Transcriptome-wide analysis of RNA m6A methylation regulation of muscle development in Queshan Black pigs.

BACKGROUND: N6-methyladenosine (m6A) refers to the methylation modification of N6 position of RNA adenine, a dynamic reversible RNA epigenetic modification that plays an important regulatory role in a variety of life processes. In this study, we used MeRIP-Seq and RNA-Seq of the longissimus dorsi (LD) muscle of adult (QA) and newborn (QN) Queshan Black pigs to screen key genes with m6A modification involved in muscle growth by bioinformatics analysis. RESULTS: A total of 23,445 and 25,465 m6A peaks were found in the whole genomes of QA and QN, respectively. Among them, 613 methylation peaks were significantly different (DMPs) and 579 genes were defined as differentially methylated genes (DMGs). Compared with the QN group, there were 1,874 significantly differentially expressed genes (DEGs) in QA group, including 620 up-regulated and 1,254 down-regulated genes. In order to investigate the relationship between m6A and mRNA expression in the muscle of Queshan Black pigs at different periods, a combined analysis of MeRIP-Seq and RNA-Seq showed that 88 genes were significantly different at both levels. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes results showed that DEGs and DMGs were mainly involved in skeletal muscle tissue development, FoxO signaling pathway, MAPK signaling pathway, insulin signaling pathway, PI3K-Akt signaling pathway, and Wnt signaling pathway. Four DEGs (IGF1R, CCND2, MYOD1 and FOS) and four DMGs (CCND2, PHKB, BIN1 and FUT2), which are closely related to skeletal muscle development, were selected as candidate genes for verification, and the results were consistent with the sequencing results, which indicated the reliability of the sequencing results. CONCLUSIONS: These results lay the foundation for understanding the specific regulatory mechanisms of growth in Queshan Black pigs, and provide theoretical references for further research on the role of m6A in muscle development and breed optimization selection.

Molecular characterization and immune efficacy of fructose-1,6-bisphosphate aldolase from Haemaphysalis longicornis (Acari: Ixodidae).

BACKGROUND: Ticks are obligate hematophagous ectoparasites that transmit a variety of pathogens to humans, wildlife and domestic animals. Vaccination is an effective and environmentally friendly method for tick control. Fructose-1,6-bisphosphate aldolase (FBA) is an important glycometabolism enzyme that is a candidate vaccine against parasites. However, the immune protection of FBA in ticks is unclear. METHODS AND RESULTS: The 1092-bp open reading frame (ORF) of FBA from Haemaphysalis longicornis (HlFBA), encoding a 363-amino acid protein, was cloned using PCR methodology. The prokaryotic expression vector pET32a(+)-HlFBA was constructed and transformed into cells of Escherichia coli BL21(DE3) strain for protein expression. The recombinant HlFBA protein (rHlFBA) was purified by affinity chromatography, and the western blot results suggested that the rHlFBA protein was immunogenic. RESULTS: Results of the enzyme-linked immunosorbent assay showed that rabbits immunized with rHlFBA produced a humoral immune response specific to rHlFBA. A tick infestation trial indicated that, compared to the ticks in the histidine-tagged thioredoxin (Trx) group, the engorged tick weight and oviposition of female ticks and egg hatching rate of those in the rHlFBA group was reduced by 22.6%, 45.6% and 24.1%, respectively. Based on the cumulative effect of the these three parameters, the overall immune efficacy of rHlFBA was estimated to be 68.4%. CONCLUSIONS: FBA is a candidate anti-tick vaccine that can significantly reduce the engorged tick weight, oviposition, and egg hatching rate. The use of enzymes involved in glucose metabolism is a new strategy in the development of anti-tick vaccines.

