NLRP3-Mediated Piezo1 Upregulation in ACC Inhibitory Parvalbumin-Expressing Interneurons Is Involved in Pain Processing after Peripheral Nerve Injury.

The anterior cingulate cortex (ACC) is particularly critical for pain information processing. Peripheral nerve injury triggers neuronal hyper-excitability in the ACC and mediates descending facilitation to the spinal dorsal horn. The mechanically gated ion channel Piezo1 is involved in the transmission of pain information in the peripheral nervous system. However, the pain-processing role of Piezo1 in the brain is unknown. In this work, we found that spared (sciatic) nerve injury (SNI) increased Piezo1 protein levels in inhibitory parvalbumin (PV)-expressing interneurons (PV-INs) but not in glutaminergic CaMKⅡ+ neurons, in the bilateral ACC. A reduction in the number of PV-INs but not in the number of CaMKⅡ+ neurons and a significant reduction in inhibitory synaptic terminals was observed in the SNI chronic pain model. Further, observation of morphological changes in the microglia in the ACC showed their activated amoeba-like transformation, with a reduction in process length and an increase in cell body area. Combined with the encapsulation of Piezo1-positive neurons by Iba1+ microglia, the loss of PV-INs after SNI might result from phagocytosis by the microglia. In cellular experiments, administration of recombinant rat TNF-α (rrTNF) to the BV2 cell culture or ACC neuron primary culture elevated the protein levels of Piezo1 and NOD-like receptor (NLR) family pyrin domain containing 3 (NLRP3). The administration of the NLRP3 inhibitor MCC950 in these cells blocked the rrTNF-induced expression of caspase-1 and interleukin-1ß (key downstream factors of the activated NLRP3 inflammasome) in vitro and reversed the SNI-induced Piezo1 overexpression in the ACC and alleviated SNI-induced allodynia in vivo. These results suggest that NLRP3 may be the key factor in causing Piezo1 upregulation in SNI, promoting an imbalance between ACC excitation and inhibition by inducing the microglial phagocytosis of PV-INs and, thereby, facilitating spinal pain transmission.

Study on the Effect of Molecular Weight on the Gut Microbiota Fermentation Properties of Blackberry Polysaccharides In Vitro.

This study aimed to investigate the effect of different molecular weights on the metabolic characteristics of blackberry polysaccharides (BBP). After degradation, three fractions, namely, BBP-8, BBP-16, and BBP-24, were obtained. During fermentation, all polysaccharide fractions were significantly degraded and utilized by the intestinal microbiota, and the lower-molecular-weight polysaccharides were easier to be fermented with higher gas production and carbohydrate consumption rates. Furthermore, the monosaccharide utilization sequence of all polysaccharides was glucose > galactose > arabinose > galacturonic acid. In addition, the lower-molecular-weight polysaccharides had a faster short-chain fatty acid (SCFA) production rate but did not affect the final SCFA yields. The fermentation of BBP promoted the increase of Bacteroidetes and the decrease of Firmicutes. The proportions of Bacteroidetes in BBP, BBP-8, BBP-16, and BBP-24 were 45.41, 47.50, 48.08, and 50.09%, respectively.

Neuroimmune Mechanisms Underlying Neuropathic Pain: The Potential Role of TNF-α-Necroptosis Pathway.

The neuroimmune mechanism underlying neuropathic pain has been extensively studied. Tumor necrosis factor-alpha (TNF-α), a key pro-inflammatory cytokine that drives cytokine storm and stimulates a cascade of other cytokines in pain-related pathways, induces and modulates neuropathic pain by facilitating peripheral (primary afferents) and central (spinal cord) sensitization. Functionally, TNF-α controls the balance between cell survival and death by inducing an inflammatory response and two programmed cell death mechanisms (apoptosis and necroptosis). Necroptosis, a novel form of programmed cell death, is receiving increasing attraction and may trigger neuroinflammation to promote neuropathic pain. Chronic pain is often accompanied by adverse pain-associated emotional reactions and cognitive disorders. Overproduction of TNF-α in supraspinal structures such as the anterior cingulate cortex (ACC) and hippocampus plays an important role in pain-associated emotional disorders and memory deficits and also participates in the modulation of pain transduction. At present, studies reporting on the role of the TNF-α-necroptosis pathway in pain-related disorders are lacking. This review indicates the important research prospects of this pathway in pain modulation based on its role in anxiety, depression and memory deficits associated with other neurodegenerative diseases. In addition, we have summarized studies related to the underlying mechanisms of neuropathic pain mediated by TNF-α and discussed the role of the TNF-α-necroptosis pathway in detail, which may represent an avenue for future therapeutic intervention.

