Cell Membrane-Anchoring Nano-Photosensitizer for Light-Controlled Calcium-Overload and Tumor-Specific Synergistic Therapy.

Poor selectivity and unintended toxicity to normal organs are major challenges in calcium ion (Ca2+ ) overload tumor therapy. To address this issue, a cell membrane-anchoring nano-photosensitizer (CMA-nPS) is constructed for inducing tumor-specific Ca2+ overload through multistage endogenous Ca2+ homeostasis disruption under light guidance, i.e., the extracellular Ca2+ influx caused by cell membrane damage, followed by the intracellular Ca2+ imbalance caused by mitochondrial dysfunction. CMA-nPS is decorated by two types of functionalized cell membranes, the azide-modified macrophage cell membrane is used to conjugate the dibenzocyclooctyne-decorated photosensitizer, and the vesicular stomatitis virus glycoprotein (VSV-G)-modified NIH3T3 cell membrane is used to guide the anchoring of photosensitizer to the lung cancer cell membrane. The in vitro study shows that CMA-nPS mainly anchors on the cell membrane, and further causes membrane damage, mitochondrial dysfunction, as well as intracellular Ca2+ overload upon light irradiation. Synergistically enhanced antitumor efficiency is observed in vitro and in vivo. This study provides a new synergistic strategy for Ca2+ -overload-based cancer therapy, as well as a strategy for anchoring photosensitizer on the cell membrane, offering broad application prospects for the treatment of lung cancer.

Iron vacancies engineering of FexC@NC hybrids toward enhanced lithium-ion storage properties.

Defect engineering have profound influence on the energy storage properties of electrode hybrids by adjusting their intrinsic electronic characteristics. For iron carbide based materials, however, the effect of defect (especially cation vacancies) toward their electrochemical performance are still unclear. Herein, the feasible and scalable synthesis of FexC@NC with 3D honeycomb-like carbon architecture and abundant Fe vacancies via template etching is reported. Such structure enable outstanding lithium-ion storage properties owing to hierarchical pores, improved intrinsic electrochemical activity, as well as the introduction of more active sites. As a result, the FexC@NC-2 presents a high reversible specific capacity of 1079 mAh g-1after 1000 cycles. Moreover, an excellent cycling stability can be achieved via maintaining a high-capacity retention (689 mAh g-1, 98.4%) over 1000 cycles at 5 A g-1. This study provides a feasible strategy for developing high-performance hybrids with hierarchical pore and rich defects structures.

[Rationality evaluation of Shuanghuanglian Injection combined with Ampicillin Sodium for Injection based on sequential analysis].

The lack of rationality evaluation method for drug combination has long restricted its clinical application. In view of this, this study took Shuanghuanglian Injection as model drug and established a "physical-chemical-biological" sequential analysis method, which is expected to provide clues for improving the safety and effectiveness of clinical drug combination. With the methods of insoluble particle testing, isothermal titration calorimetry(ITC), and real time cellular analysis(RTCA), the rationality of Shuanghuanglian Injection combined with Ampicillin Sodium for Injection was assessed. The results showed that the number of insoluble particles>10 µm in the solution of the combination met the standard of Chinese Pharmacopoeia, while the number of insoluble particles>25 µm did not meet the standard. ITC detection demonstrated that the change of Gibbs free energy(ΔG) was less than 0 during the fusion process, indicating that the process was spontaneous and enthalpy-driven reaction. Therefore, the interaction between the two was mainly chemical reaction, and the internal substances may change. RTCA found that Shuanghuanglian Injection alone and Ampicillin Sodium for Injection alone basically had no inhibitory effect on the growth of HEK293 T cells, while the combination of the two suppressed the growth of HEK293 T cells, suggesting that the combination was toxic to HEK293 T cells. This study showed that Shuanghuanglian Injection and Ampicillin Sodium for Injection reacted, yielding toxicity. This suggested that the two should not be combined for application. With the "physical-chemical-biological" sequential analysis, the molecular interaction of drugs was clarified. The method can be further applied for evaluating the rationality of other Chinese and western medicine injections.

