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Erianin, a natural product derived from Dendrobium chrysotoxum Lindl, has been proved to play antitumor activity in various cancers. However, the effects and molecular mechanisms of erianin in bladder cancer cells remain unexplored. In this study, we found that erianin triggered cell death and cell cycle arrest in bladder cancer cells. Then we demonstrated that erianin could promote the accumulation of lethal lipid-based reactive oxygen species (ROS) and the depletion of glutathione (GSH), suggesting the induction of ferroptosis. In the further study, the ferroptosis inhibitor deferoxamine (DFO), N-Acetylcysteine (NAC) and GSH but not necrostatin-1, CQ or Z-VAD-FMK rescued erianin-caused cell death, showing ferroptosis played a major role in erianin-caused cell death. In vivo, we also showed that erianin suppressed the tumor growth by inducing ferroptosis. Mechanistically, we demonstrated that nuclear factor E2-related factor 2 (NRF2) inactivation was a key determinant of ferroptosis caused by erianin. In bladder cancer cells, the compound tert-butylhydro-quinone (TBHQ), an activator of NRF2, suppressed erianin-induced ferroptosis. Whereas, NRF2 inhibition used shRNA augmented the ferroptosis response induced by erianin treatment. In conclusion, our data provide the first evidence that erianin can initiate ferroptosis-like cell death and lipid peroxidation in bladder cancer, which will hopefully become a promising anticancer compound for the treatment of bladder cancer.

Abstract Background Hepatocellular carcinoma (HCC) is a common primary malignant tumor which usually progresses to an advanced stage because of late diagnosis. Sorafenib (Sora) is a first line medicine for advanced stage HCC; however, it has been faced with enormous resistance. Simvastatin (Sim) is a cholesterol-lowering drug and has been reported to inhibit tumor growth. The present study aims to determine whether Sora and Sim co-treatment can improve Sora resistance in HCC. Methods The HCC cell line LM3 and an established Sora-resistant LM3 cell line (LM3-SR) were used to study the relationship between Sora resistance and aerobic glycolysis. Cell proliferation, apoptosis and glycolysis levels were analyzed by western blotting, flow cytometry analysis and biomedical tests. A xenograft model was also used to examine the effect of Sim in vivo. Detailed mechanistic studies were also undertaken by the use of activators and inhibitors, and lentivirus transfections. Results Our results demonstrated that the resistance to Sora was associated with enhanced aerobic glycolysis levels. Furthermore, LM3-SR cells were more sensitive to Sim than LM3 cells, suggesting that combined treatment with both Sora and Sim could enhance the sensitivity of LM3-SR cells to Sora. This finding may be due to the suppression of the HIF-1α/PPAR-γ/PKM2 axis. Conclusions Simvastatin can inhibit the HIF-1α/PPAR-γ/PKM2 axis, by suppressing PKM2-mediated glycolysis, resulting in decreased proliferation and increased apoptosis in HCC cells, and re-sensitizing HCC cells to Sora.

Dopamine (DA) and uric acid (UA) are small biological molecules involved in many important processes in the human body. Their concentrations are closely related to human health. Abnormal concentrations of these molecules lead to various diseases, such as Parkinson's and gout, so monitoring of DA and UA in blood and urine, respectively, is very meaningful in clinical analysis. Electrochemical sensor detection is a widely-used method in the field of biological analysis owing to its advantages of simple operation, high sensitivity, low cost, environmental friendliness, etc. In this paper, titanium nitride (TiN) nanomaterial with chrysanthemum morphology was synthesized by hydrothermal and reduction nitridation methods toward preparation of an effective electrochemical sensor for human testing. It was further combined with reduced graphene oxide (rGO) through the hydrothermal method to form a titanium nitride-reduced graphene oxide (TiN-rGO) composite material. The phase and morphology of the material were characterized and analyzed by scanning electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, and other test methods. The results show that the TiN-rGO composite material maintaines the three-dimensional chrysanthemum-like morphology of TiN, and the transparent and wrinkled morphology of rGO. The chrysanthemum-like TiN is uniformly coated with the layered rGO. The TiN-rGO/GCE electrochemical sensor was then prepared by modifying the glassy carbon electrode (GCE) with TiN-rGO composite material for the determination DA and UA levels in the human body. Due to the synergistic effect of TiN and rGO in the composite, the constructed electrochemical sensor exhibits excellent electrochemical performance. The detection results show that the detection limits of DA and UA for the TiN-rGO/GCE electrochemical sensor are 0.11 and 0.12 μmol·L−1, respectively, and the linear ranges are 0.5‒210 μmol·L−1 and 5‒350 μmol·L−1, respectively. TiN-rGO/GCE electrochemical sensor also has good anti-interference, reproducibility and stability, and has been successfully applied in the detection of DA and UA in real human samples.

Abstract Objective Hepatocellular carcinima is one of the most common tumors in clinic and also one of the leading causes of death from cancer worldwide. Quercetin shows significant effects on blocking the development of various cancers. Methods We used the human hepatocellular carcinoma LM3 and nude mice tumor model to assess the effects of quercetin in hepatocellular carcinoma and clarify its mechanism of action. We collected LM3 cell line treated with different doses of quercetin at different time periods and determined the vital indexes. The liver tissues of mice were collected and used for western boltting (WB), Hematoxylin and Eosin (H&E) and TUNEL staining. Results Results indicated that quercetin suppressed the Hepatocellular carcinoma (HCC) growth both in vivo and in vitro. Quercetin could disturb LM3 cells proliferation and cell cycle distribution, thus inducing apoptosis. At the same time, quercetin inhibited LM3 cells migration and invasion and promoted HCC autophagy. These effects at least partly depended on the down‐regulation of the activation of JAK2 and STAT3 by quercetin. Conclusion Quercetin inhibited hepatocellular carcinoma progression by modulating cell apoptosis, migration, invasion, and autophagy; and its effects were at least partly related with the JAK2/STAT3 signaling pathway.

