Microinvasion in hepatocellular carcinoma: predictive factor and application for definition of clinical target volume for radiotherapy.

BACKGROUND: To investigate the correlation between microinvasion and various features of hepatocellular carcinoma (HCC), and to clarify the microinvasion distance from visible HCC lesions to subclinical lesions, so as to provide clinical basis for the expandable boundary of clinical target volume (CTV) from gross tumor volume (GTV) in the radiotherapy of HCC. METHODS: HCC patients underwent hepatectomy of liver cancer in our hospital between July 2019 and November 2021 were enrolled. Data on various features and tumor microinvasion distance were collected. The distribution characteristics of microinvasion distance were analyzed to investigate its potential correlation with various features. Tumor size compared between radiographic and pathologic samples was analyzed to clarify the application of pathologic microinvasion to identify subclinical lesions of radiographic imaging. RESULTS: The average microinvasion distance was 0.6 mm, with 95% patients exhibiting microinvasion distance less than 3.0 mm, and the maximum microinvasion distance was 4.0 mm. A significant correlation was found between microinvasion and liver cirrhosis (P = 0.036), serum albumin level (P = 0.049). Multivariate logistic regression analysis revealed that HCC patients with cirrhosis had a significantly lower risk of microinvasion (OR = 0.09, 95%CI = 0.02 ~ 0.50, P = 0.006). Tumor size was overestimated by 1.6 mm (95%CI=-12.8 ~ 16.0 mm) on radiographic size compared to pathologic size, with a mean %Δsize of 2.96% (95%CI=-0.57%~6.50%). The %Δsize ranged from - 29.03% to 34.78%. CONCLUSIONS: CTV expanding by 5.4 mm from radiographic GTV could include all pathologic microinvasive lesions in the radiotherapy of HCC. Liver cirrhosis was correlated with microinvasion and were independent predictive factor of microinvasion in HCC.

Construction of a high-resolution genetic map and identification of single nucleotide polymorphism markers relevant to flower stalk height in onion.

Introduction: Onion (Allium cepa L., 2n=16) is an economically and nutritionally important vegetable crop worldwide. Construction of a high-resolution genetic map and map-based gene mining in onion have lagged behind other vegetable crops such as tomato and pepper. Methods: In this study, we constructed a high-resolution genetic map of onion using 321 F2 individuals from a cross between two double haploid lines DH-1×DH-17 and employing specific length amplified fragment (SLAF)-seq technology. The genetic map containing 10,584 polymorphic SLAFs with 21,250 single nucleotide polymorphism (SNP) markers and 8 linkage groups was developed for onion, which spanned 928.32 cM, with an average distance of 0.09 cM between adjacent markers. Results: Using this map, we carried out QTL mapping of Ms locus related to the male-fertile trait and reproduced previous mapping results, which proved that this map was of good quality. Then, four QTLs (located on LG2, LG5, and LG8) were detected for flower stalk height, explaining 26.60% of the phenotypic variance. Among them, we proposed that 20 SLAF markers (in three QTLs) of flower stalk height trait were effective favorable allelic variant markers associated with heterosis. Discussion: Overall, the genetic map was structured using SLAF-seq based on DH lines, and it is the highest-quality and highest-resolution linkage map of onion to date. It lays a foundation for the fine mapping and candidate gene identification of flower stalk height, and provides new insights into the developmental genetic mechanisms in onion breeding.

Facile synthesis of phycocyanin/polydopamine hierarchical nanocomposites for synergizing PTT/PDT against cancer.

