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Mouse reovirus type 3 (Reo-3) infection is a viral disease that is harmful for laboratory mice. No rapid and accurate detection methods are currently available for this infection. In this study, we describe a rapid, simple, closed-tube, one step, reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for Reo-3 and compare our assay with indirect enzyme-linked immunosorbent assay (ELISA). Three sets of RT-LAMP primers were designed by sequence analysis of a specific conserved sequence of the Reo-3 S1 gene. Using RS2 primer set, the RT-LAMP assay required 60 min at 65 °C to amplify the S1 gene in one step by using Reo-3 RNA template and had no cross-reactivity with the other related pathogens, such as Sendai virus (SV), pneumonia virus of mice (PVM), mouse hepatitis virus (MHV), Ectromelia virus (Ect), minute virus of mice (MVM), P. pneumotropica, B. bronchiseptica, K. pneumonia and P. aeruginosa. in our LAMP reaction system. The limit of detection (LOD) of our RT-LAMP assay is 4 fg/μL. The established RT-LAMP assay enabled visual detection when fluorescence detection reagents were added, and was demonstrated to be effective and efficient. We tested 30 clinical blood samples and five artificial positive samples from SPF mice, the concordance between the two methods for blood samples was 100% compared with indirect ELISA and RT-PCR. Considering its performance, specificity, sensitivity, and repeatability, the developed RT-LAMP could be a valuable tool to supply a more effective Reo-3 detection method in laboratory animal quality monitoring.

Counting the number of red blood cells (RBCs) in blood samples is a common clinical diagnostic procedure, but conventional methods are unable to provide the size and other physical properties of RBCs at the same time. In this work, we explore photoacoustic (PA) detection as a rapid label-free and noninvasive analysis technique that can potentially be used for single RBC characterization based on their photoabsorption properties. We have demonstrated an on-chip PA flow cytometry system using a simple microfluidic chip combined with a PA imaging system to count and characterize up to ∼60 RBCs per second. Compared with existing microfluidic-based RBC analysis methods, which typically use camera-captured image sequences to characterize cell morphology and deformation, the PA method discussed here requires only the processing of one-dimensional time-series data instead of two- or three-dimensional time-series data acquired by computer vision methods. Therefore, the PA method will have significantly lower computational requirements when large numbers of RBCs are to be analyzed. Moreover, we have demonstrated that the PA signals of RBCs flowing in a microfluidic device could be directly used to acquire the osmolarity conditions (in the range of 124 to 497 mOsm L-1) of the medium surrounding the RBCs. This finding suggests a potential extension of applicability to blood tests via PA-based biomedical detection.

The critical role of sialyltransferase (ST) enzymes in tumour cell growth and metastasis, as well as links to multi-drug and radiation resistance, has seen STs emerge as a target for potential antimetastatic cancer treatments. One promising class of ST inhibitors that improve upon the pharmacokinetic issues of previous inhibitors is the 1,2,3-triazole-linked transition-state analogues. Herein, we present the design and synthesis of a new generation of 1,2,3-triazole-linked sialyltransferase inhibitors, along with their biological evaluation demonstrating increased potency for phosphonate bearing compounds. The six most promising inhibitors presented in this work exhibited a greater number of binding modes for hST6Gal I over hST3Gal I, with K(i) ranging from 3–55 μM. This work highlights phosphonate bearing triazole-linked compounds as a promising class of synthetically accessible ST inhibitors that warrant further investigation.