Talaromyces marneffei is the only dimorphic species in its genus and causes a fatal systemic mycosis named talaromycosis. Our previous study indicated that knockdown of AcuD gene (encodes isocitrate lyase of glyoxylate bypass) of T. marneffei by RNA interference approach attenuated the virulence of T. marneffei, while the virulence of the AcuD knockout strains was not studied. In this study, T. marneffei-zebrafish infection model was successfully established through hindbrain microinjection with different amounts of T. marneffei yeast cells. After co-incubated at 28°C, the increasing T. marneffei inoculum doses result in greater larval mortality; and hyphae generation might be one virulence factor involved in T. marneffei-zebrafish infection. Moreover, the results demonstrated that the virulence of the ΔAcuD was significantly attenuated in this Zebrafish infection model.

Abstract Background The treatment for advanced primary hepatocellular carcinoma (HCC) is sorafenib (SORA), while HCC has become increasingly drug resistant with enhanced aerobic glycolysis. The present study aimed to examine the chemotherapeutic effects of a flavonoid proanthocyanidin B2 (PB2) on HCC. Methods Five kinds of HCC cell lines and LO2 were used to test the effect of PB2 on aerobic glycolysis. The proliferation, cell cycle, apoptosis and a xenograft mouse model were analyzed. Lentivirus overexpressed pyruvate kinase M2 (PKM2) or sh-PKM2 was used to verify the target of PB2. The detailed mechanism was investigated by immunofluorescence, co-immunoprecipitation, and western blotting. Results PB2 inhibited the proliferation, induced cell cycle arrest, and triggered apoptosis of HCC cells in vivo and in vitro. PB2 also suppressed glucose uptake and lactate levels via the direct inhibition of the key glycolytic enzyme, PKM2. In addition, PKM2 inhibited the nuclear translocation of PKM2 and co-localization of PKM2/HIF-1α in the nucleus, leading to the inhibition of aerobic glycolysis. Co-treatment with PB2 was also effective in enhancing the chemosensitivity of SORA. Conclusions PB2 inhibited the expression and nuclear translocation of PKM2, therefore disrupting the interaction between PKM2/HSP90/HIF-1α, to suppress aerobic glycolysis and proliferation, and trigger apoptosis in HCC via HIF-1α-mediated transcription suppression.

BackgroundBrain metastasis mainly originates from lung cancer. Napsin A and TTF-1 factors have frequently been detected in lung adenocarcinoma cases. Brain metastasis tumors with napsin A and TTF-1 positive are easily classified as lung adenocarcinoma origin. However, some thyroid cancers also exhibit these clinical features. Besides, lung is the most common metastasis of undifferential thyroid cancer. Therefore, it requires development of novel diagnostic tools to aid in distinguishing between pulmonary and thyroid origin.Patient FindingsWe reported a case that was initially diagnosed as brain metastatic lung cancer based on immunohistochemistry results. Analysis of next-generation sequencing (NGS) data from the brain lesion revealed that the cancer may have originated from the thyroid. We detected combo mutations in TERT promoter mutation, RET fusion and TP53, which are common in undifferential thyroid cancer (UTC), but rare for lung cancer. These results, coupled with identification of PAX8, indicated that this patient had UTC. Additionally, her three sons, despite being asymptomatic, were all diagnosed with papillary thyroid carcinoma.SummaryThe patient received anlotinib treatment and showed good clinical outcomes. One month after anlotinib treatment, the pulmonary nodules were found to be controlled, and the thyroid tumor drastically reduced, and tracheal compression relieved. She continued anlotinib treatment for the following two months, but died one month later because the treatment stopped owing to financial reasons. All her sons underwent total thyroidectomy with lymph node dissection.ConclusionsAlthough NGS has been reported to assist in diagnosis of the origin of some tumors, this is the first evidence of NGS for the determination of the origin of thyroid tumors. To our knowledge, this is the first time that a combination of multiple mutations has been used to help determine the origin of a tumor, compared with the previous single mutant gene. Moreover, this is the first evidence on the use of anlotinib for treatment of UTC with distant metastasis. Besides, all three sons of the patient had thyroid carcinoma in subsequent examinations, indicating high-risk for familial non-medullary thyroid cancer in UTC patients and necessity for performing thyroid ultrasound testing in other family members.

IntroductionThe numbers of retrieved lymph nodes (RLNs) and positive lymph nodes (PLNs) had a significant impact on the survival of patients with ampulla of vater cancer (AVC), but the optimal numbers of the both were controversial.ObjectiveThe cohort study aimed to explore the prognostic value and the optimal point of RLNs and PLNs for AVC.MethodsA total of 2347 AVC patients with M0 disease who underwent surgical resection and lymph node dissection from January 2004 to December 2013 were acquired from a prospective database.ResultsThe study found that the optimal cut-off values of RLNs were 18 in the N0 cohort and 16 in N1 or entire cohort due to the highest 5-year overall survival (OS) rate and disease-specific survival (DSS) rate and the separation of survival curves (all P < 0.05). In patients with RLNs ≥ 16, patients with PLN = 0 demonstrated significantly better 5-year OS and DSS rates (70.9% and 77.1%) compared to those with PLNs = 1-2 (41.6% and 44.7%; all P < 0.001), and patients with PLNs = 1-2 demonstrated significantly better 5-year OS and DSS rates (41.6% and 44.7%) compared to those with PLNs ≥ 3 (24.3% and 28.0%; all P < 0.001).ConclusionsThis article recommended that at least 16 lymph nodes will improve the prognosis of AVC patients undergoing surgery. The best cut-off values of PLNs recommended for this study were 0 and 2, which may accurately stratify patients.

