TMEM232 promotes the inflammatory response in atopic dermatitis via the nuclear factor-κB and signal transducer and activator of transcription 3 signalling pathways.

BACKGROUND: Our group previously found that the transmembrane protein 232 (TMEM232) gene was associated with atopic dermatitis (AD) by genome-wide association study and fine mapping study. However, its function is unclear so far. OBJECTIVES: To investigate the roles and mechanisms of TMEM232 in AD. METHODS: The expression of TMEM232 was investigated in skin lesions of patients with AD, the MC903-induced AD mouse model, human primary keratinocytes and immortalized human keratinocyte cell line (HaCaT) cells stimulated with different inflammatory factors. The role of TMEM232 in AD was analysed in HaCaT cells and Tmem232 knockout (Tmem232-/-) mice. Tmem232-specific small interfering RNA (siRNA) was used to evaluate its therapeutic potential in the AD mouse model. RESULTS: The expression of TMEM232 was significantly increased in skin lesions of patients with AD, the MC903-induced AD mouse model and human primary keratinocytes and HaCaT cells stimulated with different inflammatory factors compared with controls. In the presence of MC903, Tmem232-/- mice exhibited significantly reduced dermatitis severity, mast-cell infiltration in the back, and expression of T-helper (Th)1 and Th2-related inflammatory factors in skin tissue compared with wild-type mice. In vitro and in vivo experiments further showed that upregulation of TMEM232 in AD exacerbated the inflammation response through activating the pathway of nuclear factor-κB and signal transducer and activator of transcription (STAT) 3, and was regulated by the interleukin-4/STAT6 axis, which formed a self-amplifying loop. Finally, topical application of Tmem232 siRNA markedly ameliorated AD-like lesions in the AD model. CONCLUSIONS: This study is the first to outline the function of TMEM232. It is involved in regulating inflammation in AD and may be a potential target for AD treatment.

Safety and efficiency of sewage sludge and garden waste compost as a soil amendment based on the field application in woodland.

Sewage sludge (SS) and garden waste (GW) compost can be used as soil amendments to improve the soil environment. Studies done till date have been focused on the changes of harmful substances during sludge composting, but the safety and efficacy of SS and GW composting on woodland soil environment are still unclear. In the study, a field experiment was performed using to investigate the safety and efficacy of SS and GW compost as a soil amendment on woodland soil. Soil nutrients (such as nitrogen, phosphorus and potassium), organic matter and electrical conductivity were significantly increased after the addition of the SS and GW compost, while there were no significant changes in soil heavy metals content and soil enzyme activities. From these soil properties, it was found that SS and GW compost was safe and efficacious in improving the soil environment. The application of SS and GW compost had no significant effect on microbial diversity. Co-occurrence network analysis revealed that SS and GW compost efficaciously enhanced the interaction between bacterial communities, which proved that it was safe and efficacious. Furthermore, SS and GW compost enhanced ABC transporters and carbohydrate metabolism of bacterial community, while reduced the pathotroph action (such as the plant pathogen) and wood saprotrophs. Overall, these results proved the safety and efficacy of SS and GW compost as soil amendments after being added to the soil. This study contributes to the use of harmless treatments and reutilization processes of SS and GW.

Effect of hydrocolloids on gel properties and protein secondary structure of silver carp surimi.

