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Parkinson's disease (PD) is a progressive neurodegenerative disorder. The classical behavioral defects of PD patients involve motor symptoms such as bradykinesia, tremor, and rigidity, as well as non-motor symptoms such as anosmia, depression, and cognitive impairment. Pathologically, the progressive loss of dopaminergic (DA) neurons in the substantia nigra (SN) and the accumulation of α-synuclein (α-syn)-composed Lewy bodies (LBs) and Lewy neurites (LNs) are key hallmarks. Glia are more than mere bystanders that simply support neurons, they actively contribute to almost every aspect of neuronal development and function; glial dysregulation has been implicated in a series of neurodegenerative diseases including PD. Importantly, amounting evidence has added glial activation and neuroinflammation as new features of PD onset and progression. Thus, gaining a better understanding of glia, especially neuron-glia crosstalk, will not only provide insight into brain physiology events but also advance our knowledge of PD pathologies. This review addresses the current understanding of α-syn pathogenesis in PD, with a focus on neuron-glia crosstalk. Particularly, the transmission of α-syn between neurons and glia, α-syn-induced glial activation, and feedbacks of glial activation on DA neuron degeneration are thoroughly discussed. In addition, α-syn aggregation, iron deposition, and glial activation in regulating DA neuron ferroptosis in PD are covered. Lastly, we summarize the preclinical and clinical therapies, especially targeting glia, in PD treatments. [ABSTRACT FROM AUTHOR]

Mutations in leucine‐rich repeat kinase 2 (LRRK2) are the most common cause of familial and sporadic Parkinson's disease. A plethora of evidence has indicated a role for LRRK2 in endolysosomal trafficking in neurons, while LRRK2 function in glia, although highly expressed, remains largely unknown. Here, we present evidence that LRRK2/dLRRK mediates a lysosomal pathway that contributes to glial cell death and the survival of dopaminergic (DA) neurons. LRRK2/dLRRK knockdown in the immortalized microglia or flies results in enlarged and swelling lysosomes fewer in number. These lysosomes are less mobile, wrongly acidified, exhibit defective membrane permeability and reduced activity of the lysosome hydrolase cathepsin B. In addition, LRRK2/dLRRK depletion causes glial apoptosis, DA neurodegeneration, and locomotor deficits in an age‐dependent manner. Taken together, these findings demonstrate a functional role of LRRK2/dLRRK in regulating the glial lysosomal pathway; deficits in lysosomal biogenesis and function linking to glial apoptosis potentially underlie the mechanism of DA neurodegeneration, providing insights on LRRK2/dLRRK function in normal and pathological brains. Synopsis Lack of glial LRRK2/dLRRK in the immortalized microglial cells or flies results in enlarged and swelling lysosomes fewer in number. These lysosomes are less mobile, wrongly acidified, and exhibit defective membrane permeability and reduced activity of the lysosome hydrolase cathespin B. In addition, LRRK2/dLRRK depletion causes glial apoptosis, dopaminergic neurodegeneration, and locomotor deficits in an age‐dependent manner. [ABSTRACT FROM AUTHOR]

This paper uses the reconstruction-computation-quantization (RCQ)paradigm to decode low-density parity-check (LDPC) codes. RCQ facilitates dynamic non-uniform quantization to achieve good frame error rate (FER) performance with very low message precision. For message-passing according to a flooding schedule, the RCQ parameters are designed by discrete density evolution. Simulation results on an IEEE 802.11 LDPC code show that for 4-bit messages, a flooding Min Sum RCQ decoder outperforms table-lookup approaches such as information bottleneck (IB) or Min-IB decoding, with significantly fewer parameters to be stored. Additionally, this paper introduces layer-specific RCQ, an extension of RCQ decoding for layered architectures. Layer-specific RCQ uses layer-specific message representations to achieve the best possible FER performance. For layer-specific RCQ, this paper proposes using layered discrete density evolution featuring hierarchical dynamic quantization (HDQ) to design parameters efficiently. Finally, this paper studies field-programmable gate array (FPGA) implementations of RCQ decoders. Simulation results for a (9472, 8192) quasi-cyclic (QC) LDPC code show that a layered Min Sum RCQ decoder with 3-bit messages achieves more than a 10% reduction in LUTs and routed nets and more than a 6% decrease in register usage while maintaining comparable decoding performance, compared to a 5-bit offset Min Sum decoder. [ABSTRACT FROM AUTHOR]

In this work, we present a four-transistor-two-resistor (4T2R) ternary content addressable memory (TCAM) bit cell based on the resistive memory (RRAM), comprising the conventional two-transistor-two-resistor (2T2R) cell with two additional comparison transistors. It can effectively amplify the match-line signal ratio (ML-ratio), lower the leakage current of the match cell (${I}_{{\mathrm {MATCH}}}$), and suppress the read disturbance. The proposed concept is silicon verified using the 180 nm CMOS technology with transition-metal-oxide (TMO) RRAM integrated at the back-end-of-line (BEOL). It achieves a considerable ML-ratio of 1860 and a low ${I}_{{\mathrm {MATCH}}}$ of 11.15 nA on average. In a typical search operation, it shows a negligible ML drop at the match case and a large ML swing range at the mismatch case. The SPICE simulation results further show it can support a long word-length (WDL) of 256 under a clock rate of 100 MHz for search operations, which demonstrates its promise for highly parallel nonvolatile TCAM. [ABSTRACT FROM AUTHOR]

A series of near-infrared and photostable Si-oxazine fluorescent dyes was synthesized using a simple three-step procedure, and one of their reduced products, i.e. hydro-Si-oxazine HSiO3, has been utilized to sensitively detect hypochlorous acid and peroxynitrite generation by phagocytes in inflamed and pulmonary fibrosis diseased mice with emission wavelength beyond 750 nm. [ABSTRACT FROM AUTHOR]

The relationships between land use types, water and sediment parameters, and macrobenthos community structures in the upper and middle reaches of the Fenhe River and urbanization intensity were studied. Samples were collected from 23 sampling sites. Spearman rank correlation analyses were performed to assess the relationships between the percentages of impervious area or the proportions of four land uses and the water and sediment physicochemical properties, heavy metal and polycyclic aromatic hydrocarbon concentrations in water and sediment, and biological indicators of the macrobenthos communities. Some water parameters (temperature, oxidation-reduction potential, electrical conductivity, total N concentration, total P concentration, ammonia-N concentration, and nitrate-N concentration), some sediment parameters (total N concentration, total P concentration, organic matter content, percentage of particles with diameters <2 mm, and polycyclic aromatic hydrocarbon, Cd, Cr, Cu, Ni Pb, and Zn concentrations), and some macrobenthos parameters (Berger–Parker index and percentages of collectors, tolerant taxa, and Oligochaeta) significantly positively correlated with the percentage of impervious area. Some water parameters (pH and dissolved oxygen concentration), some sediment parameters (percentage of particles with diameters >2 mm), and some macrobenthos parameters (total biomass, total number of taxa, Shannon's index, N diversity index, and percentages of Ephemeroptera, Plecoptera, Trichoptera, filterers, scrapers, and sensitive taxa) significantly negatively correlated with the percentage of impervious area. The results indicate that intensification of urbanization has strongly affected the water, sediment, and macrobenthos in the Fenhe River watershed. [ABSTRACT FROM AUTHOR]