[Mechanism of Jinqi Jiangtang Tablets in treatment of pancreatic ß cell dysfunction based on network pharmacology and molecular docking technology].

The present study investigated the therapeutic efficacy and potential mechanism of Jinqi Jiangtang Tablets(JQJT) on pancreatic ß cell dysfunction based on network pharmacology and molecular docking technology. TCMSP platform was used to retrieve the chemical components and targets of the three Chinese herbal medicines of JQJT. The genes were converted to gene symbol by the UniProt, and its intersection with targets related to pancreatic ß cell function in GeneCards and CTD databases was obtained. The drugs, active components and common targets were imported into Cytoscape 3.8.2 to plot the drug-component-target network. The main effective components and targets were obtained by software analysis. The drug targets and targets related to pancreatic ß cell function were imported separately into the STRING platform for the construction of protein-protein interaction(PPI) networks. The two PPI networks were merged by Cytoscape 3.8.2 and the key targets were obtained by plug-in CytoNCA. The targets obtained from drug-component-target network and PPI networks were imported into DAVID for GO analysis and KEGG enrichment analysis. AutoDock was used to carry out molecular docking of main active components and core targets and Pymol was used to plot the molecular docking diagram. The results showed that there were 371 active components and 203 targets related to JQJT and 2 523 targets related to pancreatic ß cell damage, covering 136 common targets. The results revealed core targets(such as PTGS2, PTGS1, NOS2, ESR1 and RXRA) and effective key components(such as quercetin, kaempferol, luteolin, ß-carotene and ß-sitosterol). KEGG enrichment analysis indicated that apoptosis, inflammation, and other signaling pathways were mainly involved. Molecular docking results showed that the main active components could spontaneously bind to the targets. This study preliminarily revealed the mechanism of JQJT in improving pancreatic ß cell damage through multi-component, multi-target and multi-pathway, and provided a theoretical basis for JQJT in the treatment of pancreatic ß cell dysfunction.

Characterization of a novel theme C glycoside hydrolase family 9 cellulase and its CBM-chimeric enzymes.

In bacterial cellulase systems, glycoside hydrolase family 9 (GH9) cellulases are generally regarded as the major cellulose-degrading factors besides GH48 exoglucanase. In this study, umcel9A, which was cloned from uncultured microorganisms from compost, with the encoded protein being theme C GH9 cellulase, was heterologously expressed in Escherichia coli, and the biochemical properties of the purified enzyme were characterized. Hydrolysis of carboxylmethylcellulose (CMC) by Umcel9A led to the decreased viscosity of CMC solution and production of reducing sugars. Interestingly, cellobiose was the major product when cellulosic materials were hydrolyzed by Umcel9A. Six representative carbohydrate-binding modules (CBMs) from different CBM families (CBM1, CBM2, CBM3, CBM4, CBM10, and CBM72) were fused with Umcel9A at the natural terminal position, resulting in significant enhancement of the binding capacity of the chimeric enzymes toward four different insoluble celluloses as compared with that of Umcel9A. Catalytic activity of the chimeric enzymes against insoluble celluloses, including phosphoric acid-swollen cellulose (PASC), alkali-pretreated sugarcane bagasse (ASB), filter paper powder (FPP), and Avicel, was higher than that of Umcel9A, except for Umcel9A-CBM3. In these chimeric enzymes, CBM4-Umcel9A exhibited the highest activity toward the four tested insoluble celluloses and displayed 4.2-, 3.0-, 2.4-, and 6.6-fold enhanced activity toward PASC, ASB, FPP, and Avicel, respectively, when compared with that of Umcel9A. CBM4-Umcel9A also showed highest V max and catalytic efficiency (k cat/K M) against PASC. Construction of chimeric enzymes may have potential applications in biocatalytic processes and provides insight into the evolution of the molecular architecture of catalytic module and CBM in GH9 cellulases.

