Significance: Accurate detection of Helicobacter pylori (HP) is essential for the diagnosis of HP infection. The use of antibiotics and proton pump inhibitors (PPI) may result in false-negative rapid urease test (RUT) results. We aimed to determine the sensitivity and specificity of RUT compared with histology and assess the detection rate of combined RUT and histology for HP infection. Methodology: Retrospective data collection was performed on 192 patients who were tested for both RUT and histology at the time of upper endoscopy from 2017 to 2018. At least two gastric biopsies (1 from corpus, 1 from antrum) were taken each for RUT and histology. The endoscopy was performed by a single gastroenterologist and a single pathologist was responsible for interpreting the histology with hematoxylin and eosin (H&E) and Giemsa stain. The gold standard test for the diagnosis of HP infection was histology. Demographic profile, RUT and histology results were reviewed. Tests for diagnostic accuracy were computed using SPSSv23. Results: 192 patients were tested for RUT and histology. 52(27.1%) were males and 140(72.9%) were females with a mean age of 54±17 years. Epigastric pain was the most common indication (42.7%). 24(12.5%) patients tested positive for HP infection. Among these; 16(8.3%) tested positive for both RUT and histology(true-positive), while 8(4.2%) tested negative for RUT but had positive histology(false-negative). 6 out of 8(75%) patients with false negative results had PPI use. The sensitivity and specificity of RUT for the diagnosis of HP infection were 66.7 and 98.2%, respectively. While the positive and negative likelihood ratio were 37.3 and 0.34, respectively with a diagnostic odds ratio of 110. Conclusion The HP detection rate of RUT combined with histology increased by 33% compared with RUT alone. RUT is a highly specific test for diagnosing HP infection. Given its modest sensitivity, histology plays an important role in the diagnosis of HP infection, especially in patients taking PPIs. We recommend doing histology when RUT is negative to increase the HP detection rate.
Helicobacter Pylori ; Histology ; Azure Stains

BACKGROUND: The efficiency of leukocyte removal filter is influenced by many factors. But, filtration efficiency of leukocyte fragments was not well known. We performed this study to evaluate whether the filtration efficiency for packed red blood cells can be influenced by leukocyte fragments according to storage time. METHODS: Leukocyte fragments in packed red blood cells (three units) which were artificially made by incubation for 4 hrs at 56degrees C and each four units of packed red blood cells according to storage time (0 days, 10 days, 20 days, and 30 days) were filtered using Sepacell R-500A (Asahi medical Co, Japan). The leukocyte concentrations of the pre-leukodepleted samples were estimated using an automated hematology analyzer (XE-2100, Sysmex, Japan). The ratio between the number of normal leukocytes and leukocyte fragments on Wright Giemsa stained slide was used in the analysis. The leukocyte concentrations of the post-leukodepleted samples were performed by the conventional counting methods using Nageotte hemocytometer. RESULTS: The ratios of fragmented to total leukocytes in packed red blood cells at pre- and post leukoreduction according to storage times were 1.5% and 16.3% within 1 days, 4.5% and 30.0% at 10 days, 6.3% and 35.0% at 30 days, and 8.3% and 42.5% at 40 days, respectively. Leukoreduction efficiencies of normal leukocytes in packed red blood cells were 99.99 +/- 0.01%, 99.97 +/- 0.02%, 99.98 +/- 0.01%, and 99.86 +/- 0.09%, respectively. The 36.0% of leukocytes in packed red blood cells were changed to fragmented leukocytes, residual fragmented leukocytes ratio was 95.0% and filter efficiencies of normal leukocytes was low(99.28%, p<0.05). CONCLUSIONS: The leukodepleted efficiency for leukocyte fragments were lower than for normal leukocytes. Leukocytes fragments may be influenced to lower the leukodepleted efficiency of normal leukocytes with storage time elapse.
Azure Stains ; Erythrocytes ; Filtration ; Hematology ; Leukocytes*