Glutathione (GSH) plays crucial roles in various physiological and pathological processes. The fluctuation of the GSH level is closely associated with a variety of diseases and cellular functions. Hence, it is important to real-time monitor the fluctuation of GSH in living cells. In this work, we presented a rhodol-hybridized hemicyanine fluorophore (RdH) as a selective, rapid-response, ratiometric, and reversible fluorescent probe for intracellular GSH (t1/2 = 89 ms, Kd = 1.42 mM). The imaging assays in living cells revealed that RdH could be used to real-time monitor GSH dynamics in A549 cells under a laser scanning confocal microscope by ratiometric fluorescence changes. [ABSTRACT FROM AUTHOR]

A series of novel C -glycosyl triazolyl quinoline-based fluorescent sensors have been synthesized via click chemistry. It was found that novel sensors exhibited good selectivity for Hg 2+ over many other metal ions. The glucose framework was introduced to increase the water-solubility of the fluorescent sensors and broaden its application for the detection of Hg(II) in the water-solubility biological systems. The mechanism of the chemodosimetric behavior of the sensors has been attributed to a binding mode of triazolyl quinoline with Hg 2+ which has been characterized by a number of spectroscopic techniques. [ABSTRACT FROM AUTHOR]

Cognitive radio (CR) as a promising way to solve the spectrum scarcity allows exploitation of the shared frequency bands while guaranteeing acceptable interference to incumbent users in satellite communication systems. An improved spread slotted ALOHA (SSA) based on multi-user, multi-channel CR model applying to the satellite communications is proposed in this paper. To make full use of the detection information of satellite earth stations, a novel joint collaborative sensing method is used in the sensing phase. Moreover, a better throughput is achieved by using the improved SSA strategy in the transmission phase comparing with the traditional slotted ALOHA (SA). Theoretical analysis shows that the system performs better when SSA is adopted. Theoretical analysis and simulation results indicate that the sensing method used in this model outperforms the traditional 'hard combining' strategy in the whole sensing process. [ABSTRACT FROM AUTHOR]

Porous carbon materials have demonstrated exceptional performance in various energy and environment-related applications. Recently, research on supercapacitors has been steadily increasing, and porous carbon materials have emerged as the most significant electrode material for supercapacitors. Nonetheless, the high cost and potential for environmental pollution associated with the preparation process of porous carbon materials remain significant issues. This paper presents an overview of common methods for preparing porous carbon materials, including the carbon-activation method, hard-templating method, soft-templating method, sacrificial-templating method, and self-templating method. Additionally, we also review several emerging methods for the preparation of porous carbon materials, such as copolymer pyrolysis, carbohydrate self-activation, and laser scribing. We then categorise porous carbons based on their pore sizes and the presence or absence of heteroatom doping. Finally, we provide an overview of recent applications of porous carbon materials as electrodes for supercapacitors. [ABSTRACT FROM AUTHOR]

Purpose -- Compared with the traditional printed circuit board (PCB) drilling process, the technology of drilling IC substrate is facing more problems, such as much smaller hole diameter, more intensive hole space, thinner sheet and more complicated materials are drilled in process. Moreover, the base material of IC substrate is different from traditional PCB, more kinds of fillers added in IC substrate which make the drill worn seriously during drilling process. Micro-drills wear and micro holes quality are the most important questions when drilling IC substrate so far. Wear morphology of micro-drill, holes wall roughness and hole location accuracy are researched in this paper. The influence factors of micro-drills wear and micro holes quality are also studied in this drilling process. The paper aims to discuss these issues. Design/methodology/approach -- Two drills with same structure and different diameter are used to drill different stacks of IC substrate and drill different holes in this paper. There are four experiments made and the drilling parameters including spindle speed (n), feed rate (v[sub f]) and retraction speed (v[sub r]) are recommended by drill manufacturing company. Wear morphologies of drill are observed, holes wall roughness (R[sub max]) and holes location accuracy (C[sub pk]) are measured in this paper. Analyzing the main factors influence on drill wear, holes wall roughness and holes location accuracy through these experiments. Findings -- The micro-drills of IC substrate wear more severely compared with other material of PCB through the experimental results in this paper. Drill diameter has influence on micro-drill wear when drilling IC substrate, the smaller of drill is, the more severely of micro-drill wears. Drill diameter affect the holes wall roughness too, the holes wall roughness of larger holes is better than smaller one in a certain range. The drilled holes number also has influence on micro-drills wear, holes wall roughness and holes location accuracy. The more drilled holes, the seriously of micro-drills wear, and the worn drill would destroy the hole quality. Therefore, the more drilled holes lead the bad holes wall roughness and holes location accuracy in this paper. In addition, stacks of IC substrate affect much on the holes location accuracy, the more stacks, the worse holes location accuracy. Originality/value -- Chinese Mainland is obviously lagging behind in technology and manufacturer of IC substrate which is incompatible with the nation circumstances. There is few research of drilling IC substrate in China and research data are lacking so far. It is most necessary to improve the technology level of drilling IC substrate in China. In order to reduce the wear of micro-drills and improve the quality of micro-holes, many experimental tests about drilling IC substrate are researched in this paper. [ABSTRACT FROM AUTHOR]

The spatial distributions, sources, and source-specific risk apportionments of 26 antibiotics (5 categories) in the Fenhe River basin were determined based on sample data. The results showed that antibiotics were widely distributed in the surface water. There were significant differences between the different types of antibiotics, and the highest mean concentration was that of the sulfonamide category (33.74 ng/L), accounting for 36% of the total antibiotic concentration. Spatially, all antibiotics were mainly detected in the middle and downstream areas. The ecological risk assessment results showed that the significant risk rate of antibiotics accounted for 70% and was mainly distributed in the downstream area; however, the risks differed between the 5 categories. Quinolone antibiotics exhibited the highest significant risk rate, reaching 100%. The ecological risk associated with sulfamethoxazole was the highest among all detected antibiotics. The following five main factors influenced the antibiotic concentrations: aquaculture, pharmaceutical wastewater, livestock discharges, domestic sewage, and sewage treatment plants. Among these, pharmaceutical wastewater sources contributed the most (35%) to the total antibiotic concentration, and were distributed throughout the river. Although livestock discharges were not the main reason for the high level of ecological risk, these discharges were highest at certain sites in the midstream region. Different pollution sources posed different levels of ecological risk to the Fenhe River basin, the highest of which was pharmaceutical wastewater with a significant risk rate of 58%. [Display omitted] • Antibiotics were widely detected and mainly distributed in mid and downstream. • The overall significant risk (high, medium, and low) rate of antibiotics was 70%. • Among identified five sources, pharmaceutical wastewater contributed the most. • Risk levels of different sources were related to the concentration and toxic effects. [ABSTRACT FROM AUTHOR]

