• Korean Journal of Clinical Laboratory Science
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• v.36 no.2
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• pp.144-152
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• 2004

With technological advances in automatic hematology analyzers, primary and screening differential counts of white blood cells (WBC) are done with automatic hematology analyzers. They are using different measurement and analysis principles, so differences in WBC differentials and WBC morphology flag exist. This study was carried out to analyze WBC differential counts and WBC morphology flags comparing them with the manual method. Patient EDTA samples in Vacutainer requested for WBC differentials were analyzed with XE-2100. And those samples with suspect flags messages index over 100 were selected and were analyzed with ADVIA-120. Peripheral blood smear film was subsequently made. Three investigators counted 200 cells each (600 cells) in 111 Wright-Giemsa stained blood films. Between two automatic hematology analyzers, neutrophil, lymphocyte, eosinophil, and monocyte showed good correlations, but basophil had moderate correlation. Among automatic hematology analyzers and manual count, neutrophil, lymphocyte, and eosinophil had good correlations, but monocyte had moderate correlation. XE-2100 had higher monocyte, which was due to atypical lymphocyte and myeloblast. LUC in ADVIA-120 was not due to monocyte in XE-2100. Morphology flagging rates were 146.9% in XE-2100 and was 93.2% in ADVIA-120. Positive predictive values of morphology flag were 58.2% in XE-2100 and 54.4% in ADVIA-120. Flags such as atypical lymphocyte, immature granulocyte, and left shift had higher predictive values and those such as N-RBC, platelets clump, and blast had lower ones. Between automatic hematology analyzers, WBC differentials showed good correlations. Predictive values for morphology flags can be variable with changing criteria. Reviewing criteria for WBC differentials and morphology flags should be established in each laboratory with regards to size of laboratory and patients it serves.

• Journal of Pharmacopuncture
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• v.20 no.1
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• pp.52-56
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• 2017

Objectives: Neutrophils represent the front line of human defense against infections. Immediately after stimulation, neutrophilic enzymes are activated and produce toxic mediators such as pro-inflammatory cytokines, nitric oxide (NO) and myeloperoxidase (MPO). These mediators can be toxic not only to infectious agents but also to host tissues. Because flavonoids exhibit antioxidant and anti-inflammatory effects, they are subjects of interest for pharmacological modulation of inflammation. In the present study, the effects of rutin on stimulus-induced NO and tumor necrosis factor $(TNF)-{\alpha}$ productions and MPO activity in human neutrophils were investigated. Methods: Human peripheral blood neutrophils were isolated using Ficoll-Hypaque density gradient centrifugation coupled with dextran T500 sedimentation. The cell preparations containing > 98% granulocytes were determined by morphological examination through Giemsa staining. Neutrophils were cultured in complete Roswell Park Memorial Institute (RPMI) medium, pre-incubated with or without rutin ($25{\mu}M$) for 45 minutes, and stimulated with phorbol 12-myristate 13-acetate (PMA). Then, the $TNF-{\alpha}$, NO and MPO productions were analyzed using enzyme-linked immunosorbent assay (ELISA), Griess Reagent, and MPO assay kits, respectively. Also, the viability of human neutrophils was assessed using tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), and neutrophils were treated with various concentrations of rutin ($1-100{\mu}M$), after which MTT was appended and incubated at $37^{\circ}C$ for 4 hour. Results: Rutin at concentrations up to $100{\mu}M$ did not affect neutrophil viability during the 4-hour incubation period. Rutin significantly decreased the NO and $TNF-{\alpha}$ productions in human peripheral blood neutrophils compared to PMA-control cells (P < 0.001). Also, MPO activity was significantly reduced by rutin (P < 0.001). Conclusion: In this in vitro study, rutin had an anti-inflammatory effect due to its inhibiting NO and $TNF-{\alpha}$ productions, as well as MPO activity, in activated human neutrophils. Treatment with rutin may be considered as a therapeutic strategy for neutrophil-mediated inflammatory/autoimmune diseases.

