TLR2-hif1α-mediated glycolysis contributes to pyroptosis and oxidative stress in allergic airway inflammation.

As a pattern recognition receptor which activates innate immune system, toll-like receptor 2 (TLR2) has been reportedly mediates allergic airway inflammation (AAI), yet the underlying mechanism remains elusive. Here, in a murine AAI model, TLR2-/- mice showed decreased airway inflammation, pyroptosis and oxidative stress. RNA-sequencing revealed that allergen-induced hif1 signaling pathway and glycolysis were significantly downregulated when TLR2 was deficient, which were confirmed by lung protein immunoblots. Glycolysis inhibitor 2-Deoxy-d-glucose (2-DG) inhibited allergen-induced airway inflammation, pyroptosis, oxidative stress and glycolysis in wild type (WT) mice, while hif1α stabilizer ethyl 3,4-dihydroxybenzoate (EDHB) restored theses allergen-induced changes in TLR2-/- mice, indicating TLR2-hif1α-mediated glycolysis contributes to pyroptosis and oxidative stress in AAI. Moreover, upon allergen challenge, lung macrophages were highly activated in WT mice but were less activated in TLR2-/- mice, 2-DG replicated while EDHB reversed such effect of TLR2 deficiency on lung macrophages. Likewise, both in vivo and ex vivo WT alveolar macrophages (AMs) exhibited higher TLR2/hif1α expression, glycolysis and polarization activation in response to ovalbumin (OVA), which were all inhibited in TLR2-/- AMs, suggesting AMs activation and metabolic switch are dependent on TLR2. Finally, depletion of resident AMs in TLR2-/- mice abolished while transfer of TLR2-/- resident AMs to WT mice replicated the protective effect of TLR2 deficiency on AAI when administered before allergen challenge. Collectively, we suggested that loss of TLR2-hif1α-mediated glycolysis in resident AMs ameliorates allergic airway inflammation that inhibits pyroptosis and oxidative stress, therefore the TLR2-hif1α-glycolysis axis in resident AMs may be a novel therapeutic target for AAI.

The Effect of Tartary Buckwheat Flavonoids in Inhibiting the Proliferation of MGC80-3 Cells during Seed Germination.

Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn) is rich in functional compounds such as rutin, quercetin, d-chiro-inositol, dietary fiber, and essential amino acids. Electric field (EF) treatment before sprout germination results in physiological and chemical changes, and some alterations might lead to positive applications in plant seeds. MTT assay showed that the effect of total flavonoids on human gastric cancer cell line MGC80-3 was significantly changed after EF treatment for different germination days (3-7 days). Among them, the total flavonoids of tartary buckwheat (BWTF) on the third day had the most obvious inhibitory effect on MGC80-3 (p < 0.01). In addition, flow cytometry evidenced that different ratios of quercetin and rutin had effects on the proliferation of MGC80-3. The same content of quercetin and rutin had the best effect, reaching 6.18 ± 0.82%. The anti-cancer mechanism was mainly promoted by promoting the expression of apoptotic proteins. The expression of Bax/Bcl-2 and caspase-8 in MGC80-3 cells was mediated by BWTFs. This study has good research value for improving the biological and economic value of tartary buckwheat.

Characterizations of a pectin extracted from Premna microphylla turcz and its cold gelation with whey protein concentrate at different pHs.

In this study, the physicochemical properties of an ammonium oxalate extraction pectin (AOP) from Premna microphylla turcz was investigated. Moreover, its cold gelation with undenatured whey protein concentrate (WPC) was studied at room temperature and different pHs. Characterizations of AOP demonstrated that AOP was a linear low-methoxyl pectin rich in homogalacturonan with low branching degree of RG-I, leading to its good gelling properties. Gelation between AOP and WPC was mainly investigated by turbidity measurement, FTIR, CLSM and ITC. The results showed that an optimal complex ratio for gelation was observed at 1:5 at pH 6.0. Moreover, AOP was the backbone of the composite gel and WPC might act as crosslinking agents through electrostatic or hydrophobic interaction at different pHs. When pH was around the pHΦ of the complex, composite gel was mainly constructed by electrostatic interaction. With the increase of pH, the electrostatic interaction between AOP and WPC gradually weakened, while the hydrophobic interaction constantly increased. When pH was higher than the pHc of the complex, composite gel was mainly formed by hydrophobic interaction. The results of this study are conducive to further utilization of Premna microphylla turcz pectin to develop related food products.

