[Effect of Rate of Salinity Increase on the Performance and Microbial Community Structure of Sequencing Batch Reactors].

In this study, three sequence batch reactors were selected to evaluate the effects of salt-tolerant activated sludge acclimation. The effect of salinity increase rate on pollutant removal, physicochemical characteristics of activated sludge, and microbial community were investigated. The results showed that a rapid salinity increase to 30‰ (within 30 d) reduced removal efficiencies of COD and NH4+-N from 85.5% and 98.5% (18 d) to 72.2% and 81.7% (51 d), respectively. In comparison, a slower salinity increases to 30‰ (within 90 d) had a minor effect on COD and NH4+-N removal. During the rapid salinity increase, a stable shortcut nitrification occurred under 20‰ salinity, in which the effluent NO2--N reached 11.13 mg·L-1 and NO3--N decreased to 0.56 mg·L-1. When salinity increased to 30‰, the nitrite accumulation rate was about 90%, and the removal efficiency of total nitrogen increased to approximately 75%. The contents of polysaccharide and protein in extracellular polymer substances increased as salinity increased, and the polysaccharide content increased significantly when the salinity was higher than 15‰. High-throughput sequencing results illustrated that microbial diversity reduced as salinity increased, following the Shannon index decrease from 8.06 (0‰ salinity) to 4.34 (rapid salinity increase) and 6.17 (slower salinity increase). As salinity increased, Micropruina, Denitromonas, TM7a, and Marinicella exhibited good salt tolerance. The relative abundance of Denitratisoma, Defluviimonas, Arenimonas, and Denitromonas decreased more significantly following the rapid salinity increase compared with that after the slower salinity increase.

[Performance and Membrane Fouling Properties in an Anaerobic Membrane Bioreactor for Salty Wastewater].

In the research, an anaerobic membrane bioreactor (AnMBR) was used to treat simulated salty organic wastewater, and the effect of salinity on reactor performance and membrane fouling properties was investigated. The results indicated that when the influent salinity increased gradually but was lower than 9.1 g·L-1, the reactor ran stably and the effluent performance was good. When the salinity increased to 10 g·L-1, the COD removal-efficiency, gas production, and methane content decreased significantly; meanwhile, the sludge concentration, sludge volume index (SVI), soluble microbial products (SMP), and extracellular polymeric substance (EPS) levels became elevated at first and then declined with the rising salinity. The system developed compact flocs and a high settling ability. The hollow fiber membrane module was run for three cycles in 118 d. The membrane operating cycle was extended from 31 d to 48 d with the increasing salinity, which favored the control of membrane fouling. SEM-EDX analysis results revealed that there were similar crystalline substances in the film membrane foulants, and Na, Mg, Al, Si, Cl, K, Ca, and Fe were the main inorganic elements. Excitation emission matrix (EEM) analysis results demonstrated that proteins and humic acids were the main components of the organic membrane foulants.

Effects of sevoflurane before cardiopulmonary bypass on cerebral oxygen balance and early postoperative cognitive dysfunction.

Postoperative cognitive dysfunction (POCD) is associated with cardiopulmonary bypass (CPB). We investigated the effect of different doses of inhaled sevoflurane administered prior to CPB on cerebral oxygen supply and demand, and the incidence of associated early POCD. One hundred and twenty patients were randomly allocated into four treatment groups (n = 30, each) and administered a high- [1.5 minimum alveolar concentration (MAC)], moderate- (1.0 MAC), low- (0.5 MAC), or no- sevoflurane dose prior to CPB. Standard blood gas parameters, serum S-100 protein, and neuron-specific enolase (NSE) were measured at different time points. The mini-mental state examination (MMSE) was administered 1 day before and 24 and 72 h after surgery. The jugular bulb venous oxygen saturation (SjvO2) in the moderate- and high-dose groups at a nasopharyngeal temperature of 25-28 °C was significantly higher compared with the control group, while the arteriovenous oxygen content difference (AVDO2) and cerebral extraction of oxygen (CEO2) were significantly reduced. The serum S-100 protein and NSE concentrations of the moderate- and high-dose groups at 1 and 6 h after the cessation of CPB were significantly lower than that of the control group. The 24 h postoperative MMSE scores of the moderate- and high-dose groups were significantly higher than those of the low-dose and control groups. An inhaled optimal concentration of sevoflurane may be beneficial for cerebral oxygen balance during CPB, and may ameliorate cognitive damage. However, the effect is dose-dependent.