Preparation of core-shell MOF@MOF nanoparticle as matrix for the analysis of rhubarb anthraquinones in plasma by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

A core-shell structure UiO-66-(OH)2@UiO-66-NH2 (MOF@MOF) nanoparticle was synthesized through a simple hydrothermal method and employed as an adsorbent and laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) matrix for the quantitative analysis of rhubarb anthraquinones (RAs). The properties of the materials were characterized by field emission scanning electron microscopy, transmission electron microscopy, energy-dispersive X-ray spectroscopy, X-ray powder diffraction, Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, thermogravimetric analysis, and Brunauer-Emmett-Teller. The results indicate that MOF@MOF is regular octahedral structure with a size distribution of about 100 nm, having large BET specific surface area (920 m2/g). Using the MOF@MOF as a matrix shows lower background interference, higher sensitivity, and better storage stability than that of traditional matrices. The MOF@MOF matrix exhibits excellent salt tolerance even under a NaCl concentration of 150 mM. Then, the enrichment conditions were optimized, and the adsorption time of 10 min, adsorption temperature of 40 °C and adsorbent amount of 100 µg were selected. In addition, the possible mechanism of MOF@MOF as an adsorbent and matrix was discussed. Finally, the MOF@MOF nanoparticle was employed as a matrix for the sensitive MALDI-TOF-MS analysis of RAs in spiked rabbit plasma, and the recoveries are in the range of 88.3-101.5% with RSD ≤9.9%. In short, the novel MOF@MOF matrix has demonstrated its potential in the analysis of small-molecule compounds in biological samples.

Effects of drip irrigation frequency on the yield and nutrient utilization efficiency of tomato under long-season cultivation in solar greenhouse.

To improve the utilization efficiency of nutrients and water and determine the best drip irrigation frequency for long-season tomato cultivation in solar greenhouses, we cultivated tomato grafted seedlings in soil using an integrated water and fertilizer technology: drip irrigation under mulch. Seedlings drip-irrigated with balanced fertilizer (containing 20% N, 20% P2O5, and 20% K2O) and high-K fertilizer (containing 17% N, 8% P2O5, and 30% K2O) once every 12 days were set as control (CK) and that with water once every 12 days as CK1, while other seedling groups, drip-irrigated with a nutrient solution of Yamazaki (1978) formula for tomato, were set as treatments (T1-T4). There were four drip-irrigation frequencies, i.e., once every 2 days (T1), 4 days (T2), 6 days (T3), or 12 days (T4), who received the same total amounts of fertilizer and water over the 12 experimental days. The results showed that, with the decreases of drip irrigation frequency, tomato yield, the accumulation of N, P and K in plant dry matter, the fertilizer partial productivity, and the nutrient utilization rate first increased and then decreased, peaking at the T2 treatment. Compared with CK, under the T2 treatment, plant dry matter accumulation and the accumulation of N, P and K increased by 4.9%, 8.0%, 8.0%, 16.8%, the partial productivity of fertilizer and the utilization efficiency of water increased by 142.8% and 12.2%, the use efficiency of N, P and K was better than CK by 241.4%, 466.6% and 235.9%, respectively, and the tomato yield increased by 12.2%. Under the experimental conditions, drip irrigation with the Yamazaki nutrient solution at a frequency of 4 days could increase the tomato yield, as well as the use efficiency of nutrients and water. Under long-season cultivation, these trends would result in considerable saving of water and fertilizer. Overall, our findings provided a basis for improving the scientific management of water and fertilizers under long-season tomato cultivation in protected facilities.

A Photonics-Assisted Binary/Quaternary Phase-Coded Microwave Signal Generator Applicable to Digital I/O Interfaces.

A photonics-assisted binary/quaternary phase-coded microwave signal generator with fundamental/doubling reconfigurable carrier frequency applicable to digital I/O interfaces is proposed and has been verified by experiments. This scheme is based on a cascade modulation scheme, which is used to reconfigure fundamental/doubling carrier frequency and load the phase-coded signal, respectively. By controlling the radio frequency (RF) switch and the bias voltages of the modulator, the switching of the fundamental or doubling carrier frequency can be realized. When the amplitudes and sequence pattern of the two independent coding signals are set reasonably, binary or quaternary phase-coded signals can be realized. The sequence pattern of coding signals is applicable to digital I/O interfaces and can be directly generated through the IO interfaces of FPGA instead of an expensive high-speed arbitrary waveform generator (AWG) or other digital-to-analog conversion (DAC) systems. A proof-of-concept experiment is carried out, and the performance of the proposed system is evaluated from the aspects of phase recovery accuracy and pulse compression capability. In addition, the influence of residual carrier suppression and polarization crosstalk in non-ideal states on phase shifting based on polarization adjustment has also been analyzed.