Thymic carcinoid with multiple bone metastases: A case report.

BACKGROUND: Thymic carcinoid (TC) is a rare entity among anterior mediastinal malignancies. TCs are neuroendocrine carcinomas that constitute approximately 2%-5% of all thymic epithelial tumors. CASE SUMMARY: The study reported a rare TC with multiple bone metastases. A 77-year-old man presented with a 2-month history of lower back pain and weight loss of 5 kg. Magnetic resonance imaging scans revealed damage to the lumbar spine, sacrocaudal vertebrae and iliac crest, suggesting bone metastasis; computed tomography (CT) scan of the thorax showed a calcified anterior mediastinal mass; positron emission tomography-CT demonstrated multiple abnormal bone signals; and laboratory work-up showed no endocrine abnormalities. Fine-needle aspiration biopsy revealed predominantly single small, round to oval cells with scant cytoplasm and some loose clusters, suggesting endocrine manifestations. The pathological diagnosis was atypical carcinoid, which tend to originate from the thymus and was classified as intermediate-highly invasive. The patient underwent anlotinib-targeted therapy. Anlotinib (12 mg) was administered daily for 2 wk, after which the patient was allowed to rest for 21 d. Follow-up CT after one year demonstrated that the tumor had shrunk by approximately 29% after therapy. Treatment has a long stable disease benefit of more than 2.5 years. CONCLUSION: These findings demonstrated that anlotinib is a promising treatment regimen for patients with TC and multiple bone metastases.

Blood heparin-binding protein and neutrophil-to-lymphocyte ratio as indicators of the severity and prognosis of community-acquired pneumonia.

BACKGROUND: Community-acquired pneumonia (CAP) is particularly prevalent and has high mortality in severely ill patients, but the role of current biomarkers is limited. This study aimed to evaluate the importance of blood heparin-binding protein (HBP) and neutrophil-to-lymphocyte ratio (NLR) in assessing the severity and prognosis of CAP in adults. METHODS: The clinical information of 206 CAP patients was retrospectively analyzed. Receiver operating characteristic (ROC) curves were created, and the accuracy of the diagnosis of severe pneumonia was evaluated by the area under the curve (AUC). Univariate and multivariate Cox regression analysis was used to examine independent factors affecting the 30-day prognosis. The Kruskal-Wallis test was utilized to contrast the variations among etiology. RESULTS: Patients with severe pneumonia showed greater HBP and NLR compared to those with common pneumonia. The AUC of HBP was 0.723 (95% CI: 0.655-0.790) for the diagnosis of severe pneumonia, while NLR and HBP exhibited superior sensitivity (80.00%) and specificity (76.19%), respectively. Their combination boosted the diagnostic specificity (84.13%) while increasing the diagnostic sensitivity (86.25%) when combined with white blood cell (WBC) count. The 30-day mortality in CAP patients was independently predicted by HBP and NLR. However, there were no appreciable differences in HBP amongst patients with various etiologies. CONCLUSION: HBP and NLR were also independent predictors of 30-day death in CAP patients and grew with increasing severity in these patients. Their combination opened up new possibilities. Furthermore, there is no connection between HBP and the etiology of CAP.

Prolonged High-Fat Diet Consumption throughout Adulthood in Mice Induced Neurobehavioral Deterioration via Gut-Brain Axis.