[Levels of nerve growth factor and interleukin-4 in the induced sputum of children with cough variant asthma].

OBJECTIVE: To examine the levels of nerve growth factor (NGF) and interleukin-4 (IL-4) in the induced sputum of children with cough variant asthma (CVA), with the aim of studying the roles of NGF and IL-4 in childhood CVA. METHODS: Thirty-four children with CVA were enrolled in this study. Twenty healthy children were used as a normal control group. The induced sputum was separated into supernatant and cells. Hematoxylin and eosin staining was used to count differential cells. The expression of NGF and IL-4 in supernatant was measured using ELISA. The mRNA expression of NGF and IL-4 in cells was determined by Real-time PCR analysis. RESULTS: The percentage of eosinophils in the CVA group was significantly higher than in the control group [(13.4±3.6)% vs (2.6±1.7)%; P<0.01]. The expression of NGF and IL-4 protein and mRNA in induced sputum was significantly higher in the CVA group than in the control group (P<0.05). The expression of NGF and IL-4 protein and mRNA was positively correlated with the percentage of eosinophils (P<0.01). The expression of NGF and IL-4 protein and mRNA in induced sputum was significantly reduced in the CVA group after treatment (P<0.05). CONCLUSIONS: Eosinophils infiltration and increased expression of NGF and IL-4 play key roles in the development of childhood CVA, suggesting that they may be useful in the diagnosis and treatment of childhood CVA.

Identification of novel HLA-A*0201-restricted epitopes from anterior gradient-2 as a tumor-associated antigen against colorectal cancer.

Anterior gradient-2 (AGR2) promotes tumor growth, cell migration and cellular transformation and its enhanced expression is almost completely restricted to malignant tissues, thus making AGR2 an interesting target for the development of immunotherapeutic strategies. We investigated whether the AGR2 molecule comprises human leukocyte antigen (HLA)-A*0201-binding epitopes recognized by human cytotoxic T lymphocytes (CTLs), which could be targeted in dendritic cell (DC)-based cancer immunotherapy against colorectal cancer (CRC). We reviewed the sequence of AGR2 for peptides that could potentially bind to HLA-A*0201 with the aid of a computer-based program. Five candidate peptides with different binding scores were synthesized and tested. These peptides were then assessed for their immunogenicity to elicit specific immune responses mediated by CTLs in vitro by means of enzyme-linked immunospot assays and CTL assays. AGR2 was highly expressed in several CRC cell lines, including DK01, DLD1, KM12C, HCT-8 and HT-29. DCs pulsed with AGR2-P2 (aa 11-19; LLVALSYTL) or AGR2-P4 (aa 127-135; RIMFVDPSL) generated potent CTLs that could lyse T2 cells pulsed with AGR2-P2 or AGR2-P4 and HLA-A0201(+) AGR2-positive CRC cell lines in a strong dose-dependent and HLA-A*0201-restricted manner. In conclusion, these novel epitopes derived from AGR2 protein may be attractive candidates for DC-based immunotherapy for CRC.

Alpha-Type 1 Polarized Dendritic Cells Loaded with Apoptotic Allogeneic Breast Cancer Cells Can Induce Potent Cytotoxic T Lymphocytes against Breast Cancer.

PURPOSE: Various tumor antigens can be loaded onto dendritic cells (DCs) to induce a potent cytotoxic T lymphocyte (CTL) response in DC-based immunotherapy against breast cancer. However, in the clinical setting, obtaining a sufficient number of autologous tumor cells as a source of tumor antigens is a laborious process. We therefore investigated the feasibility of immunotherapy using breast-cancer-specific CTLs generated in vitro by use of alpha-type 1 polarized DCs (α DC1s) loaded with ultraviolet B-irradiated cells of the breast cancer cell line MCF-7. MATERIALS AND METHODS: αDC1s were induced by loading allogeneic tumor antigen generated from the MCF-7 UVB-irradiated breast cancer cell line. Antigen-pulsed αDC1s were evaluated by morphological and functional assays, and the breast-cancer-specific CTL response was analyzed by cytotoxic assay. RESULTS: The αDC1s significantly increased the expression of several molecules related to DC maturation without differences according to whether the αDC1s were loaded with tumor antigens. The αDC1s showed a high production of interleukin-12 both during maturation and after subsequent stimulation with CD40L, which was not significantly affected by loading with tumor antigens. Breast-cancer-specific CTLs against autologous breast cancer cells were successfully induced by αDC1s loaded with apoptotic MCF-7 cells. CONCLUSION: Autologous DCs loaded with an allogeneic breast cancer cell line can generate potent breast-cancer-specific CTL responses. This may be a practical method for cellular immunotherapy in patients with breast cancer.

