Knockout of the delta11-desaturase SfruDES1 disrupts sex pheromone biosynthesis, mating and oviposition in the fall armyworm, Spodoptera frugiperda.

Moth insects rely on sex pheromones for long distance attraction and searching for sex partners. The biosynthesis of moth sex pheromones involves the catalytic action of multiple enzymes, with desaturases playing a crucial role in the process of carbon chain desaturation. However, the specific desaturases involved in sex pheromone biosynthesis in fall armyworm (FAW), Spodoptera frugiperda, have not been clarified. In this study, a Δ11 desaturase (SfruDES1) gene in FAW was knocked out using the CRISPR/Cas9 genome editing system. A homozygous mutant of SfruDES1 was obtained through genetic crosses. The gas chromatography-mass spectrometry (GC-MS) analysis results showed that the three main sex pheromone components (Z7-12:Ac, Z9-14:Ac, and Z11-16:Ac) and the three minor components (Z9-14:Ald, E11-14:Ac and Z11-14:Ac) of FAW were not detected in homozygous mutant females compared to the wild type. Furthermore, behavioral assay demonstrated that the loss of SfruDES1 resulted in a significant reduction in the attractiveness of females to males, along with disruptions in mating behavior and oviposition. Additionally, in a heterologous expression system, recombinant SfruDES1 could introduce a cis double bond at the Δ11 position in palmitic acid, which resulted in the changes in components of the synthesized products. These findings suggest desaturase plays a key role in the biosynthesis of sex pheromones, and knockout of the SfruDES1 disrupts sex pheromone biosynthesis and mating behavior in FAW. The SfruDES1 could serve as tool to develop a control method for S. frugiperda.

Renal Expression of CLC-5 and Megalin/Cubilin in Dent-1 Disease With Nonsense Mutations of CLCN5 Gene.

The study aims to explore the clinicopathological features and whether the nonsense mutations of CLCN5 gene have effect on the renal expression of CLC-5 protein and megalin/cubilin complex in children with Dent-1 disease. The clinicopathological features and genetic examination of three patients with Dent-1 disease were investigated. The expression of CLC-5 and megalin/cubilin complex in renal tissues was detected by using immunohistochemistry method. Urinary albumin, α1-microglobulin, ß2-microglobulin, retinol binding protein, and calcium levels were measured by immunonephelometry. Urinary calcium and low molecular weight proteinuria (LMWP) were enhanced in three patients, and two presented with nephrotic range proteinuria. Focal glomerular obsolescence, minor tubulointerstitial injury, and focal calcification in corticomedullary junction were found in one patient. Nonsense mutations of CLCN5 gene from their mothers were identified in all three patients with Dent-1 disease; however, the expression of CLC-5 protein was not decreased in renal tubular cells. As the receptor complex of albumin and LMWP reabsorption, the expression of megalin/cubilin in the brush border of proximal tubules was decreased in Dent-1 patients. Even if the renal CLC-5 protein is expressed normally, the reduced expression of megalin/cubilin in the brush border of renal proximal tubules may be helpful to understand the physiopathology of Dent-1 disease with nonsense mutations of CLCN5 gene.

CXCR5+ CD8+ T cells potently infiltrate pancreatic tumors and present high functionality.

Despite continued improvement in conventional therapy, pancreatic cancer continues to be one of the deadliest tumors worldwide with abysmal 5-year survival rate. New immunotherapeutic strategies that aim at improving antitumor cytotoxic CD8+ T cell responses are being developed in solid tumors. To assist the development of immunotherapies, we investigated the CD8+ T cells in pancreatic cancer patients. Compared to healthy individuals, pancreatic cancer patients presented a significant enrichment in the frequency of CD8+CXCR5+ T cells. In the tumor microenvironment, the frequencies of CD8+CXCR5+ T cells were further increased. In most cases, over half of tumor-infiltrating CD8+ T cells were CD8+CXCR5+ T cells. Compared to the circulating population, the tumor-infiltrating CD8+CXCR5+ T cells expressed higher levels of PD-1 and TIM-3. Functional analyses demonstrated that upon CD3/CD28 activation, the percentages of TNF-expressing and IFN-γ-expressing cells in CD8+CXCR5+ T cells were significantly higher than that in CD8+CXCR5- T cells. CD8+CXCR5+ T cells also presented enhanced cytotoxicity than CD8+CXCR5- T cells. Upon PD-1 and TIM-3 blockade, the functions of CD8+CXCR5+ T cells were further improved. The disease-free survival of pancreatic cancer patients following tumor resection was positively correlated with the frequencies of circulating and tumor-infiltrating CD8+CXCR5+ T cells. Together, our study identified that CD8+CXCR5+ T cells were a potent subset of CD8+ T cells that were highly enriched in pancreatic cancer patients and could respond to anti-PD-1/anti-TIM-3 blockade by further upregulation in function.

