Metabolomic characterization of congenital microtia: a possible analysis for early diagnosis.

Background: Although metabolic abnormalities have been deemed one of the essential risk factors for growth and development, the relationship between metabolic abnormalities and microtia is still unclear. In this study, we aimed to establish a cell model of microtia and the changes of serum metabolites in patients with microtia. Methods: After constructing a cell model of microtia with low expression of BMP5, we performed integrative metabolomics analysis. For the altered metabolites, the content of glycerophosphocholine (PC), triacylglycerol (TG), and choline in the serum of 28 patients (15 patients with microtia and 13 controls) with microtia was verified by enzyme-linked immunosorbent assay (ELISA). Results: Detailed metabolomic evaluation showed distinct clusters of metabolites between BMP5-low expressing cells and normal control (NC) cells. The cell model of microtia had significantly higher levels of TG, PC, glycerophosphoethanolamine (PE), sphingomyelin, sulfatide, glycerophosphoglycerol, diacylglycerol, and glycosphingolipid. The main abnormal metabolites were mainly concentrated in the glycerophospholipid metabolism pathway, and PC and choline were closely related. In the serum of patients with microtia, the contents of PC, TG, and choline were significantly increased. Conclusions: The individual serum samples confirmed the different metabolites between patients with microtia and controls. In particular, we showed that a newly developed metabolic biomarker panel has a high sensitivity and specificity for separating patients with microtia from controls.

Bioactivity, physiological characteristics and efficacy of the SDHI fungicide pydiflumetofen against Sclerotinia sclerotiorum.

Sclerotinia sclerotiorum, which can cause Sclerotinia stem rot, is a devastating plant pathogen. This study aimed to assess the potential of pydiflumetofen, a new-generation succinate dehydrogenase inhibitor (SDHI) fungicide, to control Sclerotinia stem rot. Pydiflumetofen exhibited favorable bioactivity in suppressing mycelial growth, sclerotial production and morphological changes, and the myceliogenic and carpogenic germination of sclerotia. Treatment with pydiflumetofen increased the cell membrane permeability of S. sclerotiorum and decreased oxalic acid production. In addition, inoculation tests demonstrated that the protective activity of 40 µg/mL pydiflumetofen against the fungus was better than its curative activity. Under natural infection conditions, the spraying of pydiflumetofen at 200 g a.i. ha-1 significantly reduced the incidence and severity of Sclerotinia stem rot. In addition, the sensitivity baseline to pydiflumetofen was established using 171 isolates collected from various crops in China. The results showed that the frequency distribution of the EC50 values of pydiflumetofen was a unimodal curve with a mean EC50 value of 0.0095 ±â€¯0.0005 µg/mL. This study confirmed the favorable bioactivity of pydiflumetofen against S. sclerotiorum at various developmental stages and its high effectiveness under natural infection conditions, which indicates that pydiflumetofen is a promising tool for the management of Sclerotinia stem rot.

Favorable Bioactivity of the SDHI Fungicide Benzovindiflupyr Against Sclerotinia sclerotiorum Mycelial Growth, Sclerotial Production, and Myceliogenic and Carpogenic Germination of Sclerotia.

Sclerotinia sclerotiorum, which can cause Sclerotinia stem rot, is a prevalent plant pathogen. This study aims to evaluate the application potential of benzovindiflupyr, a new generation of succinate dehydrogenase inhibitor (SDHI), against S. sclerotiorum. In our study, 181 isolates collected from different crops (including eggplant [n = 34], cucumber [n = 27], tomato [n = 29], pepper [n = 35], pumpkin [n = 32], and kidney bean [n = 25]) in China were used to establish baseline sensitivity to benzovindiflupyr. The frequency distribution of the 50% effective concentration (EC50) values of benzovindiflupyr was a unimodal curve, with mean EC50 values of 0.0260 ± 0.011 µg/ml, and no significant differences in mean EC50 existed among the various crops (P > 0.99). Benzovindiflupyr can effectively inhibit mycelial growth, sclerotial production, sclerotial shape, and myceliogenic and carpogenic germination of the sclerotia of S. sclerotiorum. In addition, benzovindiflupyr showed good systemic translocation in eggplant. Using benzovindiflupyr at 100 µg/ml yielded efficacies of 71.3 and 80.5% for transverse activity and cross-layer activity, respectively, which were higher than those of acropetal and basipetal treatments (43.6 and 44.7%, respectively). Greenhouse experiments were then carried out at two experimental sites for verification. Applying benzovindiflupyr at 200 g a.i. ha-1 significantly reduced the disease incidence and severity of Sclerotinia stem rot. Overall, the results demonstrated that benzovindiflupyr is a potential alternative product to control Sclerotinia stem rot.