Uncarinic Acid C Isolated from Uncaria rhynchophylla Induces Differentiation of Th1-Promoting Dendritic Cells Through TLR4 Signaling.

Uncarinic acid C (URC) is triterpene isolated from Uncaria rhynchophylla and is a pharmacologically active substance. The induction of dendritic cells (DC) is critical for the induction of Ag-specific T lymphocyte responses and may be essential for the development of human vaccines relying on T cell immunity. DC might be a potential target for URC. We demonstrate that URC activates human DC as documented by phenotypic and functional maturation, and altered cytokine production. The expression of CD1a, CD38, CD40, CD54, CD80, CD83, CD86, HLA-DR and CCR7 on URC-primed DC was enhanced. The production of IL-12p70 by URC-primed DC was higher than that of lipopolysaccharide (LPS)-primed DC. The production of IL-12p70 by URC-primed DC was inhibited by the anti-Toll-like receptor 4 (TLR4) monoclonal antibody (mAb), but partially abolished by anti-TLR2 mAb. mRNA coding for TLR2 and TLR4 was expressed in URC-primed DC. URC-primed DC induced the NF-κB transcription factor. Naïve T cells co-cultured with URC-primed DC turned into typical Th1 cells that produced large quantities of IFN-γ depending on IL-12 secretion. URC enhanced the T cell stimulatory capacity in an allo MLR. In the cytotoxic T-lymphocyte assay (CTL) assay, DNA fragmentation assay and (51)Cr release on URC-primed DC were more augmented than that of TNF-α-primed DC. DC matured with URC had an intermediate migratory capacity towards CCL19 and CCL21. These results suggest that URC modulates DC function in a fashion that favors Th1 polarization via the activation of IL-12p70 dependent on TLR4 signaling, and may be used on DC-based vaccine for cancer immunotherapy.

Type I and II interferons enhance dendritic cell maturation and migration capacity by regulating CD38 and CD74 that have synergistic effects with TLR agonists.

The major limitation for the maturation of dendritic cells (DCs) using Toll-like receptor (TLR) agonists is their decreased ability to migrate into lymph nodes compared with conventional DCs. CD38 can be used as a multifunctional marker to modulate migration, survival and Th1 responses of DCs. CD74 has been shown to negatively regulate DC migration. The goal of this study was to investigate the combinations of TLR agonists and interferons (IFNs) that most effectively regulate CD38 and CD74 expression on DCs. Synergistic TLR agonist stimulation in combination with IFN-α and IFN-γ was the best method for regulating CD38 and CD74 expression and inducing the highest secretion of IL-12p70. An in vitro migration assay showed that DCs treated with this combination had significantly enhanced migratory ability, similar to that observed in cells expressing CD38, CD74 and CCR7. The results of this study suggest that an alternative maturation protocol in which two TLR ligands are combined with type I and II IFNs generates potent DCs that have both a high migratory capacity and high IL-12p70 production.

Epidemiological investigation of Kawasaki disease in Jilin province of China from 2000 to 2008.

OBJECTIVE: To investigate the epidemiological characteristics of Kawasaki disease in Jilin province of China and explore its clinical features. METHODS: The medical records of children with Kawasaki disease hospitalised in the First Affiliated Hospital of Jilin University and Yanbian University between January, 2000 and December, 2008 were retrospectively analysed. RESULTS: A total of 735 children with Kawasaki disease were enrolled in this study with 483 boys and 252 girls. The ratio of male to female was 1.92:1. The ages of the children at onset varied from 51 days to 12 years with a mean age of 2.8 years. The children under the age of 5 years accounted for 79.5%, but most children were 2-3 years old. Kawasaki disease occurred all the year and more frequently in both the ending of spring and the beginning of summer. Fever was the most common clinical feature and enlarged cervical lymph nodes were the smallest clinical feature. A cardiovascular lesion was found in 41.4% of these children, in whom coronary artery dilatation was the most common (26.97%). A total of 117 (18.2%) of 643 children (87.5%) receiving intravenous immunoglobulin had a non-response to gamma globulin. Of the 117 children, 66 (56.4%) had cardiovascular lesion. Kawasaki disease recurred in 19 children (2.6%). CONCLUSION: The incidence of Kawasaki disease in Jilin province has shown an increasing tendency. The age at onset is slightly higher than that described in other reports. Kawasaki disease is the most common in both the ending of spring and the beginning of summer, and the second incidence peak occurs in autumn.