The causative role of Pityrosporum ovalea(P. ovale) in the development of seborrheic dermatitis has been disputed for many decades. Using Giemsa stain, the number of P. ovale adherant to extrutidei from one keratinocyte of the scalp were counted in twenty patients with seborrheic dermatisis and twenty control persons. Ths result showed that the number of P. ovale in patient with seborrheic dermatitis were significantiy higher than in the normal control group. Furthermon not only were the number of P. ovale decreased, but the clinical lesions were also significantly improved after a 4 week trial of oral itraconazole in the seborrheic dermatitis group. From this points of view, P. ovale may be one of the important causative factor of seborrheic dermatit.is.
Azure Stains ; Dermatitis, Seborrheic* ; Humans ; Itraconazole* ; Keratinocytes ; Malassezia ; Scalp

Bacterial suspensions of Staph. albus, Staph. aureus, Es. coli and Ps. aeruginosa were added into the rabbit macrophage suspension and placed in the cubator of 37℃. After the incubation of 30, 60 and 120 minutes, percentage of macrophage with intracellular organisms, percentage of intracellular organisms, and number of bacterial cells in the macrophages were followed by microscopy of Giemsa stained preperations. Results of observation were summarized as follows. :1. In the Staph. albus-macrophage suspension; In the suspension of cell-bacterial ratio (CBR) being 1:1 or less, percentages of macrophages with intracellular organisms were small upto 120 minutes of incubation. In the suspension of CBR being about 1:100, percentage of macrophages with intracellular organisms was 40% after 30 mintes, and more than 90% after 120 minutes. Percentage of intracellular organisms was 60–80% after 30 minutes and more than 90% after 120 minutes. The more the number of organisms were present, or the longer the time of incubation elaspsed the number of intracellular organisms were greater. 2. In the Staph. aureus-macrophage suspension; Percentage of macrophages with intracellular organisms was about 25% after 30 minutes and about 85% after 120 minutes in the suspension of CBR being 1:10. In the suspension of CBR being 1:100, percentage of macrophages with intracellular organisms was 45% after 30 minutes and more than 90% after 60 minutes. 55% to 90% of organisms were located intracellularly after 30minutes and the percentages increased until 60 minutes. After 120 minutes the percentages of intracellular organisms, in the suspensions of CBR being 1:1 or more, decreased considerablly. Compared to the Staph. albus-cell suspension, more organisms were found intracellularly. 3. In the Es. coli-macrophage suspension, In the suapension of CBR being 1:100 percentage of macrophages with intracellular organisms was about 70% after 30 minutes and 99% after 120 minutes. After 30 minutes 45–70% of organism were located intracellularly and 70%–80% after 60 minutes and the percentages continued increase in the suspensions of CBR being 1:1 or less, but decreased in the suspensions of CBR being 1:10 or more. Compared to the Staphylococcus-cell suspensions, number of organisms observed intracellularly were generally smaller. 4. In the Ps. aeruginosa-macrophage suspension; About 60% of macrophages were found haboring organisms in the suspension of CBR being 1:100 after 30 minutes and 85% after 120 minutes. After 30minutes about 90% of organisms were observed intracellularly in the suspension of CBR 1:10 or more and after 60 minutes the percentage decreased. Compared to the Es. coli-macrophage suspension, the number of intracellular organisms were about the same or even less.
Azure Stains ; In Vitro Techniques ; Macrophages ; Mentha ; Microscopy ; Suspensions