Baicalin, as a natural active ingredient extracted and isolated from the traditional Chinese medicine Scutellaria baicalensis Georgi., has been potentially used in various areas for its antioxidative, antitumor, anti-inflammatory, and anti-proliferative activities. Although several studies have reported the antitumor effects of baicalin against various cancer types, its beneficial effects on lung cancer have not yet been elucidated. Therefore, the therapeutic effects and molecular mechanisms of baicalin on lung cancer cell lines H1299 and H1650 were investigated. Here, the results of its antitumor activity were shown. We found that Akt/mTOR pathway inhibition was the essential determinant in baicalin-induced cell cycle arrest. Furthermore, when the Akt Agonist SC79 or Akt plasmid transfection was performed, the antitumor effect of baicalin was significantly abrogated in both H1299 and H1650 cells. In conclusion, we found that baicalin exerted its antitumor activity mainly by inducing Akt-dependent cell cycle arrest and promoting apoptosis, which show great potential for developing a new drug for lung cancer treatment.

Hepatic ischemia and reperfusion injury is characterized by hepatocyte apoptosis, impaired autophagy, and oxidative stress. Fenofibrate, a commonly used antilipidemic drug, has been verified to exert hepatic protective effects in other cells and animal models. The purpose of this study was to identify the function of fenofibrate on mouse hepatic IR injury and discuss the possible mechanisms. A segmental (70%) hepatic warm ischemia model was established in Balb/c mice. Serum and liver tissue samples were collected for detecting pathological changes at 2, 8, and 24 h after reperfusion, while fenofibrate (50 mg/kg, 100 mg/kg) was injected intraperitoneally 1 hour prior to surgery. Compared to the IR group, pretreatment of FF could reduce the inflammatory response and inhibit apoptosis and autophagy. Furthermore, fenofibrate can activate PPAR-α, which is associated with the phosphorylation of AMPK.

Objective. The aim of this study is to confirm the hepatocellular protective functions of apigenin and the molecular mechanism on liver fibrosis in mice. Methods. Carbon tetrachloride (CCl4) and bile duct ligature (BDL) mouse fibrosis models were used to investigate the effects of apigenin on liver fibrosis. Sixty-six male C57 mice were randomly divided into eight groups, including the vehicle group, CCl4 group, CCl4+L-apigenin (20 mg/kg) group, CCl4+H-apigenin (40 mg/kg) group, sham group, BDL group, BDL+L-apigenin(20 mg/kg) group, and BDL+H-apigenin(40 mg/kg) group. Serum liver enzymes (ALT and AST), proteins associated with autophagy, and indicators linked with the TGF-β1/Smad3 and p38/PPARα pathways were detected using qRT-PCR, immunohistochemical staining, and western blotting. Results. Our findings confirmed that apigenin could decrease the levels of ALT and AST, suppress the generation of ECM, inhibit the activation of HSCs, regulate the balance of MMP2 and TIMP1, reduce the expression of autophagy-linked protein, and restrain the TGF-β1/Smad3 and p38/PPARα pathways. Conclusion. Apigenin could alleviate liver fibrosis by inhibiting hepatic stellate cell activation and autophagy via TGF-β1/Smad3 and p38/PPARα pathways.

Microbial electrosynthesis (MES) or electro-fermentation (EF) is a promising microbial electrochemical technology for the synthesis of valuable chemicals or high-value fuels with aid of microbial cells as catalysts. By introducing electrical energy (current), fermentation environments can be altered or controlled in which the microbial cells are affected. The key role for electrical energy is to supply electrons to microbial metabolism. To realize electricity utility, a process termed inward extracellular electron transfer (EET) is necessary, and its efficiency is crucial to bioelectrochemical systems. The use of electron mediators was one of the main ways to realize electron transfer and improve EET efficiency. To break through some limitation of exogenous electron mediators, we introduced the phenazine-1-carboxylic acid (PCA) pathway from Pseudomonas aeruginosa PAO1 into Escherichia coli. The engineered E. coli facilitated reduction of fumarate by using PCA as endogenous electron mediator driven by electricity. Furthermore, the heterologously expressed PCA pathway in E. coli led to better EET efficiency and a strong metabolic shift to greater production of reduced metabolites, but lower biomass in the system. Then, we found that synthesis of adenosine triphosphate (ATP), as the “energy currency” in metabolism, was also affected. The reduction of menaquinon was demonstrated as one of the key reactions in self-excreted PCA-mediated succinate electrosynthesis. This study demonstrates the feasibility of electron transfer between the electrode and E. coli cells using heterologous self-excreted PCA as an electron transfer mediator in a bioelectrochemical system and lays a foundation for subsequent optimization.

Artemisia selengensis straw is an agricultural residue with great potential as a renewable resource because it is rich in lignocellulose. In this study, A. selengensis straw was used as a source of hemicelluloses (ASH) and cellulose nanocrystals (ASCNC) to produce biodegradable films. Different content levels of ASCNC were used as additives to improve composite films with ASH and polyvinyl alcohol (PVA). Various mechanical and hydrophobic properties of the films were analyzed. The composite films enhanced by ASCNC exhibited greater strength and were more effective as a barrier to water vapor when compared to that of the control ASH/PVA film. The tensile strength of the composite film was increased 80.1% to 36.21 MPa with ASCNC loading exceeding 9%, and the water vapor transmission rate decreased 15.45% when 12% ASCNC was added. Furthermore, the ASCNC-enhanced ASH/PVA composite film reduced a greater amount of light transmission than the control film.

