1.In vitro cytotoxicity test of medical ultrasonic couplant.
Jingxian GAO ; Shasha WANG ; Meng JIN ; Xiaoli YAN
Chinese Journal of Medical Instrumentation 2013;37(3):210-212
The cytotoxicity of medical ultrasonic couplant was tested by MTT assay and agar overlay test. By MTT assay, when the inoculum density was high, the cytotoxicity level was low, or vice versa. The cytotoxicity grade tested by agar overlay was not accord to MTT assay's too. MTT assay is suitable to test the cytotoxicity of medical ultrasonic couplant because it is quantitative and more sensitive, however, the experimental condition and the preparative method of extraction should be determined.
Animals
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Cell Line
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Colorimetry
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Cytotoxicity Tests, Immunologic
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methods
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Mice
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Ultrasonics
2.Cytotoxicity evaluation of the disposable medical syringe piston.
Huahong HE ; Wei LI ; Ting WU
Chinese Journal of Medical Instrumentation 2010;34(2):123-125
When some testing institutions performed biological evaluation to the disposable medical syringe piston, cytotoxicity was found. According to the biological evaluation testing Selection Guide proposed by Ministry of Health and the Comments of Sample Provider, We performed biological evaluation to one sample by using 5 tests of basic biological evaluation. Cytotoxicity was found, which was probably caused by the residue of the lotion. This research provides reference for objective evaluation of disposable medical syringe piston and safe guarantee of the product.
Cytotoxicity Tests, Immunologic
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Disposable Equipment
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Syringes
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adverse effects
4.Immunotoxicologic assessment of genetically modified drought-resistant wheat T349 with GmDREB1.
Chun-lai LIANG ; Yong-ning LI ; Xiao-peng ZHANG ; Yan SONG ; Wei WANG ; Jin FANG ; Wen-ming CUI ; Xu-dong JIA
Chinese Journal of Preventive Medicine 2012;46(6):556-560
OBJECTIVETo assess the immunotoxicologic effects of genetically modified drought resistant wheat T349 with GmDREB1 gene.
METHODSA total of 250 female BALB/c mice (6-8 week-old, weight 18-22 g) were divided into five large groups (50 mice for each large group) by body weight randomly. In each large group, the mice were divided into five groups (10 mice for each group) by body weight randomly, which were set as negative control group, common wheat group, parental wheat group, genetically modified wheat group and cyclophosphamide positive control group, respectively. Mice in negative control and positive control group were fed with feedstuff AIN-93G, mice in common wheat group, non-genetically modified parental wheat group and genetically modified wheat group were fed with feedstuffs added corresponding wheat (proportion up to 76%) for 30 days, then body weight, organ coefficient of spleen and thymus, peripheral blood lymphocytes phenotyping, serum cytokine, serum immunoglobulin, antibody plaque-forming cell (PFC), serum 50% hemolytic value (HC50), mitogen-induced splenocyte proliferation, delayed-type hypersensitivity (DTH) reaction and phagocytic activities of phagocytes were detected respectively.
