[Clinical analysis of 48 cases of non-invasive fungal sinusitis].

Objective:To analyze the causes, imaging diagnosis, clinical characteristics and clinical effect of the endoscopic surgery of non-invasive fungal rhino-sinusitis.Method:A retrospective analysis of 48 patients diagnosed with fungal sinusitis by pathology.Result:Forty-eight cases of 40 cases are fungal sinusitis (40/48), the other 8 cases are allergic fungal rhinosinusitis. The two morphology, imaging, histopathology, treatment and outcome are not the same, more than half of the CT scan and endoscopic examination, have different degrees of anatomical abnormalities (26/28), 25 cases had a long-term frequent using of antibiotics. Postoperative follow-up 1-3 years, cured 43 cases, recurrence 5 cases, which has been cured after re-surgery and other comprehensive treatment. Conclusion: Nasal cavity anatomical abnormalities and the frequent use of antibiotics is closely related to the attack of fungal sinusitis. Sinus CT scane and nasal endoscopy is an important method for the diagnosis of fungal sinusitis, and endoscopic sinus surgery is an effective method of fungal sinusitis. The recurrence rate of allergic fungal sinusitis was higher (5/8), so comprehensive treatment was an importance measure to prevent recurrence. Caution with antibiotics and hormones is to prevent fungal sinusitis occurred in a factor that can not be ignored.

Prevalence of fimA genotypes of Porphyromonas gingivalis and periodontal health status in Chinese adults.

BACKGROUND AND OBJECTIVE: Porphyromonas gingivalis fimbriae play a key role in colonization of the oral cavity. The fimA gene, which encodes fimbrillin (FimA), can be classified into six types (I-V and Ib) according to nucleotide sequence. In the present study, we investigated the relationship between the prevalence of P. gingivalis-specific fimA genotypes and periodontal health status in Chinese adults. MATERIAL AND METHODS: One-hundred and fifteen patients with chronic periodontitis and 136 periodontally healthy adults were selected. P. gingivalis detection, determination of fimA genotypes, and the co-existence of Actinobacillus actinomycetemcomitans and Tannerella forsythia with various fimA types, were assessed by the polymerase chain reaction. Odds ratios and 95% confidence intervals were calculated for associating the fimA-specific genes with periodontitis. RESULTS: P. gingivalis was detected in 22.1% of healthy subjects and in 81.7% of the patients. A single fimA genotype was detected in most samples. In healthy adults, the most prevalent fimA genotype was type I (66.7%). However, type II was detected most frequently (43.6%) in the patient group, followed by type IV (30.9%). The frequency of co-existing A. actinomycetemcomitans and T. forsythia was highest in type II fimA-positive sites. Statistical analysis revealed that periodontitis was associated with occurrences of type I (odds ratio 0.97), Ib (odds ratio 13.26), II (odds ratio 36.62), III (odds ratio 4.57), IV (odds ratio 22.86) and V (odds ratio 1.19). CONCLUSION: P. gingivalis type II followed by type IV were considered as disease-associated strains that account for the pathogenesis of chronic periodontitis in Chinese adults.

Superinduction of IL-2 gene expression by vomitoxin (deoxynivalenol) involves increased mRNA stability.