ObjectiveColorectal cancer is a malignant tumor of the digestive system with high morbidity and mortality. 5-fluorouracil remains a widely used chemotherapeutic drug in the treatment of advanced colorectal cancer, but chemotherapy drugs are prone to develop drug resistance, p53 deletion or mutation is an important reason for the resistance of colorectal cancer cells to 5-fluorouracil. β-elemene has been proved to have the potential of reverse chemotherapy drug resistance, but the mechanism is unknown. This study aimed to investigate the effect of β-elemene to 5-fluorouracil in drug-resistant p53-deficient colorectal cancer cells HCT116p53–/–, and determine the possible molecular mechanism of β-elemene to reverse 5-fluorouracil resistance.MethodsThe effect of β-elemene on HCT116p53–/– cell activity was detected by Cell counting Kit-8. Cell proliferation was detected by monoclonal plate. The apoptosis was detected by flow cytometry and western blot. The autophagy was detected by western blot, immunofluorescence and transmission electron microscope. Determine the role of Cyclin-related protein Cyclin D3 in β-elemene reversing the resistance of HCT116p53–/– to 5-fluorouracil was detected by overexpression of Cyclin D3. The effect of β-elemene on the tumorigenic ability of p53-deficient colorectal cancer cells was detected establishing HCT116p53–/– all line xenograft model.ResultsFor p53 wildtype colorectal cancer cells, β-elemene could augment the sensitivity of 5-fluorouracil, for p53-deficient colorectal cancer cells, β-elemene significantly inhibited cell proliferation in a concentration-dependent manner, and reversed the resistance of HCT116p53–/– to 5-fluorouracil by inducing pro-death autophagy and Cyclin D3-dependent cycle arrest.Conclusionβ-elemene enhances the sensitivity of p53 wild-type cells to 5-fluorouracil, β-elemene can reverse the resistance of HCT116p53–/– to 5-fluorouracil by inducing pro-death autophagy and Cyclin D3-dependent cycle arrest in p53-deficient colorectal cancer, which will provide a new method for the treatment of p53 deletion colorectal cancer patients.

This paper presents a three-dimensional (3D) massive multiple-input and multiple-output (MIMO) antenna array model, which includes the spherical array assumption and geometric properties for future fifth generation (5G) wireless communications. A parametric approximation algorithm is developed for estimating the spatial fading correlations (SFCs) and channel capacities of the 3D massive MIMO antenna array systems under different power angular spectrum (PAS). The relationship between correlation with the spacing of antenna arrays and angular parameters was classified. The results show that the simulation values of the approximate method fit the theoretical calculation very well, thereby validating the feasibility of the proposed 3D large-scale massive MIMO model.

Document classification requires to extract high-level features from low-level word vectors. Typically, feature extraction by deep neural networks makes use of all words in a document, which cannot scale well for a long document. In this paper, we propose to tackle the long document classification task by incorporating the recurrent attention learning framework, which can produce the discriminative features with significantly less words. Specifically, the core work is to train a recurrent neural network (RNN)-based controller, which can focus its attention on the discriminative parts. Then, the glimpsed feature is extracted by a typical short text level convolutional neural network (CNN) from the focused group of words. The controller locates its attention according to the context information, which consists of the coarse representation of the original document and the memorized glimpsed features. By glimpsing a few groups, the document can be classified by aggregating these glimpsed features and the coarse representation. For our collected 11-class 10 000-word arXiv paper data set, the proposed method outperforms two subsampled deep CNN baseline models by a large margin given much less observed words.

A novel circuit theory of all-pass Negative group delay (NGD) function is investigated. The NGD function is implemented with unity direct chain feedback (UDCF) system. The active circuit is built with an operational amplifier in feedback with a lossy transmission line (TL). The UDCF topology S-parameter model is established versus TL parameters. The NGD analysis is elaborated from the frequency dependent transmission coefficient. The NGD behavior characterization is developed. The synthesis method allowing to determine the UDCF topology parameters in function of the targeted NGD values, gain and reflection coefficient is formulated. The all-pass NGD function is validated with a proof-of-concept (POC) design. Frequency and time domain simulations confirm the feasibility of the innovative all-pass NGD function. With S-parameter analysis, it was shown that the UDCF circuit exhibits NGD up to -7-ns with gain more than 0-dB and reflection coefficient -20-dB. More importantly, time-domain analysis illustrates how the transient tested voltage outputs can be in advance compared to the input.

This paper introduces a negative group delay (NGD) theory of passive topology presenting “IL”-shape geometry. The two-port topology is comprised of a coupled line with an open- and stub-loaded terminations. After introduction of the electrical equivalent circuit, the S-parameter modeling of the IL topology is established. The transmission and reflection parameters are analytically expressed. The IL topology NGD characterization is developed in function of the design parameters. The relevance of the NGD theory is verified with fully distributed and compact prototypes implemented in microstrip technology. The tested results confirm the possibility to generate bandpass NGD function with IL topology. The calculated model, simulation and measurements are in very good correlation. Validation results showing center frequencies at about 1.55 GHz and 3.55 GHz with NGD level better than -1 ns over 30 MHz bandwidth. The IL NGD prototypes present transmission and reflection coefficients better than -3 dB and -10 dB, respectively.

Cronobacter sakazakii 505108 was isolated from a sputum specimen of a neonate with severe pneumonia. C. sakazakii 505108 co-harbors 3 resistance plasmids of the IncHI2, IncX3, and IncFIB incomparability groups, respectively. These 3 plasmids have acquired several accessory modules, which carry an extremely large number of resistance genes, especially including those involved in resistance to carbapenems, aminoglycoside, tetracyclines, and phenicols and sulphonamide/trimethoprim. These plasmid-borne antibiotic resistance genes were associated with insertion sequences, integrons, and transposons, indicating that the assembly and mobilization of the corresponding accessory modules with complex chimera structures are facilitated by transposition and/or homologous recombination. This is the first report of fully sequence plasmids in clinical Cronobacter, which provides a deeper insight into plasmid-mediated multi-drug resistance in Cronobacter from hospital settings.