BRD4 expression in patients with essential hypertension and its effect on blood pressure in spontaneously hypertensive rats.

The objective of this study was to investigate BRD4 expression in patients with essential hypertension (EH) and its effects on the blood pressure in spontaneously hypertensive (SHR) rats. BRD4 expression was detected by quantitative real-time polymerase chain reaction and western blot in 163 patients with EH and its relation to systolic blood pressure and diastolic blood pressure were analyzed accordingly. In vivo, rats were divided into WKY (Wistar-Kyoto rats), SHR (spontaneously hypertensive rats), SHR + JQ1 (BRD4 inhibitor), and SHR + Vehicle control. Rats' blood pressure was measured by the tail-cuff method. The protein expressions related to inflammation and oxidative stress of rats were determined. BRD4 was higher in patients with EH than healthy controls, which was positively correlated to systolic blood pressure and diastolic blood pressure of enrolled subjects including patients with EH and healthy controls. Rats in the SHR group showed reduced food-intake, decreased body weight, and gradually increased blood pressure compared with WKY group. Besides, SHR rats were upregulated in plasma levels of Ang II, ET-1, MDA, IL-6, and TNF-α, and substantially downregulated in NO, NOS, and SOD levels. Moreover, eNOS activity in aortic tissues of SHR rats declined obviously, whereas the content of nitrite and O2-, the activity of NADPH oxidase and NADH oxidase, and the expression of p-NF-κB p65 went up statistically, which could be partially reversed by JQ1. BRD4 was highly expressed in patients with EH, and inhibiting BRD4 could reduce oxidative stress and inflammatory response, alleviate endothelial cell damage, ameliorate aortic injury, and lower blood pressure, supporting the hypothesis that BRD4 inhibition could be a potential target for the clinical treatment of patients with EH.

Giant Cellular Vacuoles Induced by Rare Earth Oxide Nanoparticles are Abnormally Enlarged Endo/Lysosomes and Promote mTOR-Dependent TFEB Nucleus Translocation.

Many nanomaterials are reported to disrupt lysosomal function and homeostasis, but how cells sense and then respond to nanomaterial-elicited lysosome stress is poorly understood. Nucleus translocation of transcription factor EB (TFEB) plays critical roles in lysosome biogenesis following lysosome stress induced by starvation. The authors previously reported massive cellular vacuolization, along with autophagy induction, in cells treated with rare earth oxide (REO) nanoparticles. Here, the authors identify these giant cellular vacuoles as abnormally enlarged and alkalinized endo/lysosomes whose formation is dependent on macropinocytosis. This vacuolization causes deactivation of mammalian target of rapamycin (mTOR), a TFEB-interacting kinase that resides on the lysosome membrane. Subsequently, TFEB is dephosphorylated at serine 142 and translocated into cell nucleus. This nucleus translocation of TFEB is observed only in vacuolated cells and it is critical for maintaining lysosome homeostasis after REO nanoparticle treatment, as knock-down of TFEB gene significantly compromises lysosome function and enhances cell death in nanoparticle-treated cells. Our results reveal that cellular vacuolization, which is commonly observed in cells treated with REOs and other nanomaterials, represents a condition of profound lysosome stress, and cells sense and respond to this stress by facilitating mTOR-dependent TFEB nucleus translocation in an effort to restore lysosome homeostasis.

