Characterization of ectomycorrhizal fungal communities associated with tree species on an iron tailings deposit undergoing restoration.

Vegetation restoration is an effective method to improve the ecological environment of mine tailings, which has a profound impact on the potential ecological functions of soil fungal communities; yet, little is known about its beneficial effect on soil ectomycorrhizal fungal community. In this study, the responses of soil characteristics and soil ectomycorrhizal fungal community diversity and structure to different revegetation, as well as the contribution of soil factors to soil ectomycorrhizal community were investigated in Liaoning Province, China. As we anticipated, the presence of vegetation significantly improved most soil properties we studied. What's more, compared to Korean pine (Pinus koraiensis Sieb. et Zucc.), Chinese poplar (Populus simonii Carr), and black locust (Robinia pseudoacacia L) could better improve soil total carbon, total nitrogen, total phosphorus, and available phosphorus. In addition, soil ectomycorrhizal fungal community diversity in black locust was greater than Korean pine and Populus simonii. Nonmetric multidimensional scaling analyses indicated that soil ectomycorrhizal community significantly differed depending on different revegetation types. Thus, these results indicated that black locust could be a suitable species for the revegetation of iron mine tailings. The study provided theoretical basis for ecological restoration of iron mine tailings using local plant species.

Fructooligosaccharides protect against OVA-induced food allergy in mice by regulating the Th17/Treg cell balance using tryptophan metabolites.

Fructooligosaccharides (FOS) can change gut microbiota composition and play a protective role in food allergy (FA). Furthermore, the protective mechanism of FOS against FA is unclear. In this study, intestinal flora and tryptophan (Trp) metabolites were investigated in a mouse model with FA supplemented with FOS. Meanwhile, we injected aryl hydrocarbon receptor antagonists (AhR-A) into a mouse model of FA supplemented with FOS to investigate whether T helper cell (Th) 17/regulatory T (Treg) cell balance was affected. Our research studies showed that dietary intake of FOS provided moderate protection from the intestinal inflammation induced by ovalbumin (OVA). This protective effect disappeared in AhR-A mice. The OVA mice manifestations had significantly lower bacterial richness, when compared to the normal control (NC) mice. Among fecal bacteria, the abundance of Akkermansiaceae (family level) and Verrucomicrobia (phylum level) increased and Ruminococcacere (phylum level) decreased in the feces of allergic mice. These changes were reversed by FOS treatment. FOS modulated the gut microbiome profiles that were altered in OVA mice, which showed an increase in the abundance of Ruminococcacere (phylum level) and a decrease in the abundance of Akkermansiaceae (family level) and Verrucomicrobia (phylum level). Liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis of Trp metabolites showed significant reductions in the level of kynurenine (kyn) in the serum of OVA mice, as compared to NC and FOS mice. Conversely, the levels of Trp and 5-hydroxytryptamine (5-HT) were significantly increased in OVA mice. Correlation analysis revealed a negative relationship between the relative abundance of Verrucomicrobiae (class level) and Akkermansiaceae (family level) with kyn, and a positive relationship with 5-HT. FOS significantly reduced interleukin-17A (IL-17A) and retinoic acid-associated nuclear orphan receptor-γt (RORγt) in FOS mice but not in AhR-A mice. FOS increased the level of interleukin-10 (IL-10) and Forkhead box P3 (Foxp3) in FOS mice but not in AhR-A mice. These findings suggest that FOS ameliorates allergic symptoms and impacts Th17/Treg balance in mice by modulating the gut microbiota composition and Trp metabolites. FOS may serve as an effective tool for the treatment of FA by regulating immune and gut microbiota.

Piper nigrum extract attenuates food allergy by decreasing Th2 cell response and regulating the Th17/Treg balance.

Piper nigrum is extensively utilized because of its antioxidation, antiallergic, antitumor, antiinflammatory, antidiarrhea, and gastrointestinal protection. We attempted to indicate whether the Piper nigrum extract (PNE) could alleviate ovalbumin (OVA)-induced food allergy, and to explore its potential mechanism. An OVA-induced food allergy mouse model was established, and different concentrations of PNE were administrated. Symptoms of food allergy, levels of immunoglobulin E (IgE), mucosal mast cell protease-1 (mMCP-1), and intestine pathological changes were assessed. Additionally, the expressions of T helper (Th) 2, Th17 and regulatory T (Treg)-associated cytokines and the proportion of Th17 and Treg cells in CD4+ T cells were measured. We found PNE attenuated symptoms of food allergy and decreased the levels of IgE and mMCP-1. In PNE group, the infiltration degree of inflammatory cells was ameliorated and the villi of small intestine were more complete. Moreover, the expressions of Th2 and Th17 cell-associated cytokines were down-regulated by PNE pretreatment, while the levels of Treg cell-associated cytokines were up-regulated. PNE decreased the number of Th17 cells, while increased the Tregs cells. PNE treatment dose-dependently improved the Th17/Treg balance. PNE plays a protective role in OVA-induced food allergy through inhibiting Th2 cell response and regulating the Th17/Treg balance.