Peli1 Contributions in Microglial Activation, Neuroinflammatory Responses and Neurological Deficits Following Experimental Subarachnoid Hemorrhage.

Early brain injury (EBI) following subarachnoid hemorrhage (SAH) is closely associated with neuroinflammation. Microglial activation is an early event that leads to neuroinflammation after SAH. Peli1 is an E3 ubiquitin ligase that mediates the induction of pro-inflammatory cytokines in microglia. Here we report Peli1 contributions in SAH mediated brain pathology. An SAH model was induced by endovascular perforation in adult male C57BL/6J mice. Peli1 was markedly induced in mice brains in a time-dependent manner and was predominantly expressed in CD16/32-positive microglia after SAH. Using genetic approaches, we demonstrated that decreased Peli1 significantly improved neurological deficits, attenuated brain edema, reduced over-expression of pro-inflammatory cytokine IL-6 and modified apoptotic/antiapoptotic biomarkers. In addition, Peli1 downregulation suppressed ERK and JNK phosphorylation levels via the downregulation of cIAP1/2 expression, subsequently reducing inducible nitric oxide synthase (iNOS) expression after SAH. Therefore, these findings demonstrate that Peli1 contributes to microglia-mediated neuroinflammation in EBI by mediating cIAP1/2 activation, thus promoting the activation of MyD88-dependent MAPK pathway after experimental SAH. Our findings also showed that Peli1 could promote the expression of M1 microglia polarization biomarker CD16/32 and iNOS after SAH. Targeting Peli1 exerts neuroprotective effects during EBI after SAH, thus could provide potential option for prevention-therapy in high-risk individuals.

Activity, Translocation, and Persistence of Isopyrazam for Controlling Cucumber Powdery Mildew.

A cotyledon bioassay was conducted to assess the activity of isopyrazam against Podosphaera xanthii (Castagne) U. Braun & N. Shishkoff, causal agent of cucumber powdery mildew. Results showed that isopyrazam has protective and curative activity against P. xanthii, with EC50 values of 0.04 and 0.05 mg liter-1, respectively. These activities are higher than those for hexaconazole, difenoconazole, pyraclostrobin, kresoxim-methyl, and azoxystrobin, fungicides currently used against cucumber powdery mildew. Isopyrazam at 0.5 mg liter-1 damaged conidiophores. Results of inoculation tests in greenhouse pots indicate that isopyrazam demonstrates a level of systemic movement in cucumber plants, especially regarding translaminar and transverse translocation. Efficacy following translaminar and transverse translocations on cotyledons and leaves treated with 60 mg liter-1 was 94.40% and 88.96%, and 95.26% and 82.83%, respectively. In addition, isopyrazam at 60 mg liter-1 exhibited a long duration of efficacy against cucumber powdery mildew, almost 2 to 3 weeks longer than that of triazoles and strobilurins. Similar trends in residual durations were observed during 2014 and 2015 greenhouse trials. Isopyrazam at 30 and 60 a.i. g ha-1 provided efficacy ranging from 83.27 to 90.83% 20 days following treatment. In conclusion, isopyrazam has translaminar and transverse translocation in cucumber leaves, and long duration of activity against cucumber powdery mildew.

Modified sequential therapy vs quadruple therapy as initial therapy in patients with Helicobacter infection.