• Parasites, Hosts and Diseases
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• v.29 no.4
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• pp.315-324
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• 1991

Each of SPF chicken(Hi-Line strain, 2-day-old males) was inoculated with 2.5 or $5\times10^4$ oocysts by stomach tube. The oocyst was the medium type of Cryptosporidium previously isolated from Korean chicken origin, and passed in 2-day-old SPF chicken. The patterns of oocyst discharge were monitored daily, and in order to observe the ultrastructure of the developmental stages, the bursa of Fabricius of the chicken was examined by transmission electron microscopy (TEM) on the 12th day postinoculation. The prepatent period for 8 chicken was 5.9 days postinoculation on the average, and the patent period was 12.9 days. The number of oocysts discharged per day for the chicken was reached peak on day 12 postinoculation on the average. A large number of oocysts was found in fecal samples obtained from inoculated chicken on days 8~14 postinoculation. The ultrastructural feature of almost every developmental stage of the medium type from chicken was very similar to that of Cryptosporidium previously isolated from mammalia including human and birds except for the attachment site of C. tsuris to the mucus cell from mammalia, but dimension of the oocysts from fecal samples of the medium type was different from those of C. meleagridis and mammalia origin. The above results reveal that the medium type of Cryptosporidium of Korean chicken origin is identified as Cryptosporidium baileyi.

• Journal of Preventive Medicine and Public Health
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• v.19 no.2 s.20
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• pp.244-251
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• 1986

Sister chromatid exchanges(SCEs) and cell cycle kinetics were proposed as a sensitive and quantitative assay for mutagenicity and cytotoxicity in short-term cultures of phytohema-gglutinin(PHA)-stimu1ated human 1ymphocytes. Therefore, this study was performed to investigate the relation between the cytotoxic effects and sister chromatid exchanges. The resultes are summarized as follows: 1) The frequency of SCEs per cell are $13.1{\pm}2.8$ in the lower concentration of $6.25{\times}10^{-9}M\;and\;75.8{\pm}8.2$ in the highest concentration of $1.00{\pm}10^{-7}M$. Mitotic index is decreased in the higher concentration of mitomycin C. The result indicates that mitomycin C led to a dose dependent increase in SCE frequency, but decease in mitotic index. 2) Chromosomal analysis was performed on metaphase cells that have divided one, two, and three or more times for cell cycle kinetics by fluorescence-plus-Giemsa(FPG) technique. According to the increased concentration of mitomycin C, the proportion of metaphase cells in the first are profoundly increased but the cells of third division are greatly decreased. 3) The frequency of SCEs per chromosome by chromosomal group are decreased gradually from A group to G group. But relationships between specific chromosomal group and SCE frequency are not found.

• Journal of Genetic Medicine
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• v.7 no.2
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• pp.156-159
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• 2010

Inversion, one of the balanced rearrangements, usually does not lead to phenotypic abnormalities; all genetic information exists in the proper amount, merely in a different order or in an abnormal location. However, offspring of an inversion carrier is at risk of chromosomal imbalance because an inversion loop can be formed during crossing-over of the paternal and the maternal chromosomes in meiosis. We report a 38-year-old woman with inversion and balanced translocation and her fetus with unusual rearrangement causing chromosomal imbalance. We performed conventional cytogenetic analysis, MLPA, and subtelomeric FISH in the cells of the embryo. The results showed that the distal portion of chromosome 13q was added to the terminal portion of chromosome 9p during crossing-over. Therefore, the final karyotype of the fetus was 46,XY,rec(9)t(9;13)(p22;q32)inv(9)(p12q13)mat, confirmed using molecular-cytogenetic analyzing tools.

• Animal Bioscience
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• v.37 no.6
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• pp.1001-1006
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• 2024

Objective: This study aimed to investigate the effect of Codonopsis pilosula polysaccharide (CPP) on the motility, mitochondrial integrity, acrosome integrity rate, and antioxidant ability of sheep sperm after preservation at 4℃. Methods: Semen from healthy adult rams were collected and divided into four groups with separate addition of 0, 200, 400, and 1,000 mg/L CPP. Sperm motility was analyzed using the Computer-Assisted Semen Analysis software after preservation at 4℃ for 24, 72, 120, and 168 h. Sperm acrosome integrity rate was analyzed by Giemsa staining at 24, 72, and 120 h, and mitochondrial membrane integrity was analyzed by Mito-Tracker Red CMXRos. The total antioxidant capacity (T-AOC) and malondialdehyde (MDA) content of spermatozoa were measured after 120 h of preservation. Results: The sperm viability and forward-moving sperm under 200 mg/L CPP were significantly higher than that in the control group at 72 h (61.28%±3.89% vs 52.83%±0.70%, 51.53%±4.06% vs 42.84%±1.14%), and 168 h (47.21%±0.85% vs 41.43%±0.37%, 38.68%±0.87% vs 31.68%±0.89%). The percentage of fast-moving sperm (15.03%±1.10% vs 11.39%±1.03%) and slow-moving sperm (23.63%±0.76% vs 20.29%±1.11%) in the 200 mg/L group was significantly higher than control group at 168 h. The mitochondrial membrane integrity of the sperm in the group with 200 mg/L CPP was significantly higher than those in the control group after storage at 4℃ for 120 h (74.76%±2.54% vs 65.67%±4.51%, p<0.05). The acrosome integrity rate in the group with 200 mg/L (87.66%±1.26%) and 400 mg/L (84.00%±2.95%) was significantly higher than those in the control group (80.65%±0.16%) after storage for 24 h (p<0.05). CPP also increased T-AOC and decreased the MDA concentration after preservation at 4℃ (p<0.05). Conclusion: Adding CPP could improve the T-AOC of sperm, inhibit lipid peroxidation, and facilitate semen preservation.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.1 no.1
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• pp.30-36
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• 1998