Deriving water quality benchmarks based on the species sensitivity distribution (SSD) is crucial for assessing the ecological risks of antibiotics. The application of extrapolation methods such as interspecies correlation estimation (ICE) and acute‐to‐chronic ratios (ACRs) can effectively supplement insufficient toxicity data for these emerging contaminants. Acute‐to‐chronic ratios can predict chronic toxicity from acute toxicity, and ICE can extrapolate an acute toxicity value from one species to another species. The present study explored the impact of two extrapolation methods on the reliability of SSDs by analyzing different scenarios. The results show that, compared with the normal and Weibull distributions, the logistic model was the best‐fitting model. For most antibiotics, SSDs derived by extrapolation have high reliability, with 82.9% of R2 values being higher than 0.9, and combining ICE and ACR methods can bring a maximum increase of 10% in R2. Based on the results of Monte Carlo simulation, the statistical uncertainty brought by ICE in SSD is 10–40 times larger than that brought by ACR, and combining the two methods could reduce uncertainty. In addition, the sensitivity test showed that whether the toxicity data came from extrapolation or actual measurement, the lower the value of toxicity endpoints was, the greater the bias caused by the corresponding species in every scenario. Combining the two aforementioned extrapolation methods could effectively increase the stability of SSD, with their bias nearly equal to 1. Environ Toxicol Chem 2023;42:191–204. © 2022 SETAC [ABSTRACT FROM AUTHOR]

A new rhodamine-aldoxime fluorescent probe was constructed, which showed a rapid fluorescence off―on response towards HClO in NIR region with ultra-low detection limit of 2.4 nM, and has successfully been utilized to distinguish cancerous from normal cells/tissues in terms of their difference in basal ROS levels. [Display omitted] • A new NIR fluorescent HClO probe was constructed by incorporating an aldoxime reaction group into rhodamine fluorophore. • The probe showed a great and rapid off-on response towards HClO in NIR region with ultra-low detection limit. • The probe showed a great and rapid off-on response towards HClO in NIR region with ultra-low detection limit. Fluorescence-based imaging technique has widely been used to discriminate between cancerous and normal cells or tissues because of its unique merits, i.e. visualization, noninvasiveness, high-sensitivity, non-ionizing radiation, and real-time bioimaging in vivo. Such fluorescent probes are commonly designed by chemically coupling fluorophores with targeting ligands specific to overexpressed surface markers of cancer cells. However, using such probes to diagnose a diverse class of cancers is difficult because of the high heterogeneity of cancer cells. Herein, inspired by the approximately ten times higher reactive oxygen species (ROS) concentration in cancer cells than normal cells, we developed a near-infrared (NIR) fluorescent ROS probe, PyOX , by incorporating an aldoxime reaction group into rhodamine fluorophore. The probe in the mimetic physiological condition showed a great, rapid, and selective fluorescence turn―on response towards hypochlorous acid (HClO, an endogenous ROS) in near-infrared (NIR) region (λ em = 680 nm) with ultra-low detection limit of 2.4 nM, due to the HClO-triggered efficient chemical transformation of aldoxime group to nitrile oxide group. Importantly, this probe has successfully been utilized to distinguish cancer from normal cells/tissues in vitro and in vivo in terms of their difference in intracellular basal ROS levels. The present work would inspire research interest in developing more and better fluorescent ROS probes for diagnosing a wide range of cancers in clinical practice. [ABSTRACT FROM AUTHOR]

Aims: Glioblastoma is the central nervous system tumor with the highest mortality rate, and the clinical effectiveness of chemotherapy is low. Curzerene can inhibit the progression of non‐small‐cell lung cancer, but its role in glioma has not been reported. The purpose of this study was to clarify the effect of curzerene on glioma progression and further explore its potential mechanism. Methods: The expression of glutathione S‐transferase A4 (GSTA4) in glioblastoma and the effect of curzerene on the expression of GSTA4 and matrix metalloproteinase 9 and the activation of the mTOR pathway were detected by Western blotting and RT‐PCR, and the effects of curzerene treatment on glioma malignant character were detected by cell biological assays. The in vivo antitumor effects of curzerene were analyzed in a nude mouse xenograft model. Results: Curzerene was found to inhibit the expression of GSTA4 mRNA and protein in U251 and U87 glioma cells, and this effect correlated with a downregulation of the proliferation of these cells in a time‐ and dose‐dependent manner. Invasion and migration were also inhibited, and curzerene treatment correlated with induction of apoptosis. Curzerene inhibited the activation of the mTOR pathway and the expression of matrix metalloproteinase 9, and it correlated with increased 4‐hydroxynonenal levels. In vivo, curzerene was found to significantly inhibit tumor growth in nude mice and to prolong the survival time of tumor‐bearing nude mice. Conclusion: In conclusion, inhibition of GSTA4 correlates with positive outcomes in glioma models, and thus, this molecule is a candidate drug for the treatment of glioma. [ABSTRACT FROM AUTHOR]

In this study, 26 typical antibiotics in the suspended matter of the Fen River basin were analyzed during the wet and dry seasons, and the main sources of antibiotic contamination were further identified. The results showed that the concentrations of antibiotics in the suspended matter varied seasonally. Sixteen antibiotics were detected in the suspended matter during the wet season with an average concentration of 463.56 ng/L. However, a total of 21 antibiotics were detected in the dry season, with an average concentration of 106.00 ng/L. The concentration of chloramphenicol antibiotics was outstanding in the wet season and dry season. The spatial distribution of the antibiotics in suspended matter showed little spatial discrepancy during the wet season. During the dry season, nevertheless, the concentration was higher upstream than midstream and downstream. The main sources of antibiotics in the Fen River Basin were livestock and poultry breeding, wastewater from wastewater treatment plants (WWTPs), agricultural drainage, domestic sewage, and pharmaceutical wastewater. Wastewater from WWTPs and domestic sewage were identified as two primary sources in the suspended matter during the wet season, with wastewater from WWTPs contributing the most accounting for 37%. While the most significant source of antibiotics in the suspended matter in the dry season was pharmaceutical wastewater, accounting for 36%. In addition, the contribution proportion of sources for antibiotics exhibited discrepant spatial distribution characteristics. In the wet season, wastewater from WWTPs dominated in the upstream and midstream, and livestock and poultry breeding was prominent in the midstream and downstream. Pharmaceutical wastewater was the main source in the midstream and downstream regions during the dry season. [Display omitted] • Spatiotemporal characteristics of antibiotics in the suspended matter were analyzed. • The higher average concentration is concentrated in upstream during the dry season. • Four main sources were identified in each season, while types varied seasonally. • Wastewater from WWTPs contributed the greatest in midstream in wet season. • Pharmaceutical wastewater dominated in midstream and downstream in dry season. [ABSTRACT FROM AUTHOR]

A seeding-free synthesis method is developed for preparation of oriented zeolite LTA membranes by using 1,4-phenylene diisocyanate (PDI) as a molecular linker. Before hydrothermal synthesis, -NCO groups are introduced with the functionalization of Al2O3 supports by PDI. A thin, well intergrown zeolite LTA membrane with a thickness of about 4.0 µm can be formed on the PDI-modified Al2O3 supports. The zeolite LTA membrane displays high pervaporation performances for dehydration of alcohols. At 90°C, the separation factor of the zeolite LTA membrane is 4480 for dehydration of 95 wt% ethanol/water mixtures, with a high water flux of 3.4 kg·m−2·h−1. [ABSTRACT FROM AUTHOR]

SDN network is a dynamic, controllable, cost-effective and adaptable system. It is suitable for communication networks with high bandwidth and high dynamic characteristics. Therefore, combining SDN ideas with the new generation of LEO satellite networks can achieve more flexible monitoring and management of the network, and can make the network expansion more convenient. Joint the Depth-First-Search (DFS) idea and Dijkstra algorithm for the huge numbers of LEO mobile satellite network based on SDN is proposed to improve the computational efficiency and the reliability of calculation result. Moreover, the communication performance of space-based network based on SDN and traditional space-based network is compared and analyzed. The simulation results show that the huge numbers of LEO mobile satellite network based on SDN breaks through the performance limitations of the traditional network architecture, and it can achieve better performance of the network. [ABSTRACT FROM AUTHOR]