Low parasitemic condition in malaria remains a diagnostic challenge; as the available diagnostic methods failed to detect. Currently, hemozoin (Hz) pigment is gaining attention in the diagnosis of malaria. The major drawback is ease of detection of Hz in routine practice. A pilot study was conducted to evaluate the role of Hz pigment and to compare the performance of quantitative buffy coat assay (QBC) and PCR in such conditions. Clinically suspected cases of malaria were examined by both Giemsa stain and immunochromatographic test (ICT). Samples positive by ICT and negative by Giemsa stain were further examined by nested PCR targeting 18S rRNA and QBC for the presence of malaria parasites and pigments. Thirty blood samples fulfilled the inclusion criteria out of which 23 were Plasmodium vivax (Pv), 4 Plasmodium falciparum (Pf), and 3 mixed (Pv and Pf) by immunochromatographic test. Twenty-one out of 30 (70%) were positive by nested PCR in comparison to 25/30 (83%) by QBC. Samples containing both malaria parasites and Hz pigment by QBC completely showed concordance with the PCR result. However, 61% of total samples containing only Hz pigment were observed positive by PCR. Hz pigment remains an important tool for malaria diagnosis. Identification of leukocytes containing pigments by QBC not only indicates recent malarial infections but also puts light on severity of the disease. QBC assay is a rapid, highly sensitive, and cost-effective method to detect malaria parasites and Hz pigment especially in low parasitemic conditions.
Azure Stains ; Diagnosis* ; Leukocytes ; Malaria ; Methods ; Parasites ; Pilot Projects ; Plasmodium falciparum ; Plasmodium vivax ; Polymerase Chain Reaction

OBJECTIVETo apply the Wright-Giemsa stain in micronucleus test and to explore the stain outcomes of Wright-Giemsa dye of various proportions and staining times.

METHODSUse Wright-Giemsa dye, Wright dye (staining time 3 min) and Giemsa dye (staining time 5 min) to stain HepG2 and then observe the staining effect. The Wright-Giemsa dye was applied under 5 different proportions (3:1-1:3) and different staining times (1, 3, 5, 10, 15 min).

RESULTSAfter stained for 3-5 min with the proportion ratio of 3:1 of Wright-Giemsa dye, the HepG2 cells showed much better staining outcomes compared with the single stain of either Wright or Giemsa.

CONCLUSIONSWright-Giemsa stain can be used in cell micronucleus test to obtain good staining outcomes.

An etiologic agent in seborrheic dermatitis is now considered to be due to Pityrosporum ovale. The connection between the yeast and the disease has been clearly dernonstrated in a number of patients treatd with antifungal agents. However, the fact, that Pityrosporum ovale (Pityrosporum orbiculare) belongs to the normal human cutaneou. flora makes it difficult, to explain the role of the organism in seborrheic dermatitis. In this clinical study, twenty eight subjects were divided into two group by treatment regimen of topical 2% ketoconazole shampoo(TKS)and 0.3% prednisolone valerate solution (PVS). Before and during the four weeks treatment periods, the number of Pityrosporum ovale on the scalp were evaluated by scrub Giemsa staining method, which estimat,ed by score scale, under direct. microscopic examination. Also clinical symptoms including erythema, scales and itching were recorded by scores every week and compared between two treatment group. Topical application of 2% ketoconazole shampoo(TKS) is a very effective treatment regimen to reduce the yeasr cell score(from 5.8+1.3 to 1.8+1.4) than PVS(from 6.4+1.3 to 3.5+1.5), significantly(p<0.05). So the results of this study acconsistent with the view that density of Pityrosgourum ovale plays a role in the cause and course of seborcheic derrnatitis on the scalp. And antifungal agent(TKS) shows favorable effects ori tnis disease clinically and mycologically.
Antifungal Agents ; Azure Stains ; Dermatitis, Seborrheic* ; Erythema ; Humans ; Ketoconazole* ; Malassezia ; Prednisolone ; Pruritus ; Scalp* ; Weights and Measures ; Yeasts