Neuropsychiatric disorders have been one of the worldwide health problems contributing to profound social and economic consequences. It is reported that consumption of an excessive high-fat diet (HFD) in middle age could induce cognitive and emotional dysfunctions, whereas the mechanisms of the effects of long-term HFD intake on brain disorders have not been fully investigated. We propose a hypothesis that prolonged HFD intake throughout adulthood could lead to neurobehavioral deterioration via gut-brain axis. In this study, the adult C57BL/6J mice consuming long-term HFD (24 weeks) exhibited more anxiety-like, depression-like, and disruptive social behaviors and poorer performance in learning and memory than control mice fed with a normal diet (ND). In addition, the homeostasis of gut microbiota was impaired by long-term HFD consumption. Changes in some flora, such as Prevotellaceae_NK3B31_group and Ruminococcus, within the gut communities, were correlated to neurobehavioral alterations. Furthermore, the gut permeability was increased after prolonged HFD intake due to the decreased thickness of the mucus layer and reduced expression of tight junction proteins in the colon. The mRNA levels of genes related to synaptic-plasticity, neuronal development, microglia maturation, and activation in the hippocampus and prefrontal cortex of HFD-fed mice were lower than those in mice fed with ND. Interestingly, the transcripts of genes related to tight junction proteins, ZO-1 and Occludin involved in blood-brain-barrier (BBB), were decreased in both hippocampus and prefrontal cortex after long-term HFD consumption. Those results indicated that chronic consumption of HFD in mice resulted in gut microbiota dysbiosis, which induced decreased expression of mucus and tight junction proteins in the colon, in turn leading to local and systemic inflammation. Those changes could further contribute to the impairment of brain functions and neurobehavioral alterations, including mood, sociability, learning and memory. In short, long-term HFD intake throughout adulthood could induce behavioral phenotypes related to neuropsychiatric disorders via gut-brain axis. The observations of this study provide potential intervention strategies to reduce the risk of HFD via targeting the gut or manipulating gut microbiota.

Characterization of Inflammatory Signals in BV-2 Microglia in Response to Wnt3a.

Activation of microglia is one of the pathological bases of neuroinflammation, which involves various diseases of the central nervous system. Inhibiting the inflammatory activation of microglia is a therapeutic approach to neuroinflammation. In this study, we report that activation of the Wnt/ß-catenin signaling pathway in a model of neuroinflammation in Lipopolysaccharide (LPS)/IFN-γ-stimulated BV-2 cells can result in inhibition of production of nitric oxide (NO), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). Activation of the Wnt/ß-catenin signaling pathway also results in inhibition of the phosphorylation of nuclear factor-κB (NF-κB) and extracellular signal-regulated kinase (ERK) in the LPS/IFN-γ-stimulated BV-2 cells. These findings indicate that activation of the Wnt/ß-catenin signaling pathway can inhibit neuroinflammation through downregulating the pro-inflammatory cytokines including iNOS, TNF-α, and IL-6, and suppress NF-κB/ERK-related signaling pathways. In conclusion, this study indicates that the Wnt/ß-catenin signaling activation may play an important role in neuroprotection in certain neuroinflammatory diseases.

Temperature-perturbed two-dimensional generalized correlation characteristic slice spectra combined with multivariate method to identify adulterated milk.

In order to improve the discrimination accuracy of adulterated milk, a detection method was proposed based on temperature-perturbed generalized two-dimensional (2D) correlation characteristic slice spectra. A total of 240 samples were prepared including three brands of 40 pure milk and 40 urea-tainted milk, respectively. The infrared attenuated total reflection spectra of each sample were collected at different temperatures. Synchronous 2D infrared correlation spectrum of each sample was calculated under the external perturbation of temperature. The characteristic slice spectra of each sample were extracted from synchronous 2D correlation spectrum at characteristic peaks of milk and adulterants. N-way partial least squares discriminant analysis (NPLS-DA) models of single brand and the fusion of three brands of adulterated milk were established based on 2D correlation characteristics slice spectra. For comparison, the discrimination models were established using synchronous 2D correlation spectra and one-dimensional (1D) infrared spectra at room temperature, respectively. For the three brand fusion models, the discrimination accuracies of unknown samples were 100%, 98.8% and 82.7% using 2D correlation characteristic slice spectra, 2D correlation spectra, and 1D spectra, respectively. The results showed that the proposed method not only compressed the data, but also effectively extracted the characteristic information, and improved the accuracy of discrimination.