Neuroinflammation in the anterior cingulate cortex: the potential supraspinal mechanism underlying the mirror-image pain following motor fiber injury.

BACKGROUND: Peripheral nerve inflammation or lesion can affect contralateral healthy structures, and thus result in mirror-image pain. Supraspinal structures play important roles in the occurrence of mirror pain. The anterior cingulate cortex (ACC) is a first-order cortical region that responds to painful stimuli. In the present study, we systematically investigate and compare the neuroimmune changes in the bilateral ACC region using unilateral- (spared nerve injury, SNI) and mirror-(L5 ventral root transection, L5-VRT) pain models, aiming to explore the potential supraspinal neuroimmune mechanism underlying the mirror-image pain. METHODS: The up-and-down method with von Frey hairs was used to measure the mechanical allodynia. Viral injections for the designer receptors exclusively activated by designer drugs (DREADD) were used to modulate ACC glutamatergic neurons. Immunohistochemistry, immunofluorescence, western blotting, protein microarray were used to detect the regulation of inflammatory signaling. RESULTS: Increased expressions of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6) and chemokine CX3CL1 in ACC induced by unilateral nerve injury were observed on the contralateral side in the SNI group but on the bilateral side in the L5-VRT group, representing a stronger immune response to L5-VRT surgery. In remote ACC, both SNI and L5-VRT induced robust bilateral increase in the protein level of Nav1.6 (SCN8A), a major voltage-gated sodium channel (VGSC) that regulates neuronal activity in the mammalian nervous system. However, the L5-VRT-induced Nav1.6 response occurred at PO 3d, earlier than the SNI-induced one, 7 days after surgery. Modulating ACC glutamatergic neurons via DREADD-Gq or DREADD-Gi greatly changed the ACC CX3CL1 levels and the mechanical paw withdrawal threshold. Neutralization of endogenous ACC CX3CL1 by contralateral anti-CX3CL1 antibody attenuated the induction and the maintenance of mechanical allodynia and eliminated the upregulation of CX3CL1, TNF-α and Nav1.6 protein levels in ACC induced by SNI. Furthermore, contralateral ACC anti-CX3CL1 also inhibited the expression of ipsilateral spinal c-Fos, Iba1, CD11b, TNF-α and IL-6. CONCLUSIONS: The descending facilitation function mediated by CX3CL1 and its downstream cascade may play a pivotal role, leading to enhanced pain sensitization and even mirror-image pain. Strategies that target chemokine-mediated ACC hyperexcitability may lead to novel therapies for the treatment of neuropathic pain.

[Preparation of a New Type of Silk Birth-canal Microecology Transporter].

OBJECTIVE: To optimize the preparation parameters of the new silk birth-canal microecology transporter (BMT) for transferring the symbiotic bacteria of the birth canal efficiently. METHODS: Birth canal microbial samples of 30 full term pregnant women at admission were collected as the control group (NC, n=30). The experimental group included 18 pregnant women terminated by Cesarean section, who were divided into 6 sub-groups （M1-M6, n=3) to complete the transfer tests of the birth-canal microecology. The new silk BMT was processed in the sterile liquid of the different osmotic pressure with the different immersion depth, and was placed in the vagina of the pregnant women for 1 h before sealed. All extracted DNA specimens were amplified in the V3-V4 region of the 16S rDNA, and were sequenced by Illumina Hiseq2500. Microbial diversity analysis was performed by Mothur, QIIME, Lefse and Metastat. Welch's t-test and Anosim nonparametric test were used to compare the difference between groups. RESULTS: The new silk BMT with 70% immersion depth could be fully covered by the solution, and had good solution preserving and adhesion. The subjects had no foreign body sensation with satisfied experience. Both of the microbes on the new BMT and the control group were lactobacillus as the dominant bacteria genus. The microbial diversity and bacteria constitution in the new BMT was similar to the control group in the condition of 0.45% NaCl solution and 70% immersion depth, and there was no significant difference between the two groups ( P>0.05). CONCLUSION: The new silk BMT can transfer the symbiotic microbes of the birth canal efficiently, and the optimal preparation parameters were 0.45% hypotonic saline solution and 70% immersion depth.