Noninvasive and specific visualization of lysosomes by fluorescence technology is critical for studying lysosomal trafficking in health and disease and for evaluating new cancer therapeutics that target tumor cell lysosomes. To date, there are two basic types of lysosomal probes whose lysosomal localization correlates with lysosomal acidity and endocytosis pathway, respectively. However, the former may suffer from pH-sensitive lysosomal localization and alkalization-induced lysosomal enzyme inactivation, and the latter need long incubation time to penetrate cell membrane due to the energy-dependency of endocytosis process. In this work, a new class of two-photon fluorescent dyes, termed amino-Si-rhodamines ( ASiR s), were developed, which possess the intrinsic lysosome-targeted ability that is independent of lysosomal acidity and endocytosis pathway. As a result, ASiR s show not only the stable lysosomal localization against lysosomal pH changes and negligible interference to lysosomal function, but also excellent cell-membrane-permeability due to the energy-independent passive diffusion pathway. These merits, coupled with their excellent two-photon photophysical properties, long-term retention ability in lysosomes, and negligible cytotoxicity, make ASiR s very suitable for real-time and long-term tracking of lysosomes in living cells or tissues without interference to normal cellular processes. Moreover, the easy functionalization via amino linker further allows the construction of various fluorescent probes for biological targets of interest based on ASiR skeleton, as indicated by the cancer-targeted fluorescent probe ASiR6 as well as a fluorescent peroxynitrite probe ASiR-P . [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) widely participate in plant growth and development. The miR396 family, one of the most conserved miRNA families, remains poorly understood in sorghum. To reveal the evolution and expression pattern of Sbi-miR396 gene family in sorghum, bioinformatics analysis and target gene prediction were performed on the sequences of the Sbi-miR396 gene family members. The results showed that five Sbi-miR396 members, located on chromosomes 4, 6, and 10, were identified at the whole-genome level. The secondary structure analysis showed that the precursor sequences of all five Sbi-miR396 potentially form a stable secondary stem–loop structure, and the mature miRNA sequences were generated on the 5′ arm of the precursors. Sequence analysis identified the mature sequences of the five sbi-miR396 genes were high identity, with differences only at the 1st, 9th and 21st bases at the 5' end. Phylogenetic analysis revealed that Sbi-miR396a, Sbi-miR396b, and Sbi-miR396c were clustered into Group I, and Sbi-miR396d and Sbi-miR396e were clustered into Group II, and all five sbi-miR396 genes were closely related to those of maize and foxtail millet. Expression analysis of different tissue found that Sbi-miR396d/e and Sbi-miR396a/b/c were preferentially and barely expressed, respectively, in leaves, flowers, and panicles. Target gene prediction indicates that the growth-regulating factor family members (SbiGRF1/2/3/4/5/6/7/8/10) were target genes of Sbi-miR396d/e. Thus, Sbi-miR396d/e may affect the growth and development of sorghum by targeting SbiGRFs. In addition, expression analysis of different tissues and developmental stages found that all Sbi-miR396 target genes, SbiGRFs, were barely expressed in leaves, root and shoot, but were predominantly expressed in inflorescence and seed development stage, especially SbiGRF1/5/8. Therefore, inhibition the expression of sbi-miR396d/e may increase the expression of SbiGRF1/5/8, thereby affecting floral organ and seed development in sorghum. These findings provide the basis for studying the expression of the Sbi-mir396 family members and the function of their target genes. [ABSTRACT FROM AUTHOR]

As a complex system, the sustainability of basins is crucial for the development and stability of human society. This study creatively combined Otsu algorithm with analytic hierarchy process to determine the relative threshold of sustainable development of the Yellow River basin system, aiming to provide a new perspective for the application of threshold theory to the study of sustainable development. The results showed that among the 24 sustainable development evaluation indicators, the Per Capita Cultivated Area (F24) had the greatest impact, while the Ecological Water use (F2) and Agricultural Water use (F19) indicators had the greatest impact range. For different subsystems, the sustainable development indices of the eco-environment subsystem in the upstream of the Yellow River exhibited more significant changes, while that of the socio-economic subsystem exhibited more prominent changes in the midstream. However, the sustainable development indices of the water-sand subsystems in the upstream, midstream, and downstream fluctuated greatly, with a maximum value occurring downstream. The sustainable development threshold was further determined to be 0.408 for the complex system in the Yellow River Basin. The sustainable development thresholds differed in upstream, midstream, and downstream, which ranged from 0.4201 to 0.4524. Based on the sustainable development threshold, the sustainable development status of the Yellow River Basin had continued to improve since 2017, and a high level of sustainability had been maintained. However, the sustainable status is a dynamic process of change, and changes in natural conditions or policies may affect the state of sustainable development. [Display omitted] • Environmental degradation affects the sustainable development of the watershed. • Relative threshold of sustainable development was determined wiht AHP and Otsu. • Relative threshold of the Basin was 0.408, while there were regional differences. • Generally, Yellow River Basin maintained a high level of sustainability since 2017. • Water conservancy projects and natural changes might affect the relative threshold. [ABSTRACT FROM AUTHOR]

To discover critical genes contributing to the stemness and maintenance of spermatogonial stem cells (SSCs) and provide new insights into the function of the leucine-rich repeat (LRR) family member Lrrc34 (leucine-rich repeat-containing 34) in SSCs from mice. Bioinformatic methods, including differentially expressed gene (DEG), gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, were used to uncover latent pluripotency-related genes. Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence analyses were utilized to verify the mRNA and protein expression levels, respectively. RNA interference of Lrrc34 using siRNA was performed to detect its transient impact on SSCs. Eight DEGs between ID4-EGFP+ (G) and ID4-EGFP+/TSPAN8High (TH), eight DEGs between G and ID4-EGFP+/TSPAN8Low (TL) and eleven DEGs between TH and TL were discovered, and eleven protein-protein interaction (PPI) modules were found to be significant in the PPI network of DEGs. One of the DEGs, Lrrc34, was selected as a potential pluripotency-related gene due to its differential expression among ID4-EGFP+ spermatogonia subsets and its interaction with fibroblast growth factor 2 in the fifth module. Immunofluorescence experiments exhibited specific expression of Lrrc34 in a subpopulation of undifferentiated spermatogonia marked by LIN28A, and RT-PCR experiments confirmed the high expression of Lrrc34 in SSCs from P7 and adult mice. The transient knockdown of Lrrc34 in SSCs resulted in reduced colony sizes and significant changes in the transcriptome and apoptotic pathways. Lrrc34 is highly expressed in mouse SSCs and is required for SSC proliferation in vitro through effects on transcriptome and signaling transduction pathways. [ABSTRACT FROM AUTHOR]

