PRMT5-mediated homologous recombination repair is essential to maintain genomic integrity of neural progenitor cells.

Maintaining genomic stability is a prerequisite for proliferating NPCs to ensure genetic fidelity. Though histone arginine methylation has been shown to play important roles in safeguarding genomic stability, the underlying mechanism during brain development is not fully understood. Protein arginine N-methyltransferase 5 (PRMT5) is a type II protein arginine methyltransferase that plays a role in transcriptional regulation. Here, we identify PRMT5 as a key regulator of DNA repair in response to double-strand breaks (DSBs) during NPC proliferation. Prmt5F/F; Emx1-Cre (cKO-Emx1) mice show a distinctive microcephaly phenotype, with partial loss of the dorsal medial cerebral cortex and complete loss of the corpus callosum and hippocampus. This phenotype is resulted from DSBs accumulation in the medial dorsal cortex followed by cell apoptosis. Both RNA sequencing and in vitro DNA repair analyses reveal that PRMT5 is required for DNA homologous recombination (HR) repair. PRMT5 specifically catalyzes H3R2me2s in proliferating NPCs in the developing mouse brain to enhance HR-related gene expression during DNA repair. Finally, overexpression of BRCA1 significantly rescues DSBs accumulation and cell apoptosis in PRMT5-deficient NSCs. Taken together, our results show that PRMT5 maintains genomic stability by regulating histone arginine methylation in proliferating NPCs.

The discovery and utilization of favorable genes in Oryza longistaminata.

Asian cultivated rice has been domesticated from ancestors of the wild rice species Oryza rufipogon. During this process, important changes have occurred in many agronomic traits, such as plant height, grain shattering, and panicle shape, and the yield has also greatly increased. However, many favored traits (e.g., stress resistance) have been lost. The genome of O. longistaminata is of the same AA type as O. sativa, harboring many genes conferring resistance to biotic and abiotic stresses, and it is considered as a potential gene pool for genetic improvement of O. sativa. In this review, we summarize the basic research on O. longistaminata, including its resistance to biotic and abiotic stresses, its rhizome traits, and other traits that are of potential application value, such as bacterial blight resistance, drought resistance, heat tolerance, self-incompatibility, nitrogen efficiency, and high yield. Furthermore, we present the current applied research progress on perennial rice breeding based on the rhizome trait of O. longistaminata. Lastly, the possibility of de novo domestication of O. longistaminata is discussed. We expect this article to provide information to enhance the basic research of O. longistaminata and accelerate the genetic improvement of cultivated rice.

Weighted Gene Co-expression Network Analysis (WGCNA) Reveals the Hub Role of Protein Ubiquitination in the Acquisition of Desiccation Tolerance in Boea hygrometrica.

Boea hygrometrica can survive extreme drought conditions and has been used as a model to study desiccation tolerance. A genome-wide transcriptome analysis of B. hygrometrica showed that the plant can survive rapid air-drying after experiencing a slow soil-drying acclimation phase. In addition, a weighted gene co-expression network analysis was used to study the transcriptomic datasets. A network comprising 22 modules was constructed, and seven modules were found to be significantly related to desiccation response using an enrichment analysis. Protein ubiquitination was observed to be a common process linked to hub genes in all the seven modules. Ubiquitin-modified proteins with diversified functions were identified using immunoprecipitation coupled with mass spectrometry. The lowest level of ubiquitination was noted at the full soil drying priming stage, which coincided the accumulation of dehydration-responsive gene BhLEA2. The highly conserved RY motif (CATGCA) was identified from the promoters of ubiquitin-related genes that were downregulated in the desiccated samples. An in silico gene expression analysis showed that the negative regulation of ubiquitin-related genes is potentially mediated via a B3 domain-containing transcription repressor VAL1. This study suggests that priming may involve the transcriptional regulation of several major processes, and the transcriptional regulation of genes in protein ubiquitination may play a hub role to deliver acclimation signals to posttranslational level in the acquisition of desiccation tolerance in B. hygrometrica.

[Aging Process of Puer Black Tea Studied by FTIR Spectroscopy Combined with Curve-Fitting Analysis].