To better understand molecular mechanisms by which the trichothecene vomitoxin (VT) superinduces cytokine gene expression, we studied the posttranscriptional effects of this mycotoxin on interleukin-2 (IL-2) gene expression in murine EL-4 thymoma cells stimulated with phorbol 12-myristate 13-acetate and ionomycin (PMA + ION). Northern analysis revealed that doses of 50 to 500 ng/ml VT superinduced IL-2 mRNA expression in a dose- and time-dependent manner in a synchronous model where VT was added at onset of PMA + ION stimulation. In accordance with the mRNA levels, IL-2 production was significantly elevated in the presence of 50 to 250 ng/ml VT. Superinduction of IL-2 mRNA was also observed in a delayed synchronous model (VT added 20 hr after PMA + ION stimulation) and an asynchronous model (VT added 20 hr after PMA + ION stimulation and removal). To assess the effects of VT (500 ng/ml) on IL-2 mRNA half-life, three transcriptional inhibitors were used in the delayed synchronous model. Actinomycin D (ActD) had a pronounced stabilizing effect on IL-2 mRNA but not on mRNA for the housekeeping gene GAPDH. VT did not affect IL-2 mRNA levels in ActD-treated cells. Although 5,6-dichloro-beta-D-ribofuranosyl-benzimidazole (DRB) also had a stabilizing effect on IL-2 mRNA, IL-2 mRNA half-life t1/2 in VT-treated cells was three times that of control. In contrast, inclusion of cyclosporin A (CsA) into the cultures specifically arrested IL-2 transcription in EL-4 cells without any stabilizing effect. VT exposure in the presence of CsA markedly prolonged the half-life of IL-2 mRNA in a dose-dependent manner. The t1/2 for IL-2 mRNA in the control culture was 2.1 hr, whereas t1/2 was 3.1, 3.4, 4.2, and 10.5 hr in cultures containing 50, 100, 250, and 500 ng/ml VT, respectively. These results suggest that VT can superinduce IL-2 at both the mRNA and the protein level and that this superinduction can be explained, in part, by posttranscriptional mechanisms such as enhanced mRNA stability.

Effects of vomitoxin (deoxynivalenol) on transcription factor NF-kappa B/Rel binding activity in murine EL-4 thymoma and primary CD4+ T cells.

The trichothecene mycotoxin vomitoxin (VT, deoxynivalenol) superinduces the gene expression of IL-2 and several other cytokines in both cellular and murine models. Because transcription factor NF-kappaB/Rel has been shown to play a crucial role in control of cytokine gene transcription, we assessed the in vitro effects of VT on NF-kappaB/Rel binding activity by electrophoretic mobility shift assay using both cloned (EL-4) and primary (CD4+) murine T cells. When EL-4 thymoma cells were stimulated with phorbol 12-myristate 13-acetate plus ionomycin in the presence of 500 ng/ml VT, DNA binding activity by NF-kappaB/Rel in nuclear extracts was increased from 2 to 48 hr when compared to controls employing no VT. VT preferentially induced a slower migrating electrophoretic band of the NF-kappaB/Rel complex particularly in later time points (8-48 hr). The band was found to contain a c-Rel/p50 heterodimer by supershift assay using antibodies specific for c-Rel and p50. NF-kappaB/Rel binding activity was enhanced by VT in a dose-dependent fashion. As little as 50 ng/ml VT was sufficient to increase NF-kappaB/Rel binding in a 1-hr EL-4 culture. Using Western blot analysis, effects on EL-4 cells were further related to VT-mediated inhibition of resynthesis of IkappaBalpha, a cytoplasmic inhibitor of NF-kappaB/Rel. Decreased IkappaBalpha levels were observed with 250-1000 ng/ml VT from 4 to 48 hr. Using primary murine CD4+ T cell cultures, elevated NF-kappaB/Rel and c-Rel/p50 binding activities were also observed at VT of 500 ng/ml from 1 to 72 hr concurrently with decreased IkappaBalpha levels. These data suggest that VT increased NF-kappaB/Rel binding activity and, in particular, the transactivating form, c-Rel. Increased NF-kappaB/Rel binding activity in the later (48 hr) stages of cell incubation may be explained, in part, by VT-mediated inhibition of resynthesis of its cytoplasmic inhibitor IkappaBalpha and by decreases in the inhibitory p50 homodimer. Elevated NF-kappaB/Rel binding activity may be involved mechanistically in VT-induced gene expression of the cytokines and resultant toxic and autoimmune effects.

Vomitoxin-mediated IL-2, IL-4, and IL-5 superinduction in murine CD4+ T cells stimulated with phorbol ester calcium ionophore: relation to kinetics of proliferation.