RESULTSAfter 30 days raise, among negative control group, common wheat group, non-genetically modified parental wheat group, genetically modified wheat group and cyclophosphamide positive control group, mice body weight were (21.0±0.3), (20.4±0.7), (21.1±1.0), (21.1±1.0), (19.4±1.0) g, respectively (F=7.47, P<0.01); organ coefficient of spleen were (0.407±0.047)%, (0.390±0.028)%, (0.402±0.042)%, (0.421±0.041)%, (0.304±0.048)%, respectively (F=12.41, P<0.01); organ coefficient of thymus were (0.234±0.032)%, (0.246±0.028)%, (0.249±0.040)%, (0.234±0.034)%, (0.185±0.039)%, respectively (F=5.58, P<0.01); the percentage of T cell in peripheral blood were (70.43±4.44)%, (68.33±5.37)%, (73.04±2.68)%, (74.42±2.86)%, (90.42±1.66)%, respectively (F=57.51, P<0.01); the percentage of B cell were (13.89±3.19)%, (15.34±4.84)%, (13.06±4.22)%, (12.93±2.36)%, (3.01±0.96)%, respectively (F=12.79, P<0.01); the percentage of Th cell were (55.87±3.80)%, (55.24±4.60)%, (57.92±3.70)%, (59.57±2.54)%, (77.37±2.31)%, respectively (F=68.58, P<0.01);the Th/Ts ratio were 4.16±0.29, 4.73±0.96, 4.19±0.78, 4.52±0.40, 6.34±0.73, respectively (F=17.57, P<0.01);the serum IgG were (1046.38±210.67), (1065.49±297.22), (1517.73±299.52), (1576.67±241.92), (1155.88±167.05) µg/ml, respectively (F=10.53, P<0.01); the serum IgM were (333.83±18.97), (327.73±27.72), (367.47±27.18), (363.42±46.14), (278.71±24.42) µg/ml, respectively (F=12.11, P<0.01); the serum IgA were (51.69±10.10), (42.40 ± 8.35), (32.11±4.22), (37.12±4.90), (41.45±8.89) µg/ml, respectively (F=8.25, P<0.01); the PFC were (29.2±14.6), (28.0±20.0), (34.8±30.9), (33.2±25.1), (4.8±5.3) per 10(6) splenocyte, respectively (F=3.33, P<0.05); the HC50 were 82.3±6.5, 79.7±4.6, 75.8±4.1, 74.9±3.6, 70.8±2.1, respectively (F=9.99, P<0.01);the LPS-induced splenocyte proliferation were 0.21±0.10, 0.21±0.14, 0.26±0.12, 0.25±0.14, 0.07±0.06, respectively (F=4.18, P<0.05).
CONCLUSIONThe genetically modified drought-resistant wheat T349 was substantially equivalent to parental wheat in the effects on immune organs and immunologic functions of mice, and it didn't show immunotoxicity.
Animals ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; Droughts ; Female ; Mice ; Mice, Inbred BALB C ; Plants, Genetically Modified ; immunology ; toxicity ; Triticum ; genetics ; immunology ; toxicity
5.Evaluation of a Korean HLA-B27 typing tray.
Pil Whan PARK ; Think You KIM ; Se Jin HWANG
Korean Journal of Clinical Pathology 1999;19(2):222-226
BACKGROUND: Routine typing for HLA-B27 has been usually accomplished by the microcytotoxicity assay in Korea because it does not require special equipment and is easily reproducible. Recently, an immunofluorescence method and the polymerase chain reaction have also been applied for HLA-B27 testing. However, the current economic crisis in Korea have led domestic manufacturers to develop a Korean HLA-B27 typing kit. The aim of this study was to assess the advantages and disadvantages of this kit and to assess the possibility of replacing the currently used foreign-made kits with this domestic one. METHODS: HLA-B27 testing by the microcytotoxicity test was performed on 116 patients during a period of 3 months in 1998. The Biotest typing tray and the Chongkundang typing tray were tested simultaneously. Results: There was no difference in results in 116 samples (positive: 39, negative: 77). The Korean typing tray showed high false positivity of the negative control well (8 point: 1 case, 6 point: 33 cases, 4 point: 47 cases, 2 point: 17 cases, 1 point: 18 cases) and 6 of the HLA-B27 negative cases showed false positivity in one of the four HLA-B27 wells. Typing of HLA-Bw4 and Bw6 revealed an inconsistency in five and six cases, respectively. CONCLUSIONS: Despite of the false positivity of the negative control in Korean panel, we believe that Korean typing tray can replace the foreign-made tray due to its low cost and adequate performance. Because placenta of Korean multiparous women was used for the kit, Chongkundang typing tray seems to correlate better with Korean HLA-B27 subtypes.
Cytotoxicity Tests, Immunologic
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Female
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Fluorescent Antibody Technique
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HLA-B27 Antigen*
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Humans
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Korea
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Placenta
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Polymerase Chain Reaction
6.Study of biological safety of scaffold material with heterologous deproteinized bone.