Identification of adulterated milk based on auto-correlation spectra.

A qualitative analysis of melamine-adulterated milk was proposed based on two-trace two-dimensional (2T2D) auto-correlation spectra. The concentration of melamine was used as external perturbation, and 40 adulterated samples of each brand with different concentrations of melamine (0.01 g/L to 1 g/L) were configured. Four brands of milk were used to configure experimental samples, including Guangming brand, Mengniu brand, Sanyuan brand and Wandashan brand. Spectroscopic data of pure milk and melamine-adulterated milk were measured by infrared (IR) (80-4000 cm-1) spectrophotometer. 2T2D auto-correlation spectral technology combined with least squares support vector machine (LS-SVM) method was used for qualitative analysis. The two strongest auto-correlation peaks in the auto-correlation spectra were selected for modeling. For Guangming brand, the intensities of auto-correlation at two wave numbers 2898 cm-1 and 2972 cm-1 were selected as independent variables. For Mengniu brand, the intensities of auto-correlation at two wave numbers 2852 cm-1 and 2920 cm-1 were selected. For Sanyuan brand, the intensities of auto-correlation at two wave numbers 2900 cm-1 and 2974 cm-1 were selected. For Wandashan brand, the intensities of auto-correlation at two wave numbers 2900 cm-1 and 2974 cm-1 were selected. For four brands fused together, the intensities of auto-correlation at two wave numbers 2900 cm-1 and 2974 cm-1 were selected. For each brand, the accuracy of qualitative analysis was 100 %. For four brands fused together, the accuracy of qualitative analysis was 99.05 %. In this way, it greatly reduced the amount of data to be processed. This study showed that 2T2D auto-correlation spectral technology combined with LS-SVM method was perfect for the discrimination of melamine-adulterated milk.

Slice spectra approach to synchronous Two-dimensional correlation spectroscopy analysis for milk adulteration discriminate.

The discrimination approach of adulterated milk was proposed combined synchronous two-trace two-dimensional (2T2D) correlation slice spectra at the characteristic wavebands of adulterant in milk with multivariate method. Two common adulterants, melamine and urea, were analyzed to demonstrate useful by the method. 2T2D (near infrared) NIR slice spectra at characteristic wavebands of adulterant were extracted from the synchronous 2T2D correlation spectra, and were input to construct the N-way partial least squares discriminant analysis (NPLS-DA) models. One-dimensional (1D) spectroscopy featuring all the present components in the samples combined with partial least squares discriminant analysis (PLS-DA) was also evaluated for comparison. The results indicated that for one kind of adulterant in model, prediction accuracies of slice spectral models were both 100% for melamine-adulterated and urea-adulterated samples discrimination. Moreover, for two kinds of adulterants in model, prediction accuracies of slice spectral models were 90.57% and 100% for melamine-adulterated and urea-adulterated discrimination, respectively, which was better than those of 1D whole models based on PLS-DA (only 81.13% and 98.15%, respectively). The comparison informs that the 2T2D slice spectra extracted at the characteristic wavebands of adulterant highlighted the adulterant spectral features and was obviously advantage to improve the discrimination accuracy. Meanwhile, the complexity of slice spectra is significantly reduced compared with the whole matrix of synchronous 2T2D correlation spectra.

Discrimination of adulterated milk using temperature-perturbed two-dimensional infrared correlation spectroscopy and multivariate analysis.