Background: Pulmonary sarcomatoid carcinoma (PSC) is a rare and poorly differentiated type of non-small cell lung cancer (NSCLC) with specific characteristics, which usually presents a challenge in clinical practice. Mesenchymal–epithelial transition (MET) gene has been identified as a promising target for treatments in the past few years. Here, we report a case of a patient with PSC harboring MET exon 14 mutation, who responded to a novel MET inhibitor – savolitinib. Case presentation: A 75-year-old male patient with symptoms of cough, dyspnea and intermittent chest pain was diagnosed with sarcomatoid carcinoma. The tumor involved the right lung, the right hilum and multiple lesions in the right pleura, indicating a clinical disease stage IV. Next-generation sequencing of lung biopsy specimen indicated a MET exon 14 skipping mutation (NM_000245:c.3028+3A>G), with a variant allele frequency of 73.9%. The patient achieved a rapid and durable partial response with the initiation of savolitinib administration (600 mg, orally, once daily). The progression-free survival in this patient was 36 weeks. There were no ≥grade 3 adverse events reported and there was no dose reduction during treatment. Following savolitinib treatment, the allele frequency of MET exon 14 mutation in plasma circulating tumor DNA decreased with the reduction in tumor size. At the time of disease progression, fibroblast growth factor receptor 1 (FGFR1), EGFR and KRAS gene amplification were newly identified in tumor biopsy sample. Conclusion: This patient with PSC harboring MET exon 14 skipping mutation achieved significant clinical benefit with savolitinib treatment. Emergence of FGFR1, EGFR and KRAS gene amplification at the time of disease progression was likely responsible for the resistance to savolitinib in this case. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa-derived pigment pyocyanin (PCN) has been proved to induce cell apoptosis mediated by the generation of reactive oxygen species (ROS), which has been studied mainly in epithelial cells and neutrophils. However, we previously found that the PCN-producing strain PA14 induces cell apoptosis in human NK cell line NK92 more effectively than in PCN-deficient strain PA14-phZ1/2 via a yet undetermined mechanism. In the current study, we found that PCN-induced NK92 cell apoptosis occurs through mitochondrial damage despite inhibiting intracellular ROS generation. Intracellular Ca2+ ([Ca2+]i) and Bcl-2 family proteins act as important "priming signals" for apoptosis. PCN treatment increased [Ca2+]i in NK92 cells more than twofold after 2 h stimulation, whereas the Ca2+-chelating agent ethylene glycol tetra-acetic acid (EGTA) inhibited apoptosis. PCN triggered the activation of Bim, Bid, Bik, Bak, and phospho-Bad in NK92 cells in a concentration-dependent manner, but these pro-apoptotic Bcl-2 family proteins were not inhibited by EGTA. In this study, we describe the function of PCN in NK92 cells and identify mitochondrial damage as the mechanism underlying the apoptosis. [Ca2+]i and pro-apoptotic Bcl-2 family proteins are novel targets for PCN-induced apoptosis. Clarification of the cytotoxic diversity of PCN provides a new therapeutic target for defense from P. aeruginosa-induced immune cell damage. [ABSTRACT FROM AUTHOR]

Chemotherapy resistance represents a major issue associated with gastric cancer (GC) treatment, and arises through multiple mechanisms, including modulation of the cell-cycle check point. Several ubiquitin kinases, including RING finger protein 138 (RNF138), have been reported to mediate the G2/M phase arrest. In this study, we investigated the role of RNF138 in the development of cisplatin resistance of two GC cell lines. We show that RNF138 levels are higher in cisplatin-resistant cell lines, compared with cisplatin-sensitive cells, and RNF138 expression was elevated during drug withdrawal following the cisplatin treatment. Using gene overexpression and silencing, we analyzed the impact of altering RNF138 level on GC cell viability, apoptosis, and cell cycle phenotypes in two isogenic cisplatin-sensitive and resistant cell lines. We show that RNF138 overexpression increased GC cell viability, decreased apoptosis and delayed cell cycle progression in the cisplatin-sensitive GC cells. Conversely, RNF138 silencing produced opposite phenotypes in the cisplatin-resistant cells. Moreover, RNF138-dependent phosphorylation of Chk1 was seen in GC cells, indicating a novel connection between cisplatin-induced DNA damage and apoptosis. Collectively, these data suggest that RNF138 modulates the cisplatin resistance in the GC cells, thus serving as a potential drug target to challenge chemotherapy failure. In addition, RNF138 can also be used as a marker to monitor the development of cisplatin resistance in GC treatment. [ABSTRACT FROM AUTHOR]

Ubiquitylation has an important role as a signal transducer that regulates protein function, subcellular localization, or stability during the DNA damage response. In this study, we show that Ring domain E3 ubiquitin ligases RNF138 is recruited to DNA damage site quickly. And the recruitment is mediated through its Zinc finger domains. We further confirm that RNF138 is phosphorylated by ATM at Ser124. However, the phosphorylation was dispensable for recruitment to the DNA damage site. Our findings also indicate that RAD51 assembly at DSB sites following irradiation is dramatically affected in RNF138-deficient cells. Hence, RNF138 is likely involved in regulating homologous recombination repair pathway. Consistently, efficiency of homologous recombination decreased observably in RNF138-depleted cells. In addition, RNF138-deficient cell is hypersensitive to DNA damage insults, such as IR and MMS. And the comet assay confirmed that RNF138 directly participated in DNA damage repair. Moreover, we find that RAD51D directly interacted with RNF138. And the recruitment of RAD51D to DNA damage site is delayed and unstable in RNF138-depleted cells. Taken together, these results suggest that RNF138 promotes the homologous recombination repair pathway. [ABSTRACT FROM AUTHOR]

Numerous investigations have focused on the detection of antisperm antibodies, which have a naturally occurring impact on male and female fertility. In this study, spermatogenic glyceraldehyde-3-phosphate dehydrogenase (GAPDHS) was considered to be a candidate biomarker of immune infertility. The concentrations of anti-GAPDHS antibodies in the sera of sterile individuals and fertile couples were measured by enzyme-linked immunosorbent assay. Sera were collected from immune infertile ( n = 175) and fertile ( n = 237) individuals and were screened by tray agglutination tests (TAT). Infertile sera were further divided into two groups according to the serum titers obtained by TAT (titers ≤ 1:8, n = 58; titers > 1:8, n = 117). The concentrations of anti-GAPDHS antibodies were significantly higher in the immune infertile group than in the fertile group and were much higher with regard to the increased degrees of sperm agglutination (titers > 1:8). Surprisingly, we found statistically significantly higher concentrations of antibodies in the sera of infertile men than in those of infertile women, and a similar statistical result was obtained in the sera when primary infertility was compared with secondary infertility. Thus, anti-GAPDHS antibodies seem to be a sensitive parameter in immune infertile detection and might be one of the main factors causing immune infertility. This factor might be valuable as an indicator in the clinical diagnosis and monitoring treatment of infertility. [ABSTRACT FROM AUTHOR]

Apoptosis is a regulated cellular suicide program that is critical for the development and maintenance of healthy tissues. Previous studies have shown that small kinetochore associated protein (SKAP) cooperates with kinetochore and mitotic spindle proteins to regulate mitosis. However, the role of SKAP in apoptosis has not been investigated. We have identified a new interaction involving SKAP, and we propose a mechanism through which SKAP regulates cell apoptosis. Our experiments demonstrate that both overexpression and knockdown of SKAP sensitize cells to UV-induced apoptosis. Further study has revealed that SKAP interacts with Pre-mRNA processing Factor 19 (Prp19). We find that UV-induced apoptosis can be inhibited by ectopic expression of Prp19, whereas silencing Prp19 has the opposite effect. Additionally, SKAP negatively regulates the protein levels of Prp19, whereas Prp19 does not alter SKAP expression. Finally, rescue experiments demonstrate that the pro-apoptotic role of SKAP is executed through Prp19. Taken together, these findings suggest that SKAP promotes UV-induced cell apoptosis by negatively regulating the anti-apoptotic protein Prp19. [ABSTRACT FROM AUTHOR]