Properties of L-type amino acid transporter 1 in epidermal ovarian cancer.

HYPOTHESIS: To investigate the expression and the functional properties of L-type amino acid transporter 1 (LAT1) in human epithelial ovarian cancer to provide a basis for potential new therapies to control the growth and the metastasis of ovarian cancer. METHODS: The material used comprised 63 surgically resected specimens obtained from female patients undergoing gynecologic surgery at Kyorin University School of Medicine (Tokyo, Japan). The expression of LAT1 in 53 cases of ovarian cancers was determined by Western blot and immunohistochemical staining, and results were compared with those of normal ovarian tissues (5 cases) and benign ovarian tumors (5 cases). Furthermore, we examined the effect of 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH), the classic inhibitor of system L on the survival, the migration, and the uptake of l-leucine by human epithelial ovarian cancer cell line (OVCAR-3). RESULTS: The LAT1 was significantly up-regulated in various human epithelial ovarian cancers that was localized predominantly on their plasma membrane and in the plasma membrane of the ovarian cancer cell line in conjunction with 4F2hc via disulfide bonds. The BCH inhibited the proliferation and the migration of the OVCAR-3 cells and the uptake of [14C]l-leucine by these cells in a dose-dependent manner. The OVCAR-3 cells did not express LAT2, and the uptake of [14C]l-leucine by these cells was Na-independent and almost completely inhibited by BCH. Thus, our findings indicated that most l-leucine uptake in OVCAR-3 cells was mediated by LAT1. CONCLUSIONS: The LAT1 plays significant roles in nutrition, proliferation, and migration of ovarian cancer. Then, LAT1 inhibition would be useful for anticancer therapy in suppressing tumor growth without affecting normal tissues.

Alpha-type 1-polarized dendritic cells loaded with apoptotic allogeneic myeloma cell line induce strong CTL responses against autologous myeloma cells.

To induce a potent cytotoxic T lymphocyte (CTL) response, various tumor antigens should be loaded onto dendritic cells (DCs). In multiple myeloma (MM), it is difficult to obtain a sufficient number of autologous tumor cells as a source of tumor antigens in the clinical setting. We investigated the feasibility of immunotherapy in patients with MM, using myeloma-specific CTLs generated in vitro by alpha-type 1-polarized DCs (alphaDC1s) loaded with the ultraviolet B-irradiated allogeneic myeloma cell line, ARH77. alphaDC1s significantly increased the expression of several costimulatory molecules without differences in loading with tumor antigens. alphaDC1s showed a high production of interleukin-12 during maturation and after subsequent stimulation with CD40L but were not significantly affected by loading tumor antigens. Myeloma-specific CTLs against autologous myeloma cells from MM patients were induced by alphaDC1s pulsed with apoptotic ARH77 cells. Our data indicate that autologous DCs loaded with an allogeneic myeloma cell line can generate potent myeloma-specific CTL responses against autologous myeloma cells and might provide a practical method for cellular immunotherapy in patients with MM.

All-trans retinoic acid inhibits the differentiation, maturation, and function of human monocyte-derived dendritic cells.

All-trans retinoic acid (ATRA) affects on the function of antigen presenting cells with somewhat controversies. We investigated the effects of ATRA on differentiation, maturation and function of human monocyte-derived dendritic cells (DCs). Low dose (10(-14)M) or high dose (10(-6)M) of ATRA was added either when monocytes were differentiated into immature DCs (imDCs) or mature DCs (mDCs) were induced. Apoptotic cell populations were dramatically increased in imDCs or mDCs with increasing concentration of ATRA. The productions of IL-12p40 and IL-12p70 were significantly suppressed in imDCs or mDCs induced by the addition of ATRA in the dose-dependent manner, whereas IL-10 was increased. DCs cultured with ATRA induced the differentiation of naïve T cells towards a helper T cell type 2 (Th2) response and expansion of CD4(+)CD25(+)Foxp3(+) regulatory T cells. Allostimulatory capacity of DCs was suppressed with increasing concentration of ATRA. These findings suggest that ATRA inhibits the effects on the differentiation, maturation and function of human monocyte-derived DCs in vitro and also enhance the differentiation of naïve T cell toward the Th2 type.

Induction of myeloma-specific cytotoxic T lymphocytes ex vivo by CD40-activated B cells loaded with myeloma tumor antigens.