The nutritional value of cow milk mainly depends on its fatty acid content and protein composition. The identification of genes controlling milk production traits and their regulatory mechanisms is particularly important for accelerating genetic progress in the breeding of dairy cows. On the basis of mammary gland transcriptome analyses, in this study we identified an miRNA, bta-miR-125a, that could control bovine milk-fat production by targeting the 3′ untranslated region (UTR) of the serum amyloid A-1 (SAA1) mRNA. The presence of synthetic bta-miR-125a (i.e., an miR-125a mimic) significantly down-regulated the expression of luciferase from mRNAs containing the binding sequence for bta-miR-125a in the 3′-UTR in a dual-luciferase reporter assay. Furthermore, the presence of the miR-125a mimic decreased the steady-state level of the SAA1 protein, but increased the accumulation of triglycerides and cholesterol content in bovine mammary epithelial cells (MAC-Ts). Blocking the function of bta-miR-125a using a specific inhibitor decreased the level of triglycerides and cholesterol content in the cells. These results indicate that bta-miR-125a can serve as a positive regulator of lipid synthesis in mammary epithelial cells, which acts by targeting SAA1 gene expression.

BODIPY dyes, though widely explored, have not been pursued as chromophore reaction based chemical probes. Here, the authors synthesize a meso-naked BODIPY core flanked with two electron-withdrawing groups, which undergoes a reversible change in conjugated structure in the presence of base and functions as a dual signal and ultrahigh turn-on ratio chemical probe.

Physiological (metabolite analysis) and molecular (gene expression) approaches were used to understand the mechanism underlying russet formation in response to the application of GA3 and CPPU (Forchlorfenuron) in a Japanese table grape cultivar ‘Shine Muscat’. Several different concentrations of GA3 and GA3 + CPPU [25 mg L−1 GA3 (A), 25 mg L−1 GA3 + 5 mg L−1 CPPU (B), 25 mg L−1 GA3 + 10 mg L−1 CPPU (C), and 25 mg L−1 GA3 + 15 mg L−1 CPPU (D)] were applied to grape berry clusters at two weeks after flowering (WAF). No russet was observed on the berries treated with the ‘C’ combination. Lower levels of phenylalanine ammonia-lyase (PAL) activity was observed in the treated samples, relative to the untreated material. Reduced peroxide (POD) activity was also observed in response to different treatments, while the expression of Peroxidase 17 and Phenylalanine ammonia-lyase G1 genes mirrored lignin content. Increased activity of 4-coenzyme A ligase (4CL) may contribute to decreasing the level of russet and help to improve grape berry quality.

Abstract Background Understanding the bioprocess limitations is critical for the efficient design of biocatalysts to facilitate process feasibility and improve process economics. In this study, a proline hydroxylation process with recombinant Escherichia coli expressing l-proline cis-4-hydroxylase (SmP4H) was investigated. The factors that influencing the metabolism of microbial hosts and process economics were focused on for the optimization of cis-4-hydroxy-l-proline (CHOP) production. Results In recombinant E. coli, SmP4H synthesis limitation was observed. After the optimization of expression system, CHOP production was improved in accordance with the enhanced SmP4H synthesis. Furthermore, the effects of the regulation of proline uptake and metabolism on whole-cell catalytic activity were investigated. The improved CHOP production by repressing putA gene responsible for l-proline degradation or overexpressing l-proline transporter putP on CHOP production suggested the important role of substrate uptake and metabolism on the whole-cell biocatalyst efficiency. Through genetically modifying these factors, the biocatalyst activity was significantly improved, and CHOP production was increased by twofold. Meanwhile, to further improve process economics, a two-strain coupling whole-cell system was established to supply co-substrate (α-ketoglutarate, α-KG) with a cheaper chemical l-glutamate as a starting material, and 13.5 g/L of CHOP was successfully produced. Conclusions In this study, SmP4H expression, and l-proline uptake and degradation, were uncovered as the hurdles for microbial production of CHOP. Accordingly, the whole-cell biocatalysts were metabolically engineered for enhancing CHOP production. Meanwhile, a two-strain biotransformation system for CHOP biosynthesis was developed aiming at supplying α-KG more economically. Our work provided valuable insights into the design of recombinant microorganism to improve the biotransformation efficiency that catalyzed by Fe(II)/α-KG-dependent dioxygenase.

Multilayer colloidal crystal has been prepared by the layer-by-layer deposition of silica microspheres on a glass slide. Each layer is a slab consisting of a fcc close-packed colloidal arrays. By properly choosing the sizes of spheres, the whole spectral feature of multilayer colloidal crystal can be tuned. Here, we engineered a multilayer superlattice structure with an effective passband between two stop bands. This gives a strong narrowing effect on emission spectrum. With the stop bands at the shortwave and longwave edges of emission spectrum, the passband in the central wavelength region can be regarded as a strong decrease of suppression effect and enhancement of a narrow wavelength region of emission. The spectral narrowing modification effect of suitably engineered colloidal crystals shows up their importance in potential application as optical filters and lasing devices.DOI: http://dx.doi.org/10.5755/j01.ms.23.3.16320

Abstract Background This study aims to characterize genetically related class 1 integrons In1069, In893 and In1287 to In1290, and to further propose a scheme of stepwise integration or excision of individual gene cassettes (GCs) to generation of these integron variations. Methods Six of 139 non-redundant Enterobacteriaceae strains were studied by bacterial antimicrobial susceptibility testing, detection of carbapenemase activity, and integron sequencing and sequence comparison. Results Six novel class 1 integrons, In0, In1069, and In1287 to In1290, together with the previously characterized In893, were determined from the above strains. An unusual bla KPC-2-carrying In0 and the bla IMP-30-carrying In1069 coexists in a single isolate of Escherichia coli. In0 contains a PcH1 promoter and a truncated aacA4’-3 gene cassette (GCaacA4’-3), as well as a bla KPC-2-containing region of Tn6296 integrated between PcH1 and GCaacA4’-3. In1069 carries GCbla IMP-30 and GCaacA4’-3 in this order. The other five integrons, In893 and In1287 to In1290, are genetically related to In1069, and all possess a core GCaacA4’-3. The integration or excision of one or more individual gene cassettes, such as GCbla IMP-30, GCaadA16, GCcatB3, GCarr3 and GCdfrA27, upstream or downstream of GCaacA4’-3 generates various gene cassettes arrays among these five integrons. Conclusions These findings provide the insight into stepwise and parallel evolution of In1069-associated integron variations likely under antibiotic selection pressure in clinical settings.