For better determination of the chemical components in the Puer black tea, Fourier transform infrared spectroscopy was used for obtaining vibrational spectra of Puer black tea at different aging time. Fourier transform infrared (FTIR) spectra indicated that the chemical components had change in Puer black tea at different aging time. The leaf of Puer black tea was a complex system, its Fourier transform infrared spectrum showed a total overlap of each absorption spectrum of various components. Each band represented an overall overlap of some characteristic absorption peaks of functional groups in the Puer black tea. In order to explore the change of characteristic absorption peaks of functional groups with aging time, the prediction positions and the number of second peaks in the range of 1900-900 cm(-1) were determined by Fourier self-deconvolution at first, and later the curve fitting analysis was performed in this overlap band. At different aging time of Puer black tea, the wave number of second peaks of amide II, tea polyphenol, pectin and polysaccharides at overlap band were assigned by curve fitting analysis. The second peak at 1520 cm(-1) was characteristic absorption band of amide II, the second peaks of tea polyphenol and pectin appeared at 1278 and 1103 cm(-1) respectively. Two second peaks at 1063 and 1037 cm(-1), corresponds mainly to glucomannan and arabinan. The relative area of these second peaks could be indicated the content of protein, tea polyphenol, pectin and polysaccharides in the Puer black tea. The results of curve fitting analysis showed that the relative area of amide II was increasing first and then decreasing, it indicated the change of protein in Puer black tea. At the same time, the content of tea polyphenol and pectin were decreased with the increase of aging time, but the glucomannan and arabinan were increased in reverse. It explained that the bitter taste was become weak and a sweet taste appeared in the tea with the increase of aging time. The present study suggested that Fourier transform infrared spectroscopy combined with curve-fitting analysis could reveal the biochemical changed of Puer black tea with the increase of aging time, and it was evidence for evaluation the quality of Puer black tea.

[Identification of Pummelo Cultivars Based on Hyperspectral Imaging Technology].

Existing methods for the identification of pummelo cultivars are usually time-consuming and costly, and are therefore inconvenient to be used in cases that a rapid identification is needed. This research was aimed at identifying different pummelo cultivars by hyperspectral imaging technology which can achieve a rapid and highly sensitive measurement. A total of 240 leaf samples, 60 for each of the four cultivars were investigated. Samples were divided into two groups such as calibration set (48 samples of each cultivar) and validation set (12 samples of each cultivar) by a Kennard-Stone-based algorithm. Hyperspectral images of both adaxial and abaxial surfaces of each leaf were obtained, and were segmented into a region of interest (ROI) using a simple threshold. Spectra of leaf samples were extracted from ROI. To remove the absolute noises of the spectra, only the date of spectral range 400~1000 nm was used for analysis. Multiplicative scatter correction (MSC) and standard normal variable (SNV) were utilized for data preprocessing. Principal component analysis (PCA) was used to extract the best principal components, and successive projections algorithm (SPA) was used to extract the effective wavelengths. Least squares support vector machine (LS-SVM) was used to obtain the discrimination model of the four different pummelo cultivars. To find out the optimal values of σ2 and γ which were important parameters in LS-SVM modeling, Grid-search technique and Cross-Validation were applied. The first 10 and 11 principal components were extracted by PCA for the hyperspectral data of adaxial surface and abaxial surface, respectively. There were 31 and 21 effective wavelengths selected by SPA based on the hyperspectral data of adaxial surface and abaxial surface, respectively. The best principal components and the effective wavelengths were used as inputs of LS-SVM models, and then the PCA-LS-SVM model and the SPA-LS-SVM model were built. The results showed that 99.46% and 98.44% of identification accuracy was achieved in the calibration set for the PCA-LS-SVM model and the SPA-LS-SVM model, respectively, and a 95.83% of identification accuracy was achieved in the validation set for both the PCA-LS-SVM and the SPA- LS-SVM models, which were built based on the hyperspectral data of adaxial surface. Comparatively, the results of the PCA-LS-SVM and the SPA-LS-SVM models built based on the hyperspectral data of abaxial surface both achieved identification accuracies of 100% for both calibration set and validation set. The overall results demonstrated that use of hyperspectral data of adaxial and abaxial leaf surfaces coupled with the use of PCA-LS-SVM and the SPA-LS-SVM could achieve an accurate identification of pummelo cultivars. It was feasible to use hyperspectral imaging technology to identify different pummelo cultivars, and hyperspectral imaging technology provided an alternate way of rapid identification of pummelo cultivars. Moreover, the results in this paper demonstrated that the data from the abaxial surface of leaf was more sensitive in identifying pummelo cultivars. This study provided a new method for to the fast discrimination of pummelo cultivars.