The discrimination method for adulterated milk is proposed based on temperature-perturbed two-dimensional (2D) infrared correlation spectroscopy and N-way partial least squares discriminant analysis (NPLS-DA). Two brands of pure and adulterated milk samples were prepared. The mid-infrared spectra of all samples were obtained from 30 â„ƒ to 55 â„ƒ with an interval of 5 â„ƒ. Under the perturbation of temperature, synchronous 2D correlation spectra were calculated to build discrimination models of pure milk and adulterated milk. In comparison, the NPLS-DA models were built based on three-dimensional (3D) stacked map (sample × temperature × wavenumber variable). For the NPLS-DA models of two brands of milk, the discrimination accuracy of unknown samples in the prediction set is 100% using temperature-perturbed 2D infrared correlation spectra, versus 77.8% using conventional 3D stacked map. The proposed method can be used as an alternative way for classifying pure and adulterated milk.

Therapeutic Effect and Safety of Tripterygium Glycosides Combined With Western Medicine on Type 2 Diabetic Kidney Disease: A Meta-Analysis.

PURPOSE: Tripterygium glycosides (TG) are widely used for the treatment of kidney disease in China. However, the application of TG in clinical practice is limited, as the therapeutic window is narrow, and the therapeutic dose is close to the toxic dose. In addition, the therapeutic effect of TG combined with Western medicine has not been fully elucidated. This study sought to explore standardized treatment, efficacy, and safety of TG combined with Western medicine for patients with type 2 diabetic kidney disease (T2DKD). METHODS: The PubMed, EMBASE, Cochrane Library, Chinese National Knowledge Infrastructure, Chinese Biomedical Literature, Chinese Scientific Journal, and Wan Fang databases were searched for randomized controlled trials of TG combined with Western medicine on T2DKD from their inception until May 4, 2021. A random effects model was used to explore heterogeneity of studies. FINDINGS: A total of 33 studies with 2034 patients were included in the current meta-analysis. The findings showed that TG combined with Western medicine effectively reduced urinary albumin excretion rates (standardized mean difference, -2.55; 95% CI, -4.70 to -0.40; P = 0.02), 24-hour urinary protein level (mean difference, -0.79; 95% CI, -1.22 to -0.36; P = 0.0003), and serum creatinine level (mean difference, -8.23; 95% CI, -14.48 to -1.99; P = 0.01) and increased albumin level (mean difference, 4.70; 95% CI, 3.27 to 6.13; P < 0.00001) in patients with T2DKD. No serious adverse reactions occurred, and the incidence of adverse events in the TG combined with Western medicine treatment group was slightly higher than in the control group (8.14% vs 2.65%). The results were stable, and a significant publication bias was not detected (P > 0.05). IMPLICATIONS: Based on our results, TG combined with Western medicine may be an effective and safe therapy for T2DKD; the best treatment duration may be 3 to 6 months. Nevertheless, larger, longer multicenter studies should be conducted for clinical application of the regimen to patients in more countries and regions. PROSPERO registration number: CRD42021259466.

Myeloid-derived suppressor cell: A crucial player in autoimmune diseases.

Myeloid-derived suppressor cells (MDSCs) are identified as a highly heterogeneous group of immature cells derived from bone marrow and play critical immunosuppressive functions in autoimmune diseases. Accumulating evidence indicates that the pathophysiology of autoimmune diseases was closely related to genetic mutations and epigenetic modifications, with the latter more common. Epigenetic modifications, which involve DNA methylation, covalent histone modification, and non-coding RNA-mediated regulation, refer to inheritable and potentially reversible changes in DNA and chromatin that regulate gene expression without altering the DNA sequence. Recently, numerous reports have shown that epigenetic modifications in MDSCs play important roles in the differentiation and development of MDSCs and their suppressive functions. The molecular mechanisms of differentiation and development of MDSCs and their regulatory roles in the initiation and progression of autoimmune diseases have been extensively studied, but the exact function of MDSCs remains controversial. Therefore, the biological and epigenetic regulation of MDSCs in autoimmune diseases still needs to be further characterized. This review provides a detailed summary of the current research on the regulatory roles of DNA methylation, histone modifications, and non-coding RNAs in the development and immunosuppressive activity of MDSCs, and further summarizes the distinct role of MDSCs in the pathogenesis of autoimmune diseases, in order to provide help for the diagnosis and treatment of diseases from the perspective of epigenetic regulation of MDSCs.