To determine if the effect of manipulative acupuncture monitored by laser speckle contrast imaging (LSCI) can improve facial blood perfusion in patients with severe Bell’s palsy. This randomized, single-blind, controlled trial included 120 newly diagnosed patients (within 14 days) with severe Bell’s palsy (House–Brackmann grading system (HBGS) ≥ grade IV). The patients were randomized (1 : 1) to receive either acupoints acupuncture combined with manipulations of twirling, lifting, and thrusting treatments (manipulative acupuncture) or acupoints acupuncture therapy alone (simple acupuncture). These treatments consisted of a total of 24 sessions, three times per week, and each treatment lasted for 30 min. Following 8 weeks of treatment and 6 months after the initial onset of facial palsy, facial nerve functioning was scored (HBGS) and clinical efficacy was measured. The patients’ facial blood perfusion significantly improved following manipulative acupuncture assisted by LSCI compared with that at baseline ( P < 0.01 ). At the conclusion of the 8-week treatment, both groups showed improvement; however, the recovery rate was significantly different (manipulative acupuncture 53.3% vs. simple acupuncture 33.9%, P < 0.05 ). Follow-up analysis at 6 months after the onset of facial palsy revealed a significantly higher recovery rate (91.7% vs. 78.0%; P < 0.05 ). In addition, the number of treatments in the observation group was less than that in the simple acupuncture therapy group ( P < 0.05 ). Compared with simple acupuncture therapy, manipulative acupuncture therapy led to a more significant recovery rate in the treatment of severe Bell’s palsy and required a shorter course of treatment. This trial was registered with ChiCTR1800019463.

Super-resolution imaging using microspheres has attracted tremendous scientific attention recently because it has managed to overcome the diffraction limit and allowed direct optical imaging of structures below 100 nm without the aid of fluorescent microscopy. To allow imaging of specific areas on the surface of samples, the migration of the microspheres to specific locations on two-dimensional planes should be controlled to be as precise as possible. The common approach involves the attachment of microspheres on the tip of a probe. However, this technology requires additional space for the probe and could not work in an enclosed environment, e.g., in a microfluidic enclosure, thereby reducing the range of potential applications for microlens-based super-resolution imaging. Herein, we explore the use of laser trapping to manipulate microspheres to achieve super-resolution imaging in an enclosed microfluidic environment. We have demonstrated that polystyrene microsphere lenses could be manipulated to move along designated routes to image features that are smaller than the optical diffraction limit. For example, a silver nanowire with a diameter of 90 nm could be identified and imaged. In addition, a mosaic image could be constructed by fusing a sequence of images of a sample in an enclosed environment. Moreover, we have shown that it is possible to image Escherichia coli bacteria attached on the surface of an enclosed microfluidic device with this method. This technology is expected to provide additional super-resolution imaging opportunities in enclosed environments, including microfluidic, lab-on-a-chip, and organ-on-a-chip devices.

Background: Late-life depression often coexists with vascular cognitive impairment and affects the quality of life for elders. However, little is known about cortical morphometric interactions between subcortical vascular mild cognitive impairment (svMCI) and concomitant mild depressive symptoms at the early stage. Objective: We aimed to investigate cortical alterations of svMCI with and without depressive symptoms and determine whether these parameters are associated with depression symptoms and/or cognitive impairments. Methods: Surface based morphometry was performed on 18 svMCI patients with depressive symptoms (svMCI + D), 16 svMCI patients without depressive symptoms (svMCI–D), and 23 normal controls (NC). Results: Compared to NC, both svMCI + D and svMCI–D patients exhibited significantly decreased surface area (SA) in many cortical areas. Interestingly, svMCI + D patients showed significantly increased rather than decreased SA in right lateral occipital gyrus (LOG.R), and a consistent trend of increased SA in these areas compared to svMCI–D. In addition, the svMCI + D showed increased gray matter volume of left pericalcarine (periCAL.L) than svMCI–D, whereas svMCI–D showed decreased gray matter volume of periCAL.L than NC. Further correlation analyses revealed that the SA of left superior temporal gyrus (STG.L) and right lateral orbital part of frontal gyrus (lorbFG.R) were significantly correlated with Hamilton depression rating scale of svMCI + D. Conclusion: In conclusion, these results extend our insight into svMCI and add weight to reevaluation of concomitant early stage depressive symptoms. Moreover, we suggest that LOG.R∖periCAL.L∖STG.L∖lorbFG.R might serve as sensitive and trait-dependent biomarkers to detect concomitant depressive symptoms in svMCI patients.

2,5-Dimethylpyrazine (2,5-DMP) is an indispensable additive for flavoring in the food industry and an important substrate for producing hypoglycemic and antilipolytic drugs. However, 2,5-DMP is pro...