[Determination of photosynthetic pigments in citrus leaves based on hyperspectral images datas].

The effective region was segmented from the hyperspectral image of citrus leaf by threshold method with the average spectrum extracted and used to describe the corresponding leaf. Based on the different spectral pre-processing methods, the prediction models of three photosynthetic pigments (i. e., chlorophyll a, chlorophyll b, and carotenoid) were calibrated by partial least squares (PLS), BP neural network (BPNN) and least square support vector machine (LS-SVM). The LS-SVM model for chlorophyll a was established based on multiplicative scatter correction (MSC), and the correlation coefficient (Rp) and the root mean square error of prediction (RMSEP) were 0.898 3 and 0.140 4, respectively. The LS-SVM model for chlorophyll b with Rp = 0.912 3 and RMSEP = 0.042 6, was established based on standard normal variable (SNV). The PLS model for carotenoid was established with Rp = 0.712 8 and RMSEP = 0.062 4 based on moving average smoothing (MAS), but the result was no better than the other two. The results illustrated that these three photosynthetic pigments could be nondestructively and real time estimated by hyperspectral image.

Widely targeted metabolomics analysis of Sanghuangporus vaninii mycelia and fruiting bodies at different harvest stages.

Sanghuangprous vaninii is a medicinal macrofungus cultivated extensively in China. Both the mycelia and fruiting bodies of S. vaninii have remarkable therapeutic properties, but it remains unclear whether the mycelia may serve as a substitute for the fruiting bodies. Furthermore, S. vaninii is a perennial fungus with therapeutic components that vary significantly depending on the growing year of the fruiting bodies. Hence, it is critical to select an appropriate harvest stage for S. vaninii fruiting bodies for a specific purpose. With the aid of Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), metabolomics based on ultra-high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UHPLC-QQQ-MS) was used to preliminarily determine 81 key active metabolites and 157 active pharmaceutical metabolites in S. vaninii responsible for resistance to the six major diseases. To evaluate the substitutability of the mycelia and fruiting bodies of S. vaninii and to select an appropriate harvest stage for the fruiting bodies of S. vaninii, we analyzed the metabolite differences, especially active metabolite differences, among the mycelia and fruiting bodies during three different harvest stages (1-year-old, 2-year-old, and 3-year-old). Moreover, we also determined the most prominent and crucial metabolites in each sample of S. vaninii. These results suggested that the mycelia show promise as a substitute for the fruiting bodies of S. vaninii and that extending the growth year does not necessarily lead to higher accumulation levels of active metabolites in the S. vaninii fruiting bodies. This study provided a theoretical basis for developing and using S. vaninii.

Nanoscale coordination polymers enabling antioxidants inhibition for enhanced chemodynamic therapy.

Reactive oxygen species (ROS) generation to induce cell death is an effective strategy for cancer therapy. In particular, chemodynamic therapy (CDT), using Fenton-type reactions to generate highly cytotoxic hydroxyl radical (•OH), is a promising treatment modality. However, the therapeutic efficacy of ROS-based cancer treatment is still limited by some critical challenges, such as overexpression of enzymatic and non-enzymatic antioxidants by tumor cells, as well as the low tumor targeting efficiency of therapeutic agents. To address those problems, biomimetic CuZn protoporphyrin IX nanoscale coordination polymers have been developed, which significantly amplify oxidative stress against tumors by simultaneously inhibiting enzymatic and non-enzymatic antioxidants and initiating the CDT. In this design, cancer cell membrane camouflaged nanoparticle exhibits an excellent homotypic targeting effect. After being endocytosed into tumor cells, the nanoparticles induce depletion of the main non-enzymatic antioxidant glutathione (GSH) by undergoing a redox reaction with GSH. Afterward, the redox reaction generated cuprous ion (Cu+) works as a CDT agent for •OH generation. Furthermore, the released Zn protoporphyrin IX strongly inhibits the activity of the typical enzymatic antioxidant heme oxygenase-1. This tetra-modal synergistic strategy endows the biomimetic nanoparticles with great capability for anticancer therapy, which has been demonstrated in both in vitro and in vivo studies.