Access to the nucleus and functional association with c-Myc is required for the full oncogenic potential of ΔEGFR/EGFRvIII.

ΔEGFR is a potent glioblastoma oncogene which has been studied primarily as a plasma membrane kinase. Using intracranial xenograft studies in mice, we show that blocking ΔEGFR access to the nucleus attenuates its tumorigenicity and, conversely, that promoting nuclear accumulation enhances this, providing the first in vivo evidence that the nuclear actions of ΔEGFR contribute strongly to its oncogenic function. Nuclear actions of ΔEGFR include regulation of gene expression by participation in chromatin-bound complexes, and genome-wide mapping of these sequences by chromatin immunoprecipitation and massively parallel sequencing identified 2294 peaks. Bioinformatic analysis showed enrichment of the E-box motif in the dataset, and c-Myc and ΔEGFR were corecruited to the promoters of and transcriptionally activated a subset of nuclear ΔEGFR chromatin targets. Knockdown of c-Myc decreased the expression of these targets and diminished ΔEGFR-stimulated anchorage-independent colony formation. We conclude that transcriptional regulation of target genes by association with gene regulatory chromatin in cooperation with c-Myc by nuclear ΔEGFR makes a unique contribution to its oncogenicity and propose that this venue provides new targets for therapeutic intervention.

Removal performance and dissolved organic matter biodegradation characteristics in advection ecological permeable dam reactor.

In this present study, an advection ecological permeable dam (AEPD) based on a biofilm reactor was established to investigate pollution control performance and dissolved organic matter (DOM) bio-degradation. The AEPD achieved optimal efficiency-chemical oxygen demand, 6-53 mg/L; total nitrogen concentration, 1.47-6.89 mg/L; total phosphorus concentration, 0.53-3.93 mg/L, and increases in values for ultraviolet-visible parameters-SUVA254, from 0.392 to 0.673-1.438; E4/E6, from 1.09 to 1.11-1.26; A240-400, from 12.06 to 13.09-19.95; and A253-203, from 0.03 to 0.04-0.23. This showed that DOM degradation promoted its humification, aromatisation, and unsaturation as well as increased the number of polar functional groups in the organic aromatic rings of DOM. Synchronous fluorescence and parallel factor analyses indicated that AEPD could effectively degrade tyrosine-like and tryptophan-like compounds, which showed the most significant decrease in fluorescence intensity. Additionally, AEPD displayed some stable dominant bacterial genera (e.g. Proteobacteria_unclassified, Bacteroidetes_unclassified, Gemmobacter, Pseudofulvimonas, Flavobacterium, Pseudomonas, and Nitrospira), although their relative abundance differed under variable hydraulic loading rates. This research provided further technical support for the application of AEPD in the treatment of water environment pollution.

Dissecting beta-ring assembly pathway of the mammalian 20S proteasome.

The 20S proteasome is the catalytic core of the 26S proteasome. It comprises four stacked rings of seven subunits each, alpha(1-7)beta(1-7)beta(1-7)alpha(1-7). Recent studies indicated that proteasome-specific chaperones and beta-subunit appendages assist in the formation of alpha-rings and dimerization of half-proteasomes, but the process involved in the assembly of beta-rings is poorly understood. Here, we clarify the mechanism of beta-ring formation on alpha-rings by characterizing assembly intermediates accumulated in cells depleted of each beta-subunit. Starting from beta2, incorporation of beta-subunits occurs in an orderly manner dependent on the propeptides of beta2 and beta5, and the C-terminal tail of beta2. Unexpectedly, hUmp1, a chaperone functioning at the final assembly step, is incorporated as early as beta2 and is required for the structural integrity of early assembly intermediates. We propose a model in which beta-ring formation is assisted by both intramolecular and extrinsic chaperones, whose roles are partially different between yeast and mammals.

