION: We suggest that serotype-specific nested PCR is a simple, rapid and precise diagnostic method, and useful for early diagnosis of tsutsugamushi disease. Furthermore, we might detect the sequence variations of serotype-specific DNA sequences encoding 56-kDa antigen among strains.