We collected paired samples of tumor and adjacent normal colorectal tissues from 22 patients with colorectal carcinoma to compare the differences in the expression of lysine specific demethylase 1 (LSD1) in these two tissues. The results showed that in 19 paired samples (86.4%), LSD1 is more highly expressed in tumor tissue than in normal tissue. To explore the role of LSD1 in colorectal tumorigenesis, we used somatic cell gene targeting to generate an LSD1 knockout (KO) HCT 116 human colorectal cancer cell line as a research model. The analysis of phenotypic changes showed that LSD1 KO colorectal cancer cells are less tumorigenic, both in vivo and in vitro. The differential expression analysis of the cells by mRNA sequencing (RNA-Seq) yielded 2,663 differentially expressed genes, and 28 of these genes had highly significant differences (Q <0.01). We then selected the 4 colorectal cancer-related genes ADM, DKK1, HAS3 and SMURF2 for quantitative real-time PCR verification. The results showed that the differences in the expression of ADM, DKK1 and HAS3 were consistent with those measured using the RNA-Seq data. As DKK1 was the gene with the most significant differential expression, we analyzed the key proteins of the DKK1-related Wnt/β-catenin signaling pathway and found that, after knocking out LSD1, the amount of free β-catenin translocated to the nucleus was significantly reduced and that the transcription of the signaling pathway target gene c-Myc was down-regulated. Our studies show that LSD1 activates the Wnt/β-catenin signaling pathway by down-regulating the pathway antagonist DKK1, which may be one of the mechanisms leading to colorectal tumorigenesis. [ABSTRACT FROM AUTHOR]

Highlights: [•] RHBDD1 could induce the specific activation of AP-1 in a dose-dependent manner. [•] RHBDD1 overexpression increased c-Jun activity, and the activity was visibly inhibited when RHBDD1 was knocked down. [•] RHBDD1 functioned as an antiapoptotic protein by regulating Bcl-3, which is the downstream target of AP-1. [ABSTRACT FROM AUTHOR]

Objective: To demonstrate that capacitation in mouse spermatozoa involves alterations in the expression and localization of ACTL7a.Design: Determine the alteration in the expression level and localization of ACTL7a in the induction of capacitation in mouse spermatozoa.Setting: The Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, Peking Union Medical College, People's Republic of China.Animal(s): ICR (Institute of Cancer Research) mice.Intervention(s): None.Main Outcome Measure(s): Western blot, indirect immunostaining.Result(s): The expression of ACTL7a is upregulated via the PKA pathway and undergoes remodeling during the early period of capacitation in mouse spermatozoa.Conclusion(s): ACTL7a is an essential component of capacitation in mouse spermatozoa. The alteration in the expression and localization of ACTL7a may be the primary biochemical event in the induction of capacitation in mouse spermatozoa. [ABSTRACT FROM AUTHOR]

Dipeptide permease (Dpp), which belongs to an ABC transport system, imports peptides consisting of two or three L-amino acids from the matrix to the cytoplasm in microbes. Previous studies have indicated that haem competes with dipeptides to bind DppA in vitro and in vivo and that the Dpp system can also translocate haem. Here, the crystal structure of DppD, the nucleotide-binding domain (NBD) of the ABC-type dipeptide/oligopeptide/nickel-transport system from Thermoanaerobacter tengcongensis, bound with ATP, Mg2+ and a [4Fe-4S] iron-sulfur cluster is reported. The N-terminal domain of DppD shares a similar structural fold with the NBDs of other ABC transporters. Interestingly, the C-terminal domain of DppD contains a [4Fe-4S] cluster. The UV-visible absorbance spectrum of DppD was consistent with the presence of a [4Fe-4S] cluster. A search with DALI revealed that the [4Fe-4S] cluster-binding domain is a novel structural fold. Structural analysis and comparisons with other ABC transporters revealed that this iron-sulfur cluster may act as a mediator in substrate (dipeptide or haem) binding by electron transfer and may regulate the transport process in Dpp ABC transport systems. The crystal structure provides a basis for understanding the properties of ABC transporters and will be helpful in investigating the functions of NBDs in the regulation of ABC transporter activity. [ABSTRACT FROM AUTHOR]

Abstract: The E3 ubiquitin ligase activity and subcellular localisation of human TRIM69 (hTRIM69) gene were studied. It was found that hTRIM69 mediated ubiquitination in an E2 conjugating enzyme selective fashion in vitro and an intact RING finger domain was indispensible for the process. Further evidences showed that hTRIM69 could mediate ubiquitination in vivo, which could be enhanced by a proteasome inhibitor. hTRIM69 was found to localise in both the cytoplasm and the nucleus in a speckled aggregating pattern, which also required an intact RING finger domain. Collectively, hTRIM69 is a novel E3 ubiquitin ligase identified from human testis and may function to ubiquitinate its particular substrates during spermatogenesis. [Copyright &y& Elsevier]

Abstract: Di- and oligopeptide- binding protein OppAs play important roles in solute and nutrient uptake, sporulation, biofilm formation, cell wall muropeptides recycling, peptide-dependent quorum-sensing responses, adherence to host cells, and a variety of other biological processes. Soluble OppA from Thermoanaerobacter tengcongensis was expressed in Escherichia coli. The protein was found to be >95% pure with SDS–PAGE after a series of purification steps and the purity was further verified by mass spectrometry. The protein was crystallized using the sitting-drop vapour-diffusion method with PEG 400 as the precipitant. Crystal diffraction extended to 2.25Å. The crystal belonged to space group C2221, with unit-cell parameters of a =69.395, b =199.572, c =131.673Å, and α=β=γ=90°. [Copyright &y& Elsevier]

Abstract: Spermatogenesis is an extremely intricate process that is tightly regulated and orchestrated by a series of well-coordinated gene expression programmes. Nemo-like kinase (NLK) is an evolutionarily conserved serine/threonine kinase that functions in a wide variety of developmental events. Nevertheless, the function of NLK in spermatogenesis has not been investigated. In this study, we found that the distribution of NLK in mice exhibited a dynamic change during testicular development and gradually became concentrated in the acrosomes of elongated spermatids. NLK overexpression promoted etoposide-induced apoptosis of male germ cell-derived GC-1 cells, while knockdown of NLK by RNA interference (RNAi) attenuated etoposide-induced apoptosis. Our findings suggest that NLK plays an important role in etoposide-induced germ cell apoptosis and may be associated with spermatogenesis. [Copyright &y& Elsevier]

Abstract: MicroRNAs (miRNAs) are posttranscriptional modulators of gene expression that play important roles in various biological processes. Spermatogenesis is a highly regulated process in which diploid spermatogonia eventually differentiate into haploid spermatozoa. In this study, we identified four differentially expressed miRNAs between two premeiotic male germ cells, made predictions about their putative targets, and confirmed cyclin T2 (Ccnt2) as a direct target of miR-15a. We also report that miR-15a inhibited muscle differentiation at least in part by targeting Ccnt2, which represents a novel interaction. Subsequently, miR-15a and Ccnt2 were profiled in developing mice testes to observe their inverse correlations in the postnatal 3-week period to understand their roles in spermatogenesis. [Copyright &y& Elsevier]