AIM: To evaluate the efficacy and safety of modified sequential therapy and to compare modified sequential therapy with standard quadruple therapy for Helicobacter pylori (H. pylori) eradication. METHODS: In total, 200 consecutive patients who were diagnosed with H. pylori-infected chronic gastritis by electronic endoscopy and rapid urease testing from December 2012 to October 2013 were enrolled in this study. The patients had not previously received H. pylori eradication treatment, and were randomized into two groups. The patients in Group A (n = 101) were treated with ilaprazole + bismuth potassium citrate + amoxicillin and clavulanate potassium + levofloxacin, and the patients in Group B (n = 99) were administered a modified sequential therapy composed of ilaprazole at 5 mg bid and amoxicillin and clavulanate potassium at 914 mg for the first five days followed by ilaprazole at 5 mg bid, furazolidone at 100 mg bid and levofloxacin at 500 mg qid for the next five days. Four to six weeks after the end of treatment, a 14C-urea breath test was performed for all the subjects to confirm the eradication of H. pylori. The intention-to-treat and per-protocol eradication rates were determined. RESULTS: A total of 190 of the 200 patients completed the study. All 200 patients were included in the intention-to-treat analysis, whereas 190 patients were included in the per-protocol analysis. In the intention-to-treat analysis, the rates of H. pylori eradication in Groups A and B were 85.15% (86/101) and 81.82% (81/99), respectively. In the per-protocol analysis, the H. pylori eradication rates in Groups A and B were 88.66% (86/97) and 87.09% (81/93), respectively. No significant difference was observed (χ(2) = 0.109, P = 0.741) in the eradication rate between Groups A and B. The rates of adverse effects observed in the groups were similar at 6.19% (6/97) for Group A and 7.53% (7/93) for Group B (P > 0.05). No mortality or major morbidities were observed in any of the patients. Symptomatic improvements in the presentation of stomachache, acid regurgitation, and burning sensation were not significantly different between the two groups. CONCLUSION: Ilaprazole-based 10-d standard quadruple therapy does not offer an incremental benefit over modified sequential therapy for the treatment of H. pylori infection, as both treatment regimens appear to be effective, safe, and well-tolerated as initial treatment options.

Inhibitory effect of sihuangxiechai decoction on ovalbumin-induced airway inflammation in Guinea pigs.

The aim of this study was to investigate the effect of sihuangxiechai decoction on asthmatic Guinea pig model which was sensitized by intraperitoneal (i.p.) injection of ovalbumin (OVA) and challenged by OVA inhalation to induce chronic airway inflammation. Differential cell counts of cytospins were performed after staining with Giemsa solution. The quantity of leukocytes and its classification in bronchoalveolar lavage fluid (BALF) and blood were evaluated by blood cell analyzer and microscope. Histological analysis of the lung was performed by hematoxylin and eosin (H&E) staining. The levels of interleukin-4 (IL-4) and tumor necrosis factor-alpha (TNF-α) in BALF and serum were detected by radioimmunoassay (RIA). The total number of leukocytes in BALF and blood has no significant difference between Sihuangxiechaitang decoction treated group and dexamethasone (DXM) treated group but was significantly lower than those of asthma group. The percentage of eosinophils in lung tissues of sihuangxiechai decoction treated group was significantly lower than that of asthma group. The results demonstrated that the levels of IL-4 and TNF-α in the sihuangxiechai decoction treated group were significantly reduced compared with the asthma group. In conclusion, these findings demonstrate that sihuangxiechai decoction has a protective effect on OVA-induced asthma in reducing airway inflammation and airway hyperresponsiveness (AHR) in a Guinea pig model and may be useful as an adjuvant therapy for the treatment of bronchial asthma.

[Co-expression of CD99/MIC2 and ALK in anaplastic large-cell lymphoma tissues and its significance].

