Complement and the Alternative Pathway Play an Important Role in LPS/D-GalN-Induced Fulminant Hepatic Failure

Fulminant hepatic failure (FHF) is a clinically severe type of liver injury with an extremely high mortality rate. Although the pathological mechanisms of FHF are not well understood, evidence suggests that the complement system is involved in the pathogenesis of a variety of liver disorders. In the present study, to investigate the role of complement in FHF, we examined groups of mice following intraperitoneal injection of LPS/D-GalN: wild-type C57BL/6 mice, wild-type mice treated with a C3aR antagonist, C5aR monoclonal antibody (C5aRmAb) or CR2-Factor H (CR2-fH, an inhibitor of the alternative pathway), and C3 deficient mice (C3−/− mice). The animals were euthanized and samples analyzed at specific times after LPS/D-GalN injection. The results show that intraperitoneal administration of LPS/D-GalN activated the complement pathway, as evidenced by the hepatic deposition of C3 and C5b-9 and elevated serum levels of the complement activation product C3a, the level of which was associated with the severity of the liver damage. C3a receptor (C3aR) and C5a receptor (C5aR) expression was also upregulated. Compared with wild-type mice, C3−/− mice survived significantly longer and displayed reduced liver inflammation and attenuated pathological damage following LPS/D-GalN injection. Similar levels of protection were seen in mice treated with C3aR antagonist,C5aRmAb or CR2-fH. These data indicate an important role for the C3a and C5a generated by the alternative pathway in LPS/D-GalN-induced FHF. The data further suggest that complement inhibition may be an effective strategy for the adjunctive treatment of fulminant hepatic failure

Transcriptome Analysis Reveals Regulatory Networks and Hub Genes in the Flavonoid Metabolism of <i>Rosa roxburghii</i>

Rosa roxburghii Tratt, the most popular fruit that blooms in the southwest of China, has high antioxidant properties and is rich in different flavonoids. However, the regulatory network and critical genes that regulate the flavonoid biosynthesis of R. roxburghii are still unknown. In this study, HPLC analysis revealed that total flavonoids, anthocyanins, and catechin were enriched in mature fruits, flowers, and leaves, respectively. Differentially expressed genes (DEGs) between five organs of R. roxburghii involved in flavonoid metabolism were obtained by transcriptome sequencing. A total of 1130 DEGs were identified, including 166 flavonoid pathway biosynthesis genes, 622 transcription factors (TFs), 301 transporters, and 221 cytochrome P450 proteins. A weighted gene co-expression network analysis (WGCNA) of the DEGs was conducted to construct co-expression networks. Regarding enzymes in the biosynthesis of flavonoids, cytochrome P450 CYP749A22 and CYP72A219 were highlighted in the regulation of total flavonoids of mature fruits. Anthocyanin 3-O-glucosyltransferase and F3′H were the top two critical enzymes for anthocyanin accumulation in flowers. By contrast, caffeic acid 3-O-methyltransferase, 4-coumarate-CoA ligase, and shikimate O-hydroxycinnamoyltransferase were essential for catechin accumulation in leaves. Additionally, we analyzed the eigengene network of the “black” module, which had high correlations with total flavonoids (r = 0.9, p = 5 × 10−6). There were 26 eigengenes in the “black” module, consisting of 6 flavonoid biosynthesis, 14 TFs, and 6 transporters. Among them, the transcription factors RrWRKY45 (DN142829_c1_g5), RrTCP20 (DN146443_c1_g1), and RrERF118 (DN141507_c3_g2) were screened as the hub genes, which significantly correlated with total flavonoids in R. roxburghii. The present biochemical and transcriptomic data provide insights into functional genomics for breeding R. roxburghii with flavonoid accumulation

Transcriptome Analysis Reveals Regulatory Networks and Hub Genes in the Flavonoid Metabolism of Rosa roxburghii