Regulation of apoptosis at various stages of differentiation plays an important role in spermatogenesis. Therefore, the identification and characterisation of highly expressed genes in the testis that are involved in apoptosis is of great value to delineate the mechanism of spermatogenesis. Here, we reported that Fank1, a novel gene highly expressed in testis, functioned as an anti-apoptotic protein that activated the activator protein 1 (AP-1) pathway. We found that Jab1 (Jun activation domain-binding protein 1), a co-activator of AP-1, specifically interacted with Fank1. Reporter analyses showed that Fank1 activated AP-1 pathway in a Jab1-dependent manner. Fank1 overexpression also increased the expression and activation of endogenous c-Jun. Further study showed that Fank1 inhibited cell apoptosis by upregulating and activating endogenous c-Jun and its downstream target, Bcl-3. This process was shown to be Jab1 dependent. Taken together, our results indicated that by interacting with Jab1, Fank1 could suppress cell apoptosis by activating the AP-1-induced anti-apoptotic pathway. [ABSTRACT FROM AUTHOR]

Abstract: cAMP response element modulator (CREM)-mediated gene expression is an essential regulatory mechanism for germ cell differentiation. CREM and its coactivator in testis, ACT, activate the transcription of many essential genes for spermatogenesis. Sperm associated antigen 8 (SPAG8) is a testis-specific component that is expressed during germ cell differentiation. In this study, we found the pattern of SPAG8 expression largely overlapped with that of ACT during spermatogenesis and verified the association of SPAG8 with ACT. Furthermore, we showed that SPAG8 enhanced the transcriptional activation of ACT-mediated CREMτ by strengthening the binding of ACT to CREMτ. These results indicate that SPAG8 acts as a regulator of ACT and plays an important role in CREM-ACT-mediated gene transcription during spermatogenesis. Structured summary: MINT-7892874: CREM tau (uniprotkb:P27699-1) physically interacts (MI:0915) with ACT (uniprotkb:Q9WTX7) by anti tag coimmunoprecipitation (MI:0007) MINT-7892809: ACT (uniprotkb:Q9WTX7) physically interacts (MI:0915) with SPAG8 (uniprotkb:B9EKF1) by pull down (MI:0096) MINT-7892820, MINT-7892840, MINT-7892987, MINT-7892854: ACT (uniprotkb:Q9WTX7) physically interacts (MI:0915) with SPAG8 (uniprotkb:B9EKF1) by anti tag coimmunoprecipitation (MI:0007) [Copyright &y& Elsevier]

Abstract: The adhesive domain of SdrD from Staphylococcus aureus was solubly expressed in Escherichia coli in high yield. After a series of purification steps, the purified protein was >95% pure, which was SdrD from S. aureus identified by SDS–PAGE and MALDI-TOF MS. Crystals were grown at 18°C using 25% polyethylene glycol 3350 as precipitant. Diffraction by the crystal extends to 1.65Å resolution, and the crystal belongs to the space group C2, with the unit cell parameters a =133.3, b =58.3, c =112.3Å, α =90.00, β =111.14, γ =90.00. [Copyright &y& Elsevier]

Objective: To examine the relationship between an antibody against GAPDH-2, a sperm-specific protein, and infertility of female mice. Design: Basic research. Setting: National Research Institute for Family Planning Beijing, World Health Organization Collaboration Center of Human Reproduction. Animal(s): New Zealand rabbit, NIH and ICR mice. Intervention(s): None. Main Outcome Measure(s): Enzyme-linked immunoabsorbent assay, Western blot and indirect immunostaining assays, standard fertility assay, and sperm agglutination assay. Result(s): Antibodies against the full-length GAPDH-2 were raised. Its specificity was assessed by immunoblotting and indirect immunostaining assays. The antibody immunoreacted with human sperm GAPDH-2 and the mouse homolog GAPDS but did not cross-react with GAPDH. Treatment of female mice with IP injection of anti-GAPDH-2 serum significantly reduced their fertility. Anti-GAPDH-2 serum caused the agglutination of normal mice sperm in vitro. The anti-GAPDH-2 antibody was detectable in the sera and uterine fluid of the mice immunized with GAPDH-2. Conclusion(s): These results show that GAPDH-2 should be further evaluated as a promising candidate in the development of an antifertility immunogen. Detecting anti-GAPDH-2 antibodies in the bodily fluid of subjects afflicted with indeterminate infertility may be a new diagnostic index. [Copyright &y& Elsevier]

Abstract: Applying the method of segmentation of seminiferous tubules combined with DDRT-PCR and cDNA library screening, a novel DnaJ homologue, rDJL was identified in rat testis. The reading frame encodes a protein of 223 amino acid residues containing J domain in the NH2 terminal region. rDJL gene is expressed mainly in testis and rDJL protein was immunolocalized notably in the acrosome region of spermatozoa. Immunoprecipitation experiments showed that rDJL interacted with Hsc70 and clathrin protein. When CHO cells were treated with EGF, rDJL and clathrin protein were found to be colocalized and be concentrated as endosome vesicles. The present findings suggest that rDJL functions as co-chaperone to Hsc70, participates in vesicular trafficking and may play an important role in acrosomogenesis. [Copyright &y& Elsevier]

The expression of stage-specific genes during spermatogenesis was determined by isolating two segments of rat seminiferous tubule at different stages of the germinal epithelium cycle delineated by transillumination-delineated microdissection, combined with differential display polymerase chain reaction to identify the differential transcripts formed. A total of 22 cDNAs were identified and accepted by GenBank as new expressed sequence tags. One of the expressed sequence tags was radiolabeled and used as a probe to screen a rat testis cDNA library. A novel full-length cDNA composed of 2228 bp, designated as RSD-3 (rat sperm DNA no.3, GenBank accession no. AF094609) was isolated and characterized. The reading frame encodes a polypeptide consisting of 526 amino acid residues, containing a number of DNA binding motifs and phosphorylation sites for PKC, CK-II, and p34cdc2. Northern blot of mRNA prepared from various tissues of adult rats showed that RSD-3 is expressed only in the testis. The initial expression of the RSD-3 gene was detected in the testis on the 30th postnatal day and attained adult level on the 60th postnatal day. Immunolocalization of RSD-3 in germ cells of rat testis showed that its expression is restricted to primary spermatocytes, undergoing meiosis division I. A human testis homologue of RSD-3 cDNA, designated as HSD-3.1 (GenBank accession no. AF144487) was isolated by screening the Human Testis Rapid-Screen arrayed cDNA library panels by RT-PCR. The exon-intron boundaries of HSD-3.1 gene were determined by aligning the cDNA sequence with the corresponding genome sequence. The cDNA consisted of 12 exons that span approximately 52.8 kb of the genome sequence and was mapped to chromosome 14q31.3. [ABSTRACT FROM AUTHOR]

