Molecular Characterization of Phospholipase C in Infection Structure Differentiation Induced by Pear Fruit Surface Signals, Stress Responses, Secondary Metabolism, and Virulence of Alternaria alternata.

Fungal pathogens use plant surface physiochemical signals to trigger specific developmental processes. To assess the role of phospholipase C (PLC) in mediating plant stimuli sensing of Alternaria alternata, the function of three PLC genes was characterized by constructing ΔAaPLC mutants. Here we showed that fruit wax-coated surfaces significantly induced appressorium formation in A. alternata and mutants. Germination of ΔAaPLC mutants did not differ from the wild type. Deletion of AaPLC1 led to the decrease of appressorium formation and infected hyphae, but the degree of reduction varies between the different types of waxes, with the strongest response to pear wax. Appressorium formation and infected hyphae of the ΔAaPLC1 mutant on dewaxed onion epidermis mounted with pear wax (θ4) were reduced by 14.5 and 65.7% after 8 h incubation, while ΔAaPLC2 and ΔAaPLC3 formed the same infection hyphae as wild type. In addition, AaPLC1 mutation caused pleiotropic effects on fungal biological function, including growth deficiency, changes in stress tolerance, weakening of pathogenicity to the host, as well as destruction of mycotoxin synthesis. Both AaPLC2 and AaPLC3 genes were found to have some effects on stress response and mycotoxin production. Taken together, AaPLC genes differentially regulate the growth, stress response, pathogenicity, and secondary metabolism of A. alternata.

Damage to Trichothecium roseum caused by sodium silicate is independent from pH.

Trichothecium roseum is one of the most important postharvest pathogens in arid and semiarid regions. Sodium silicate (NaSi) and environmental pH have significant inhibitory effects on fungal growth. However, no study has addressed the relationship of NaSi and pH in combination and the effects on T. roseum. In this work, we showed that spore germination, germ tube elongation, and mycelial growth of T. roseum were significantly inhibited by various NaSi concentrations, which had corresponding increasing pHs. Furthermore, these NaSi solutions showed a much greater impact than did pH treatments alone. The pathogenicity of NaSi-treated conidia on a model assay (conidia-inoculated apple fruit) was dramatically reduced, whereas no changes of pathogenicity were evident for the corresponding pH (various sodium hydroxide (NaOH) solutions) treatments. Fluorescent microscopy, using propidium iodide staining, showed damage of the plasma membranes of T. roseum conidia treated with both NaSi and NaOH, although the damage was more severe with NaSi. Leakage of proteins and sugars was significantly higher in NaSi-treated and NaOH-treated conidia than in untreated controls. In addition, serious damage was observed in the conidia exposed to NaSi for longer periods of time. Ultrastructural observations showed that treatment with either NaSi or NaOH caused a plasmolysis state and disorganized organelles. Taken together the results show that NaSi has inhibitory effects on T. roseum and that the inherent higher pH of NaSi solutions of higher concentrations simply acts as an enhancer of the inhibitory effects of NaSi.

Activation of phenylpropanoid pathway and PR of potato tuber against Fusarium sulphureum by fungal elicitor from Trichothecium roseum.

The induced resistance of potato tuber (Solanum tuberosum cv. Xindaping) tissue against Fusarium sulphureum by a fungal elicitor from the incompatible pathogen Trichothecium roseum and its possible mechanism were studied. The results showed that the lesion development of the wound-inoculated potato tuber was significantly reduced by treatment with the fungal elicitor from T. roseum (P < 0.05). Inoculation with F. sulphureum on the 16th day after treatment with the fungal elicitor80 at 15.0 µg/ml had the best resistant effect in the potato tuber, with the diameter being only reduced by 47 % that of the control. In addition, the results also showed that the potato tuber treated with the fungal elicitor80 could systemically induce lignin deposition, total phenolic content, flavonoid content and defense enzymes, including three keys phenylpropanoid pathway (PAL, 4CL and C4H) and pathogenesis-related (GLU and CHT) enzymes. The fungal elicitor80 also enhanced the up-regulation of the transcription and expression of PAL, C4H, 4CL, GLU and CHT genes. The treatment with the fungal elicitor80 + F. sulphureum caused the marked and/or prompt enhancement of all indexes when compared to treatment with the fungal elicitor80 or inoculation with the pathogen alone. The results suggested that the fungal elicitor of T. roseum could significantly enhance defense responses in potato tuber against dry rot mainly due to the up-regulation of the transcription and expression of resistance-related genes as well as increasing the activity of resistance-related enzymes and antifungal compounds.