Rosa roxburghii Tratt, the most popular fruit that blooms in the southwest of China, has high antioxidant properties and is rich in different flavonoids. However, the regulatory network and critical genes that regulate the flavonoid biosynthesis of R. roxburghii are still unknown. In this study, HPLC analysis revealed that total flavonoids, anthocyanins, and catechin were enriched in mature fruits, flowers, and leaves, respectively. Differentially expressed genes (DEGs) between five organs of R. roxburghii involved in flavonoid metabolism were obtained by transcriptome sequencing. A total of 1130 DEGs were identified, including 166 flavonoid pathway biosynthesis genes, 622 transcription factors (TFs), 301 transporters, and 221 cytochrome P450 proteins. A weighted gene co-expression network analysis (WGCNA) of the DEGs was conducted to construct co-expression networks. Regarding enzymes in the biosynthesis of flavonoids, cytochrome P450 CYP749A22 and CYP72A219 were highlighted in the regulation of total flavonoids of mature fruits. Anthocyanin 3-O-glucosyltransferase and F3&prime;H were the top two critical enzymes for anthocyanin accumulation in flowers. By contrast, caffeic acid 3-O-methyltransferase, 4-coumarate-CoA ligase, and shikimate O-hydroxycinnamoyltransferase were essential for catechin accumulation in leaves. Additionally, we analyzed the eigengene network of the &ldquo;black&rdquo; module, which had high correlations with total flavonoids (r = 0.9, p = 5 &times; 10&minus;6). There were 26 eigengenes in the &ldquo;black&rdquo; module, consisting of 6 flavonoid biosynthesis, 14 TFs, and 6 transporters. Among them, the transcription factors RrWRKY45 (DN142829_c1_g5), RrTCP20 (DN146443_c1_g1), and RrERF118 (DN141507_c3_g2) were screened as the hub genes, which significantly correlated with total flavonoids in R. roxburghii. The present biochemical and transcriptomic data provide insights into functional genomics for breeding R. roxburghii with flavonoid accumulation

A novel compound mutation in GLRA1 cause hyperekplexia in a Chinese boy- a case report and review of the literature

Abstract Background The pathogenesis of hereditary hyperekplexia is thought to involve abnormalities in the glycinergic neurotransmission system, the most of mutations reported in GLRA1. This gene encodes the glycine receptor α1 subunit, which has an extracellular domain (ECD) and a transmembrane domain (TMD) with 4 α-helices (TM1–TM4). Case presentation We investigated the genetic cause of hyperekplexia in a Chinese family with one affected member. Whole-exome sequencing of the 5 candidate genes was performed on the proband patient, and direct sequencing was performed to validate and confirm the detected mutation in other family members. We also review and analyse all reported GLRA1 mutations. The proband had a compound heterozygous GLRA1 mutation that comprised 2 novel GLRA1 missense mutations, C.569C > T (p.T190 M) from the mother and C.1270G > A (p.D424N) from the father. SIFT, Polyphen-2 and MutationTaster analysis identified the mutations as disease-causing, but the parents had no signs of hyperekplexia. The p.T190 M mutation is located in the ECD, while p.D424N is located in TM4. Conclusions Our findings contribute to a growing list GLRA1 mutations associated with hyperekplexia and provide new insights into correlations between phenotype and GLRA1 mutations. Some recessive mutations can induce hyperekplexia in combination with other recessive GLRA1 mutations. Mutations in the ECD, TM1, TM1-TM2 loop, TM3, TM3-TM4 loop and TM4 are more often recessive and part of a compound mutation, while those in TM2 and the TM2-TM3 loop are more likely to be dominant hereditary mutations

A first CLN6 variant case of late infantile neuronal ceroid lipofuscinosis caused by a homozygous mutation in a boy from China: a case report

Abstract Background Neuronal ceroid lipofuscinosis (NCLs) are lysosomal storage disorders characterized by seizures, motor impairment, and loss of vision. Ceroid lipofuscinosis (CLN) gene mutations are the cause, but NCL cases arising from CLN6 mutations have not been described in China to date. The CLN6 protein, which plays a role in lysosomal function, is an endoplasmic reticulum (ER) membrane protein with seven transmembrane (TM) domains. It has a cytosolic-facing amino terminal domain and a luminal-facing carboxyl terminal domain, with six loops between the TM domains. Case presentation Here we report a case involving a Chinese boy whose suspected diagnosis was a hereditary leukoencephalopathy, based on brain MRI imaging and epilepsy symptoms, language articulation disorders, ataxia, and unstable gait. The electroencephalogram showed epileptic discharges, and the brain MRI scan showed high signal intensity adjacent to the bilateral posterior horns of the lateral ventricles on T2-weighted images, along with cerebellar atrophy. Using next-generation sequencing for the genes in a panel for hereditary leukoencephalopathies, we detected a homozygous missense point mutation c.892G > A(p.Glu298Lys) in CLN6, and the variant was interpreted as pathogenic on in silico analysis. Absence of this mutation was confirmed in 259 controls. Late infantile NCL and secondary epilepsy were diagnosed, and oral sodium valproate was prescribed. The epilepsy was not well controlled, however, and the other signs had not improved at the 6-month follow-up. We also analyzed the loci of 31 CLN6 missense mutations, including those previously reported and the current one. We found that 22.6% (7/31) of the mutations are in the cytoplasmic domains, about 32.2% (10/31) are in the TM domains, and about 45.2% (14/31) are in the luminal domains. These mutations were mostly located in the TM3-TM4 loop (6/31), TM1-TM2 loop (4/31), and C-terminus (4/31), with none found in the TM4-TM5 loop, TM5-TM6 loop, or TM7. Conclusions We report the first case in China of NCL caused by a CLN6 mutation, expanding the genotype options for NCLs. In practice, NCLs generally are not the initial suspected diagnosis for such cases. Use of a gene sequencing panel for investigating unexplained seizures or leukoencephalopathies can help confirm the diagnosis