Background: TP53 is one of the most frequently mutated genes among all cancer types, and TP53 mutants occur more than 60% in colorectal cancer (CRC). Among all mutants, there are three hot spots, including p53-R175H, p53-R248W and p53-R273H. Emerging evidence attributes cancer carcinogenesis to cancer stem cells (CSCs). Long noncoding RNAs (lncRNAs) play crucial roles in maintaining the stemness of CSCs. However, it is unknown if mutant p53-regulated lncRNAs are implicated in the maintenance of CSC stemness. Methods: RNA-sequencing (RNA-seq) and ChIP-sequencing (ChIP-seq) were used to trace the lncRNA network regulated by p53-R273H in HCT116 endogenous p53 point mutant spheroid cells generated by the somatic cell knock-in method. RT-qPCR was used to detect lncRNA expression patterns, verifying the bioinformatics analysis. Transwell, spheroid formation, fluorescence activated cell sorter (FACS), xenograft nude mouse model, tumor frequency assessed by extreme limiting dilution analysis (ELDA), Western blot assays and chemoresistance analysis were performed to elucidate the functions and possible mechanism of lnc273–31 and lnc273–34 in cancer stem cells. Results: p53-R273H exhibited more characteristics of CSC than p53-R175H and p53-R248W. RNA-seq profiling identified 37 up regulated and 4 down regulated differentially expressed lncRNAs regulated by p53-R273H. Combined with ChIP-seq profiling, we further verified two lncRNAs, named as lnc273–31 and lnc273–34, were essential in the maintenance of CSC stemness. Further investigation illustrated that lnc273–31 or lnc273–34 depletion dramatically diminished colorectal cancer migration, invasion, cancer stem cell self-renewal and chemoresistance in vitro. Moreover, the absence of lnc273–31 or lnc273–34 dramatically delayed cancer initiation and tumorigenic cell frequency in vivo. Also, lnc273–31 and lnc273–34 have an impact on epithelial-to mesenchymal transition (EMT). Finally, lnc273–31 and lnc273–34 were significantly highly expressed in CRC tissues with p53-R273H mutation compared to those with wildtype p53. Conclusions: The present study unveiled a high-confidence set of lncRNAs regulated by p53-R273H specific in colorectal CSCs. Furthermore, we demonstrated that two of them, lnc273–31 and lnc273–34, were required for colorectal CSC self-renewal, tumor propagation and chemoresistance. Also, the expression of these two lncRNAs augmented in colorectal cancer patient samples with p53-R273H mutation. These two lncRNAs may serve as promising predictors for patients with p53-R273H mutation and are vital for chemotherapy. [ABSTRACT FROM AUTHOR]

Background: Our previous work revealed that rhomboid domain-containing protein 1 (RHBDD1) participates in the modulation of cell growth and apoptosis in colorectal cancer cells. This study aimed to investigate the function of RHBDD1 in regulating breast cancer progression and its underlying molecular basis. Methods: Immunohistochemistry was performed to evaluate RHBDD1 expression in 116 breast cancer tissue and 39 adjacent normal tissue and expression of RHBDD1, phospho-Akt (p-Akt) and cyclin-dependent kinase 2 (CDK2) in the same 84 breast cancer specimens. RHBDD1-knock-out cells were established using breast cancer cell lines. In vitro studies were carried out to estimate the function of RHBDD1 in cell proliferation, migration and invasion. Fluorescence microscopy assay and flow cytometric analysis were used to measure apoptosis and cell cycle regulation. RNA sequencing and western blot analysis were used to investigate the molecular mechanisms of RHBDD1. Results: RHBDD1 was highly up-regulated in breast cancer tissue compared with that in normal tissue and associated with pathological tumor (pT) stage, pathological tumor-node-metastasis (pTNM) stage and estrogen receptor (ER) expression. RHBDD1 up-regulation was associated with poor prognosis in several subtypes of breast cancer. Deletion of RHBDD1 promoted apoptosis and suppressed proliferation, migration and invasion in breast cancer cells. RHBDD1 deletion suppressed Akt activation and decreased CDK2 protein level via proteasome pathway, thus inhibited cell cycle progression and G1/S phase transition. Moreover, the protein level of RHBDD1, p-Akt and CDK2 was significantly positively correlated in breast cancer tissue. Conclusions: Our study reveals that RHBDD1 promotes breast cancer progression by regulating p-Akt and CDK2 protein levels, and might be a potential biomarker and prognostic indicator for breast cancer patients. [ABSTRACT FROM AUTHOR]

In the publication of this article [1], there is an error in the Methods section at paragraph Cell culture and reagents and the Additional file2: Fig. S1 was erroneous linked. [ABSTRACT FROM AUTHOR]

Background: 40–50% of colorectal cancer (CRC) patients develop metastatic disease; the presence of metastasis hinders the effective treatment of cancer through surgery, chemotherapy and radiotherapy, which makes 5-year survival rate extremely low; therefore, studying CRC metastasis is crucial for disease therapy. In the present study, we investigated the role of rhomboid domain containing 1 (RHBDD1) in tumor metastasis of CRC. Methods: The expression of RHBDD1 was analyzed in 539 colorectal tumor tissues for its correlation with lymphatic metastasis and distal metastasis. Transwell assay in vitro and pleural metastasis analysis in vivo were performed to determine the functions of RHBDD1 during CRC cells metastasis. RNA-seq analysis, TOP/FOP flash reporter assay, western blot and transwell assay were performed to investigate the underlying mechanism for the function of RHBDD1 on Wnt signaling pathway. Bioinformatics analysis was conducted to investigate epithelial-mesenchymal transition (EMT) and stemness in HCT-116 cells. Tissue microarray analysis, Q-PCR and western blot were performed to determine the correlation of RHBDD1 and Zinc Finger E-Box Binding Homeobox 1 (ZEB1). Results: In this study, we found that RHBDD1 expression was positively correlated with lymphatic metastasis and distal metastasis in 539 colorectal tumor tissues. RHBDD1 expression can promote CRC cells metastasis in vitro and in vivo. RNA-Seq analysis showed that the Wnt signaling pathway played a key role in this metastatic regulation. RHBDD1 mainly regulated ser552 and ser675 phosphorylation of β-catenin to activate the Wnt signaling pathway. Rescuing ser552 and ser675 phosphorylation of β-catenin resulted in the recovery of signaling pathway activity, migration, and invasion in CRC cells. RHBDD1 promoted EMT and a stem-like phenotype of CRC cells. RHBDD1 regulated the Wnt/β-catenin target gene ZEB1, a potent EMT activator, at the RNA and protein levels. Clinically, RHBDD1 expression was positively correlated with ZEB1 at the protein level in 71 colon tumor tissues. Conclusions: Our findings therefore indicated that RHBDD1 can promote CRC metastasis through the Wnt signaling pathway and ZEB1. RHBDD1 may become a new therapeutic target or clinical biomarker for metastatic CRC. [ABSTRACT FROM AUTHOR]

Through exomic sequencing of ten hepatitis C virus (HCV)-associated hepatocellular carcinomas (HCC) and subsequent evaluation of additional affected individuals, we discovered novel inactivating mutations of ARID2 in four major subtypes of HCC (HCV-associated HCC, hepatitis B virus (HBV)-associated HCC, alcohol-associated HCC and HCC with no known etiology). Notably, 18.2% of individuals with HCV-associated HCC in the United States and Europe harbored ARID2 inactivation mutations, suggesting that ARID2 is a tumor suppressor gene that is relatively commonly mutated in this tumor subtype. [ABSTRACT FROM AUTHOR]