Lei LIU ; Kang-lai TANG ; Liu YANG ; Yue-kui JING ; Qi-hong LI
Chinese Journal of Traumatology 2006;9(4):234-237
OBJECTIVETo evaluate the biological safety of manufactured heterologous deproteinized bone and to provide an experimental basis for clinical applications.
METHODSDeproteinized bone (10 mm) and leaching liquor were made from pig ribs with a series of physical and chemical methods, then were evaluated through acute and subacute toxicity test, hemolysis test, pyrogen test, intracutaneous test, intramuscular implantation test and cytotoxity test.
RESULTSNo obvious toxicity, hemolysis, pyrogenic characteristics, skin irritation, inflammatory reaction after intramuscular implantation and cytotoxity were observed.
CONCLUSIONSThe heterologous deproteinized bone has good biological safety and meets all the demands of scaffold material for tissue engineering.
Animals ; Bone Transplantation ; Cytotoxicity Tests, Immunologic ; Hemolysis ; Mice ; Rabbits ; Tissue Engineering ; Transplantation, Heterologous
7.A cell-based detection of ciguatoxin using sodium fluorescence probe.
Jian-hui YUAN ; Hui YANG ; Huan-wen TANG ; Wei HUANG ; Xin-yun XU ; Jian-jun LIU ; Yue-bin KE ; Jin-quan CHENG ; Zhi-xiong ZHUANG
Journal of Southern Medical University 2011;31(4):653-655
OBJECTIVETo establish a cell-based detection method of ciguatoxin using fluorescence assay.
METHODSMouse neuroblastoma N-2A cells were exposed to ouabain and veratridine and different concentrations of standard ciguatoxin samples (P-CTX-1) to establish the curvilinear relationship between the toxin dosage and fluorescence intensity using the sodium fluorescence probe CoroNaTM Green. The toxicity curvilinear relationship was also generated between the toxin dosage and cell survival using CCK-8 method. Based on these standard curves, the presence of ciguatoxin was detected in 33 samples of deep-sea coral fish.
RESULTSA correlation was found between the detection results of cell-based fluorescence assay and cytotoxicity assay, whose detection limit reached 103 g/ml and 1012 g/ml, respectively. The cell-based fluorescent assay sensitivity showed a higher sensitivity than cytotoxicity assay with a 2-4 h reduction of the detection time.
CONCLUSIONSThe cell-based fluorescent assay can quickly and sensitively detect ciguatoxin and may serve as a good option for preliminary screening of the toxin.
Animals ; Cell Line, Tumor ; Ciguatoxins ; toxicity ; Cytotoxicity Tests, Immunologic ; methods ; Fishes ; Fluorescent Dyes ; Mice ; Sodium
8.Natural Killer Cell Activity in Rheumatoid Arthritis Measured by a Single Cell Cytotoxicity Assay.
Gye Sung KIM ; Jung Koo YOUN ; Joo Deuk KIM ; Nam Hyun KIM
Yonsei Medical Journal 1988;29(2):160-165
The natural killer(NK) cell activity of mononuclear cells (MNC) from peripheral blood (PB) and synovial fluid (SF) of 40 rheumatoid arthritis(RA) patients was investigated by employing 51-chromium-(51Cr) release microcytotoxicity and single cell cytotoxicity assays against K562 target cells. It has been revealed that SF-MNC from RA patients showed a significantly lower NK activity than PB-MNC from the same patients and this might be due to an impaired target binding capacity of the effector cells and not due to a deficiency of active NK cells.
Adolescent
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Adult
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Aged
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Arthritis, Rheumatoid/*immunology
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Chromium Radioisotopes/diagnostic use
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Comparative Study
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Cytotoxicity Tests, Immunologic/methods
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*Cytotoxicity, Immunologic
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Female
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Human
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In Vitro
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Killer Cells, Natural/*immunology
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Male
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Middle Age
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Support, Non-U.S. Gov't
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Synovial Fluid/immunology
9.Effects of polypeptides from HCV core region on the function of cytotoxic T cells.
Qiaoyu MA ; Yuming WANG ; Fei HAO
Chinese Journal of Hepatology 2002;10(6):426-428
OBJECTIVETo investigate the pathogenesis of cytotoxic T cell (CTL) dysfunction in patients with HCV infection.