[Chemical Characteristics and Source Apportionment for VOCs During the Ozone Pollution Episodes and Non-ozone Pollution Periods in Qingdao].

The ambient concentration of ozone is high in Qingdao, and ozone pollution episodes occur frequently in summer. The refined source apportionment of ambient volatile organic compounds (VOCs) and their ozone formation potential (OFP) during ozone pollution episodes and non-ozone pollution periods can play an important role in effectively reducing air ozone pollution in coastal cities and continuously improving ambient air quality. Therefore, this study applied the online VOCs monitoring data with hourly resolution in summer (from June to August) in 2020 in Qingdao to analyze the chemical characteristics of ambient VOCs during the ozone pollution episodes and non-ozone pollution periods and conducted the refined source apportionment of ambient VOCs and their OFP using a positive matrix factorization (PMF) model. The results showed that the average mass concentration of ambient VOCs in Qingdao in summer was 93.8 µg·m-3, and compared with that during the non-ozone pollution period, the mass concentration of ambient VOCs during the ozone pollution episodes increased by 49.3%, and the mass concentration of aromatic hydrocarbons increased by 59.7%. The total OFP of ambient VOCs in summer was 246.3 µg·m-3. Compared with that in the non-ozone pollution period, the total OFP of ambient VOCs in the ozone pollution episodes increased by 43.1%; that of alkanes increased the most, reaching 58.8%. M-ethyltoluene and 2,3-dimethylpentane were the species with the largest increase in OFP and its proportion during the ozone pollution episodes. The main contributors of ambient VOCs in Qingdao in summer were diesel vehicles (11.2%), solvent use (4.7%), liquefied petroleum gas and natural gas (LPG/NG) (27.5%), gasoline vehicles (8.9%), gasoline volatilization (26.6%), emissions of combustion- and petrochemical-related enterprises (16.4%), and plant emissions (4.8%). Compared with that in the non-ozone pollution period, the contribution concentration of LPG/NG in the ozone pollution episodes increased by 16.4 µg·m-3, which was the source category with the largest increase. The contribution concentration of plant emissions increased by 88.6% in the ozone pollution episodes, which was the source category with the highest increase rate. In addition, emissions from combustion- and petrochemical-related enterprises were the largest contributor to the OFP of ambient VOCs in summer in Qingdao, with its OFP and contribution proportion being 38.0 µg·m-3and 24.5%, respectively, followed by that of LPG/NG and gasoline volatilization. Compared with the non-ozone pollution period, the total contributions of LPG/NG, gasoline volatilization, and solvent use to the increase in OFP for ambient VOCs in the ozone pollution episodes were 74.1%, which were the main contribution source categories.

[Source Apportionment of Ambient VOCs in Qingdao Based on Photochemical Loss Correction].