The exceptional biocompatibility and biosafety of natural proteins have made them a popular choice for tumor therapy in recent years, but their therapeutic effectiveness is severely constrained by factors including physiological instability, insufficient delivery, limited accumulation in tumor cells, etc. Here, a novel Mn-doped phycocyanin (Pc)/polydopamine (PDA) hierarchical nanostructure (MnPc@P) with excellent optical absorption, photothermal conversion, and photodynamic performances, is first designed and fabricated by a simply one-pot reaction, which not only successfully encapsulates natural protein Pc with intact activity in the nanostructure of MnPc@P but also gives them better biocompatibility. Upon laser irradiation, PDA-mediated hyperthermia and Pc-induced ROS elevation in tumor cells have been demonstrated, leading to drastic tumor cell death via combined PTT/PDT effect, greater than single PTT or PDT. In general, the expert fusion of Pc and PDA into a single nanomedicine opens fascinating perspectives in the delivery of natural proteins and tumor therapy.

A weakened recurrent circuit in the hippocampus of Rett syndrome mice disrupts long-term memory representations.

Successful recall of a contextual memory requires reactivating ensembles of hippocampal cells that were allocated during memory formation. Altering the ratio of excitation-to-inhibition (E/I) during memory retrieval can bias cell participation in an ensemble and hinder memory recall. In the case of Rett syndrome (RTT), a neurological disorder with severe learning and memory deficits, the E/I balance is altered, but the source of this imbalance is unknown. Using in vivo imaging during an associative memory task, we show that during long-term memory retrieval, RTT CA1 cells poorly distinguish mnemonic context and form larger ensembles than wild-type mouse cells. Simultaneous multiple whole-cell recordings revealed that mutant somatostatin expressing interneurons (SOM) are poorly recruited by CA1 pyramidal cells and are less active during long-term memory retrieval in vivo. Chemogenetic manipulation revealed that reduced SOM activity underlies poor long-term memory recall. Our findings reveal a disrupted recurrent CA1 circuit contributing to RTT memory impairment.

Ruthenium-catalyzed C-H amination of aroylsilanes.

Acylsilane represents a valuable synthon in synthetic chemistry. We report on ruthenium(ii)-catalyzed ortho-C-H amination of aroylsilanes to provide facile access to synthetically useful imidobenzoylsilanes and tosyl-amidobenzoylsilanes. The protocols, with broad substrate scope and excellent functional group tolerance, are enabled with the weak chelation-assistance of acylsilane via C-H cyclometallation.

MeCP2 Levels Regulate the 3D Structure of Heterochromatic Foci in Mouse Neurons.

Methyl-CpG binding protein 2 (MeCP2) is a nuclear protein critical for normal brain function, and both depletion and overexpression of MeCP2 lead to severe neurodevelopmental disease, Rett syndrome (RTT) and MECP2 multiplication disorder, respectively. However, the molecular mechanism by which abnormal MeCP2 dosage causes neuronal dysfunction remains unclear. As MeCP2 expression is nearly equivalent to that of core histones and because it binds DNA throughout the genome, one possible function of MeCP2 is to regulate the 3D structure of chromatin. Here, to examine whether and how MeCP2 levels impact chromatin structure, we used high-resolution confocal and electron microscopy and examined heterochromatic foci of neurons in mice. Using models of RTT and MECP2 triplication syndrome, we found that the heterochromatin structure was significantly affected by the alteration in MeCP2 levels. Analysis of mice expressing either MeCP2-R270X or MeCP2-G273X, which have nonsense mutations in the upstream and downstream regions of the AT-hook 2 domain, respectively, showed that the magnitude of heterochromatin changes was tightly correlated with the phenotypic severity. Postnatal alteration in MeCP2 levels also induced significant changes in the heterochromatin structure, which underscored importance of correct MeCP2 dosage in mature neurons. Finally, functional analysis of MeCP2-overexpressing mice showed that the behavioral and transcriptomic alterations in these mice correlated significantly with the MeCP2 levels and occurred in parallel with the heterochromatin changes. Taken together, our findings demonstrate the essential role of MeCP2 in regulating the 3D structure of neuronal chromatin, which may serve as a potential mechanism that drives pathogenesis of MeCP2-related disorders.SIGNIFICANCE STATEMENT Neuronal function is critically dependent on methyl-CpG binding protein 2 (MeCP2), a nuclear protein abundantly expressed in neurons. The importance of MeCP2 is underscored by the severe childhood neurologic disorders, Rett syndrome (RTT) and MECP2 multiplication disorders, which are caused by depletion and overabundance of MeCP2, respectively. To clarify the molecular function of MeCP2 and to understand the pathogenesis of MECP2-related disorders, we performed detailed structural analyses of neuronal nuclei by using mouse models and high-resolution microscopy. We show that the level of MeCP2 critically regulates 3D structure of heterochromatic foci, and this is mediated in part by the AT-hook 2 domain of MeCP2. Our results demonstrate that one primary function of MeCP2 is to regulate chromatin structure.