[Allocation of rice photosynthates in plant-soil system in response to elevated CO2 and nitrogen fertilization.] / æ°´ç¨»å ‰åˆç¢³åœ¨æ¤ç‰©-åœŸå£¤ç³»ç»Ÿä¸­çš„åˆ†é åŠå ¶å¯¹CO2升高和施氮的响应.

To examine the allocation of rice photosynthates and its response to the elevated CO2 (800 µL·L-1) and N fertilization (100 mg·kg-1) at both tillering stage and booting stage in plant-soil system, rice was continually labelled with 13CO2. The results showed that the rice root biomass at the tillering stage and the shoot biomass at the booting stage were significantly increased under elevated CO2. Elevated CO2 increased the rice biomass and root-shoot ratio at tillering stage, but reduced it at booting stage. Under elevated CO2, N fertilization promoted shoot biomass during rice growth, but significantly decreased the root biomass at booting stage. Elevated CO2 significantly increased the allocation of assimilated 13C to the soil at the booting stage. N fertilization did not promote the elevated CO2-induced stimulation of assimilated 13C allocated to the soil, and it even decreased the proportion of assimilated 13C in the soil. In summary, elevated CO2 increased the photosynthetic C allocation into soil and promoted the turnover of soil organic carbon in paddy soil. N fertilization enhanced rice shoot biomass but decreased the belowground allocation of photosynthetic C.

Identification and Pathogenicity of Fungal Pathogens Causing Black Point in Wheat on the North China Plain.

Fungi associated with black point were isolated from three highly susceptible wheat genotypes in the North China Plain. The 21 isolates represented 11 fungal genera. The most prevalent genera were Alternaria (isolation frequency of 56.7%), Bipolaris (16.1%), and Fusarium (6.0%). The other eight genera were Curvularia, Aspergillus, Cladosporium, Exserohilum, Epicoccum, Nigrospora, Penicillium, and Ulocladium; their isolation frequencies ranged from 0.8 to 4.8%. The pathogenicity of the isolates was individually assessed in the greenhouse by inoculating wheat plants with spore suspensions. Ten of the 21 isolates caused significantly higher incidences of black point than that the controls. These isolates belonged to eight fungal species (A. alternata, B. sorokiniana, B. crotonis, B. cynodontis, C. spicifera, F. equiseti, E. rostratum, and E. sorghinum) based on morphological traits and phylogenetic analysis. The average incidences of black point in the eight fungal species were 32.4, 54.3, 43.0, 41.9, 37.2, 38.8, 50.1, and 34.1%, respectively. B. sorokiniana and A. alternata were determined to be the most important pathogens in the North China Plain based on fungal prevalence and symptom severity. This study is the first to identify E. rostratum as a major pathogen causing black point in wheat.

[Characteristics and Influencing Factors of Biologically-based Phosphorus Fractions in the Farmland Soil].

A suitable fractionation method of phosphorus (P) is a key to effective assessment of soil P componential features. Here a new biologically-based P (BBP) method was used to evaluate the P fractions in the upland and paddy soils across large-scale area in China. The soil P was divided into four components:① soluble or rhizosphere-intercepted (CaCl2-P), ② organic acid activated and inorganic weakly bound (Citrate-P), ③ enzyme mineralization of organic P (Enzyme-P), ④ potential activation of inorganic P (HCl-P). Then, the relationships between biologically-based P fractions and standard Olsen-P were investigated, and driving factors of P fractions were identified. The results showed that P content was in order of HCl-P>Citrate-P>Enzyme-P>CaCl2-P. All P components of upland soil displayed higher levels than those of paddy soil. Moreover, the P components were highly positively correlated with the Olsen-P, suggesting that each P component contributed to soil P availability. However, it was found that Olsen-P was most highly correlated with CaCl2-P and Enzyme-P (R2=0.359; R2=0.386) in upland soil, while Olsen-P was most highly with Citrate-P (R2=0.788) in paddy soil. This result indicated that available P of upland soil was mainly from organic P mineralization and soluble P, and available P in paddy soil was mainly from inorganic P activation. Redundancy analysis (RDA) showed that the P components were mainly affected by soil pH and silt content, which suggested that it could enhance the P availability via regulating soil pH in the agricultural activities.