Ozone was one of the major pollutants affecting the environmental air quality in China. The accurate apportionment of key sources and their contributions of ambient ozone and its precursor VOCs played an important role in the effective prevention and control of ozone pollution. Therefore, this study utilized the photochemical-age-based parameterization method to estimate the initial concentrations of ambient VOCs data collected from January 1 to February 28, 2021 in Jiaozhou, Qingdao and corrected the photochemical losses of ambient VOC species. The positive matrix factorization(PMF) and ozone formation potential(OFP) models were used to conduct source apportionment of ambient VOCs and their OFPs so as to provide data support for the prevention and control of ozone pollution in Qingdao. The results showed that the average values of ambient ρ(TVOCs) and OFP in Qingdao during the study period were 65.9 µg·m-3 and 176.7 µg·m-3, respectively. Propane had the highest concentration(12.4 µg·m-3) and percentage(18.9%), whereas m/p-xylene had the highest OFP(24.6 µg·m-3) and percentage(13.9%). The mean initial concentration of TVOCs during the study was 153.1 µg·m-3, and its photochemical loss rate reached 63.8%. Alkenes were the VOC species with the highest photochemical loss rate(92.1%), and the photochemical loss rate of isoprene reached 98.6%, which was substantially higher than that of other VOC species. According to the source apportionment results of initial concentrations(IC-PMF), liquefied petroleum gas(24.2%), solvent use(17.8%), natural gas and petrochemical-related enterprises(16.6%), gasoline volatilization(13.2%), combustion and gasoline vehicle emissions(12.2%), biogenic emissions(8.6%), and diesel vehicle emissions(7.4%) were the main contributing sources of the ambient VOCs in Jiaozhou. Compared with the apportioned results of IC-PMF, the contribution of biogenic emissions was underestimated by 38.9% in the apportioned results based on observed concentrations(OC-PMF), and the contribution of natural gas and petrochemical-related enterprises was underestimated by 28.5%, and the underestimations of their contributions were substantially higher than those of other sources. Compared with that before the Spring Festival, the contribution of gasoline volatilization to ambient VOCs increased markedly during the Spring Festival, whereas the contributions of solvent use, combustion, and gasoline vehicle emissions to ambient VOCs increased most significantly after the Spring Festival. The main contributing sources of ambient ozone during the study period were solvent use(31.3%), natural gas and petrochemical-related enterprises(16.1%), biogenic emissions(14.5%), and combustion and gasoline vehicle emissions(13.2%). The primary contributors of ambient ozone in different Spring Festival periods showed substantial differences. Before the Spring Festival, solvent use had the highest contribution(71.1 µg·m-3), and gasoline volatilization was the highest contributor during the Spring Festival(34.4 µg·m-3), whereas biogenic emissions after the Spring Festival were the highest contributor(39.1 µg·m-3).

[Application of Qu single abdominal aorta clamping for bloodless hepatectomy and Pringle hepatectomy in 118 cases of liver tumors].

OBJECTIVE: To compare the results of application of Qu single abdominal aorta clamping for bloodless hepatectomy and Pringle hepatectomy in 118 cases of liver tumors. METHODS: The clinical data of 118 patients, including 59 patients undergoing Qu single abdominal aorta clamping for bloodless hepatectomy (Group QG) and 59 patients undergone Pringle first hepatic portal clamping hepatectomy (Group PG) since March 2009 in the Ningbo Tumor Hospital and Jiangxi Provincial Hospital were retrospectively reviewed. The changes of blood pressure, oxygen saturation, urine volume, intravenous fluid volume, amount of bleeding, time of abdominal aorta (or first hepatic portal) clamping, duration of operation and anesthesia, and other intraoperative indexes of the two groups were compared, and the changes of peritoneal drainage, blood tests, liver functions, etc. before operation and 1, 3, 7, 14 days after the hepatectomy in the two groups were also analyzed. RESULTS: After taking appropriate measures for intraoperative blood pressure control, only small fluctuations of blood pressure, which could be safely adjusted and controlled with stable vital signs, was observed in the group QG. The amount of intraoperative bleeding in the group QG was (96.25 ± 18.45) ml, significantly less than (536.25 ± 35.65) ml in the group PG (P < 0.05). In the group QG, both the duration of operation time [(227.58 ± 28.20) min] and duration of anesthesia [(249.48 ± 31.35) min] were significantly shorter than that [(261.46 ± 32.12) min and (286.58 ± 35.62) min, respectively] in the group PG (both P < 0.05). The postoperative liver dysfunction in the group QG was also milder than that in the group PG (P < 0.05). CONCLUSIONS: For liver tumor patients, Qu single abdominal aorta clamping for bloodless hepatectomy can basically achieve the goal of bloodless hepatectomy. This surgical operation is simple and safe, worthy of recommendation to skillful liver surgeons in hospitals there are some difficulties of blood supply.