The effects of the trichothecene vomitoxin (VT) on the kinetics of cell proliferation and cytokine production were evaluated in murine CD4+ T cells. The CD4+ cultures were stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin to activate protein kinase C and increase cytoplasmic free calcium, respectively, in a range of VT concentrations. Total and viable cell counts at 3, 5, 7, 9, and 11 days revealed delayed or impaired cell proliferation in cultures containing between 50 and 100 ng/ml VT, with complete inhibition being observed at 250 and 500 ng/ml of VT. The VT concentration required to inhibit protein synthesis in a 3-day culture by 50% in this model was 40 ng/ml. When enzyme-linked immunosorbent assay (ELISA) was used to quantitate cytokines, IL-2 levels in control cultures were highest at Day 1 and declined rapidly thereafter, whereas, in VT groups, IL-2 levels were highest at Day 3 and remained elevated up to 11 days. IL-2 levels were elevated by continuous exposure to 100-500 ng/ml of VT with more than 100-fold differences being observed between control and 250 ng/ml VT from Days 5 to 11. When IL-2 levels were expressed on a per viable cell basis, increases were even more marked with as much as 6 log differences being observed between the treatments at 250-500 ng/ml VT and control cultures at Day 7. Supernatant IL-4 and IL-5 levels were also elevated by 100 and 250 ng/ml VT in a dose- and time-dependent fashion compared to control cultures, whereas 500 ng/ml VT was inhibitory. When relative IL mRNA abundance was analyzed during the first 3 days of culture by reverse transcriptase-polymerase chain reaction (RT-PCR) in conjunction with Southern hybridization analysis, IL-2 mRNA levels in Days 1, 2 and 3 in cultures containing 100 and 250 ng/ml VT were greater than corresponding controls. IL-2 mRNA abundance in both control and VT-treated cultures was maximal at Day 1 and decreased rapidly thereafter in controls, whereas much slower rates of IL-2 disappearance were noted in 100 and 250 ng/ml of VT. IL-4 and IL-5 mRNA levels at VT doses of 50 and 100 ng/ml were also elevated compared to controls. Pulsed VT (8 to 48 hr) or cycloheximide (4 to 48 hr) exposure of CD4+ cells enhanced supernatant levels of IL-2 but not IL-4 upon incubation for 24 hr in fresh medium. This effect was not persistent. Taken together, VT enhanced and/or delayed peak IL-2, IL-4, and IL-5 gene expression and secretion in CD4+ T cells stimulated with PMA and ionomycin. Remarkably, cytokine superinduction occurred simultaneously with partial or maximal inhibition of cell proliferation.

Effects of trichothecene structure on cytokine secretion and gene expression in murine CD4+ T-cells.

The effects of trichothecene structure on cytokine secretion and gene expression were assessed in primary CD4+ T-cells from murine spleen. CD4+ T-cells were stimulated with concanavalin A (Con A) for 2 or 7 days in the presence of various concentrations of the trichothecenes, vomitoxin (VT or deoxynivalenol), nivalenol (NIV), 15-acetyl deoxynivalenol (15-ADON), 3-acetyl deoxynivalenol (3-ADON), T-2 toxin (T-2) and verrucarin A (Ver A). Culture supernatants were subsequently analyzed for interleukin (IL)-2, IL-4 and IL-5 by ELISA. At day 2, all trichothecenes were found to have inhibited production of IL-2, IL-4, and IL-5. However, at day 7, supernatant IL-2 was significantly increased (2-5.5-fold) in cultures containing VT, NIV, 3-ADON, and 15-ADON at 250, 250, 2500, and 1000 ng/ml doses, respectively, when compared to control Con A-stimulated cultures; significant increases in IL-2 were not observed with T-2 and Ver-A. Similarly, at day 7, IL-4 and IL-5 were significantly increased in the presence of VT (100 ng/ml), NIV (100 ng/ml), 3-ADON (1000 ng/ml), 15-ADON (500 ng/ml), T-2 (1 ng/ml), and Ver A (50 pg/ml, only IL-5) when compared to control cultures. IL production was inhibited at trichothecene concentrations exceeding the aforementioned optima. When total RNA of 2-day cultures was assessed by reverse transcriptase polymerase chain reaction (RT-PCR) in conjunction with Southern analysis, IL-2 mRNA was also found to be superinduced by VT (50 and 100 ng/ml), NIV (50, 100 and 250 ng/ml), 3-ADON (1500 ng/ml), 15-ADON (100 ng/ml), T-2 (0.5 ng/ml) and Ver A (25, 50 and 100 pg/ml); IL-4 mRNA by VT (50 ng/ml), NIV (50 ng/ml), and Ver A (25, 50 and 100 pg/ml); IL-5 mRNA by VT (50 ng/ml); and IL-6 mRNA by 15-ADON (100 ng/ml) and Ver A (50 pg/ml). As the trichothecene concentration increased from these levels, inhibition of mRNA transcript levels was also observed for many of the interleukins. Taken together, the results suggest that trichothecenes as a group can either inhibit or superinduce both IL secretion and mRNA levels in CD4+ T-cells. Superinduction exhibited a rank order of macrocyclic > type A > type B trichothecenes and was dependent on acylation of the trichothecene nucleus.