Expression profile and bioinformatics analysis of COMMD10 in BALB/C mice and human.

COMMD10, a member of COMMD protein, has been proved to target p65 NF-kappaB (nuclear factor-kappaB) subunit and reduce its nuclear translocation, thereby leading to the inactivation of NF-kappaB pathway and suppression of colorectal cancer invasion and metastasis. The aim of this study is to explore its expression pattern and tissue distribution in human normal tissues and other tumor tissues and to investigate the relevant mechanism. We firstly provided the expression profile and histological distribution of COMMD10 in various BALB/c mice tissues and identified the biological distribution of COMMD10 in different kinds of human normal and tumor tissues. We verified the expression profile of COMMD10 using TCGA database. The interacting genes of COMMD10 were predicted by using STRING using. Finally, we performed database, and the microRNAs targeting COMMD10 were predicted using miRDB, miRWalk, TargetScan and microRNA. GO and KEGG pathway analyses were performed to predict the biological function of COMMD10 and its interacting genes. mRNA expression of COMMD10 showed the highest level in the lung and spleen, and the lowest level in the heart and brain. Immunohistochemistry detection revealed that COMMD10 was expressed in different tissues with different degrees and was was located mainly in the cytoplasm. Subsequently, we showed that COMMD10 displayed various degrees of expression in different human normal tissues that mainly located in cytoplasm, while COMMD10 of liver cells resided in both nucleus and cytoplasm. All the tumor tissues except breast small cell carcinoma, breast phyllodes tumor, lung adenocarcinoma, thymoma, cervical cancer and bladder urothelial carcinoma showed that COMMD10 was positive staining in cytoplasm. Kaplan-Meier plotter indicated that renal clear cell carcinoma patients with increased expression level of COMMD10 exhibited longer survival. STRING database revealed that COMMD10 had 41 interacting genes, and data from 4 different databases indicated that hsa-miR-590-3p may be the potential regulator of COMMD10. GO analysis demonstrated that COMMD10 and its interacting genes were mainly enriched in Cullin-RING ubiquitin ligase complexes, binding and transport of copper ions, the transport and steady-state maintenance of copper ions, transcription, translation and transport of proteins, and negatively regulate the activity of NF-kappaB transcription factors. KEGG pathway showed that COMMD10 and its interacting genes were mainly involved in renal cell carcinoma, HIF-1 signaling pathways, ubiquitination-mediated proteolysis, endocytosis and mineral absorption. COMMD10 may play a tumor suppressive role in renal clear cell carcinoma through the miR-590-3p-COMMD10-Cul2-RBX1-NF-κB/HIF/NRF2 pathway and regulate the chemotherapy resistance of various tumor cells to cisplatin.

Bidentate auxiliary-directed alkenyl C-H allylation via exo-palladacycles: synthesis of branched 1,4-dienes.

An alkenyl C-H allylation by an exo-palladacycle intermediate is demonstrated, employing unactivated (Z)-alkenes and allyl carbonates. With the use of an 8-aminoquinoline (AQ) derived amide as the directing group, the N,N-bidentate-chelation-assisted C-H activation protocol proceeded under mild and oxidant-free conditions with excellent selectivity. The utility of this approach is demonstrated by the preparative scale, selective conversion of inseparable Z/E alkenes and ready removal of the amide auxiliary to provide the corresponding ester.