BACKGROUND: Hydrocolloids are the most commonly used additive in the processing of surimi products. However, the effect of hydrocolloids on surimi protein conformation has not been reported, and the level of hydrocolloids may be a key factor influencing the quality of surimi. Therefore, this study investigated the effect of curdlan, xanthan gum, κ-carrageenan, and gelatin at various levels on gel properties and protein conformation of surimi from silver carp. RESULTS: Addition of curdlan, κ-carrageenan, or gelatin at lower level could significantly promote gel strength, textural profiles, and water holding capacity (WHC) of the surimi gels. However, gel strength and WHC gradually decreased with increasing amount of xanthan gum added. The addition of curdlan or κ-carrageenan remarkably increased the whiteness of surimi gel, but the whiteness decreased when the concentration of κ-carrageenan reached 5 g kg-1 . Along with the increase of curdlan, κ-carrageenan, or gelatin concentration, the index of hydrophobic interaction and hydrogen bonds first increased and then decreased, whereas index of ionic bonds first decreased and then increased. According to Raman spectroscopy data, a small content of curdlan or κ-carrageenan promoted the conformational transition of surimi protein from α-helix to ß-sheet, leading to the changes in gel properties of surimi gels. Scanning electron microscopy photographs showed surimi gels added with 4 g kg-1 curdlan or 2 g kg-1 κ-carrageenan had a finer and denser network structure. CONCLUSION: Curdlan or κ-carrageenan at an appropriate concentration is a potential modifier to effectively improve the quality of surimi products. © 2020 Society of Chemical Industry.

A novel matrix protein PNU5 facilitates the transformation from amorphous calcium carbonate to calcite and aragonite.

For both nacre formation and biomineralization in mollusks, understanding the molecular mechanism is imperative. Biomineralization, especially shell formation, is dedicatedly regulated by multiple matrix proteins. However, ACC conversion to stable crystals still lacks positive factors. In this research, we found a novel matrix protein named PNU5 in Pinctada fucata that plays a regulatory role in both prismatic layer and nacreous layer formation. Functional studies in vivo and in vitro have shown that it might be involved in shell formation in a positive manner. RT-qPCR analysis showed that pnu5 was highly expressed in mantle pallial and participated in shell repairing and regeneration. RNAi-mediated repression of pnu5 could affect the normal structure of prismatic layer and nacreous layer. The recombinant protein rPNU5 significantly enhanced the precipitation rate of CaCO3 both in the calcite and aragonite crystallization systems, as well as altering the morphology of the crystals. Based on ACC transition experiments, the recombinant protein rPNU5 facilitated amorphous calcium carbonate (ACC) transformation into stable calcite or aragonite. This study could provide us with a better understanding of how positive regulatory mechanisms contribute to biomineralization.

The regulatory effect of NF-κB signaling pathway on biomineralization and shell regeneration in pearl oyster, Pinctada fucata.

Pinctada fucata is an important pearl production shellfish in aquaculture. The formation of shells and pearls is a hot research topic in biomineralization, and matrix proteins secreted by the mantle tissues play the key role in this process. However, upstream regulatory mechanisms of transcription factors on the matrix protein genes remain unclear. Previous studies have shown that NF-κB signaling pathway regulated biomineralization process through expression regulation of specific matrix proteins, including Nacrein, Prismalin-14 and MSI60. In this study, we systematically investigated the regulatory effect of the NF-κB signaling pathway key factor Pf-Rel and inhibitory protein poI-κB on the biomineralization and shell regeneration process. We applied RNA interference and antibody injection assays to study in vivo function of transcription factor Pf-Rel and characterized shell morphology changes using scanning electron microscopy and Raman spectroscopy. We found that transcription factor Pf-Rel plays a positive regulatory role in the growth regulation of the prismatic and nacreous layers, while the function of inhibitory protein poI-κB is to prevent excessive growth and accumulation of both layers. RNA-seq was conducted based on RNA interference animal model to identify potential regulatory genes by transcription factor Pf-Rel. Shell damage repair experiments were performed to simulate shell regeneration process, and observations of newly formed shells revealed that NF-κB signaling pathway had different functions at different times. This study provides us with a more macroscopic perspective based on transcription factors to investigate biomineralization and shell regeneration.

Combined metabolomic and transcriptomic analysis evidences the interaction between sugars and phosphate in rice.