Abstract This study aimed to genetically characterize two fully-sequenced novel IncFII-type multidrug resistant (MDR) plasmids, p0716-KPC and p12181-KPC, recovered from two different clinical Klebsiella pneumoniae isolates. p0716-KPC and p12181-KPC had a very similar genomic content. The backbones of p0716-KPC/p12181-KPC contained two different replicons (belonging to a novel IncFII subtype and the Rep_3 family), the IncFIIK and IncFIIY maintenance regions, and conjugal transfer gene sets from IncFIIK-type plasmids and unknown origins. p0716-KPC and p12181-KPC carried similar three accessory resistance regions, namely ΔTn6209, a MDR region, and the bla KPC-2 region. Resistance genes bla KPC-2, mph(A), strAB, aacC2, qacEΔ1, sul1, sul2, and dfrA25, which are associated with transposons, integrons, and insertion sequence-based mobile units, were located in these accessory regions. p0716-KPC carried two additional resistance genes: aphA1a and bla TEM-1. Together, our analyses showed that p0716-KPC and p12181-KPC belong to a novel IncFII subtype and display a complex chimeric nature, and that the carbapenem resistance gene bla KPC-2 coexists with a lot of additional resistance genes on these two plasmids.

IntroductionThe American Joint Committee on Cancer (AJCC) recommended retrieval of at least 12 lymph nodes and firstly classified N category by the number of positive lymph nodes (PLNs) for Distal Cholangiocarcinoma (DCC).ObjectiveThe end of this cohort study was to explore the optimal cut-off values of the number of examined lymph nodes (ELNs) and PLNs to better stratify patients by utilizing a population-based database.MethodsA number of 758 patients with DCC from the Surveillance, Epidemiology, and End Results (SEER) database were enrolled in the study and comparing by the survival analysis.ResultsSurvival analysis found that patients with ELNs < 5 had a lower 3-year disease-specific survival rate than ELNs ≥ 5 in N0M0 cohort (35.3% vs. 53.0%, P = 0.001) and in M0 cohort (42.7% vs. 32.8%, P = 0.006); survival curves between patients with ELNs < 12 and ELNs ≥ 12 were overlapped in N0M0 cohort (P = 0.256) and in M0 cohort (P = 0.233). Among patients with ELNs ≥ 5, using the optimal cut-off value of the number of PLNs (0, 2) could accurately stratify patients, but the recommendation of the number of PLNs (0, 3) by the AJCC could not.ConclusionsThis study recommended examining at least 5 lymph nodes and defining PLNs = 1-2 as the N1 category and PLNs ≥ 3 as the N2 category, which may better stratify distal cholangiocarcinoma patients and improve the accuracy of the eighth edition AJCC staging.

As a core driving force of the most recent round of industrial transformation, artificial intelligence has triggered significant changes in the world economic structure, profoundly changed our life and way of thinking, and achieved an overall leap in social productivity. This paper aims to examine the effect of knowledge transfer performance on the artificial intelligence industry innovation network and the path artificial intelligence enterprises can take to promote sustainable development through knowledge transfer in the above context. First, we construct a theoretical hypothesis and conceptual model of the innovation network knowledge transfer mechanism within the artificial intelligence industry. Then, we collect data from questionnaires distributed to Chinese artificial intelligence enterprises that participate in the innovation network. Moreover, we empirically analyze the impact of innovation network characteristics, organizational distance, knowledge transfer characteristics, and knowledge receiver characteristics on knowledge transfer performance and verify the hypotheses proposed in the conceptual model. The results indicate that innovation network centrality and organizational culture distance have a significant effect on knowledge transfer performance, with influencing factors including network scale, implicit knowledge transfer, receiver's willingness to receive, and receiver's capacity to absorb knowledge. For sustainable knowledge transfer performance on promoting Chinese artificial intelligence enterprises innovation, this paper finally delivers valuable insights and suggestions.

Liver fibrosis is a pathological process involving diffuse extracellular matrix (ECM) deposition in the liver. It is typical of many chronic liver diseases, including cirrhosis, and effective drugs are needed. In this study, we explored the protective effect of bergenin on liver fibrosis induced by carbon tetrachloride and bile duct ligation. A variety of molecular biological methods (qRT-PCR, western blotting, and immunohistochemistry) were employed to confirm the increased degree of hepatocyte injury and ECM formation in the disease model, consistent with autophagy and activation of the TGF-β pathway. Bergenin activated PPAR-γ and inhibited TGF-β and autophagy and decreased liver fibrosis by inhibiting hepatocyte necrosis and ECM formation in a dose-dependent manner. The results suggest that bergenin may be a promising drug candidate for the treatment of liver fibrosis.

Mobile gene elements play an important role in the continuous evolution of the prophage DNA of bacteria, promoting the emergence of new gene structures. This study explored the evolution of four strains of Klebsiella pneumoniae harboring prophages, 19051, 721005, 911021, and 675920, and 16 genomes of K. pneumoniae from GenBank. The results revealed a wide range of genetic variation in the prophage DNA inserted into the sap sites of K. pneumoniae chromosomes. From analysis and comparison of the sequences of the 20 prophage DNAs determined from the four strains and the 16 GenBank genomes of K. pneumoniae using high-throughput sequencing and antimicrobial susceptibility tests, we identified a novel transposon, Tn6556. We also identified at least nine large genetic structures with massive genetic acquisitions or losses and five hotspot sites showing a tendency to undergo insertion of gene elements such as IS1T, IS1R, IS26, ISKpn26, ISKpn28, Tn6556, MDR, and In27-related regions as variable regions; however, the only highly conserved core genes were int and umuCD among the 20 prophage DNAs. These findings provide important insights into the evolutionary diversity of bacteriophage DNA contained in K. pneumoniae.