The Interaction Between StCDPK14 and StRbohB Contributes to Benzo-(1, 2, 3)-Thiadiazole-7-Carbothioic Acid S-Methyl Ester-Induced Wound Healing of Potato Tubers by Regulating Reactive Oxygen Species Generation.

Reactive oxygen species (ROS) production is essential for both physiological processes and environmental stress in diverse plants. Previous studies have found that benzo-(1, 2, 3)-thiadiazole-7-carbothioic acid S-methyl ester (BTH)-inducible ROS were associated with wound healing of potato tubers. Calcium-dependent protein kinases (CDPKs), the important calcium receptors, are known to play a crucial part in plant development and adaptation to abiotic stresses. However, whether CDPK-mediated ROS generation induced by BTH is involved in wound healing is elusive. In this study, we measured Solanum tuberosum CDPKs (StCDPKs) expression using real-time PCR, and it was found that the transcriptional levels of StCDPKs from BTH-treated tissues were significantly induced, among which StCDPK14 presented the most increased level. Subcellular localization results showed that StCDPK14 is located in the nucleus and membrane. The transgenic potato plants and tubers were developed using interference-expression of StCDPK14 by Agrobacterium tumefaciens-mediated transformation. The St respiratory burst oxidase homologs (StRbohs) expression showed a remarkable decrease in StCDPK14 transgenic tubers, notably, H2O2 content and suberin deposition were also significantly declined. To confirm the relationship between StCDPK14 and StRbohB, yeast-two-hybrid and bimolecular fluorescence complementation were used to examine the interaction, and it was shown that StCDPK14 interacted with the specific Ca2 + -binding motif (helix-loop-helix, called EF-hand) of StRbohB N-terminus. The above results unraveled that StCDPK14 functions in ROS generation via interacting with StRbohB during wound healing of potato tubers.

Inflammatory myofibroblastic tumor of the spleen: a case report and review of the literature.

Inflammatory myofibroblastic tumor of the spleen (IMTS) is an extremely rare, intermediate malignant tumor with unclear etiology, and is most frequently detected incidentally. IMTS presents with nonspecific symptoms that pose a diagnostic challenge to clinicians or presents with the similar appearances to splenetic malignant neoplasms that pose a misdiagnose prior to surgery. Histopathology of the resected specimen remains the gold standard for diagnosing these rare splenic tumors. But these tumors may be misdiagnosed if pathologists are not familiar with the histologic pattern of their variations. In this paper we report a rare case of IMTS in a 55-year-old female admitted to the Xingtai People's Hospital affiliated to Hebei Medical University, with a mass of the spleen. The mass was identified incidentally two months ago and was initially diagnosed as a splenic lymphoma. The patient underwent laparoscopic splenectomy and the histologic study of the specimen revealed findings consistent with IMTS. Histological examination of the nodular growth revealed spindle cells in a hyalinized stroma with inflammatory infiltration of predominantly plasma cells and lymphocytes, coupled with lymphoid follicle structures. Immunohistochemical staining was performed to confirm the diagnosis of IMTS. Splenectomy is both diagnostic and curative for IMTS, and prognosis is generally favorable following the procedure. Our case report of IMTS adds to pathologists' knowledge of diagnosis. Meanwhile, the description and the review of features of IMTS, based on published cases, should help to improve the understanding and diagnosis level of this rare disease.

Improvement in the neural stem cell proliferation in rats treated with modified "Shengyu" decoction may contribute to the neurorestoration.