Inhibiting the Notch signaling pathway suppresses Th17-associated airway hyperresponsiveness in obese asthmatic mice.

Notch signaling is crucial for the regulation of asthma and obesity. The interleukin (IL)-17-expressing CD4+ T cell (Th17 cell) response and airway hyperresponsiveness (AHR) are critical features of both asthma and obesity. We previously demonstrated that inhibiting the Notch signaling pathway alleviates the Th17 response in a mouse model of asthma. However, obese asthmatic individuals show increased Th17 responses and AHR, with the underlying mechanism not currently understood. We aimed to assess the function of Notch signaling in obese mice with asthma and to determine the impact of a γ-secretase inhibitor (GSI), which inhibits the Notch signaling pathway, on the regulation of the Th17 response and AHR. C57BL/6 mice were administered ovalbumin (OVA) to induce asthma, while a high-fat diet (HFD) was used to induce mouse diet-induced obesity (DIO). GSI was then administered intranasally for 7 days in DIO-OVA-induced mice. The results showed increased Notch1 and hes family bHLH transcription factor 1 (Hes1) mRNA levels and Notch receptor intracellular domain (NICD) protein levels in obese asthmatic mice. Furthermore, these mice showed an increased proportion of Th17 cells, serum IL-17A, IL-6, and IL-1ß levels, mucin 5AC (MUC5AC) mRNA level, retinoic acid-related orphan receptor-γt (RORγt) mRNA and protein levels, and increased AHR severity. Interestingly, GSI treatment resulted in reduced Notch1 and Hes1 mRNA and NICD protein levels in DIO-OVA-induced mice, with a decreased Th17 cell proportion and IL-17A quantity and alleviated AHR. These data strongly indicate that the Notch pathway is critical in obese asthmatic mice. In addition, inhibiting the Notch pathway ameliorates AHR and the Th17 response in obese mice with asthma.

Protective effect of curcumin on acute airway inflammation of allergic asthma in mice through Notch1-GATA3 signaling pathway.

Curcumin, a natural product derived from the plant Curcuma longa, has been found to have anti-inflammatory, antineoplastic and antifibrosis effects. It has been reported that curcumin attenuates allergic airway inflammation in mice through inhibiting NF-κB and its downstream transcription factor GATA3. It also has been proved the antineoplastic effect of curcumin through down-regulating Notch1 receptor and its downstream nuclear transcription factor NF-κB levels. In this study, we aimed to investigate the anti-inflammatory effect of curcumin on acute allergic asthma and its underlying mechanisms. 36 male BALB/c mice were randomly divided into four groups (normal, asthma, asthma+budesonide and asthma+curcumin groups). BALF (bronchoalveolar lavage fluid) and lung tissues were analyzed for airway inflammation and the expression of Notch1, Notch2, Notch3, Notch4 and the downstream transcription factor GATA3. Our findings showed that the levels of Notch1 and Notch2 receptors were up-regulated in asthma group, accompanied by the increased expression of GATA3. But the expression of Notch2 receptor was lower than Notch1 receptor. Curcumin pretreatment improved the airway inflammatory cells infiltration and reversed the increasing levels of Notch1/2 receptors and GATA3. Notch3 receptor was not expressed in all of the four groups. Notch4 receptor protein and mRNA expression level in the four groups had no significant differences. The results of the present study suggested that Notch1 and Notch2 receptor, major Notch1 receptor, played an important role in the development of allergic airway inflammation and the inhibition of Notch1-GATA3 signaling pathway by curcumin can prevent the development and deterioration of the allergic airway inflammation. This may be a possible therapeutic option of allergic asthma.

[Clinical study of strengthening pi and nourishing shen therapy combined with Western medicine on patients with glucocorticoid resistant myasthenia gravis].

OBJECTIVE: To study the effect of strengthening Pi and nourishing Shen therapy (SPNST) in treating patients with glucocorticoid resistant myasthenia gravis (GR-MG). METHODS: Twenty-seven patients with MG were enrolled, who were relapse cases after treated by cholinesterase inhibitor with systemic glucocorticoid treatment and showed resistance to glucocorticoid. All were treated by Western medicines, methylprednisolone (MP) and pyridostigmine bromide (PSB), together with Chinese medicine (CM) given according to their syndrome types, namely, for the 15 patients of Pi-Shen qi-yin deficiency type, Buzhong Yiqi Pill and Liuwei Dihuang Pill, and for the 12 patients of Pi-Shen yang-deficiency type, Buzhong Yiqi Pill and Zishen Yutai Pill. The dosages of medicines were reduced gradually in MP-PSB-CM order along with the progressing of the therapy in 4 stages (symptom curing, choline receptor restoration, immune regulation, and functional strengthening). Muscle strength and overall state of patients were re-examined before and after each of the 4 stages. RESULTS: After 1-year treatment, the therapeutic effect in 9 patients was judged as completely remitted; in 7 as remitted with continuous medication; in 5, 1 and 3 as significantly improved, moderately improved and unchanged respectively, while 2 patients died. CONCLUSION: Integrative medicine shows definite effects in treating GR-MG, and it is worthy of further studying.