Effect of cloprostenol on luteolysis and comparison of different estrus synchronization protocols in jennies.

Estrus synchronization is necessary for intensive donkey farming. Studies on estrus synchronization in jennies are, however, scarce. We aimed to investigate the susceptibility of the donkey corpus luteum to cloprostenol and design a successful estrus synchronization protocol. Firstly, the effects of different cloprostenol doses and the timing effect of cloprostenol treatment on estrous cycle was investigated. The time from treatment to luteolysis, the ovulation interval, pre-ovulatory diameter, and ovulation rates were compared between groups. Secondly, to identify the best protocol, eight estrus synchronization protocols from three categories were examined. In the first category, jennies in groups A (n = 55) and B (n = 30) received a progesterone releasing intra-vaginal device (JVID®) and cloprostenol treatment. In the second category (group C to F), jennies were pretreated with deslorelin, and then treated with JVID and cloprostenol, including groups C (n = 50), D (n = 50), E (n = 70), and F (n = 65). In the third category, jennies were treated with deslorelin and cloprostenol, including groups G (n = 40) and H (n = 40). Comparisons were made among groups regarding the degree of synchronization, ovulation, and pregnancy rates. Treatment with 0.4 mg cloprostenol on the third day following ovulation minimized the length of the luteal phase and estrous cycle. Synchronization rate varied from 60.0% to 88.6% among groups and was highest in group E. Pregnancy rates did not differ among the eight protocols. In conclusion, cloprostenol effectively induced luteolysis in jennies and a treatment protocol combining deslorelin, cloprostenol, and JVID is efficient for estrus synchronization in donkeys.

Transabdominal ultrasound alleviates LPS-induced neuroinflammation by modulation of TLR4/NF-κB signaling and tight junction protein expression.

AIM: Inflammatory bowel disease (IBD) may be a risk factor in the development of brain inflammation. It has been demonstrated noninvasive neuromodulation through sub-organ ultrasound stimulation. The purpose of this study was to investigate whether abdominal low-intensity pulsed ultrasound (LIPUS) alleviates lipopolysaccharide (LPS)-induced cortical inflammation via inhibition of colonic inflammation. MATERIALS AND METHODS: Colonic and cortical inflammation was induced in mice by LPS (0.75 mg/kg, i.p. injection) for 7 days, followed by application of LIPUS (0.5 and 1.0 W/cm2) to the abdominal area for 6 days. Biological samples were collected for Western blot analysis, gelatin zymography, colon length measurement, and histological evaluation. KEY FINDINGS: LIPUS treatment significantly attenuated LPS-induced increases in IL-6, IL-1ß, COX-2, and cleaved caspase-3 expression in the colon and cortex of mice. Moreover, LIPUS significantly increased the levels of tight junction proteins in the epithelial barrier in the mouse colon and cortex with LPS-induced inflammation. Compared to the group treated only with LPS, the LIPUS-treated groups showed decreased muscle thickness and increased crypt length and colon length. Furthermore, LIPUS treatment reduced inflammation by inhibiting the LPS-induced activation of TLR4/NF-κB inflammatory signaling in the brain. SIGNIFICANCE: We found that LIPUS alleviated LPS-induced colonic and cortical inflammation through abdominal stimulation of mice. These results suggest that abdominal LIPUS stimulation may be a novel therapeutic strategy against neuroinflammation via enhancement of tight junction protein levels and inhibition of inflammatory responses in the colon.

Developing a Gadolinium(III) Compound Based on Apoferritin for Targeted Magnetic Resonance Imaging and Dual-Modal Therapy of Cancer.