A Simultaneous Design and Optimization Framework for the Reaction and Distillation Sections of Methanol to Olefins Process

The reaction and separation sections are the keys to the methanol-to-olefins (MTO) chemical processes, and they should be optimized to reduce the cost of production. This work develops a framework for the simultaneous design and optimization of the reaction and distillation sections. An optimization model with shortcut and rigorous methods combined is established for distillation columns to improve accuracy and efficiency. With the auxiliary devices and the selection of utilities considered, the reaction and distillation sections are integrated to maximize profits. The genetic algorithm targets the optimal parameters, including the catalyst’s coke content and reaction temperature, each column’s operating pressure, and the allocation of utilities and auxiliary devices. For the studied MTO process, the optimal reaction temperature and catalyst’s coke content were identified to be 496 °C and 7.8%, respectively. The maximum profit is 15.3% greater than that identified with only the separation section optimized, and the minimum total annual cost (TAC) of the separation section is 3.73% less

Genome-wide identification and expression analyses of the pectate lyase (PL) gene family in Fragaria vesca

Abstract Background Pectate lyase (PL, EC 4.2.2.2), as an endo-acting depolymerizing enzyme, cleaves α-1,4-glycosidic linkages in esterified pectin and involves a broad range of cell wall modifications. However, the knowledge concerning the genome-wide analysis of the PL gene family in Fragaria vesca has not been thoroughly elucidated. Results In this study, sixteen PLs members in F. vesca were identified based on a genome-wide investigation. Substantial divergences existed among FvePLs in gene duplication, cis-acting elements, and tissue expression patterns. Four clusters were classified according to phylogenetic analysis. FvePL6, 8 and 13 in cluster II significantly contributed to the significant expansions during evolution by comparing orthologous PL genes from Malus domestica, Solanum lycopersicum, Arabidopsis thaliana, and Fragaria×ananassa. The cis-acting elements implicated in the abscisic acid signaling pathway were abundant in the regions of FvePLs promoters. The RNA-seq data and in situ hybridization revealed that FvePL1, 4, and 7 exhibited maximum expression in fruits at twenty days after pollination, whereas FvePL8 and FvePL13 were preferentially and prominently expressed in mature anthers and pollens. Additionally, the co-expression networks displayed that FvePLs had tight correlations with transcription factors and genes implicated in plant development, abiotic/biotic stresses, ions/Ca2+, and hormones, suggesting the potential roles of FvePLs during strawberry development. Besides, histological observations suggested that FvePL1, 4 and 7 enhanced cell division and expansion of the cortex, thus negatively influencing fruit firmness. Finally, FvePL1-RNAi reduced leaf size, altered petal architectures, disrupted normal pollen development, and rendered partial male sterility. Conclusion These results provide valuable information for characterizing the evolution, expansion, expression patterns and functional analysis, which help to understand the molecular mechanisms of the FvePLs in the development of strawberries

Preparation and Characterization of High Thermal Conductivity and Low CTE Polyimide Composite Reinforced with Diamond Nanoparticles/SiC Whiskers for 3D IC Interposer RDL Dielectric

Low thermal conductivity and large coefficient of thermal expansion (CTE) are the most serious disadvantages of the polymer dielectric for the interposer redistribution layer (RDL). In this paper, a high thermal conductivity and low CTE composite with polyimide (PI) matrix and diamond nanoparticles/SiC whiskers reinforcement is proposed. The preparation and characterization of the composite film are presented and the effects of the composite on the improvement of the interposer properties are investigated. With 10 wt% diamond-nanoparticles and 7 wt% SiC-whiskers, the composite film has a thermal conductivity of 1.63 W/m&#183;K and a CTE of 16.7 ppm/&#176;C (compared with 0.19 W/m&#183;K and 55.6 ppm/&#176;C of the PI). Interposers with PI RDL dielectric and the composite RDL dielectric are fabricated, respectively. The simulation result shows that the composite dielectric can significantly enhance the properties of the interposer compared with the PI dielectric. The thermal resistance of the interposer decreases from 8.04 &#176;C/W to 1.15 &#176;C/W. The maximum von Mises stress decreases from 72.8 MPa to 16.9 MPa and the warpage decreases from 1.13 &#956;m to 0.15 &#956;m. Thermal distribution tests are performed as well. The results show that the maximum temperature of the interposer decreases from 64 &#176;C to 45.1 &#176;C. The composite developed in this study can reduce the temperature and enhance the reliability of the chips with interposers. It has the potential to expand the application of the interposers in high thermal density integration and high reliability devices