Analysis for global characteristics of Lyapunov exponents in vehicle plane motion system.

In the field of vehicle system dynamics, it is significant to propose an appropriate quantitative indicator for system stability or dynamics characteristics. Lyapunov exponents method is an excellent quantitative indicator for analysing nonlinear system characteristics. It was used to studied the stable region of nonlinear vehicle plane motion system. However, the effect of Lyapunov exponents method in revealing the global dynamics characteristics has not been fully studied. Aiming at this key problem, this paper analyses the global characteristics of Lyapunov exponents with different degrees of freedom nonlinear models. The results show that Lyapunov exponents can well reflect the global layer characteristics for vehicle system. However, the value characteristic under different DOF models is not unified. The research and analysis in this paper supplement the quantitative analysis theory for vehicle system dynamics.

Kinetic study on degradation of micro-organics by different UV-based advanced oxidation processes in EfOM matrix.

Effluent organic matter (EfOM) contains a large number of substances that are harmful to both the environment and human health. To avoid the negative effects of organic matter in EfOM, advanced treatment of organic matter is an urgent task. Four typical oxidants (H2O2, PS, PMS, NaClO) and UV-combined treatments were used to treat micro-contaminants in the presence or absence of EfOM, because the active radical species produced in these UV-AOPs are highly reactive with organic contaminants. However, the removal efficiency of trace contaminants was greatly affected by the presence of EfOM. The degradation kinetics of two representative micro-contaminants (benzoic acid (BA) and para chlorobenzoic acid (pCBA)) was significantly reduced in the presence of EfOM, compared to the degradation kinetics in its absence. Using the method of competitive kinetics, with BA, pCBA, and 1,4-dimethoxybenzene (DMOB) as probes, the radicals (HO·, SO4-·, ClO·) proved to be the key to reaction species in advanced oxidation processes. UV irradiation on EfOM was not primarily responsible for the degradation of micro-contaminants. The second-order rate constants of the EfOM with radicals were determined to be (5.027 ± 0.643) × 102 (SO4-·), (3.192 ± 0.153) × 104 (HO·), and 1.35 × 106 (ClO·) (mg C/L)-1 s-1. In addition, this study evaluated the production of three radicals based on the concept of Rct, which can better analyze its reaction mechanism.

MiRNA-21 Regulates Bronchial Epithelial Cell Proliferation by Activating Tgfß1/Smad Signaling Pathway and Its Correlation with Asthma Severity in Children.

BACKGROUND: This research was designed to probe into the role of miRNA-21 in the pathogenesis of childhood asthma and its correlation with the severity. METHODS: Fifty-four children with bronchial asthma admitted to the Third Affiliated Hospital of Qiqihar Medical University from Jun 2018 to Dec 2019 were included. Forty nine healthy children underwent physical examination at this time period were also enrolled. The miR-21 expression in peripheral blood serum was analyzed by qRT-PCR. The relationship between the expression and severity of asthma in children was explored by Spearman correlation analysis and ROC curve. Bronchial epithelial cell lines were cultured in vitro and divided into blank control group, negative control group and miR-21 inhibition and activation group. The changes of cell proliferation after treatment were detected by CCK-8 test in different groups. The expression of TGF-ß1/Smad signaling pathway protein in cells was assessed by Western blot (WB). RESULTS: Compared with that of healthy children, the miR-21 expression in peripheral blood serum of asthmatic children was higher (P<0.001). MiR-21 expression was positively correlated with the severity of illness (r=0.853, P<0.001). The results of cell experiments in vitro signified that miR-21 can promote the proliferation of bronchial epithelial cells, and may be involved in regulating the expression of TGF-ß1/Smad3 signaling pathway, thus affecting cell proliferation. CONCLUSION: miRNA-21 regulates the proliferation of bronchial epithelial cells by activating TGFß1/Smad signaling pathway. And it is positively correlated with the severity of asthma in children.

Highly selective suppression of melanoma cells by inducible DNA cross-linking agents: bis(catechol) derivatives.