Transcriptome Analysis for Abnormal Spike Development of the Wheat Mutant dms.

BACKGROUND: Wheat (Triticum aestivum L.) spike development is the foundation for grain yield. We obtained a novel wheat mutant, dms, characterized as dwarf, multi-pistil and sterility. Although the genetic changes are not clear, the heredity of traits suggests that a recessive gene locus controls the two traits of multi-pistil and sterility in self-pollinating populations of the medium plants (M), such that the dwarf genotype (D) and tall genotype (T) in the progeny of the mutant are ideal lines for studies regarding wheat spike development. The objective of this study was to explore the molecular basis for spike abnormalities of dwarf genotype. RESULTS: Four unigene libraries were assembled by sequencing the mRNAs of the super-bulked differentiating spikes and stem tips of the D and T plants. Using integrative analysis, we identified 419 genes highly expressed in spikes, including nine typical homeotic genes of the MADS-box family and the genes TaAP2, TaFL and TaDL. We also identified 143 genes that were significantly different between young spikes of T and D, and 26 genes that were putatively involved in spike differentiation. The result showed that the expression levels of TaAP1-2, TaAP2, and other genes involved in the majority of biological processes such as transcription, translation, cell division, photosynthesis, carbohydrate transport and metabolism, and energy production and conversion were significantly lower in D than in T. CONCLUSIONS: We identified a set of genes related to wheat floral organ differentiation, including typical homeotic genes. Our results showed that the major causal factors resulting in the spike abnormalities of dms were the lower expression homeotic genes, hormonal imbalance, repressed biological processes, and deficiency of construction materials and energy. We performed a series of studies on the homeotic genes, however the other three causal factors for spike abnormal phenotype of dms need further study.

[Expression of BRAF V600E mutation in different thyroid lesions].

OBJECTIVE: To evaluate the expression of BRAF V600E mutation in 240 Chinese patients with thyroid lesions. METHODS: Two hundred and forty Chinese patients with thyroid lesions, including 129 papillary thyroid carcinomas (PTC), 12 follicular carcinomas, 4 medullary carcinomas, 30 adenomas, 30 nodular goiters, and 35 papillary hyperplasia. DNA was extracted from thyroid biopsy and paraffin embedded thyroid tissues, and the expression of BRAF V600E mutation was detected by polymerase chain reaction and DNA sequencing assays. RESULTS: The presence of BRAF V600E mutation was found in 61 of the total group of 240 cases (25.4%). It was only detected in PTC (47.3%), and not detected in other types of malignant and benign thyroid lesions. There was a statistically significant difference between the expression of BRAF V600E mutation in classic type PTC (49.6%) and in follicular type PTC (12.5%,P < 0.05), but statistical data did not show any correlation between BRAF V600E mutation and clinicopathologic parameters in PTC (P > 0.05). CONCLUSIONS: BRAF V600E mutation has a significant correlation with PTC and the detection of BRAF V600E mutation may be used as an important prognostic marker of PTC. Our new method of DNA extraction from paraffin embedded tissues is efficient and inexpensive.

[Effect of ginsenoside Rg1 on angiogenesis after neonatal hypoxia ischemia brain damage in rats].