To integrate targeted diagnosis and treatment of cancer, we proposed to develop a gadolinium (Gd) agent based on the properties of apoferritin (AFt). To this end, we not only optimized a series of Gd(III) 8-hydroxyquinoline-2-carboxaldehyde-thiosemicarbazone compounds to obtain a Gd(III) compound (C4) with remarkable T1-weighted magnetic resonance imaging (MRI) performance and cytotoxicity to cancer cells in vitro but also constructed an AFt-C4 nanoparticle (NP) delivery system. Importantly, AFt-C4 NPs improved the targeting ability of C4 in vivo and showed enhanced MRI performance and tumor growth inhibition ratio relative to C4 alone. Furthermore, we confirmed that C4 and AFt-C4 NPs inhibited tumor growth through apoptosis, ferroptosis, and ferroptosis-induced immune response.

CircREOS suppresses lipid synthesis and osteosarcoma progression through inhibiting HuR-mediated MYC activation.

MYC proto-oncogene (MYC) is a transcription factor among the most commonly activated oncoproteins, playing vital roles in lipid metabolism and tumor aggressiveness with broad effects. However, it is still largely unknown about the regulating mechanisms of MYC in osteosarcoma (OS). In this study, we identify a circRNA with Reduced Expression in OS (termed as circREOS) generated from MYC gene, as a novel regulator of MYC and OS progression. CircREOS is down-regulated in OS cells and localized in the nucleus. CircREOS suppresses MYC expression, lipid metabolism and growth, invasion in OS cells. Mechanically, circREOS physically interacts with HuR (human antigen R) protein, and subsequently restrains its binding and activation on the 3'-UTR (untranslated region) of MYC mRNA, resulting in down-regulation of MYC and inhibition of OS. Moreover, circREOS serves as a tumor suppressor via targeting lipid metabolism. CircREOS reduces FASN expression and lipid accumulation through inhibiting MYC-facilitated FASN regulation. Taken together, these results indicate that circREOS suppress lipid synthesis and OS progression through inhibiting HuR-mediated MYC activation, providing a potential therapeutic target for OS.

Anisotropic Growth of One-Dimensional Carbides in Single-Walled Carbon Nanotubes with Strong Interaction for Catalysis.

Tungsten and molybdenum carbides have shown great potential in catalysis and superconductivity. However, the synthesis of ultrathin W/Mo carbides with a controlled dimension and unique structure is still difficult. Here, inspired by the host-guest assembly strategy with single-walled carbon nanotubes (SWCNTs) as a transparent template, we reported the synthesis of ultrathin (0.8-2.0 nm) W2C and Mo2C nanowires confined in SWCNTs deriving from the encapsulated W/Mo polyoxometalate clusters. The atom-resolved electron microscope combined with spectroscopy and theoretical calculations revealed that the strong interaction between the highly carbophilic W/Mo and SWCNT resulted in the anisotropic growth of carbide nanowires along a specific crystal direction, accompanied by lattice strain and electron donation to the SWCNTs. The SWCNT template endowed carbides with resistance to H2O corrosion. Different from normal modification on the outer surface of SWCNTs, such M2C@SWCNTs (M = W, Mo) provided a delocalized and electron-enriched SWCNT surface to uniformly construct the negatively charged Pd catalyst, which was demonstrated to inhibit the formation of active PdHx hydride and thus achieve highly selective semihydrogenation of a series of alkynes. This work could provide a nondestructive way to design the electron-delocalized SWCNT surface and expand the methodology in synthesizing unusual 1D ultrathin carbophilic-metal nanowires (e.g., TaC, NbC, ß-W) with precise control of the anisotropy in SWCNT arrays.

Redox Electrolytes-Assisting Aqueous Zn-Based Batteries by Pseudocapacitive Multiple Perovskite Fluorides Cathode and Charge Storage Mechanisms.