[Fertility and Environmental Impacts of Urban Scattered Human Feces Used as Organic Granular Fertilizer for Leaf Vegetables].

The disposal of urban scattered human feces has become a difficult problem for the management of modern city. In present study, the scattered human feces underwent the collection, scum removal, flocculation and dehydration, finally became the granular fertilizer; the effects of the ratio of fertilizer to soil on the growth of the pakchoi and the quality of soil and leaching water were evaluated, and the feasibility of granular fertilizer manuring the pakchoi was discussed by pot experiments. The results showed that the granular fertilizer significantly enhanced the production of the pakchoi which were not polluted by the intestinal microorganisms under the experiment conditions; meanwhile, at the proper ratio of fertilizer to soil, the concentration of these microorganisms in the leaching water was lower than that in the control check. Chemical analyses of soil revealed that the nutrient content of nitrogen, phosphorus, potassium and organic matters in soil became much richer in all treatments. In addition, the granular fertilizer improved the physical- chemical properties of soil, including raising the level of soil porosity and reducing the volume weight of soil. Application of granular fertilizer won't pollute the soil or leaching water; instead, it can also prevent nitrogen, potassium and intestinal microorganisms from leaching inio ground water at the proper ratio of granular fertilizer to soil.

Synchronous bilateral pleomorphic adenomas of the parotid gland.

Bilateral pleomorphic adenomas of the parotid gland are extremely rare, accounting for less than 0.2% of all parotid gland tumors. We present a rare case of a patient with synchronous bilateral pleomorphic adenomas of the parotid gland. A 27-year-old woman presented with a 5-month history of a slowly-growing, painless swelling of the right parotid gland. After a thorough bilateral parotid examination and magnetic resonance imaging evaluation, bilateral tumors of the parotid gland were found. The patient underwent surgical excision of the bilateral tumors in one surgical setting. Histopathological examination showed that both tumors were pleomorphic adenomas. The patient has been followed for 24 months without recurrence of tumors. Careful preoperative examination and radiological evaluation of bilateral parotid glands might be necessary for the early diagnosis of synchronous bilateral tumors. We suggest that treatment be individualized depending on the sizes and locations of tumors, and the surgical and neurological risks.

[Morphology and function of platelets stored in modified platelet additive solution at low temperature].

The aim of study was to evaluate the function of modified platelet additive solution (PAS-IIIM) with trehalose as a substitute of plasma for the storage of platelet concentrates at low temperature (10 degrees C). Apheresis platelets from 6 donors were divided and added with different media (group A: 100% plasma; group B: 70% PAS-IIIM/30% plasma; group C: 100% plasma/trehalose). Groups A, B, C were stored at 10 degrees C, 22 degrees C and -85 degrees C separately. In addition, group D (platelet concentrates stored with 100% plasma at 4 degrees C) was set up as control group for scan electronmicroscopy. The samples of each platelets were collected on day 0, 1, 5, 7 and 9 after storage respectively, while samples of platelets stored at -85 degrees C (group C) were collected on day 20 after storage. CD62p, hypotonic shock response (HSR), platelet aggregation, lactic dehydrogenase (LDH) and morphology of platelets were evaluated. The results showed that the expressions of CD62p in groups A and B increased in a time-dependent manner, but HSR and platelet aggregations decreased. The expression of CD62p, LDH release, and platelet aggregation in group A were significant higher than that in group B (p < 0.05). HSR in group A was significant lower than that in group B (p < 0.05). LDH release was significant high in samples of group C and the expression of CD62p was lower than that in other two groups (p < 0.05). It is concluded that the protective effects of 70% PAS-IIIM/30% plasma (10 degrees C) and plasma platelets (22 degrees C) on morphology of platelets are similar, but better than those of plasma platelets (4 degrees C) and plasma/trehalose (-85 degrees C). In short, PAS-IIIM serves as a good substitute of plasma for platelet storage, and protects the chilled platelets.