Pinus massoniana bark extract (PMBE) is a natural product developed in China, with flavonoids, especially proanthocyanidins, as its main components. Flavonoids are shown to inhibit the abnormally high expression of cathepsin B (CatB) in some tumour cells. CatB is normally located in lysosomes and plays a role in the autophagy process. However, if the cells turn malignant, CatB may secrete and take part in the metastasis of cancer cells. Some researchers have reported that PMBE can inhibit the migration of HeLa cells. Thus, this research aimed to examine whether PMBE inhibits the invasion of HeLa cells. We first confirmed the metastasis capability of HeLa cells and then explored the effects of PMBE on the expression of CatB. Our results proved that HeLa cells can metastasise across a Matrigel® matrix and this invasion capability can be significantly inhibited by PMBE, possibly by down-regulating the expression of CatB, mainly at the transcriptional level.

Glutarate is an important C5 platform chemical produced during the catabolism of L-lysine through 5-aminovalerate (5-AMV) pathway. Here, we first established a whole-cell biocatalysis system for the glutarate production from L-lysine with the engineered Escherichia coli (E. coli) that co-expressed DavAB and GabDT. However, the accumulation of intermediate 5-AMV was identified as one important factor limiting glutarate production. Meanwhile, the negative interaction of co-expressing DavAB and GabDT in a single cell was also confirmed. Here, we solved these problems through engineering a microbial consortium composed of two engineered E. coli strains, BL21-22AB and BL21-YDT, as the whole-cell biocatalysts, each of which contains a part of the glutarate pathway. After the optimization of bioconversion conditions, including temperature, metal ion additives, pH, and cell ratio, 17.2 g/L glutarate was obtained from 20 g/L L-lysine with a yield of 95.1%, which was improved by 19.2% compared with that in a single cell. Little accumulation of 5-AMV was detected. Even at the high substrate concentration, the reduced 5-AMV accumulation and increased glutarate production were achieved. This synthetic consortium produced 43.8 g/L glutarate via a fed-batch strategy, the highest titer reported to date.

This study presents three novel integrons In1394, In1395, and In1443, three novel unit transposons Tn6392, Tn6393, and Tn6403, one novel conjugative element (ICE) Tn6413, and the first sequenced IncP-7 resistance plasmid p1160-VIM from clinical Pseudomonas aeruginosa. Detailed sequence comparison of p1160-VIM (carrying Tn6392 and Tn6393) and Tn6413 (carrying Tn6403) with related elements were performed. Tn6392, Tn6393, and Tn6403 were generated from integration of In1394 (carrying blaVIM–24), In1395 and In1443 (carrying blaVIM–4) into prototype Tn3-family unit transposons Tn5563, Tn1403, and Tn6346, respectively. To the best of our knowledge, this is the first report of a blaVIM–24-carrying P. aeruginosa isolate.

IncI-complex plasmids can be divided into seven subgroups IncI1, IncI2, IncIγ, IncB/O, IncK1, IncK2, and IncZ. In this study, a replicon-based scheme was proposed for typing IncI-complex plasmids into four separately clustering subgroups IncI2, IncI1/B/O, IncIγ/K1 and IncK2/Z, the last three of which were combined from IncI1 and IncB/O, IncIγ and IncK1, and IncK2 and IncZ, respectively. Four IncIγ/K1 plasmids p205880-NR2, p14E509-CTXM, p11011-CTXM and p61806-CTXM were fully sequenced and compared with IncIγ/K1 reference pCT, IncI2 reference R721, IncI1/B/O reference R64 and IncK2/Z reference pO26-CRL-125. These plasmids shared conserved gene organization in the replication and conjugal transfer regions, but displaying considerable sequence diversity among different subgroups. Remarkable modular differences were observed in the maintenance and transfer leading regions. p205880-NR2 contained no resistance genes or accessory modules, while the other seven plasmids acquired one or more accessory modules, which harbored mobile elements [including unit transposons, insertion sequence (IS)-based transposition units and individual IS elements] and associated resistance markers (especially including those involved in resistance to β-lactams, aminoglycosides, tetracyclins, phenicols, streptomycins, trimethoprims, sulphonamides, tunicamycins and erythromycins). Data presented here provided a deeper insight into diversification and evolution of IncI-complex plasmids.

Hepatic ischemia reperfusion (IR) injury (IRI) occurs during liver transplantation, hepatectomy, and hemorrhagic shock. Oleanolic acid (OA) is a natural compound with antioxidant and anti-inflammatory activity that has been used to treat liver disorders in clinical practice for several years. Here, we investigated the effects and underlying mechanisms of OA in hepatic IRI. A 60-minute partial (70%) hepatic, warm, ischemic reperfusion model was established in BALB/c mice, and two doses (30 and 60 mg/kg) of OA were administered intragastrically for 7 consecutive days prior to hepatic IR. Orbital blood and liver specimens were collected at 2, 8, and 24 h after IR. The results showed that OA preconditioning significantly alleviated hepatic injury, as evidenced by decreased alanine aminotransferase and aspartate aminotransferase levels; improved histology, inhibition of JNK phosphorylation, and high mobility group box 1 (HMGB1); and tumor necrosis factor-α downregulation in hepatic IR mice. OA upregulated Bcl-2 and downregulated caspase-3, caspase-9, Bax, Beclin 1, and LC3, which play crucial roles in the regulation of apoptosis and autophagy. These findings highlighted the protective effects of OA against hepatic IRI mediated by the inhibition of apoptosis and autophagy and the release of HMGB1, which acted as a late inflammatory mediator in hepatic IRI.