[The experimental study on bone marrow stem cell transplantition combinated with bushen fang therapy for DMD].

OBJECTIVE: To investigate the effect on bone marrow stem cell transplantition (BMST) combinated with Bushen fang therapy in mdx mice. METHODS: Cultured the bone marrow cells of C56BL/6 in vitro and tranplante these cells to mdx mice after irradiation. Bushen fang was used to cure the mdx mice after BMST. The survivel rate, symptom of Graft Versus Host Disease (GVHD) and motor function of model mice following irradiation and transplantation were observed 4 or 8 months after BMST, prepared the freeze-splice of muscles of each group, and made HE staining. The dystrophin (DYS) was detected by SABC immunofluorescene. RESULTS: Bushen fang could decrease the rate of centrally nucleated fibers (CNF), enhance expressional rate of DYS in different level and relieve the symptom of GVHD in transplanted mice. CONCLUSION: Bushen fang can improve the therapeutic efficacy of BMST.

[Eosinophils apoptosis, fas mRNA and bcl-2 mRNA expressions in asthma model of young rat and effects of achyranthes bidentata polysaccharides].

OBJECTIVE: Asthma is a chronic respiratory tract disorder characterized by airway hyperreaction (AHR), persistent airway inflammation, high serum IgE, overproduction of IL-4, IL-5 and IL-13 by allergen-specific Th2 cells. The morbidity and mortality of asthma have continued to increase despite the use of currently available therapeutic agents. The reputed effects of traditional Chinese medicines (TCMs) have led to increasing use of TCMs for treatment of asthma throughout the world. The aims of this study were to investigate in asthma model of young rat the mRNA expressions of apoptotic gene fas and bcl-2, eosinophils (EOS) apoptosis in airway, and effects of achyranthes bidentata polysaccharides (ABPS), a group of polysaccharides extracted from TCM Achyranthes bidentata blume, on treatment of asthma. METHODS: Fifty Sprague-Dawley (SD) rats were divided into five groups, 10 rats per group. Asthma in rats was induced by intraperitioneal sensitization and challenge with nebulized ovalbumin (OVA). A pretreatment with ABPS [50 mg/(kg x d)] was done according to three different schedules: consecutively 3 days at sensitization (T1), at challenge (T2) or both of the two periods (T3). Sham-treated rats (A) and naive rats (C) served as controls. The animals were sacrificed 24 hours after the last challenge. The mRNA expression of bcl-2 and fas in eosinophils presenting in airway and the apoptosis of eosinophils in airway were assessed by using in situ hybridization with oligonucleotide probe and TUNEL methods, respectively. RESULTS: (1) Twenty-four hours after the last antigen challenge, the mRNA expression of fas in eosinophils presenting in airway significantly decreased in group A [(43.4 +/- 10.0)%] compared with that in group C [(73.2 +/- 11.9)%] (P < 0.01). ABPS could increase the fas mRNA expression significantly in all the three groups [(59.0 +/- 8.1)%, (57.5 +/- 9.6)%, (76.2 +/- 2.7)%], compared with that in group A (P < 0.05, P < 0.05, P < 0.01, respectively). The expression of the bcl-2 mRNA in group C was (47.9 +/- 8.7)%, it was elevated to (67.4 +/- 7.3)% in group A (P < 0.01). The expression of the bcl-2 mRNA in ABPS treated T1 and T3 groups was significantly lowered [(57.7 +/- 12.7)%, (57.3 +/- 6.8)%, P < 0.05], but not in T2 group [(72.4 +/- 6.7)%]. (2) In group A, the EOS presenting in the airway increased significantly, but there were few apoptotic EOS; the percentage of apoptotic eosinophil was distinctly lower in group A than that in group C [(5.3 +/- 2.2)% vs. (15.9 +/- 2.4)%, P < 0.01]. Compared with that in group A, the eosinophil apoptosis ratio in those ABPS treated groups T1, T3 was evidently elevated [(8.7 +/- 2.9)%, (9.8 +/- 2.2)%, P < 0.05, P < 0.05], but ABPS treated at challenge (T2) could not change the eosinophil apoptosis ratio significantly (P > 0.05). CONCLUSION: (1) In asthmatic rat, the expressions of the genes fas and bcl-2 mRNA in EOS were changed evidently and the ratio of EOS apoptotosis reduced greatly. (2) ABPS could enhance the apoptosis of EOS by upregulating the expression of the genes fas and bcl-2 mRNA.