Iridium-Catalyzed Cross-Coupling Reactions of Alkenes by Hydrogen Transfer.

A range of Ru-, Rh-, or Pd-catalyzed vinylic C-H/C-H cross-coupling reactions of olefins have been demonstrated to provide 1,3-dienes, using a quantitative amount of metal oxidants. Although transfer hydrogenation and C-H alkenylation are two important areas that evolved independently, we herein report the first iridium-catalyzed cross-coupling reactions of alkenes by integration of directed C(alkenyl)-H alkenylation and transfer hydrogenation to obviate the usage of a metal oxidant, employing a hydrogen acceptor such as inexpensive chloranil.

Prognosis and modulation mechanisms of COMMD6 in human tumours based on expression profiling and comprehensive bioinformatics analysis.

BACKGROUND: The Copper Metabolism MURR1 (COMM) domain family has been reported to play important roles in tumorigenesis. As a prototype for the COMMD family, the expression pattern and biological function of COMMD6 in human tumours remain unknown. METHODS: COMMD6 expression in BALB/c mice and human tissues was examined using real-time PCR and immunohistochemistry. Kaplan-Meier analysis was applied to evaluate the prognosis of COMMD6 in tumours. Competing endogenous RNA (ceRNA) and transcriptional regulation network were constructed based on differentially expressed mRNAs, microRNAs and long non-coding RNAs from the cancer genome atlas database. GO and KEGG enrichment analysis were used to explore the bioinformatics implication. RESULTS: COMMD6 expression was widely observed in BALB/c mice and human tissues, which predicted prognosis of cancer patients. Furthermore, we shed light on the underlying tumour promoting role and mechanism of COMMD6 by constructing a TEX41-miR-340-COMMD6 ceRNA network in head and neck squamous cell carcinoma and miR-218-CDX1-COMMD6 transcriptional network in cholangiocarcinoma. In addition, COMMD6 may modulate the ubiquitination and degradation of NF-κB subunits and regulate ribonucleoprotein and spliceosome complex biogenesis in tumours. CONCLUSIONS: This study may help to elucidate the functions and mechanisms of COMMD6 in human tumours, providing a potential biomarker for tumour prevention and therapy.

Cobalt-Catalyzed Olefinic C-H Alkenylation/Alkylation Switched by Carbonyl Groups.

The first cobalt-catalyzed cross-couplings between olefins has been demonstrated to provide C(alkenyl)-H alkenylation and alkylation products, using complex [Cp*Co(CO)I2]. While coupling partner acrylates afforded conjugated dienoates, α,ß-unsaturated ketones led to γ-alkenyl ketones completely, representing a switchable C-H functionalization controlled by different carbonyl groups.

Iridium-catalyzed alkenyl C-H allylation using conjugated dienes.

An iridium-catalyzed C-H allylation of acrylamides with conjugated dienes was developed, using NH-Ts amide as the directing group. The ligand- and additive-free protocol provided a convenient and atom economic synthesis of branched 1,4-diene skeletons, enabling the tolerance of a wide scope of functionalities such as OMe, F, Cl, Br and CF3. The utility of this protocol is also demonstrated by a preparative scale, as well as C-H functionalization of artemisic amide. Furthermore, NH-Ts amide was efficiently removed by methylation and hydrolysis procedures to provide 1,4-dienoic acid.

Additive- and Ligand-Free Cross-Coupling Reactions between Alkenes and Alkynes by Iridium Catalysis.

Although a range of transition-metal-catalyzed cross-coupling reactions of alkenes and alkynes have been developed to produce valuable conjugated dienes, extension of these reactions to iridium catalysis has yet to be demonstrated. The first iridium-catalyzed alkene-alkyne cross-coupling reactions have been realized under ligand- and additive-free conditions. A wide range of acrylamides and alkynes could be used as coupling partners, providing branched ( Z, Z)-butadiene skeletons with excellent site- and stereoselectivities. The utility of this approach is also demonstrated by preparative-scale and C-H functionalization of perillic and artemisic amides.