We investigated to establish CD40-activated B cells (CD40-B cells) as alternative antigen-presenting cells (APCs) for the induction of myeloma-specific cytotoxic T lymphocytes (CTLs). To generate CD40-B cells, peripheral blood mononuclear cells were co-cultured with CD40L-transfected J558 cells in the presence of IL-4, insulin, transferrin, and cyclosporine for 14 days, and pulsed with myeloma lysates. The CD40-B cells consistently expressed high levels of CD80, CD86, CD54, CCR7, and HLA-DR. The CD40-B cells produced IL-12, IFN-gamma, and IL-6 during the culture period, but not IL-10. In addition, the CD40-B cells showed potent allogeneic T-cell stimulatory capacities that depended on the dose ratio and had the potential to polarize naïve T cells into Th1 subsets. The CD40-B cells loaded with tumor lysates induced strong target-specific CTLs, based on large numbers of IFN-gamma secreting cells and higher cytotoxic activity against target cells compared to the CD40-B cells without the tumor lysates. These results suggest that CD40-B cells loaded with myeloma lysates might provide alternative APCs for cellular immunotherapy in patients with myeloma.

The dysfunction and abnormal signaling pathway of dendritic cells loaded by tumor antigen can be overcome by neutralizing VEGF in multiple myeloma.

We investigated whether dendritic cells (DCs) from multiple myeloma (MM) patients were affected by loading tumor antigens and whether the defective DC function associated with MM could be overcome by the neutralization of VEGF. MM-specific DCs were generated by loading tumor lysates from myeloma cells at diagnosis or relapsed/progressive state, respectively. DCs loaded with tumor lysates showed lower phenotypic maturation, less T cell stimulatory capacity, less cytotoxic T lymphocyte activities, and highly abnormal cytokine secretions of IL-6 and IL-12, compared to myeloma lysate-unloaded DCs. The levels of VEGF, phospho-STAT3 and phospho-ERK1/2 in DCs were significantly higher with loading myeloma lysates. After the neutralization of VEGF activity, the DC function, signal transduction and cytokine production returned to normal. The defective function of DC in patients with MM is significantly affected by loading tumor antigens, correlating with abnormal STAT3 and the NF-kappaB signaling pathway, and neutralization of VEGF can overcome this DC dysfunction through the elimination of abnormal signal transduction.

Plasma urate level is directly regulated by a voltage-driven urate efflux transporter URATv1 (SLC2A9) in humans.

Hyperuricemia is a significant factor in a variety of diseases, including gout and cardiovascular diseases. Although renal excretion largely determines plasma urate concentration, the molecular mechanism of renal urate handling remains elusive. Previously, we identified a major urate reabsorptive transporter, URAT1 (SLC22A12), on the apical side of the renal proximal tubular cells. However, it is not known how urate taken up by URAT1 exits from the tubular cell to the systemic circulation. Here, we report that a sugar transport facilitator family member protein GLUT9 (SLC2A9) functions as an efflux transporter of urate from the tubular cell. GLUT9-expressed Xenopus oocytes mediated saturable urate transport (K(m): 365+/-42 microm). The transport was Na(+)-independent and enhanced at high concentrations of extracellular potassium favoring negative to positive potential direction. Substrate specificity and pyrazinoate sensitivity of GLUT9 was distinct from those of URAT1. The in vivo role of GLUT9 is supported by the fact that a renal hypouricemia patient without any mutations in SLC22A12 was found to have a missense mutation in SLC2A9, which reduced urate transport activity in vitro. Based on these data, we propose a novel model of transcellular urate transport in the kidney; urate [corrected] is taken up via apically located URAT1 and exits the cell via basolaterally located GLUT9, which we suggest be renamed URATv1 (voltage-driven urate transporter 1).

Mouse organic anion transporter 2 (mOat2) mediates the transport of short chain fatty acid propionate.

In this study, we have elucidated that propionate, one of the short chain fatty acids (SCFAs), is the transport substrate for murine organic anion transporter 2 (mOat2), which is expressed in the kidneys and the liver. When expressed in Xenopus oocytes, mOat2-mediated [(3)H]PGE(2) transport was inhibited by three- to five-carbon SCFAs (C3 to C5). Among the SCFAs tested, propionate (3-carbon SCFA) was transported by mOat2 in a time-dependent manner. Since propionate is a potent glucogenic compound, Oat2 may be involved in the regulation of cellular metabolism through the transport of these metabolites in the kidneys and the liver.

[Effects of different co-substrates on degradation of nitrophenols using upflow anaerobic sludge bed (UASB) reactors].