OBJECTIVE: To investigate co-expression of CD99/MIC2 and anaplastic lymphoma kinase (ALK) protein in anaplastic large-cell lymphoma (ALCL) tissues and Karpas 299 cells and its significance. METHODS: Clinical prognoses and ALK protein expressions of 25 cases of ALCL were reviewed retrospectively, the median duration of survival was analyzed for patients with ALK(+) ALCL and ALK(-) ALCL. Histological and immunohistochemical staining were applied to other 25 cases of ALCL and paraffin-embedded tissue from human anaplastic large-cell lymphoma Karpas 299 cells to detect the protein of CD99 and ALK. RESULTS: Of former 25 cases of ALCL, median duration of survival for ALK(+) patients was 59 months, whereas 20 months for ALK(-) patients. The prognosis of ALK(+) group was better than that of ALK(-) group, survival curves of these two groups showed statistically significant (P < 0.05). CD99 was positive in 18 cases (72.0%) while negative in 7 cases (28.0%) of the latter 25 ALCL, ALK was positive in 19 cases (76.0%) while negative in 6 cases (24.0%); Of 19 ALK(+) ALCL, 16 (84.2%) cases co-expressed CD99-ALK; and in 6 ALK(-) ALCL, 2(33.3%) were CD99-ALK double negative, the expression of CD99 protein strongly correlated with that of ALK protein (P < 0.05). ALK and CD99 protein expressed in Karpas 299 cells with diffuse distribution. CONCLUSIONS: CD99 highly expressed in ALCL, and showed high rate of co-expression with ALK. CD99 protein expression could be considered as a helpful diagnostic and prognostic factor of ALCL, especially for ALK(+) ALCL.

[Expression of miR-9 in B lymphocytes and B cell lymphomas cell lines and its significance].

OBJECTIVE: To investigate the expression of miR-9 in B lymphocytes, B cell lymphoma and classical Hodgkin's lymphoma (cHL) cell lines and its significance. METHODS: CD19(+) B lymphocytes were sorted from normal lymph node by magnetic beads. Total cellular micro-RNA was extracted from cHL cell line L428, B cell lymphoma cell lines Ly1 and Ly10 (diffuse large B cell lymphoma), Raji cells (Burkitt's lymphoma) and CD19(+) B lymphocytes, respectively. These micro-RNAs were separately transformed into cDNA by reverse transcription. The expression levels of miR-9 were measured by fluorescence quantitative PCR. In situ hybridization was used to detect the expression of miR-9 in cell lines. RESULTS: The expression of miR-9 was high in L428 cells (104.44 ± 1.61), and low in cell lines of B cell lymphoma (Ly1: 2.17 ± 0.38; Ly10: 1 ± 0.015; Raji: 2.65 ± 0.89), and extremely low in CD19(+) B lymphocytes (0.0026 ± 0.00040). Compared with that in the other cell lines, the expression of miR-9 in L428 cells was statistically significant (P < 0.05). miR-9 localized in the cytoplasm diffusely and strongly in L428, but scattered and slightly with some prominent distribution around the nuclear membranes in Ly1 and Ly10, and only weakly in Raji. CONCLUSIONS: miR-9 highly expressed in cHL cell line and might be a molecular marker for diagnosis and treatment of cHL.

Correlation of seven biological factors (Hsp90a, p53, MDM2, Bcl-2, Bax, Cytochrome C, and Cleaved caspase3) with clinical outcomes of ALK+ anaplastic large-cell lymphoma.

OBJECTIVE: To explore correlation of seven apoptosis-related proteins (Hsp90a, p53, MDM2, Bcl-2, Bax, Cytochrome C, and Cleaved caspase3) with clinical outcomes of ALK+ anaplastic large-cell lymphoma (ALCL). METHODS: Using immunohistochemistry and immunofluorescence double staining methods, the expressions of these seven apoptosis-associated proteins were studied to clarify their relationship with clinical outcomes of 36 ALK+ and 25 ALK-systemic ALCL patients enrolled between 1996 and 2006. The relationship of these apoptosis-regulating proteins with NPM-ALK status was also evaluated with the tyrosine inhibitor herbimycin A (HA) in vitro by immunocytochemistry, Western blotting and flow cytometric assays. RESULTS: The presence of Hsp90α-, MDM2-, Bax-, Cytochrome C, and Cleaved caspase3-positive tumor cells was found significantly different in ALK+ and ALK-ALCLs, which was correlated with highly favorable clinical outcome. The Bcl-2- and p53-positive tumor cells were found in groups of patients with unfavorable prognosis. Inhibition of NPM-ALK by HA could reactivate the p53 protein and subsequent apoptosis-related proteins and therefore induced apoptosis in ALK+ ALCL cells. CONCLUSION: Our results suggest that these seven proteins might be involved in apoptosis regulation and associated with clinical outcome of ALK+ systemic ALCLs. We also reveal a dynamic chain relation that NPM-ALK regulates p53 expression and subsequent apoptosis cascade in ALK+ ALCLs.