Acylsilane directed aromatic C-H alkenylations by ruthenium catalysis.

A ruthenium-catalyzed C-H alkenylation of aroylsilanes with electron-deficient alkenes was developed, using acylsilane as the directing group. The mild reaction conditions enable the tolerance of a wide scope of functionalities such as OMe, F, Cl, Br and CF3, providing a convenient and highly effective method for the synthesis of styrene derivatives bearing acylsilane. Steroid and heterocycles such as furan and thiophene were also well tolerated. Furthermore, acylsilane was efficiently converted to the corresponding aldehyde or carboxylic acid.

Conditioned medium from overly excitatory primary astrocytes induced by La3+ increases apoptosis in primary neurons via upregulating the expression of NMDA receptors.

Lanthanum (La) can accumulate in the brain and impair learning and memory. However, the underlying mechanism of La-induced neurotoxicity has remained elusive. Under physiological conditions, it has been reported that moderately excitatory astrocytes play an important role in the regulation of neuronal signals and synaptic plasticity. However, under pathological conditions, overly excitatory astrocytes can release excess excitatory transmitters, such as glutamate (Glu) and d-serine, and induce the over-activation of NMDA receptors (NMDAR) in neurons, ultimately leading to neuronal excitotoxicity. To date, limited work has been performed with respect to whether La can induce neuronal excitotoxicity by inducing astrocytes to become overexcited. In this study, in vitro models of primary culture rat cortical astrocytes and neurons were established. First, the astrocytes were treated with 0.125 mM, 0.25 mM and 0.5 mM lanthanum chloride (LaCl3) for 24 h, and the supernatants were collected as a conditioned medium (CM) which is denoted as CM (La3+); then, the neurons were treated with CM (La3+) for 48 h. The results illustrate that LaCl3 treatment significantly upregulated the mRNA and protein expression levels of metabotropic glutamate receptor 5 (mGLUR5), phospholipase C (PLC), connexin 43 (Cx43) and Cx30, increased the concentrations of inositol trisphosphate (IP3) and [Ca2+]i, and promoted the synthesis and release of Glu and d-serine in astrocytes. Moreover, the CM (La3+) could increase the mRNA and protein expression levels of NMDAR subunits (NR1, NR2A, NR2B), the concentration of [Ca2+]i and the rate of apoptosis in neurons. Furthermore, after removal of La, CM (La-free) had a similar effect on neurons which could be antagonized by MK-801, DCKA and DAAO. These results suggest that the neuron apoptosis induced by La is closely related to the excessive release of Glu and d-serine from overly excitatory astrocytes.

Microstructure, Tensile, and Creep Behaviors of Ti-22Al-25Nb (at.%) Orthorhombic Alloy with Equiaxed Microstructure.

This article investigates the tensile and creep behaviors of the Ti-22Al-25Nb (at.%) alloy with equiaxed microstructure. The experimental results show that the equiaxed microstructures are formed by isothermal forging in the α2 + B2 + O phase region, and then heat treating in α2 + B2 + O and B2 + O phase regions. The equiaxed particles are determined by isothermal forging and solution heat treating, and the acicular O phase is obtained by adjusting the aging temperature. The strengths of the alloy are sensitive to the thickness of the secondary acicular O phase. Increase in aging temperature improves strength and reduces the ductility. Deformation of the alloy mainly depends on the volume fraction and deformability of the B2 phase. During the high-temperature tensile deformation, the flow stress decreases with the increasing deformation temperature and increases with the increasing strain rate. The microstructure obtained by higher aging temperature (HT-840) has better creep resistance, due to the coarsening of the secondary acicular O phase.

Genome-wide distribution of linker histone H1.0 is independent of MeCP2.