OBJECTIVE: To investigate effects of ginsenoside Rg1 on angiogenesis in neonatal rats with hypoxia ischemia brain damage (HIBD), and explore the possible mechanism. METHODS: Fifty-four of 10-day-old SD rats were randomly divided into sham-operation group (n = 6), hypoxia-ischemia brain damage group (n = 24) and ginsenoside Rg1 treatment group (n = 24). SD rats in HIBD group and Rg1 group were treated by separation and ligation of right common carotid artery (CCA) and subsequently exposed to hypoxia for 2.5 hours, and those in sham group were treated by only separation of right CCA, without ligation or exposure to hypoxia. Intraperitoneal injection of 0.1 mL normal saline (NS) containing 40 mg/kg Rg1 was performed immediately after operation in Rg1 group, and such process was repeated every 24 h for 3 days. Intraperitoneal injection of 0.1 mL pure NS was performed in both HIBD group and sham group, in the same way as that of in Rg1 group. General state of SD rats after operation was monitored, 4, 8, 24 and 72 hours after HIBD, animals were executed and the right side of brain tissue was separated for further process. Protein expression of both hypoxia inducible factor 1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) were detected by both Western blot and immunohistochemistry. Immunohistochemistry for von willebrand factor (vwf) was used to labeling micro vessels. RESULTS: All rats survived to the end of the study and neurological dysfunction was observed in both HIBD group and Rgl group, but not in sham group. Expression of HIF-1alpha protein in HIBD group was increased at 4, 8, 24 and 72 h, compared to that in sham group (P < 0.05). Expression of HIF-1alpha protein in Rg1 group was increased compared to that in HIBD group at the same time points (P < 0.05). Expression of VEGF protein in HIBD group was increased at 4, 8, and 24 h, compared to that in sham group (P < 0.05). Expression of VEGF protein in Rg1 group was increased at 24 and 72 h compared to that in HIBD group at the same time point (P < 0.05). Number of vwf-positive cells at 24 and 72 h in HIBD group was increased compared to that in sham group (P < 0.05), and number of vwf-positive cells at 72 h in Rg1 group was increased compared to that in HIBD group at the same time point (P < 0.05). CONCLUSION: Rg1 could facilitate angiogenesis after HIBD in Neonatal rats by strengthening and stabilizing HIF-1alpha/VEGF signaling pathway.

Transgenic barley with overexpressed PTrx increases aluminum resistance in roots during germination.

A transgenic barley line (LSY-11-1-1) with overexpressed Phalaris coerulescens thioredoxin gene (PTrx) was employed to measure the growth, protein oxidation, cell viability, and antioxidase activity in barley roots during germination on the presence of 2 mmol/L AlCl(3) on filter paper. The results show that (1) compared with the non-transgenic barley, LSY-11-1-1 had enhanced root growth, although both were seriously inhibited after AlCl(3) treatment; (2) the degree of protein oxidation and loss of cell viability in roots of LSY-11-1-1 were much less than those in roots of non-transgenic barley, as reflected by lower contents of protein carbonyl and Evans blue uptakes in LSY-11-1-1; (3) activities of catalase (CAT), glutathione peroxidase (GPX), ascorbate peroxidase (APX), and glutathione reductase (GR) in LSY-11-1-1 root tips were generally higher than those in non-transgenic barley root tips, although these antioxidase activities gave a rise to different degrees in both LSY-11-1-1 and non-transgenic barley under aluminum stress. These results indicate that overexpressing PTrx could efficiently protect barley roots from oxidative injury by increasing antioxidase activity, thereby quenching ROS caused by AlCl(3) during germination. These properties raise the possibility that transgenic barley with overexpressed PTrx may be used to reduce the aluminum toxicity in acid soils.

Hepatic angiomyolipoma: dynamic computed tomography features and clinical correlation.

AIM: To study the dynamic computed tomography (CT) features of hepatic angiomyolipoma (AML) in patients with or without tuberous sclerosis complex (TSC). METHODS: The clinical information, CT findings and histopathological results of hepatic AML were analyzed retrospectively in 10 patients. RESULTS: Hepatic AML was prone to occur in female patients (7/10), and most of the patients (8/10) had no specific symptoms. All tumors presented as well-defined, unenveloped nodules in the liver. Six patients with sporadic hepatic AML had a solitary hepatic nodule with a definite fat component. Non-fat components of the hepatic lesions were enhanced earlier and persistently. Prominent central vessels were noted in the portal venous phase in three patients. In four patients with hepatic AML and TSC, most of the nodules were within the peripheral liver. Seven fat-deficient nodules were found with earlier contrast enhancement and rapid contrast material washout in two patients. Lymphangioleiomyomatosis was found in one patient. CONCLUSION: Imaging features of hepatic AML are characteristic. Correct diagnosis preoperatively can be made in combination with clinical features.