ETHNOPHARMACOLOGICAL RELEVANCE: "Shengyu" decoction, a traditional Chinese medicine, has been used to treat diseases with deficit in "qi" and "blood". The modified "Shengyu" decoction (MSD) used in the present study was designed to treat traumatic brain injury (TBI) on the basis of the "Shengyu" decoction, in which additional four herbs were added. Many ingredients in these herbs have been demonstrated to be effective for the treatment of brain injury. The present study was performed to evaluate the neurorestorative effect and the underlying mechanisms of MSD on the rat brain after a TBI. MATERIALS AND METHODS: TBI was induced in the right cerebral cortex of adult rats using Feeney's weight-drop method. Intragastrical administration of MSD (1.0 ml/200 g) was begun 6h after TBI. The neurological functions and neuronal loss in the cortex and hippocampus were determined. The levels of nerve growth-related factors GDNF, NGF, NCAM, TN-C, and Nogo-A and the number of GFAP(+)/GDNF(+), BrdU(+)/nestin(+), BrdU(+)/NeuN(+) immunoreactive cells in the brain ipsilateral to TBI were also measured. Moreover, the influences of MSD on these variables were observed at the same time. RESULTS: We found that treatment with MSD in TBI rats ameliorated the neurological functions and alleviated neuronal loss. MSD treatment elevated the expression of GDNF, NGF, NCAM, and TN-C, and inhibited the expression of Nogo-A. Moreover, MSD treatment increased the number of GFAP(+)/GDNF(+), BrdU(+)/nestin(+), and BrdU(+)/NeuN(+) immunoreactive cells in the cortex and hippocampus. CONCLUSION: The present results suggest that MSD treatment in TBI rats could improve the proliferation of neural stem/progenitor cells and differentiation into neurons, which may facilitate neural regeneration and tissue repair and thus contribute to the recovery of neurological functions. These effects of modified "Shengyu" decoction may provide a foundation for the use of MSD as a prescription of medicinal herbs in the traditional medicine to treat brain injuries in order to improve the neurorestoration.

The neuroprotective effect of modified "Shengyu" decoction is mediated through an anti-inflammatory mechanism in the rat after traumatic brain injury.

ETHNOPHARMACOLOGICAL RELEVANCE: "Shengyu" decoction, a traditional Chinese medicine, has been used to treat diseases with deficit in "qi" and "blood" induced frequently by profound loss of blood or by long sores with heavy pus, in which a potential anti-inflammatory effect is implied. The modified "Shengyu" decoction (MSD) used in the present study was designed on the basis of the "Shengyu" decoction, additional four herbs were added in. Many ingredients in these herbs have been demonstrated to be anti-inflammatory and thus MSD may be used for the treatment of traumatic brain injury (TBI). To evaluate the neuroprotective effect and the underlying mechanisms of MSD on the rat brain after TBI. MATERIALS AND METHODS: TBI was induced in the right cerebral cortex of male adult rats using Feeney's weight-drop method. The rats were administered a gavage of MSD (0.5, 1.0 or 2.0 ml/200 g) 6h after TBI. The neurological functions, brain water content, contusion volume, and neuron loss were determined. The levels of TNF-α, IL-1ß, IL-6, and IL-10 and the number of GFAP- and Iba1-positive cells in the brain ipsilateral to TBI were also measured. Moreover, the influence of MSD on these variables was observed at the same time. RESULTS: The neurological deficits, brain water content, and neuron loss were significantly reduced after 1.0 or 2.0 ml/200 g of MSD treatment but not after 0.5 ml/200 g. In addition, treatment with MSD (1.0 ml/200 g) significantly increased the level of IL-10 and reduced the level of TNF-α and IL-1ß and the number of GFAP- and Iba1-positive cells after TBI. However, the contusion volume of brain tissue and the expression of IL-6 were not significantly changed. CONCLUSION: MSD may be a potential therapeutic for the treatment of TBI because MSD alleviated secondary brain injury induced by TBI. In addition, MSD inhibited the inflammatory response through reducing the expression of inflammatory cytokines and the activation of microglial cells and astrocytes in the brain tissue of rats after TBI. Therefore, a potential anti-inflammatory mechanism of the "Shengyu" decoction was confirmed, which may be one of the main reasons of "Shengyu" decoction used to treat diseases with obvious inflammatory responses.

[Therapeutic efficacy of hyperbaric oxygen on traumatic brain injury in the rat and the underlying mechanisms].