[Identification of different Citrus sinensis (L.) Osbeck trees varieties using Fourier transform infrared spectroscopy and hierarchical cluster analysis].

Researched on diversity of the spring leaf samples of seven different Citrus sinensis (L.) Osbeck varieties by Fourier transform infrared (FTIR) spectroscopy technology, the results showed that the Fourier transform infrared spectra of seven varieties leaves was composited by the absorption band of cellulose and polysaccharide mainly, the wave number of characteristics absorption peaks were similar at their FTIR spectra. However, there were some differences in shape of peaks and relatively absorption intensity. The conspicuous difference was presented at the region between 1 500 and 700 cm(-1) by second derivative spectra. Through the hierarchical cluster analysis (HCA) of second derivative spectra between 1 500 and 700 cm(-1), the results showed that the clustering of the different varieties of Citrus sinensis (L.) Osbeck varieties was classification according to genetic relationship. The results showed that FTIR spectroscopy combined with hierarchical cluster analysis could be used to identify and classify of citrus varieties rapidly, it was an extension method to study on early leaves of varieties orange seedlings.

NLRP3 inflammasome activation in neutrophils directs early inflammatory response in murine peritonitis.

NLR family pyrin domain containing 3 (NLRP3) inflammasome mediates caspase-1-dependent processing of inflammatory cytokines such as IL-1ß, an essential endothelial activator, and contributes to the pathology of inflammatory diseases. To evaluate the role of NLRP3 in neutrophils in endothelial activation, which is still elusive, we used the thioglycollate-induced peritonitis model characterized by an early neutrophil influx, on Nlrp3-/- and Nlrp3+/+ mice. Nlrp3-/- mice recruited fewer neutrophils than Nlrp3+/+ into the peritoneum and showed lower IL-1ß in peritoneal lavage fluid. The higher production of IL-1ß in Nlrp3+/+ was neutrophil-dependent as neutrophil depletion prevented the IL-1ß production. The Nlrp3+/+ neutrophils collected from the peritoneal fluid formed significantly more filaments (specks) than Nlrp3-/- neutrophils of ASC (apoptosis-associated speck-like protein containing a caspase activating and recruitment domain), a readout for inflammasome activation. Intravital microscopy revealed that leukocytes rolled significantly slower in Nlrp3+/+ venules than in Nlrp3-/-. Nlrp3-/- endothelial cells isolated from mesenteric vessels demonstrated a lower percentage of P-selectin-positive cells with lower intensity of surface P-selectin expression than the Nlrp3+/+ endothelial cells evaluated by flow cytometry. We conclude that neutrophils orchestrate acute thioglycollate-induced peritonitis by producing IL-1ß in an NLRP3-dependent manner. This increases endothelial P-selectin expression and leukocyte transmigration.

Activation of CD81+ skin ILC2s by cold-sensing TRPM8+ neuron-derived signals maintains cutaneous thermal homeostasis.

As the outermost barrier tissue of the body, the skin harbors a large number of innate lymphoid cells (ILCs) that help maintain local homeostasis in the face of changing environments. How skin-resident ILCs are regulated and function in local homeostatic maintenance is poorly understood. We here report the discovery of a cold-sensing neuron-initiated pathway that activates skin group 2 ILCs (ILC2s) to help maintain thermal homeostasis. In stearoyl-CoA desaturase 1 (SCD1) knockout mice whose skin is defective in heat maintenance, chronic cold stress induced excessive activation of CCR10-CD81+ST2+ skin ILC2s and associated inflammation. Mechanistically, stimulation of the cold-sensing receptor TRPM8 expressed in sensory neurons of the skin led to increased production of IL-18, which, in turn, activated skin ILC2s to promote thermogenesis. Our findings reveal a neuroimmune link that regulates activation of skin ILC2s to support thermal homeostasis and promotes skin inflammation after hyperactivation.