Enhancing the Production of Pinene in Escherichia coli by Using a Combination of Shotgun, Product-Tolerance and I-SceI Cleavage Systems.

Tolerance breeding through genetic engineering, sequence and omics analyses, and gene identification processes are widely used to synthesize biofuels. The majority of related mechanisms have been shown to yield endogenous genes with high expression. However, the process was time-consuming and labor-intensive, meaning there is a need to address the problems associated with the low-throughput screening method and significant time and money consumption. In this study, a combination of the limit screening method (LMS method) and product-tolerance engineering was proposed and applied. The Escherichia coli MG1655 genomic DNA library was constructed using the shotgun method. Then, the cultures were incubated at concentrations of 0.25%, 0.5%, 0.75% and 1.0% of pinene with different inhibitory effects. Finally, the genes acrB, flgFG, motB and ndk were found to be associated with the enhanced tolerance of E. coli to pinene. Using the I-SceI cleavage system, the promoters of acrB, flgFG and ndk genes were replaced with P37. The final strain increased the production of pinene from glucose by 2.1 times.

Lipophagy: A New Perspective of Natural Products in Type 2 Diabetes Mellitus Treatment.

Autophagy has been reported to involve in the pathogenesis of type 2 diabetes mellitus (T2DM), which protects the insulin target tissues and pancreatic ß-cells. However, autophagy is inhibited when the cells are lipid overload. That, in turn, increases the accumulation of fat. Lipotoxicity caused by excessive lipid accumulation contributes to pathogenesis of T2DM. Therefore, it is undeniable to break the vicious circles between lipid excess and autophagy deficiency. Lipophagy, a selective form of autophagy, is characterized by selective breakdown of lipid droplets (LDs). The nutritional status of cells contributes to the way of autophagy (selective or non-selective), while selective autophagy helps to accurately remove excess substances. It seems that lipophagy could be an effective means to decrease abnormal lipid accumulation that leads to insulin resistance and ß-cell impairment by removing ectopic LDs. Based on this process, many natural compounds have been reported to decrease lipid accumulation in tissues through autophagy-lysosomal pathway, which gradually highlights the significance of lipophagy. In this review, we focus on the mechanisms that lipophagy improves T2DM and natural products that are applied to induce lipophagy. It is also suggested that natural herbs with rich contents of natural products inducing lipophagy would be potential candidates for alleviating T2DM.

Long noncoding RNA DUXAP8 contributes to the progression of hepatocellular carcinoma via regulating miR-422a/PDK2 axis.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most deadly cancer worldwide. Multiple long noncoding RNAs (lncRNAs) are recently identified as crucial oncogenic factors or tumor suppressors. In this study, we explored the functon and mechanism of lncRNA double homeobox A pseudogene 8 (DUXAP8) in the progression of HCC. METHODS: Expression levels of DUXAP8 in HCC tissue samples were measured using qRT-PCR. The association between pathological indexes and the expression of DUXAP8 was also analyzed. Human HCC cell lines SMMC-7721 and QSG-7701 were used in in vitro studies. CCK-8 assay was used to assess the effect of DUXAP8 on HCC cell line proliferation. Scratch healing assay and Transwell assay were conducted to detect the effect of DUXAP8 on migration and invasion. Furthermore, dual-luciferase reporter assay was used to confirm targeting relationship between miR-422a and DUXAP8. Additionally, Western blot was used to detect the regulatory function of DUXAP8 on pyruvate dehydrogenase kinase 2 (PDK2). RESULTS: DUXAP8 expression HCC clinical samples was significantly increased and this was correlated with unfavorable pathological indexes. High expression of DUXAP8 was associated with shorter overall survival time of patients. Its overexpression remarkably facilitated the proliferation, metastasis, and epithelial-mesenchymal transition of HCC cells. Accordingly, knockdown of it suppressed the malignant phenotypes of HCC cells. Overexpression of DUXAP8 significantly reduced the expression of miR-422a by sponging it, but enhanced the expression of PDK2. CONCLUSIONS: DUXAP8 was a sponge of tumor suppressor miR-422a in HCC, enhanced the expression of PDK2 indirectly, and functioned as an oncogenic lncRNA.