Protective role of exogenous nitric oxide against oxidative-stress induced by salt stress in barley (Hordeum vulgare).

To probe into the potential of relieving the oxidative damage of salt stress, we investigated the protective role of nitric oxide on barley under salt stress. Salt stress resulted in increased ion leakage, lipid peroxidation and protein oxidation in barley leaves. Simultaneous treatments of barley leaves with 50 microM sodium nitroprusside, a nitric oxide donor, alleviated the damage of salt stress, reflected by decreased ion leakage, and malendialdehyde (MDA), carbonyl, and hydrogen peroxide content in barley leaves. The presence of the nitric oxide donor increased the activities of superoxide dismutases (SOD), ascorbate peroxidases (APX), and catalases (CAT). Meantime, sodium nitroprusside addition increased accumulation of ferritin at the protein level, indicating that nitric oxide directly regulated ferritin accumulation. These results suggested that nitric oxide can effectively protect seedlings from salt stress damage by enhancing activities of antioxidant enzymes to quench the excessive reactive oxygen species caused by salt stress and inducing the increase of ferritin accumulation to chelate larger number of ferrous ion. Information from this study can be used to improve soil management practices for sustainable use of salt-affected soils in the future.

Bis(5-aminotetrazole-1-acetato-kappaO)tetraaquacobalt(II) and catena-poly[[cadmium(II)]-bis(mu-5-aminotetrazole-1-acetato-kappa3N4:O,O')].

The Co(II) atom in bis(5-aminotetrazole-1-acetato)tetraaquacobalt(II), [Co(C3H4N5O2)2(H2O)4], (I), is octahedrally coordinated by six O atoms from two 5-aminotetrazole-1-acetate (atza) ligands and four water molecules. The molecule has a crystallographic centre of symmetry located at the Co(II) atom. The molecules of (I) are interlinked by hydrogen-bond interactions, forming a two-dimensional supramolecular network structure in the ac plane. The Cd(II) atom in catena-poly[[cadmium(II)]-bis(mu-5-aminotetrazole-1-acetato], [Cd(C3H4N5O2)2]n, (II), lies on a twofold axis and is coordinated by two N atoms and four O atoms from four atza ligands to form a distorted octahedral coordination environment. The Cd(II) centres are connected through tridentate atza bridging ligands to form a two-dimensional layered structure extending along the ab plane, which is further linked into a three-dimensional structure through hydrogen-bond interactions.

Bis[bis(3,5-dimethylpyrazol-1-yl)methane]bis(isothiocyanato)cadmium(II) and catena-poly[[di-mu-dicyanamidato-bis{[bis(3,5-dimethylpyrazol-1yl)methane]cadmium(II)}]-di-mu-dicyanamidato].

The Cd atom in bis[bis(3,5-dimethylpyrazol-1-yl)methane]bis(isothiocyanato)cadmium(II), [Cd(NCS)2(C11H16N4)2], is octahedrally coordinated by four N atoms from two bis(3,5-dimethylpyrazolyl)methane (dmpzm) ligands and two isothiocyanate ligands. The molecule has a crystallographic center of symmetry located at the Cd atom. There are two intramolecular C-H...N interactions, each of which is formed between the methylene group of a dmpzm ligand and the N atom of an isothiocyanate ligand. On the other hand, in catena-poly[[di-mu-dicyanamidato-bis{[bis(3,5-dimethylpyrazol-1-yl)methane]cadmium(II)}]-di-mu-dicyanamidato], [Cd2(C2N3)4(C11H16N4)2]n, each Cd atom is octahedrally coordinated by two N atoms from one dmpzm ligand and four N atoms from four bridging dicyanamide (dca) anions. Two Cd atoms are bridged by a pair of dca anions, forming a dimeric [Cd2(mu-dca)2(dmpzm)2] unit. Another two pairs of dca anions further link this unit with neighboring units to form a brick-wall layer parallel to (100). A C-H...N interaction between the methylene group of one dmpzm ligand in one layer and a coordinating N atom of a dca ligand in an adjacent layer completes a three-dimensional network.