A series of bis(catechol) quaternary ammonium derivatives were designed and synthesized. We investigated their ability to cross-link DNA induced by tyrosinase and found that the o-quinone is key intermediate in the process by using the nucleophile 3-methyl-2-benzothiazolinone hydrazone (MBTH) in the tyrosinase assay. Their cytotoxicities to B16F1, Hela, and CHO cells were tested by MTT assays. The specific and potent abilities to kill the tyrosinase-efficient melanoma cells kindled our interest in exploring the relationship between their abilities of cross-linking DNA and their selective cytotoxicities to cells. Through an integrated approach including intracellular imaging for detection of the dihydroxyphenyl groups, alkaline comet assays, and γ-H2AX immunofluorescence assays, the speculation was confirmed. The bis(catechol) quaternary ammonium derivatives showed notable cell selectivity because they displayed cytotoxicities after being oxidized by tyrosinase, and they were able to target the DNA efficiently in the tyrosinase-efficient melanoma cells, forming both alkylated and cross-linked species.

HBcAg induces interleukin-10 production, inhibiting HBcAg-specific Th17 responses in chronic hepatitis B patients.

T-helper (Th) 17 cells have been shown to have an important role in host defense against viral infection. However, little is known about the regulation of Th17 cells in hepatitis B virus (HBV) infections. Peripheral blood mononuclear cells (PBMCs) isolated from patients with chronic hepatitis B (CHB) were stimulated with anti-interleukin (IL)-10 antibody or recombinant IL-10. The frequency of hepatitis B core antigen (HBcAg)-specific Th17 cells was characterized and produced cytokines were determined by flow cytometry. A low frequency of Th17 cells and a high frequency of Th1 cells were detected in CHB patients. HBcAg stimulation promoted IL-17A, IL-22, IL-23, IL-6, transforming growth factor (TGF)-ß and IL-10 production by PBMCs from CHB patients, but not from healthy controls. Furthermore, endogenous IL-10 inhibited HBcAg-stimulated production of IL-17A, IL-22, IL-6 and IL-23, but not TGF-ß. Treatment with IL-10 inhibited the HBcAg-stimulated activation of Th17 cells, whereas anti-IL-10 antibody significantly increased the frequency of Th17 and Th1 cells, but not that of CD4(+)CD25(+) regulatory T cells, associated with upregulating RORγt expression in CD4(+) T cells. HBcAg stimulated the production of IL-10, which negatively regulated HBcAg-specific Th17 cell responses in CHB patients. Our findings may represent one evasion strategy for HBV to subvert specific antiviral responses in humans.

Nonalcoholic Fatty Liver Disease: Pathogenesis and Treatment in Traditional Chinese Medicine and Western Medicine.

Nonalcoholic Fatty Liver Disease (NAFLD) is one of the most important causes of liver disease worldwide and probably destined to become the leading cause of end-stage liver disease in the coming decades, affecting both adults and children. Faced with the severe challenges for the prevention and control of NAFLD, this article discusses the understanding and mechanism of NAFLD from Chinese and Western medicine. Moreover, the progress regarding its treatment in both Chinese and Western medicine is also summarized. Both Chinese medicine and Western medicine have their own characteristics and clinical efficacy advantages in treating diseases. The purpose of this article is to hope that Chinese and Western medicine have complementary advantages, complementing each other to improve clinical NAFLD therapy prevention and treatment methods to receive more and more attention throughout the global medical community.

In-depth Analysis of the Lid Subunits Assembly Mechanism in Mammals.

The 26S proteasome is a key player in the degradation of ubiquitinated proteins, comprising a 20S core particle (CP) and a 19S regulatory particle (RP). The RP is further divided into base and lid subcomplexes, which are assembled independently from each other. We have previously demonstrated the assembly pathway of the CP and the base by observing assembly intermediates resulting from knockdowns of each proteasome subunit and the assembly chaperones. In this study, we examine the assembly pathway of the mammalian lid, which remains to be elucidated. We show that the lid assembly pathway is conserved between humans and yeast. The final step is the incorporation of Rpn12 into the assembly intermediate consisting of two modular complexes, Rpn3-7-15 and Rpn5-6-8-9-11, in both humans and yeast. Furthermore, we dissect the assembly pathways of the two modular complexes by the knockdown of each lid subunit.