Aqueous Zn-based batteries (AZBs) have attracted intensive attention. However, to explore advanced cathode materials with in-depth elucidation of their charge storage mechanisms, improve energy storage capacity, and construct novel cell systems remain a great challenge. Herein, a new pseudocapacitive multiple perovskite fluorides (ABF3 ) cathode is designed, represented by KMF-(IV, V, and VI; M = NiCoMnZn/-Mg/-MgFe), and constructed Zn//KMF-(IV, V, and VI) AZBs and their flexible devices. Ex situ tests have revealed a typical bulk phase conversion mechanism of KMF-VI electrode for charge storage in alkaline media dominated by redox-active Ni/Co/Mn species, with transformation of ABF3 nanocrystals into amorphous metal oxide/(oxy)hydroxide nanosheets. By employing single or bipolar redox electrolyte strategies of 20 mm [Fe(CN)6 ]3- or/and 10 mm SO3 2- /Cu[(NH3 )4 ]2+ acting on KMF-(IV, V, and VI) cathode and Zn anode, the AZBs show an improved energy storage owing to additional capacity contribution of redox electrolytes. The as-designed Zn//polyvinyl alcohol (PVA)-KOH-K3 [Fe(CN)6 ]//KMF-(IV, V, and VI) redox gel electrolytes-assisting flexible AZBs (RGE-FAZBs) exhibit remarkable performance under different bending angles because of slight dissolution corrosion of zinc anode compared with liquid electrolytes. Overall, the work demonstrates the novel idea of conversion-type multiple ABF3 cathode for redox electrolytes-assisting AZBs (RE-AZBs) and their flexible systems, showing great significance on electrochemical energy storage.

Novel Bis-pyrazoline Fluorescent Probe for Cu2+ and Fe3+ Detection and Application in Cell Imaging.

A fluorescent probe Y((1,1'-([1,1'-biphenyl]-4,4'-diylbis(3-(2-hydroxyphenyl)-4,5-dihydro-1H-pyrazole-5,1-diyl)) bis(ethan-1-one))) was designed and synthesized, which could be used to Cu2+ and Fe3+ sensors. Through the study of optical properties, the probe Y shows good selectivity and sensitivity to Cu2+ and Fe3+ in aqueous tetrahydrofuran solution [10.0 mM HEPES, pH 7.4, THF-H2O = 9:1(v/v)] with has excellent anti-interference performance, and its detection limits were 0.931 uΜ for Cu2+ and 0.401uΜ for Fe3+. The coordination mechanism of probe Y with Cu2+ and Fe3+ was speculated and verified at DFT level and HRNM. By Hela cytotoxicity and imaging tests, probe Y not only has good biocompatibility, but also can be used for sensing Cu2+ in cells.

Interferon-lambda: New role in intestinal symptoms of COVID-19.

The tremendous public health and economic impact of coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has become a huge challenge globally. There is increasing evidence that SARS-CoV-2 induces intestinal infections. Type III interferon (IFN-λ) has an antiviral role in intestinal infection, with focused, long-lasting, and non-inflammatory characteristics. This review presents a summary of the structure of SARS-CoV-2, including its invasion and immune escape mechanisms. Emphasis was placed on the gastrointestinal impact of SARS-CoV-2, including changes to the intestinal microbiome, activation of immune cells, and inflammatory responses. We also describe the comprehensive functions of IFN-λ in anti-enteric SARS-CoV-2 infection, and discuss the potential application of IFN-λ as a therapeutic agent for COVID-19 with intestinal symptoms.

Extremely hair follicle density is associated with a significantly different cecal microbiota in rex rabbits.

It has become increasingly clear that gut microbiota and skin are interconnected since the discovery of the 'gut-brain-skin' axis. Hair follicles (HFs) are skin microorganisms, but few studies have investigated their relationship to gut microbiota. Hence, we hypothesize that HFs have a close relationship with the gut, similarly to what was reported for the skin. Using rex rabbits as an animal model, one hundred healthy half-sibling rex rabbits were selected for the experiment, and 16 s rRNA gene sequencing was performed on the cecal microbiota of nine rabbits with the extremely high (HS) and low (LS) hair density (n = 9 per group) to determine differences between the composition and function of these communities. In comparison with the LS group, several alpha diversity index values were significantly lower in the HS group, although the higher variation in species composition in the HS group. Additionally, species diversity and abundance differed significantly in the cecum microbiota of HS and LS rabbits. Further, primary and secondary HF density was significantly correlated with the families Muribaculaceae and Bacteroidaceae, and genera Blautia, Bacteroides and Desulfovibrio. In particular, Muribaculaceae, Bacteroidaceae, Blautia and Bacteroides may support the development of HFs. Moreover, the expression of WNT4, WNT10a, WNT10b, CTNNB1 (ß-catenin) and LEF1 in the skin was significantly higher in the HS group compared with the LS group. Altogether, the results of this study suggest that the extremely high density of HF in rabbits is associated with a significantly different microbiota diversity and community structure, and the Wnt/ß-catenin signalling pathway was activated in the HS group. Thus, key bacteria may promote the development of HF.