Neutrophil stimulation with citrullinated histone H4 slows down calcium influx and reduces NET formation compared with native histone H4.

Peptidylarginine deiminase 4 (PAD4) catalyzes posttranslational modification of many target proteins through converting protein arginine or mono-methylarginine to citrulline. Neutrophil extracellular trap (NET) formation is the most dramatic manifestation of PAD4-mediated hypercitrullination reaction in neutrophils, which is characterized by the release of nuclear chromatin to form a chromatin network in the extracellular space. Histones H4, one of the major protein components of chromatin, is released into the extracellular space during sepsis, trauma, and ischemia-reperfusion injury and can also be released during the process of NET formation, along with its citrullinated form. The present study showed that histone H4 can induce NET formation in a calcium and PAD4 dependent manner. Histone H4 caused permeabilization of the neutrophil membrane and sustained rise in intracellular calcium that is necessary for activation of PAD4. In comparison, citrullinated histone H4 induced less calcium influx compared with its native form, leading to reduced NET formation. These studies suggest that citrullinated histone H4 could serve as a brake in the pathology of NETs, slowing down the vicious circle between histone H4 and NETs.

NLRP3 Inflammasome Assembly in Neutrophils Is Supported by PAD4 and Promotes NETosis Under Sterile Conditions.

Neutrophil extracellular trap formation (NETosis) and the NLR family pyrin domain containing 3 (NLRP3) inflammasome assembly are associated with a similar spectrum of human disorders. While NETosis is known to be regulated by peptidylarginine deiminase 4 (PAD4), the role of the NLRP3 inflammasome in NETosis was not addressed. Here, we establish that under sterile conditions the cannonical NLRP3 inflammasome participates in NETosis. We show apoptosis-associated speck-like protein containing a CARD (ASC) speck assembly and caspase-1 cleavage in stimulated mouse neutrophils without LPS priming. PAD4 was needed for optimal NLRP3 inflammasome assembly by regulating NLRP3 and ASC protein levels post-transcriptionally. Genetic ablation of NLRP3 signaling resulted in impaired NET formation, because NLRP3 supported both nuclear envelope and plasma membrane rupture. Pharmacological inhibition of NLRP3 in either mouse or human neutrophils also diminished NETosis. Finally, NLRP3 deficiency resulted in a lower density of NETs in thrombi produced by a stenosis-induced mouse model of deep vein thrombosis. Altogether, our results indicate a PAD4-dependent formation of the NLRP3 inflammasome in neutrophils and implicate NLRP3 in NETosis under noninfectious conditions in vitro and in vivo.

[Comparative proteomic approach in differentiating multicentric occurrence and intrahepatic metastasis in multinodular hepatocellular carcinomas].

BACKGROUND AND OBJECTIVE: Multinodular hepatocellular carcinoma(HCC) might originate from multicentric occurrence (MO) or intrahepatic metastasis(IM). This study was to find out proteins which play important roles in clonal origin of multinodular hepatocellular carcinoma bt screening the differentially expressed proteins between the MO and IM tissues using comparative proteomic analysis. METHODS: Total protein extracted was separated by two-dimensional gel electrophoresis. Comparative analyses of the 2-DE protein patterns between the two groups were carried out using computerized imaging techniques. Proteins exhibiting significant alternations were subsequently isolated and identified by mass spectrometry. RESULTS: A total 1025+/-52 and 900+/-98 spots were detected in the protein profile in IM and MO, respectively. Twenty-five protein spots were statistically different at expression levels between the two groups. Twenty of them were identified by MALDI-TOF-MS and bioinformatics. CONCLUSIONS: The protein profile of MO HCC tissues is different from that in IM HCC tissues. The twenty differentially expressed proteins might play a key role in the carcinogenesis and progression of multinodular HCC. These newly identified proteins might be potential and valuable biomarkers for identifying the multinodular HCC of clonal origin.

[Research on zinc content in leaf of Olinda Valencia orange using visible near infrared spectroscopy model].