Purpose: It is well known that duodenal ulcer disease does not relapse if H. pylori is cleared from the gastric mucosa. Little is known about the recurrence of duodenal ulcer in children. The purpose of this study was to evaluate the effect of the eradication of H. pylori in duodenal ulcer in children upon the duodenal ulcer recurrence. Methods: 105 patients (M:F=78:27) diagnosed as duodenal ulcer by endoscopy in 1987~1995 were reviewed clinically, and were parted into two groups. The two treatment groups were ranitidine/antacid (RAN/ANT) and ranitidine/amoxicillin/denol (RAN/AMX/D). The latter was for H. pylori-positive children with duodenal ulcer who were diagnosed by serology and/or antral biopsies for histology, culture, and urease testing. The recurrence rates were compared between the two groups. Results: 1) 30 patients with primary duodenal ulcer underwent endoscopy for H. pylori and 27 (90.0%) of them were positive for H. pylori. 2) 27 of H. pylori-positive children received RAN/AMX/D. 23(85.2%) of them showed cure of duodenal ulcer and eradication of H. pylori. 3) The duodenal ulcer recurrence rate in RAN/ANT group was 65.3% and the rate in RAN/AMX/D was 4.3% by a year. Conclusions: There is a strong correlation between the duodenal ulceration and H. pylori infection in children, and the eradication of H. pylori in duodenal ulcer patients reduces the recurrence of the ulcer. Because of the low incidence of duodenal ulcers in children, a multicenter prospective study is required to determine the effect of treating H. pylori infetion on the long term natural history of duodenal ulcer disease.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.12
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• pp.4773-4780
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• 2014

Background: To investigate the effects of small interference RNA (siRNA) targeting BCR/ABL mRNA on proliferation and apoptosis in the K562 human chronic myeloid leukemia (CML) cell line and to provide a theoretical rationale and experimental evidence for its potential clinical application for anti-CML treatment. Materials and Methods: The gene sequence for BCR/ABL mRNA was found from the GeneBank. The target gene site on the BCR/ABL mRNA were selected according to Max-Planck-Institute (MPI) and rational siRNA design rules, the secondary structure of the candidate targeted mRNA was predicted, the relevant thermodynamic parameters were analyzed, and the targeted gene sequences were compared with BLAST to eliminate any sequences with significant homology. Inhibition of proliferation was evaluated by MTT assay and colony-formation inhibiting test. Apoptosis was determined by flow cytometry (FCM) and the morphology of apoptotic cells was identified by Giemsa-Wright staining. Western blotting was used to analyze the expression of BCR/ABL fusion protein in K562 cells after siRNA treatment. Results: The mRNA local secondary structure calculated by RNA structure software, and the optimal design of specific siRNA were contributed by bioinformatics rules. Five sequences of BCR/ABL siRNAs were designed and synthesized in vitro. Three sequences, siRNA1384, siRNA1276 and siRNA1786, which showed the most effective inhibition of K562 cell growth, were identified among the five candidate siRNAs, with a cell proliferative inhibitory rate nearly 50% after exposure to 12.5nmol/L~50nmol/L siRNA1384 for 24,48 and 72 hours. The 50% inhibitory concentrations ($IC_{50}$) of siRNA1384, siRNA1276 and siRNA1786 for 24hours were 46.6 nmol/L, 59.3 nmol/L and 62.6 nmol/L, respectively, and 65.668 nmol/L, 76.6 nmol/L, 74.4 nmol/L for 72 hours. The colony-formation inhibiting test also indicated that, compared with control, cell growth of siRNA treated group was inhibited. FCM results showed that the rate of cell apoptosis increased 24 hours after transfecting siRNA. The results of annexinV/PI staining indicated that the rate of apoptosis imcreased (1.53%, 15.3%, 64.5%, 57.5% and 21.5%) following treamtne with siRNAs (siRNA34, siRNA372, siRNA1384, siRNA1276 and siRNA1786). Morphological analysis showed td typical morphologic changes of apoptosis such as shrunken, fragmentation nucleus as well as "apoptotic bodies" after K562 cell exposure to siRNA. Western blot analysis showed that BCR/ABL protein was reduced sharply after a single dose of 50nmol/L siRNA transfection. Conclusions: Proliferation of K562 cells was remarkbly inhibited by siRNAs (siRNA1384, siRNA1276 and siRNA1786) in a concentration-dependent manner in vitro, with effective induction of apoptosis at a concentration of 50 nmol/L. One anti-leukemia mechanism in K562 cells appeared that BCR/ABL targeted protein was highly down-regulated. The siRNAs (siRNA1384, siRNA1276 and siRNA1786) may prove valuable in the treatment of CML.