OBJECTIVE: To investigate the effects of hyperbaric oxygen (HBO) treatment on the activation of astrocytes and the expression of glia-derived neurotrophic factor (GDNF) and nerve growth factor (NGF) in the brain after traumatic brain injury (TBI). METHODS: 54 male SD rats were randomly divided into three groups (n = 18): sham-operated, TBI and HBO treatment groups. TBI was induced with Feeney's method, bone window was opened without strike on the brain tissue in the sham-operated group. HBO group rats received HBO treatment for 60 min in the hyperbaric chamber containing O2 100% at 3 ATA. When neurological functions were measured 48 h after TBI, rats were decapitated, the brain water content of 18 rats was measured, 18 brains were sliced for the morphological observation after Nissl staining and for the immunohistochemistry staining of astrocyte markers glial fibrillary acidic protein (GFAP), vimentin and S100, and the other 18 brains of injured side were used for Western blot analysis of GDNF and NGF. RESULTS: HBO treatment reduced the neurological deficit, brain water content and hippocampal neuronal loss. In the observed cortex and hippocampal area astrocytes were activated, the cell number of positive expression of astrocyte markers GFAP, vimentin and S100 was increased, and the expression of GDNF and NGF was elevated after TBI. However, these indices were all enhanced further after the HBO treatment. CONCLUSION: It is suggested that HBO may be an effective therapy for TBI and upregulation of the expression of GDNF and NGF may underly the effect of HBO.

Free-radical scavenger edaravone treatment confers neuroprotection against traumatic brain injury in rats.

Traumatic brain injury (TBI) is one of the leading causes of neurological disability in young adults. Edaravone, a novel synthetic small-molecule free-radical scavenger, has been shown to have a neuroprotective effect in both animal models of cerebral ischemia and stroke patients; however, the underlying mechanism is poorly understood. In this report, we investigated the potential mechanisms of edaravone treatment in a rat model of TBI. TBI was induced in the right cerebral cortex of male adult rats using Feeney's weight-drop method. Edaravone (0.75, 1.5, or 3 mg/kg) or vehicle (normal saline) was intravenously administered at 2 and 12 h after TBI. Edaravone treatment significantly decreased hippocampal CA3 neuron loss, reduced oxidative stress, and decreased neuronal programmed cell death compared to vehicle treatment. The protective effects of edaravone treatment were also related to the pathology of TBI on non-neuronal cells, as edaravone decreased astrocyte and glial activation. Lastly, edaravone treatment significantly reduced the presence of inflammatory cytokines, cerebral edema, blood-brain barrier (BBB) permeability, and, importantly, neurological deficits following TBI. Our results suggest that edaravone exerts a neuroprotective effect in the rat model of TBI. The likely mechanism is via inhibiting oxidative stress, leading to a decreased inflammatory response and glial activation, and thereby reducing neuronal death and improving neurological function.

Neuroprotective effects of hyperbaric oxygen treatment on traumatic brain injury in the rat.

This study was designed to evaluate the potential benefits of hyperbaric oxygen (HBO) in the treatment of traumatic brain injury (TBI). The right cerebral cortex of rats was injured by the impact of a 20-g object dropped from a predetermined height. The rats received HBO treatment at 3 ATA for 60 min after TBI. Neurological behavior score, brain water content, neuronal loss in the hippocampus, and cell apoptosis in brain tissue surrounding the primary injury site were examined to determine brain damage severity. Three and six hours after TBI, HBO-treated rats displayed a significant reduction in brain damage. However, by 12 h after TBI, the efficacy of HBO treatment was considerably attenuated. Furthermore, at 24, 48, and 72 h after TBI, the HBO treatment did not show any notable effects. In contrast, multiple HBO treatments (three or five times in all), even when started 48 h after TBI, remarkably reduced neurology deficit scores and the loss of neuronal numbers in the hippocampus. Although multiple treatments started at 48 h significantly improved neurological behaviors and reduced brain injury, the overall beneficial effects were substantially weaker than those seen after a single treatment at 6 h. These results suggest that: (1) HBO treatment could alleviate brain damage after TBI; (2) a single treatment with HBO has a time limitation of 12 h post-TBI; and (3) multiple HBO treatments have the possibility to extend the post-TBI delivery time window. Therefore, our results clearly suggest the validity of HBO therapy for the treatment of TBI.