Olinda valencia orange leaves dry powder-like were taken as sample, and chemical analysis combined with technology of visible near-infrared spectroscopy (Vis/NIRS) was used, through the treatment process of second derivative spectrum of samples of the original spectrum and denoising (Noise). Meanwhile, method of partial least squares (PLS) and cross-validation were used to establish maths model of Zn concentration which applying band combination composited by 400-500 and 1201-1300 nm of characteristic wavelength band. The coefficient of establishing models is 0.9975, while the coefficient of correlation coefficient of prediction is 0.9920. The root mean square error of prediction (RMSEP) of cross-validation is 0.5868. Therefore, the means using visible near-infrared spectroscopy (Vis/NIRS) and the methods of cross-validation and PLS to establish the spectral correction model reflecting the Zn content in leaves and characteristic wavelength bands can detect the Zn content in citrus leaves quantitatively and quickly.

[Study on synchronous correlation between fruit characteristic spectrum and the parameter of internal quality for Hamlin sweet orange fruit].

The purpose of our study is to study the relationship between the reflective spectrum of fruit and the internal quality of the orange fruit and find the suitable mark spectrum that can synchronously measure the several fruit quality index at same time to lay a foundation for the development of a rapid and nondestructive field fruit quality analysis technique by analyzing the visible-near infrared spectrum. Mature Hamlin orange (Citrus sinensis (L)cv. Hamlin sweet orange) fruits were individually mensurated for their reflective spectrum by using FieldSpec-HH spectrometer and for their contents of total soluble solid (TSS), citric acid and vitamin C (Vc) by chemistry analysis. The experiment results showed that the fruit reflectivity values (x) at 988 nm was significantly correlated to both TSS (y)(r = 0.387* *, y = 13. 957x + 5.405), TSS/acid ratio(y)(r = 0. 440* *, y = 75.120x + 37.256), and Vc(r = 0.309*). Both of TSS and Vc contends were positively correlated with the second derivatives of the reflective spectrum at 943 nm, with correlation coefficients of 0.339* and 0.355*. TSS/acid ratio was positively correlated only with the reciprocal log values of the reflective spectrum at 944 nm (r = 0.304*). The results in this study indicated that fruit quality indexes TSS, Vc and TSS/acid can be synchronously, rapidly and nondestructively field measured at the same time by the 988 or 429 nm reflective spectrum test and special regress equation operation.

Endogenous PAD4 in Breast Cancer Cells Mediates Cancer Extracellular Chromatin Network Formation and Promotes Lung Metastasis.

Peptidyl arginine deiminase 4 (PAD4/PADI4) is a posttranslational modification enzyme that converts protein arginine or mono-methylarginine to citrulline. The PAD4-mediated hypercitrullination reaction in neutrophils causes the release of nuclear chromatin to form a chromatin network termed neutrophil extracellular traps (NET). NETs were first described as antimicrobial fibers that bind and kill bacteria. However, it is not known whether PAD4 can mediate the release of chromatin DNA into the extracellular space of cancer cells. Here, we report that murine breast cancer 4T1 cells expressing high levels of PADI4 can release cancer extracellular chromatin networks (CECN) in vitro and in vivo. Deletion of Padi4 using CRISPR/Cas9 abolished CECN formation in 4T1 cells. Padi4 deletion from 4T1 cells also reduced the rate of tumor growth in an allograft model, and decreased lung metastasis by 4T1 breast cancers. DNase I treatment, which degrades extracellular DNA including CECNs, also reduced breast to lung metastasis of Padi4 wild-type 4T1 cells in allograft experiments in the Padi4-knockout mice. We further demonstrated that DNase I treatment in this mouse model did not alter circulating tumor cells but decreased metastasis through steps after intravasation. Taken together, our genetic studies show that PAD4 plays a cell autonomous role in cancer metastasis, thus revealing a novel strategy for preventing cancer metastasis by inhibiting cancer cell endogenous PAD4. IMPLICATIONS: This study shows that PADI4 can mediate the formation of CECNs in 4T1 cells, and that endogenous PADI4 plays an essential role in breast cancer lung metastasis. VISUAL OVERVIEW: http://mcr.aacrjournals.org/content/molcanres/18/5/735/F1.large.jpg.