Phosphorus is one of the macro-elements required by plants, but phosphate (Pi), the only form that can be absorbed by plants, is always limited for plant growth and development. To adapt to Pi deficiency, plants have evolved a complex regulatory system to improve Pi acquisition and utilization efficiency. In this study, metabolomic and transcriptomic analyses were performed to exam the global metabolites and gene expressions profiles responding to Pi deficiency in rice. A total of 23 metabolites were co-changed in leaves and roots after Pi deficiency, with sucrose, trehalose and melibiose significant accumulated. A total of 779 genes were co-changed in these leaves and roots. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that differentially expressed genes and differentially accumulated metabolites were co-enriched in galactose metabolism. Further exogenous sugars supply with rice roots could induce Pi starvation responsiveness and the expression of OsPHR2, which codes the central regulator for Pi starvation responsiveness in rice. This work revealed the interaction between sugars and phosphate in rice, and the importance of OsPHR2 in this interaction.

Engineer chimeric Cas9 to expand PAM recognition based on evolutionary information.

Although Cas9 nucleases are remarkably diverse in microorganisms, the range of genomic sequences targetable by a CRISPR/Cas9 system is restricted by the requirement of a short protospacer adjacent motif (PAM) at the target site. Here, we generate a group of chimeric Cas9 (cCas9) variants by replacing the key region in the PAM interaction (PI) domain of Staphylococcus aureus Cas9 (SaCas9) with the corresponding region in a panel of SaCas9 orthologs. By using a functional assay at target sites with different nucleotide recombinations at PAM position 3-6, we identify several cCas9 variants with expanded recognition capability at NNVRRN, NNVACT, NNVATG, NNVATT, NNVGCT, NNVGTG, and NNVGTT PAM sequences. In summary, we provide a panel of cCas9 variants accessible up to 1/4 of all the possible genomic targets in mammalian cells.

[Effects of electroacupuncture at "Zusanli" (ST 36) on expression of mitophagy-related proteins in skeletal muscle in rats with spleen deficiency syndrome].

OBJECTIVE: To observe the expression change of mitophagy-related proteins in skeletal muscle in rats with spleen deficiency syndrome and to explain the partial action mechanism of acupuncture at Zusanli (ST 36) for spleen deficiency syndrome. METHODS: Forty male SD rats, after normal feeding, were randomly divided into a normal group, a spleen deficiency group, a Zusanli group and a non-acupoint group, ten rats in each group. Except the normal group, the three factors modeling method was used for 14 days to establish the model of spleen deficiency syndrome on the other 3 groups. The rats in the Zusanli group were treated with EA at bilateral "Zusanli" (ST 36), while the rats in the non-acupoint group were treated with EA at bilateral non acupoint (dense-sparse wave, frequency of 2 Hz/100 Hz, 20 min per treatment, once a day for 10 days). The rats in the normal group and spleen deficiency group were treated with immobilization for 20 min per day, and no EA was given. The HPLC method was applied to measure the content of adenosine triphosphate (ATP) and adenosine monophosphate (AMP) in skeletal muscle. The Western blotting method was applied to measure the expression of adenosine monophosphate activated protein kinase (AMPK), p-AMPK, ULK1, p-ULK1，LC3-Ⅰand LC3-Ⅱ in skeletal muscle. RESULTS: The ATP content in the spleen deficiency group was significantly lower than that in the normal group (P<0.01); the ATP content in the Zusanli group was significantly higher than that in the spleen deficiency group (P<0.05) but lower than that in the normal group (P<0.05), there was no significant difference between the non-acupoint group and the spleen deficiency group (P>0.05). Compared with the normal group, the AMP/ATP in the spleen deficiency group and the Zusanli group were significantly up-regulated (P<0.01, P<0.05). The differences of p-AMPK/AMPK between the spleen deficiency group and the normal group was not significant (P>0.05). Compared with the normal group and spleen deficiency group, the p-AMPK/AMPK in the Zusanli group was significantly up-regulated (both P<0.05). The p-ULK1/ULK1 and LC3-Ⅱ/LC3-Ⅰin the Zusanli group was higher than those in the normal group and spleen deficiency group (all P<0.01). CONCLUSION: EA at "Zusanli" (ST 36) might activate AMPK and produce stable ULK1/AMPK compound and increase the mitochondrial autophagy, which could regulate spleen-stomach and treat spleen deficiency.