• Clinical and Experimental Pediatrics
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• v.51 no.5
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• pp.492-499
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• 2008

Purpose : Helicobacter pylori infection is one of the most common gastrointestinal infections worldwide; it almost invariably causes chronic gastritis. Pediatric studies may provide important insights into the mucosal immune response of H. pylori-infection, as children are not submitted to environmental factors such as alcohol, tobacco and anti-inflammatory medication. The aim of the present study was to investigate the mucosal immune response against H. pylori in clinically well-defined groups: H. pylori-positive (divided into peptic ulcer disease and gastritis) and H. pylori-negative control. Methods : Antral biopsies were obtained from 45 children undergoing an upper GI endoscopy for dyspeptic symptoms. T cells (CD3+, CD4+, CD8+) and B cells (CD20+) were analyzed by quantitative immunohistochemistry. The correlation of lymphocyte subsets of gastric mucosa with histology was evaluated. Results : T cells (CD3+, CD4+, CD8+) and B cells (CD20+) were significantly increased in the lamina propria of H. pylori-positive group (P<0.01). CD8+ T cells were significantly increased in the lamina propria of the H. pylori-positive peptic ulcer disease (P<0.01). Within the epithelium, only CD4+ T cells were significantly increased in the H. pylori-positive group (P<0.01). Gastric histological parameters had a closer correlation with lymphocytes in the lamina propria than intraepithelial lymphocytes. Conclusion : This study suggests that both T cells and B cells in the lamina propria play important roles in the local immune response of H. pylori-infected children. Furthermore, it remains to be elucidated whether CD8+ T cells in the lamina propria may contribute to peptic ulcer formation in H. pylori-infected children.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.3 no.2
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• pp.151-159
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• 2000

Purpose: Recently, a wide application of gastrofiberscopy in the pediatric group have revealed that nodular duodenitis is not an uncommon disease in children and is suspected to be associated with H. pylori infection. The aim of this retrospective study was to investigate the clinical and histopathologic features in children with nodular duodenitis, and to assess the correlations beween both. Methods: During a period of 5 years (Jan. 1995~Dec. 1999), we investigated clinical, endoscopic and histopathologic features of 39 consecutive patients diagnosed as having nodular duodenitis at Pediatric department of Seoul Red Cross Hospital. In 35 children with nodular duodenitis endoscopic biopsy specimens were stained with Hematoxylin & Eosin and Giemsa's stain, and were graded according to the criteria outlined by Triadafilopoulos, Whitehead et al., and Prieto et al.. Statistical analyses were performed with Graph PAD InStat. Results: The prevalence rate of nodular duodenitis was 17.1% and the most frequent chief complaint was abdominal pain (69.2%). Endoscopically grade 1 was the most common (45.7%) and nodular gastritis was coexistent in 28.3%. The most common histology of the duodenum was grade 2 (54.3%), and the most common histologic score of the stomach was 2 (42.9%). H. pylori was found in the duodenum in 37.1%, and in the stomach in 31.4%. The correlation coefficient between the endoscopic grade and the histologic grade of nodular duodenitis was 0.3983 (p=0.0178). And the correlation coefficient between the histologic grade and the grade of H. pylori colonization in the duodenum was 0.5154 (p=0.0018). Conclusion: There was significant correlation between the endoscopic grade and the histologic grade of nodular duodenitis, and was also significant correlation between the histologic grade and the grade of H. pylori colonization in the duodenum. Therfore H. pylori infection should be regarded as an etiologic factor of nodular duodenitis.