Design and Implementation of Three-Channel Drainage Pipeline Ground Penetrating Radar Device.

In order to solve the current problems that conventional video inspection can only detect, as an internal pipeline defect and drainage pipeline radar inspection device detects in a single direction and at radar frequency in water pipeline defect detection, a three-channel drainage pipeline ground penetrating radar (GPR) inspection device was designed and developed, the assembly and commissioning of the device prototype were completed, and an actual engineering test application was carried out. Focusing on the problem that the detection direction and depth of the single-channel detection device are limited, a three-channel drainage pipeline GPR inspection device is designed to realize the synchronous detection of the inside of the pipeline, the pipeline body, and the external environment of the pipeline, improving the detection depth and efficiency. According to the design scheme of the three-channel drainage pipeline GPR inspection device, the assembly of the device prototype was completed. The device contains three radar channels, the top of the main frequency of the antenna is 1.4 GHz, the two sides are 750 MHz, the video camera has a pixel count of 4 million, and the positioning accuracy is less than 1 mm, the waterproof grade is IP68, the detection accuracy of pipe deformation (slope) is 0.1°, the detection depth outside the pipe is 1.2 m, and the detection accuracy of corrosion thickness is 15 mm. In a practical application of the device, the Jianguomenqiao sewage pipeline in Beijing, China, was tested, resulting in the discovery of 87 defects, including 39 loose soil areas at the bottom of the pipe exterior, 40 void areas, and 8 cavities.

Indocyanine green inhalation visualizes lung tumour during video-assisted thoracoscopic surgery.

OBJECTIVES: Accurate intraoperative identification of small lung tumours is crucial for precise resection of these lesions during video-assisted thoracoscopic surgery (VATS). This study aimed to evaluate the feasibility and safety of indocyanine green (ICG) inhalation for intraoperative visualization of lung tumours. METHODS: From January 2022 to May 2022, 43 patients with lung nodules were included into this study. All patients received intraoperative ICG inhalation for visualization of lung tumours under near-infrared (NIR) imaging. The primary outcomes of this trial were the detection rate and background-tumour-ratio (BTR) of lung nodules, and the secondary objectives were time to search for nodules and operative time to nodules excision. RESULTS: A total of 50 pulmonary nodules in 43 patients were identified and completely resected. And 44 lung nodules were detected during intraoperative fluorescent exploration with a median inhaled ICG dose of 18.8 mg. In vivo, the median BTR was 7.10. The median detection time of nodules was 100 seconds and the median operative time to nodules excision was 18 min. Quantification analysis showed that the fluorescence intensity of postoperative sputum declined to about 10% of the first fluorescent sputum within 20 hours. No adverse events attributed to ICG inhalation were recorded during the follow-up period. CONCLUSIONS: Intraoperative inhalation of ICG was a feasible and safe method for detection of lung tumours at low dose of ICG. This technique could be a remedial measure for identification of unpalpable lung nodules without preoperative localization. TRIAL REGISTRATION: Chinese Clinical Trial Registry, Identifier: ChiCTR2100053708.

Prevalence and Characterization of Serratia marcescens Isolated from Clinical Bovine Mastitis Cases in Ningxia Hui Autonomous Region of China.

