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 ; Cell Line ; Colorimetry ; Cytotoxicity Tests, Immunologic ; methods ; Mice ; Ultrasonics

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 ; Disposable Equipment ; Syringes ; adverse effects

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

In a previous report, it was felt that the rat tumor cell line, T-333, was a mixture of heterogeneous cells with different characteristics with respect to karyotype, tumorigenicity, and response to Rolls Sarcoma virus (RSV) infection. These characteristics of hetero-geneous cell subpopulations could be selected by use of different culture substrates. In this experiment, diversity of the cells in response to complement mediated cytolysis employing syngeneic rat anti-sera was studied. More than 50% of the glass grown cells were lysed while only 19% of the plastic grown cells were lysed by the specific immune sera of syngeneic rats. This finding suggests that growth in plastic culture wares selects cells with resistance to complement mediated cytolysis. It seems likely that the previously reported enhanced tumorigenicity of plastic grown T-333 cells is due to clonal selection of cell subpopulations which can better tolerate at least one arm of the in vivo immune surveillance system.
Animal ; Cells, Cultured ; Cytotoxicity, Immunologic ; Neoplasms, Experimental/immunology* ; Plastics ; Rats ; Rats, Inbred Strains

Cellular immunity is an important component of human immune system and plays a crucial role in the fight against tumor cell or invasive pathogens. Researches on cell-based immunotherapy have long been focused on eliciting tumor-specific CD8+ cytotoxic T lymphocytes (CTL) because of their potent killing activity and their ability to reject transplanted organs. However, the resulting treatments have been surprisingly poor at inducing complete tumor rejection, in both the experimental models and clinical trials. The CD4+ T cells has been studied mainly for their role as helpers for CD8+ CTL, even suggesting that the tumor-specific CD4 T regulatory cells could act as suppressors of antitumor responses. Recent studies indicated that CD4+T cells are not a pure cell lineage with single function, but a cell population with complex functions. Moreover CD4+ T cells may not only be helper cells, but also act as potent effector cells or partners with NK cells that can clear a wide variety of tumors. In a word, the antitumor potential of effector CD4+ T cells might have been underestimated. In this article, the classification and differentiation of CD4+ T cells, the function and secreted cytokines of CD4+ T cells, the CD4+ T cells and tumor immune, the tumor-immuno regulatory effects of CD4+ T cells, and clinical researches of CD4+ T cells are reviewed.
CD4-Positive T-Lymphocytes ; classification ; cytology ; immunology ; Cytokines ; metabolism ; Cytotoxicity, Immunologic ; Humans ; Immunity, Cellular

Natural killer (NK) cells are important innate immune effector cells with broad applications in killing the tumor cells and pathogens due to its cytotoxicity without prior immune sensitization. Unfortunately, in humans, the activity of NK cells against fungi is poorly characterized. Insight progress in the fields of NK cells activating, pattern recognition receptors, signal modulating and correlated cell factors (IFN-γ, GM-CSF, IL-10 and so on) has revolutionized understanding of the selective killing fungi by NK cells. Different morphotypes also can affect the immune status of NK cells. This article reviews the mechanism of fungi immune reaction, and the interaction between NK cells and fungi, and provides some new ideas for further study on pathogenesis of fungus and other infectious diseases and NK cell adoptively transferred immunotherapy.
Cytotoxicity, Immunologic ; Fungi ; immunology ; Host-Pathogen Interactions ; Humans ; Killer Cells, Natural ; immunology

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

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 ; Female ; Fluorescent Antibody Technique ; HLA-B27 Antigen* ; Humans ; Korea ; Placenta ; Polymerase Chain Reaction

OBJECTIVETo establish a novel method for ex vivo expansion of natural killer cells from human umbilical blood, so as to provide the basis for NK cell therapy.

METHODSMononucleated cells from human umbilical blood were harvested and suspended in a serum-free medium containing 5% autologous plasma, recombinant human IL-15 (50 ng/ml) and hydrocortisone sodium succinate (5×10 mol/L) at a concentration of 1.5×10/ml, then the cells were seeded into flasks pre-coated with heparin sodium (100 U/cm) or/and anti-human CD16 antibody (1 µg/cm). After culture for 2 weeks, the cells were harvested and counted. Ratios of CD3/CD56 of the cells were determined by flow cytometry. MTT test was performed to assess the cytotoxicity against K562 cells with graded ratios of effector/target cells.

RESULTSIn contrast to the cells in flasks without pre-coating, the attached colonies appeared predominantly within 1 week of culture from heparin- and antibody-coated groups. The cell numbers from the pre-coated groups were significantly higher than that of uncoated one after culture for 2 weeks. Furthermore, the ratios of CD3/CD56 cells were much higher in pre-coated groups, and that of the cells from flasks pre-coated with heparin and antibody were the highest (all the P values <0.01). MTT test showed that the cytotoxic activity of the cells stimulated by precoating were much more potent than that of the cells without the stimulation.

CONCLUSIONAdvantageous expansion of NK cells can be achieved by precoating with heparin and anti-CD16 antibody, and also by supplement with IL-15 and hydrocortisone into the media, so the umbilical NK cells with high purity and potent cytotoxicity can be obtained.

CD56 Antigen ; Cells, Cultured ; Cytotoxicity, Immunologic ; Fetal Blood ; Heparin ; Humans ; Killer Cells, Natural

This study was conducted to compare the hemostatic efficacy of three ferric subsulfate- and chitosan-based styptics as a powder and a gel containing ferric subsulfate and chitosan (FSC-PO and FSC-G, respectively) and a soaked pad containing ferric subsulfate and lidocaine (FSL-SP) using a rat tail bleeding model. The cytotoxicity of the styptics against L-929 mouse fibroblasts was also evaluated using a cell counting kit-8 assay. Four groups of 10 rats each were assigned to the three different styptics and a non-treated control groups. Rat tail tips were transected, after which styptics were applied with pressure. The wounds were observed for hemostasis for 3 min, then irrigated with saline to check for recurrent hemorrhage. L-929 mouse fibroblasts were exposed to extracts of the styptics (100 mg/mL) and their dilutions (1:10, 1:100, and 1:1,000). FSC-PO and FSC-G more effectively controlled initial hemorrhage than FSL-SP (p = 0.033). Additionally, FSC-PO and FSC-G more effectively maintained hemostasis than the control group (p = 0.02 and p < 0.01, respectively). However, all styptics showed enhanced cytotoxicity against L-929 cells in a dose-dependent manner. Therefore, although FSC-PO and FSC-G would be recommended to control hemorrhage, the benefits of styptics must be balanced against the clinical significance of their cytotoxicity.
Animals ; Cell Count ; Chitosan ; Cytotoxicity, Immunologic ; Fibroblasts ; Hemorrhage ; Hemostasis ; Hemostatics ; Lidocaine ; Mice ; Rats ; Tail ; Wounds and Injuries