Effects of vomitoxin (deoxynivalenol) and cycloheximide on IL-2, 4, 5 and 6 secretion and mRNA levels in murine CD4+ cells.

The effects of continuous in vitro exposure to the trichothecene, vomitoxin (VT) or another protein synthesis inhibitor, cycloheximide (CHX), on interleukin (IL) secretion and mRNA levels were evaluated in murine splenic CD4+ cells. Significant increases were seen in supernatant IL-2, IL-4 and IL-5 obtained from 7 day Concanavalin A (Con A)-stimulated CD4+ cultures containing VT concentrations of 250, 100 and 100 ng/ml, respectively, compared with controls run in the absence of VT. The effect of VT on CD4+ cell proliferation was also assessed after culturing for 3, 5 and 7 days with Con A. Although total cell numbers were not affected at day 3, cultures at day 5 with 50 or more ng VT/ml and at day 7 with 100 or more ng VT/ml had significantly lower cell numbers than controls. In addition, viable cell number was unaffected at day 3, but was significantly decreased at day 5 by VT concentrations of 12.5 ng or more ml and at day 7 by 100 or more ng VT/ml. Elevations in IL-2, IL-4 and IL-5 were also observed in 7-day Con A-stimulated CD4+ cell cultures containing CHX at 50-100, 50 and 10 ng/ml, respectively. When CD4+ cells were stimulated with Con A in the absence of inhibitors and then subjected to reverse transcriptase-polymerase chain reaction coupled with Southern analysis, maximal IL-2, IL-4 and IL-6 mRNA levels were induced at 48 hr whereas peak IL-5 mRNA was observed at 72 hr. Superinduction of IL-2 mRNAs was observed in the presence of VT at 50-100 ng/ml and CHX at 50-250 ng/ml. IL-4 and IL-5 mRNAs were superinduced by VT at 100 ng/ml and CHX at 50 ng/ml. The results suggest that VT and CHX could superinduce both interleukin secretion and mRNA transcript levels in CD4+ cell cultures and that, for VT, these effects occurred concurrently with inhibition of cell proliferation.

Effects of high erucic acid rapeseed oil on fatty acid oxidation in rat liver.

The effects of high erucic acid rapeseed oil (HER) on fatty acid oxidation in rat liver compared with low erucic acid rapeseed oil (LER) were studied. Weanling male SD rats were fed diets containing 20% HER or LER for 1 week or 4 weeks, or 5% HER diet for 4 weeks. The hepatic oxidation capacity of butyric acid or palmitic acid was determined by titrating the propanone produced by their oxidation. The results showed that feeding HER to rats led to an increase in the weight of liver and a decrease in the hepatic oxidation capacity of palmitic acid. Hepatic oxidation of butyric acid was not influenced by the intake of HER. The inhibitory action of HER on the oxidation of long-chain fatty acids probably resulted from the incorporation of erucic acid into mitochondrial membranes, interfering the fatty acyl-CoA transferring system on the membranes, but not from the beta-oxidation enzyme system in mitochondria being directly inhibited.