Previous studies suggested that MeCP2 competes with linker histone H1, but this hypothesis has never been tested in vivo. Here, we performed chromatin immunoprecipitation followed by sequencing (ChIP-seq) of Flag-tagged-H1.0 in mouse forebrain excitatory neurons. Unexpectedly, Flag-H1.0 and MeCP2 occupied similar genomic regions and the Flag-H1.0 binding was not changed upon MeCP2 depletion. Furthermore, mild overexpression of H1.0 did not alter MeCP2 binding, suggesting that the functional binding of MeCP2 and H1.0 are largely independent.

Environmentally relevant concentrations of mercury exposure alter thyroid hormone levels and gene expression in the hypothalamic-pituitary-thyroid axis of zebrafish larvae.

Mercury (Hg) is one of the most toxic heavy metals that can cause severe damage to fish. Studies have demonstrated that Hg has a specific affinity for the endocrine system, but little is known about the effects of Hg on thyroid endocrine system in fish. In this study, zebrafish embryos were exposed to environmentally relevant concentrations of 1, 4, and 16 µg/L Hg2+ (added as HgCl2) from 2 h post-fertilization (hpf) to 168 hpf. Thyroid hormone (TH) levels and mRNA expression levels of genes involved in the hypothalamus-pituitary-thyroid (HPT) axis were determined. The results showed that exposure to 16 µg/L Hg2+ increased the whole-body thyroxine (T4) and triiodothyronine (T3) levels. The transcription levels of corticotrophin releasing hormone (crh) and thyroid stimulating hormone (tshß) were up-regulated by Hg2+ exposure. Analysis of the mRNA levels of genes related to thyroid development (hhex, nkx2.1, and pax8) and THs synthesis (nis and tg) revealed that exposure to higher Hg2+ concentrations markedly up-regulated hhex, nkx2.1, nis, and tg expression, while had no significant effect on the transcripts of pax8. For the transcription of two types of deiodinases (deio1 and deio2), deio1 showed no significant changes in all the treatments, whereas deio2 was significantly up-regulated in the 16 µg/L Hg2+ group. In addition, Hg2+ exposure up-regulated thyroid hormone receptor ß (trß) mRNA level, while the transcription of trα was not changed. Overall, our study indicated that environmentally relevant concentrations of Hg2+ exposure could alter TH levels and the transcription of related HPT-axis genes, disturbing the normal processes of TH metabolism.

Effects of inorganic mercury exposure on histological structure, antioxidant status and immune response of immune organs in yellow catfish (Pelteobagrus fulvidraco).

Mercury (Hg) is well recognized as a highly toxic substance to fish. Nevertheless, little is known about the toxic effects of Hg on immune organs. In this study, we investigated histology, antioxidant status and immune response of the spleen and head kidney in yellow catfish following 6 weeks of exposure to environmentally relevant concentrations of inorganic Hg (2 and 10 µg l-1 Hg2+ ). As expected, Hg accumulation and histological injury in both tissues were observed. Meanwhile, Hg2+ exposure induced oxidative stress, which increased antioxidant enzyme (superoxide dismutase, catalase and glutathione peroxidase) activities, glutathione content, anti-hydroxyl radical capacity and the expression of genes associated with antioxidant (sod1, cat, gpx1, nrf2 and mt) and stress (hsp70) responses in dose- and tissue-specific manners. In the spleen, the mRNA levels of immune-related genes (il-1ß, il-8, tnf-α, il-10, tgf-ß, lys and c3) were upregulated by Hg2+ exposure. However, in the head kidney, upregulation of tnf-α, il-10 and tgf-ß mRNAs and downregulation of il-1ß and lys expressions were observed, while transcriptions of il-8 and c3 were remarkably upregulated only in the 2 µg l-1 group. Overall, our study indicated that Hg2+ exposure could result in Hg accumulation and thereby induced histological impairment, oxidative stress and immunotoxicity in immune organs of yellow catfish, but some enzymes and/or genes involved in antioxidant and immune systems would be activated to resist Hg2+ -induced damage.