Functions of TNF-α1 and TNF-α2 in large yellow croaker (Larimichthys crocea) in monocyte/macrophage activation.

Tumor necrosis factor-α (TNF-α) plays crucial roles in cell development, proliferation, apoptosis, inflammation, and immunity. TNF-α genes have been identified in various fish species, however, their biological functions remain to be further clarified. In this study, we identified a novel TNF-α homologue (LcTNF-α2) from large yellow croaker (Larimichthys crocea), which shares a low amino acid sequence identity to the previously reported large yellow croaker TNF-α (LcTNF-α1). The open reading frame of LcTNF-α2 is 714 nucleotides long, encoding a peptide of 237 amino acids (aa). The deduced LcTNF-α2 protein contains a 23-aa transmembrane region, a TACE restriction site at residues T71/L72, a TNF family signature (I108- F135), and two conserved cysteine residues (C39 and C179), as found in other known TNF-α sequences. Both LcTNF-α1 and LcTNF-α2 genes were constitutively expressed in all examined tissues and significantly up-regulated in the spleen and head kidney by Vibrio alginolyticus. Their transcripts were also detected in primary head kidney monocytes/macrophages (MO/Mϕs), lymphocytes (PKLs), granulocytes (PKGs), and large yellow croaker head kidney (LYCK) cell line and significantly increased in these cell types by inactivated Vibrio alginolyticus. Recombinant LcTNF-α1 and LcTNF-α2 proteins (rLcTNF-α1 and rLcTNF-α2) produced in Pichia pastoris not only significantly increased the production of reactive oxygen species (ROS), but also promoted the expression of proinflammatory cytokines (IL-1ß, IL-6,IL-8, and TNF-α1) in MO/Mϕs from large yellow croaker. Even more, after stimulation with rLcTNF-α1 and rLcTNF-α2, the production of nitrogen oxide (NO) and the expression of inducible NO synthase (iNOS) gene were significantly up-regulated. However, only rLcTNF-α1 remarkedly enhanced the phagocytosis of MO/Mϕs and increased the expression of TNF-α2 in MO/Mϕs. These results therefore indicated that LcTNF-α1 and LcTNF-α2 both play roles in promoting activation of head kidney MO/Mϕs.

Genome assembly and transcriptome analysis provide insights into the antischistosome mechanism of Microtus fortis.

Microtus fortis is the only mammalian host that exhibits intrinsic resistance against Schistosoma japonicum infection. However, the underlying molecular mechanisms of this resistance are not yet known. Here, we perform the first de novo genome assembly of M. fortis, comprehensive gene annotation analysis, and evolution analysis. Furthermore, we compare the recovery rate of schistosomes, pathological changes, and liver transcriptomes between M. fortis and mice at different time points after infection. We observe that the time and type of immune response in M. fortis are different from those in mice. M. fortis activates immune and inflammatory responses on the 10th day post infection, such as leukocyte extravasation, antibody activation, Fc-gamma receptor-mediated phagocytosis, and the interferon signaling cascade, which play important roles in preventing the development of schistosomes. In contrast, an intense immune response occurrs in mice at the late stages of infection and could not eliminate schistosomes. Infected mice suffer severe pathological injury and continuous decreases in cell cycle, lipid metabolism, and other functions. Our findings offer new insights into the intrinsic resistance mechanism of M. fortis against schistosome infection. The genome sequence also provides the basis for future studies of other important traits in M. fortis.