The cytogenetic analysis of mesenchymal stromal cells (MSCs) is essential for verifying the safety and stability of MSCs. An in situ technique, which uses cells grown on coverslips for karyotyping and minimizes cell manipulation, is the standard protocol for the chromosome analysis of amniotic fluids. Therefore, we applied the in situ karyotyping technique in MSCs and compared the quality of metaphases and karyotyping results with classical G-banding and chromosomal abnormalities with fluorescence in situ hybridization (FISH). Human adipose- and umbilical cord-derived MSC cell lines (American Type Culture Collection PCS-500-011, PCS-500-010) were used for evaluation. The quality of metaphases was assessed by analyzing the chromosome numbers in each metaphase, the overlaps of chromosomes and the mean length of chromosome 1. FISH was performed in the interphase nuclei of MSCs for 6q, 7q and 17q abnormalities and for the enumeration of chromosomes via oligo-FISH in adipose-derived MSCs. The number of chromosomes in each metaphase was more variable in classical G-banding. The overlap of chromosomes and the mean length of chromosome 1 as observed via in situ karyotyping were comparable to those of classical G-banding (P=0.218 and 0.674, respectively). Classical G-banding and in situ karyotyping by two personnel showed normal karyotypes for both cell lines in five passages. No numerical or structural chromosomal abnormalities were found by the interphase-FISH. In situ karyotyping showed equivalent karyotype results, and the quality of the metaphases was not inferior to classical G-banding. Thus, in situ karyotyping with minimized cell manipulation and the use of less cells would be useful for karyotyping MSCs.
Azure Stains ; Chromosome Banding/*methods ; Humans ; In Situ Hybridization, Fluorescence/*methods ; Karyotyping/*methods ; Mesenchymal Stromal Cells/*cytology

PURPOSE: The aim of the present study is to detect cagA and vacA genes of Helicobacter pylori(H. pylori) strain in gastric biopsy specimens and to evaluate its association with gastric inflammation in children. METHODS: The cagA and vacA genes were detected by a direct polymerase chain reaction(PCR) assay of gastric biopsy specimens in 22 patients who were found to be H. pylori positive by histological detection with modified Giemsa stain, rapid urease test(CLO; Delta West Pty Ltd, Australia) and PCR using ureC primer in gastric biopsy specimens. RESULTS: The cagA gene was detected in 16(72.7%) of 22 patients. Eleven patients(50%) had both the cagA and vacA gene. Five patients had only the vacA gene. Twenty one patients(95.5%) had the cagA or vacA gene. The cagA gene was detected in 66.7% of gastritis and in 87.5% of peptic ulcer patients. The association of the cagA gene with peptic ulcer or the higher degree of inflammation did not reach statistical significance. The histological H. pylori density of antrum was significantly correlated with gastric inflammation(P<0.05). CONCLUSION: These findings suggest that the antral density of H. pylori was associated with the gastric inflammation. The association of the cagA gene with peptic ulcer or the higher degree of imflammation was not significant.
Azure Stains ; Biopsy* ; Child* ; Gastritis ; Helicobacter pylori* ; Helicobacter* ; Humans ; Inflammation* ; Peptic Ulcer ; Polymerase Chain Reaction* ; Urease

Since the culture of M. furfur is impossible, the KOH wet mount and various staining techniques have been applied for identification of the M. furfur. However, these methods still have many disputed points. Practically, the KOH wet mount method is in common use but. there are many difficulties in identifying the fungi. The author intended to suggest an easy and simple method for identification of the fungi, using the KOH and various other staining solutions, and comparing this with many known methods. At the same time, by applying the best method of identification which the author was able to develop, distribution of the fungi in the horny layer and the viability of the fungi during treatment were abserved In identifying the fungi, 1% toluidine blue was most excellent, but hematoxylin, eosin, cotton blue, Giemsa stain, and Wright stain were not so satisfactory. 2. After staining with l% toluidine blue to the skin lesion scotch tape was applied to the lesion briefly and then examined under direct microscope. This was most easy and convenient method. 3. Repeated scotch taping from ] to 12 times produced no change in the distribution of fungi in the horny layer, but after 28 applications there was remarkable reduction of the amount of the fungi and no fungi was demonstrated in groups taped more than 46 times. 4. No influence was noted in the distribution of fungi after repeated irradiation of Ultra-violet light once daily for 18 days. 5. Continous daily application of 25% sodium thiosulfate solution for average 3. 9 days, caused the disappearance of tungi and no fungal elements were found following EO days of successive observation. 6. Application of 2.5% selenium sulfide on alternate day for average 6.2 days, caused the disappearance of fungi and no fungal elements were found following 55- 62 days of successive observation.
Azure Stains ; Eosine Yellowish-(YS) ; Fungi ; Hematoxylin ; Malassezia* ; Selenium ; Skin ; Sodium ; Tinea Versicolor* ; Tinea* ; Tolonium Chloride