Purpose: This study aimed to investigate the prevalence and genetic characterization of Serratia marcescens isolates from clinical bovine mastitis in Ningxia Hui Autonomous Region of China. Methods: S. marcescens was identified by the polymerase-chain reaction of 16S rRNA gene and sequencing. Antimicrobial susceptibility was tested by the disk diffusion method. Genes of resistance and virulence were determined by the PCR. Results: Overall, S. marcescens were confirmed from 32 of 2897 (1.1%) mastitis milk samples. These isolates showed high resistance to cefazolin (30/32, 93.8%) and chloramphenicol (28/32, 87.5%). A 12.5% (4/32) of the isolates displayed multidrug resistance (MDR). The most prevalent resistant genes found in S. marcescens were TEM (32/32, 100%) and CTX-M (24/32, 75.0%; CTX-M-15, 14/32, 43.8%; CTX-M-14, 8/32, 25.0%; CTX-M-65, 2/32, 6.3%) for extended-spectrum beta-lactamase, cmlA (28/32, 87.5%) and floR (16/32, 50.0%) for chloramphenicol resistance, SIM-1 (2/32, 6.3%) for carbapenemases, and sdeB (28/32, 87.5%), sdeY (26/32, 81.3%), sdeR (26/32, 81.3%) and sdeD (20/32, 62.5%) for efflux pumps. Moreover, all isolates carried virulence genes flhD, entB, and kpn, and most of them contained mrkD (30/32, 93.8%), ycfM (26/32, 81.3%), bsmB (26/32, 81.3%), pigP (26/32, 81.3%), kfu (24/32, 75.0%) and shlB (24/32, 75.0%). Conclusion: To our knowledge, this is the first report of genetic determinants for antimicrobial resistance and virulence in S. marcescens isolated from bovine mastitis cases in China. These findings are useful for developing strategies for prevention and treatment of bovine mastitis caused by S. marcescens in China.

TRAF4-mediated nonproteolytic ubiquitination of androgen receptor promotes castration-resistant prostate cancer.

Castration-resistant prostate cancer (CRPC) poses a major clinical challenge with the androgen receptor (AR) remaining to be a critical oncogenic player. Several lines of evidence indicate that AR induces a distinct transcriptional program after androgen deprivation in CRPCs. However, the mechanism triggering AR binding to a distinct set of genomic loci in CRPC and how it promotes CRPC development remain unclear. We demonstrate here that atypical ubiquitination of AR mediated by an E3 ubiquitin ligase TRAF4 plays an important role in this process. TRAF4 is highly expressed in CRPCs and promotes CRPC development. It mediates K27-linked ubiquitination at the C-terminal tail of AR and increases its association with the pioneer factor FOXA1. Consequently, AR binds to a distinct set of genomic loci enriched with FOXA1- and HOXB13-binding motifs to drive different transcriptional programs including an olfactory transduction pathway. Through the surprising upregulation of olfactory receptor gene transcription, TRAF4 increases intracellular cAMP levels and boosts E2F transcription factor activity to promote cell proliferation under androgen deprivation conditions. Altogether, these findings reveal a posttranslational mechanism driving AR-regulated transcriptional reprogramming to provide survival advantages for prostate cancer cells under castration conditions.

Low Pi stress enhances the sensitivity of hepatocellular carcinoma to sorafenib.

Sorafenib is a tyrosine kinase inhibitor for the treatment of advanced-stage HCC; however, clinical trials of sorafenib failed to demonstrate long-term survival benefits due to drug resistance. Low Pi stress has been shown to inhibit tumor growth and the expression of multidrug resistance-associated proteins. In this study, we investigated the sensitivity of HCC to sorafenib under conditions of low Pi stress. As a result, we found that low Pi stress facilitated sorafenib-mediated suppression of migration and invasion of HepG-2 and Hepa1-6 cells by decreasing the phosphorylation or expression of AKT, Erk and MMP-9. Angiogenesis was inhibited due to decreased expression of PDGFR under low Pi stress. Low Pi stress also decreased the viability of sorafenib-resistant cells by directly regulating the expression of AKT, HIF-1a and P62. In vivo drug sensitivity analysis in the four animal models showed a similar tendency that low Pi stress enhances sorafenib sensitivity in both the normal and drug-resistant models. Altogether, low Pi stress enhances the sensitivity of hepatocellular carcinoma to sorafenib and expands the indications for sevelamer.