The removal efficiencies of 3-nitrophenol (3-NP) and 2, 6-dinitrophnol (2, 6-DNP) were investigated in two lab-scale upflow anaerobic sludge bed (UASB) reactors using two different co-substrates. Initially, glucose was used as co-substrate and followed by sodium acetate. The results showed that glucose was found to be a better co-substrate for 3-NP degradation compared to sodium acetate. While for the degradation of 2,6-dinitrophenol, sodium acetate was better. For the study of 3-NP degradation, input COD concentration was kept as 2,500 mg/L and hydraulic retention time (HRT) was kept as 26 h with glucose as co-substrate. Maximum 3-NP concentration was 254.6 mg/L and 3-NP removal efficiencies were always more than 99.0%. While HRT was 30 h with sodium acetate as co-substrate, maximum 3-NP concentration was 71.6 mg/L and over 90.0% 3-NP removal efficiencies could be obtained. For the study of 2,6-DNP degradation, HRT was 35 h using the same input COD concentration as 3-NP degradation. The maximum 2,6-DNP concentration was 170.0 mg/L and 2,6-DNP removal efficiencies were always more than 98.0% using glucose as co-substrate. While HRT was 30 h with sodium acetate as co-substrate, maximum 2,6-DNP concentration was 189.5 mg/L and over 99.2% 2,6-DNP removal efficiencies could be obtained.

[Anaerobic degradation kinetics of 2,4-dinitrophenol].

With glucose as co-substrate, anaerobic degradation kinetics of 2,4-dinitrophenol (2,4-DNP) were investigated in batch culture. The results show that 2,4-DNP and glucose can be degraded by the bacteria simultaneously. Although the effect of COD on 2,4-DNP degradation was minimal, addition of 2,4-DNP effected COD degradation obviously. The rate of 2,4-DNP degradation increased with increasing initial 2,4-DNP concentrations up to 225 mg/L. Further increase in initial 2,4-DNP concentrations caused decrease in the rate of degradation because of substrate inhibition. Uncompetitive inhibition equation is proposed to describe the degradation of 2,4-DNP. With non-linear regression technology, the kinetic model parameter q(max), K(s) and K(i) are found to be 3.24 mg/(h x g), 196.23 mg/L and 165.91 mg/L respectively. The experimental data verification for the model equation is satisfactory.

[Anaerobic biodegradation of nitrophenols with glucose as co-substrate].

A batch anaerobic test was conducted to examine the biodegradation of 2-nitrophenol, 4-nitrophenol, 2,4-dinitrophenol and 2,6-dinitrophenol through measuring accumulative methane production. The Relative activity values were used to judge the inhibition level of nitrophenols on methanogenic bacteria. The test conditions was as follow: glucose was used as co-substrate and the temperature is 35 degrees C. It didn't cause inhibition when concentrations of 2-nitrophenol, 4-nitrophenol, 2,6-dinitrophenol and 2,4-dinitrophenol were below 24mg/L, 20mg/L, 12mg/L and 4mg/L respectively. Slight inhibition was caused when concentrations of 4-nitrophenol and 2,6-dinitrophenol were 24mg/L and 16mg/L-24mg/L respectively. Middle inhibition when concentrations of 2,4-dinitrophenol were 8mg/L-24mg/L was observed. The inhibition level was 2,4-dinitrophenol > 2,6-dinitrophenol > 4-nitrophenol > 2-nitrophenol.

[Relationship between programmed cell death mechanisms and neuronal necrosis induced by seizures].

OBJECTIVE: To clarify if programmed cell death mechanisms induced by seizures take part in the necrotic process of neurons. METHODS: Seizure was induced by pilocarpine (P) in Sprague-Dawley adult rats which were allowed to recover for 24 or 72 hours before perfusion-fixation. Neuronal death was assessed by light microscopy with the hematoxylin-eosin (HE) staining and with in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Bax and Bcl-2 protein expression were examined by histochemistry. RESULTS: Twenty-four and 72 hours after seizures, neuronal death in hippocampus CA1 region was morphologically necrotic. TUNEL-positive and morphologically necrotic cells increased in the hippocampal CA1 region at 72 hours after seizures, there was significant difference compared with controls (P < 0.001). Bax expression was also increased in the hippocampal CA1 region at 72 hours after seizures (P < 0.001), but Bcl-2 expression did not increase, while Bcl-2/Bax ratio decreased. CONCLUSION: Seizures induced late-onset neuronal necrosis was accompanied by programmed cell death mechanisms.