Objective. Liver fibrosis is a consequence of wound-healing responses to chronic liver insult and may progress to liver cirrhosis if not controlled. This study investigated the protection against liver fibrosis by isorhamnetin. Methods. Mouse models of hepatic fibrosis were established by intraperitoneal injection of carbon tetrachloride (CCl4) or bile duct ligation (BDL). Isorhamnetin 10 or 30 mg/kg was administered by gavage 5 days per week for 8 weeks in the CCl4 model and for 2 weeks in the BDL model. Protein and mRNA expressions were assayed by western blotting, immunohistochemistry, and quantitative real-time polymerase chain reaction. Results. Isorhamnetin significantly inhibited liver fibrosis in both models, inhibiting hepatic stellate cell (HSC) activation, extracellular matrix (ECM) deposition, and autophagy. The effects were associated with downregulation of transforming growth factor β1 (TGF-β1) mediation of Smad3 and p38 mitogen-activated protein kinase (MAPK) signaling pathways. Conclusion. Isorhamnetin protected against liver fibrosis by reducing ECM formation and autophagy via inhibition of TGF-β1-mediated Smad3 and p38 MAPK signaling pathways.

Forty-five KPC-producing Enterobacteriaceae strains were isolated from multiple departments in a Chinese public hospital from 2014 to 2015. Genome sequencing of four representative strains, namely Proteus mirabilis GN2, Serratia marcescens GN26, Morganella morganii GN28, and Klebsiella aerogenes E20, indicated the presence of blaKPC-2-carrying IncX6 plasmids pGN2-KPC, pGN26-KPC, pGN28-KPC, and pE20-KPC in the four strains, respectively. These plasmids were genetically closely related to one another and to the only previously sequenced IncX6 plasmid, pKPC3_SZ. Each of the plasmids carried a single accessory module containing the blaKPC-2/3-carrying ΔTn6296 derivatives. The ΔTn6292 element from pGN26-KPC also contained qnrS, which was absent from all other plasmids. Overall, pKPC3_SZ-like blaKPC-carrying IncX6 plasmids were detected by PCR in 44.4% of the KPC-producing isolates, which included K. aerogenes, P. mirabilis, S. marcescens, M. morganii, Escherichia coli, and Klebsiella pneumoniae, and were obtained from six different departments of the hospital. Data presented herein provided insights into the genomic diversity and evolution of IncX6 plasmids, as well as the dissemination and epidemiology of blaKPC-carrying IncX6 plasmids among Enterobacteriaceae in a hospital setting.

In this paper, a distributed “Win-Win” reciprocal-selection-based medium access scheme (DWWRS-MAS) is designed for a cooperative spectrum leasing system hosting multiple licensed transmission pairs and multiple unlicensed transmission pairs. Based on the proposed DWWRS-MAS, the primary transmitter (PT) intends to lease its spectral resources to an appropriate secondary transmitter (ST) in exchange for cooperative transmission assistance for the sake of minimizing its transmit power and simultaneously satisfying its transmit rate requirement. The ST has an incentive to collaborate with the best PT for the sake of minimizing the ST's transmit power under the constraint of its Quality of Service (QoS) requirement, while simultaneously winning a transmission opportunity for its own traffic. Moreover, based on the matching theory and queueing theory, we analyze the algorithmic stability and the queueing stability of the cooperative spectrum leasing system exploiting our DWWRS-MAS, respectively. Simulation results demonstrate that our DWWRS-MAS is capable of providing both considerable energy savings and substantial rate improvements for the cooperative spectrum leasing system hosting multiple licensed transmission pairs and multiple unlicensed transmission pairs.

The codewords of a low-density parity-check (LDPC) convolutional code (LDPC-CC) are characterised into structured and non-structured. The number of the structured codewords is dominated by the size of the polynomial syndrome former matrix H T ( D ) , while the number of the non-structured ones depends on the particular monomials or polynomials in H T ( D ) . By evaluating the relationship of the codewords between the mother code and its super codes, the low weight non-structured codewords in the super codes can be eliminated by appropriately choosing the monomials or polynomials in H T ( D ) , resulting in improved distance spectrum of the mother code.

In dynamic propagation environments, beamforming algorithms may suffer from strong interference, steering vector mismatches, a low convergence speed and a high computational complexity. Reduced-rank signal processing techniques provide a way to address the problems mentioned above. This paper presents a low-complexity robust data-dependent dimensionality reduction based on an iterative optimization with steering vector perturbation (IOVP) algorithm for reduced-rank beamforming and steering vector estimation. The proposed robust optimization procedure jointly adjusts the parameters of a rank reduction matrix and an adaptive beamformer. The optimized rank reduction matrix projects the received signal vector onto a subspace with lower dimension. The beamformer/steering vector optimization is then performed in a reduced dimension subspace. We devise efficient stochastic gradient and recursive least-squares algorithms for implementing the proposed robust IOVP design. The proposed robust IOVP beamforming algorithms result in a faster convergence speed and an improved performance. Simulation results show that the proposed IOVP algorithms outperform some existing full-rank and reduced-rank algorithms with a comparable complexity.

Orthogonal frequency-division multiplexing (OFDM) systems over rapidly time varying channels may suffer from significant inter-carrier interference (ICI), which destroys the orthogonality between subcarriers and degrades the detection performance. Without sufficient ICI suppression, OFDM systems usually experience an error floor. According to the approximate matched filter bound (AMFB), the error floor in a coded OFDM system is not irreducible. In this work, we introduce novel multiple feedback matched filter (MBMF)-based ICI cancellation receivers. Based on the output of a novel MBMF scheme, the approach employs a multiple ICI cancellation strategy with or without signal-to-interference-plus-noise-ratio (SINR) ordering. The developed schemes can significantly improve the performance and remove the error floor with a negligible complexity increase. Given the multiple cancellation approach, we compare the SINR performance of the MBMF outputs with that employing single feedback and show that the SINR performance with multiple cancellation candidates is improved over that with a single one at practical SNR values. Additionally, for time-varying channels, we exploit partial fast Fourier transform (PFFT) by splitting one OFDM symbol into multiple segments; the channel state is separately estimated by least-squares (LS) methods without inserting more pilots. Simulation results demonstrate the superiority of the proposed methods over serial and block equalizers and the robustness to the Doppler effects compared to conventional single-segment method.