METHODSBALB/c mice were immunized by subcutaneous injection of polypeptides from HCV core region, and the CTL activity of mouse spleen cells was detected by the LDH release test. Two polypeptides which can enhance CTL function and two polypeptides which can inhibit CTL function were selected and cross-combined. BALB/c mice were immunized using the combined polypeptides and the CTL activities were detected afterwards.
RESULTSCTL activity was inhibited by CPA9 (39-74 amino acids), CPB7 (67-76 amino acids) and CPB8 (71-80 amino acids), and promoted by CPA10 (5-23 amino acids), CPB6 (63-72 amino acids) and CPB2 (131-140 amino acids). Using single factor analysis of variance, the CTL activity in the mice could be enhanced by polypeptides from the HCV core region, CPB2+CPB8, CPB6+CPB8, respectively. There was no obvious difference between CPB2+CPB7, CPB6+CPB7 and negative control.
CONCLUSIONSCPA9, CPB7, and CPB8, the 3 polypeptides from HCV core region play an inhibition role and CPA10, CPB6, and CPB2 play an enhancement role in CTL activity in mice. The inhibition and enhancement functions of the polypeptides from HCV core region interact each other.
Animals ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; Mice ; Mice, Inbred BALB C ; Peptide Fragments ; administration & dosage ; immunology ; Spleen ; cytology ; drug effects ; immunology ; T-Lymphocytes, Cytotoxic ; cytology ; immunology ; Viral Core Proteins ; administration & dosage ; chemistry ; immunology
10.Characteristics of peripheral NK cells in hepatocellular carcinoma patients.
Lin ZHOU ; Lun CAI ; Zheng ZHANG ; Yong-ping YANG ; Fu-sheng WANG
Chinese Journal of Hepatology 2010;18(2):136-139
OBJECTIVEFunctional defects in NK cells have been proposed to be responsible for the impairment of anti-tumor immune responses. However, it remained unclear whether the function of NK cells were impaired in patients with hepatocellular carcinoma. To address this issue, we analyzed the frequency and function of peripheral NK cell subsets in hepatocellular carcinoma (HCC) patients.
METHODS35 HCC patients and 24 healthy controls (HC) were enrolled in the study. Peripheral NK frequency was analyzed using flow cytometry. In addition, the capacity of NK cells to produce IFN gamma and to lyse K562 cells was evaluated.
RESULTSIn contrast with the healthy controls, the frequency of peripheral NK cells in hepatocellular carcinoma patients was decreased (12.19%+/-10.85% vs 24.01%+/-8.78%, u = 4.01, probability value less than 0.01), while the frequency of CD56(bright)CD16(neg) NK cells was increased (0.62%+/-0.39% vs 0.48%+/-0.28%, u = 1.96, probability value less than 0.05), and the frequency of CD56(dim)CD16(pos) NK cells was significantly decreased (11.59%+/-7.49% vs 22.66%+/-8.84%, u = 3.92, probability value less than 0.01). In addition, peripheral NK cells from HCC patients exhibited decreased capacity to produce IFN gamma (effective cells 13.31%) and to lyse K562 cells (mixed ratio 30:1, 10:1, 1:1, effective cells 16.72%+/-7.33% vs 26.29%+/-12.36%, u = 2.52, P less than 0.05, 8.01%+/-4.40% vs 13.09%+/-5.03%, u = 3.32, probability value less than 0.05, 3.51%+/-2.82% vs 3.42%+/-1.64%, u = 1.56, probability value more than 0.05, respectively) as compared with healthy subjects.
CONCLUSIONAnti-tumor activity of NK cells in HCC patients was impaired.
Adult ; CD56 Antigen ; immunology ; Carcinoma, Hepatocellular ; immunology ; Case-Control Studies ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; Female ; Flow Cytometry ; Humans ; Interferon-gamma ; metabolism ; K562 Cells ; Killer Cells, Natural ; immunology ; Liver Neoplasms ; immunology ; Lymphocyte Subsets ; immunology ; Male ; Middle Aged ; Receptors, IgG ; immunology