Synthesis and evaluation of cationic phthalocyanine derivatives as potential inhibitors of telomerase.

A series of water-soluble cationic phthalocyanine derivatives (1-10) were designed and synthesized to develop novel and potent telomerase inhibitors. These phthalocyanine derivatives as inhibitors of telomerase were investigated via modified telomerase repeat amplification protocol (TRAP) assay. The TRAP assay indicates that these cationic compounds had strong telomerase inhibitory activity (IC(50)<1.65 microM). To determine whether the phthalocyanine derivatives binding to G-quadruplex enhance the block to DNA synthesis, primer extension reactions were carried out in the presence of phthalocyanines. The interaction of the G-quadruplex of telomerase DNA with these molecules was examined by CD melting and PCR stop assay. These cationic phthalocyanine derivatives can stabilize G-quadruplex, which is demonstrated by the increased T(m) values. All these results indicate that the phthalocyanine derivatives might be potential lead compounds for the development of new telomerase inhibitor.

[Clinical research of plasma exchange with continuous veno-venous hemofiltration in treating mid- and late-stage chronic severe viral hepatitis B patients].

OBJECTIVE: To evaluate the clinical efficacy and study the mechanism of combining plasma exchange and continuous veno-venous hemofiltration in treating patients with chronic severe viral hepatitis B in their mid- and late stages. METHODS: 94 patients suffering from chronic severe viral hepatitis B were divided into three groups. 29 patients were treated with plasma exchange plus continuous veno-venous hemofiltration (group A). 31 patients were treated with plasma exchange alone (group B). 34 patients received routine treatment (group C). The efficacy of treatment and survival rate of the three groups was investigated. Before and after artificial liver support system treatment the levels of cytokine were examined. RESULTS: In group A, hyponatremia improved, the levels of interleukin 8 (IL-8) obviously decreased, the level of IL-10 increased, 5 of the 10 patients in coma regained normal consciousness (50.0%) and their survival rate was 48.3%. In group B, hyponatremia did not change, the level of IL-8 and IL-10 did not change. 2 of 11 patients in coma regained normal consciousness (18.2%) while survival rate was 22.6%. In group C, 1 of 11 patients in coma regained normal consciousness (9.1%) while survival rate was 20.6%. CONCLUSIONS: It shows that plasma exchange with continuous veno-venous hemofiltration in treating patients with mid- and late stage chronic severe viral hepatitis B can increase the survival rate. IL-8 can be significantly removed, IL-10 significantly increased. This combined therapy is easy to practice, and should be used as an artificial liver support system.

Assembly mechanisms of specialized core particles of the proteasome.

The 26S proteasome has a highly complicated structure comprising the 20S core particle (CP) and the 19S regulatory particle (RP). Along with the standard CP in all eukaryotes, vertebrates have two more subtypes of CP called the immunoproteasome and the thymoproteasome. The immunoproteasome has catalytic subunits ß1i, ß2i, and ß5i replacing ß1, ß2, and ß5 and enhances production of major histocompatibility complex I ligands. The thymoproteasome contains thymus-specific subunit ß5t in place of ß5 or ß5i and plays a pivotal role in positive selection of CD8+ T cells. Here we investigate the assembly pathways of the specialized CPs and show that ß1i and ß2i are incorporated ahead of all the other ß-subunits and that both ß5i and ß5t can be incorporated immediately after the assembly of ß3 in the absence of ß4, distinct from the assembly of the standard CP in which ß-subunits are incorporated in the order of ß2, ß3, ß4, ß5, ß6, ß1, and ß7. The propeptide of ß5t is a key factor for this earlier incorporation, whereas the body sequence seems to be important for the earlier incorporation of ß5i. This unique feature of ß5t and ß5i may account for preferential assembly of the immunoproteasome and the thymoproteasome over the standard type even when both the standard and specialized subunits are co-expressed.

Synthesis and biological activities of quinazoline derivatives with ortho-phenol-quaternary ammonium salt groups.

One phenol-quaternary ammonium salt derivative with a flexible linker and three derivatives with a quinazoline moiety are present. Their binding affinities for DNA are discussed and it is clearly demonstrated that this class of phenol-quaternary ammonium salt derivatives could inhibit DNA transcription effectively.