Recently, the role of long non-coding RNAs (lncRNAs) in regulating multiple cancer types has attracted increasing interest because of their involvement in cell metastasis in different cancer types. Previous studies indicated that LINC00657 may work as an oncogene in gastric and colon cancer. However, the functional role and mechanistic action of LINC00657 on colorectal cancer (CRC) remains unknown. Therefore, in this study, the role of LINC00657 in CRC was evaluated. Our results showed that LINC00657 was enriched in CRC stem-like cells (CSCs) and significantly promoted CSCs invasion ability. LINC00657 expression resulted frequently up-regulated in CRC patient tissue, and high expression of LINC00657 was correlated with an advanced clinical stage, lymph node metastasis, distant metastasis and poor overall survival of CRC patients. Furthermore, LINC00657 worked as a competing endogenous RNA (ceRNA) for miR-203a, antagonizing its function as a tumor suppressor and leading to the de-repression of CSCs invasion. Collectively, our observations revealed that LINC00657 is involved in CRC invasion by acting as a competing endogenous RNA. Thus, LINC00657 may serve as a potential prognostic factor and/or therapeutic target for CRC.

Objective. To evaluate the quality of methodologies used in previous systematic reviews (SRs) and compare efficacy of different acupuncture therapies for allergic rhinitis. Methods. Seven electronic databases were searched for systematic reviews (SRs) performed on different acupuncture therapies for allergic rhinitis from inception to 15 November 2019. The AMSTAR2 instrument was employed to assess the methodological quality of included SRs. Eligible randomized controlled trials (RCTs) were selected from the included systematic reviews. We also included recent RCTs published by 15 November 2019. Cochrane risk of bias tool was utilized to determine risk of bias of the included RCTs. Pairwise meta-analyses were performed using the random-effects model. Network meta-analysis of the included RCTs was carried out using frequentist framework. Results. We identified 2 SRs with low quality and 18 SRs with very low quality, both of which contained 33 eligible RCTs (n = 3769). Most of these studies had unclear risk of bias. On the basis of ranking probability, NMA analysis showed that acupuncture at the sphenopalatine ganglion acupoint (OR: 1.31, 95% CI 1.07 to 1.61) had the highest probability of improving global allergic rhinitis symptoms, followed by San-Fu-Tie (OR: 1.17, 95% CI 1.08 to 1.27), manual acupuncture (OR:1.15, 95% CI 1.07 to 1.24) compared with conventional western medicine treatment. Moreover, direct comparison of the follow-up period showed that the clinical outcomes of acupuncture and related therapies at three-month (OR:1.34, 95% CI 1.17 to 1.55), six-month (OR: 1.31, 95% CI 1.10 to 1.57), and twelve-month (OR: 1.30, 95%CI 1.11 to 1.53) follow-up were better than those of traditional western medicine. Conclusion. These results indicate that for patients with allergic rhinitis who are unresponsive to conventional western medicine or cannot tolerate the side effects, acupuncture at the sphenopalatine ganglion acupoint is an effective alternative therapy. Further studies are advocated to deeply explore methodological quality of SRs by incorporating high-quality RCTs.

This study was conducted to investigate whether dietary supplementation of black soldier fly (Hermetia illucens Linnaeus) pulp (BSFP) affects growth performance, antioxidant capacity and intestinal health of juvenile mirror carp (Cyprinus carpio var. specularis). A total of 270 juvenile mirror carp (13.68 ± 0.02 g) were randomly allotted to five dietary treatments, BSFP0, BSFP25, BSFP50, BSFP75 and BSFP100, in which BSFP was included in the basal diet at 0, 43.7, 87.3, 131 and 174.7 g/kg, respectively. Then, fish were fed to apparent satiation for 8 weeks. Fish growth performance and nutrient utilization were not different among the five groups (p > .05). Increasing BSFP dietary content significantly decreased whole‐body lipid content but increased kidney index (p .05). In conclusion, this study indicated that BSFP at the level below 131 g/kg could be added in diet of juvenile mirror carp without any negative effect on growth performance and intestinal health. Dietary BSFP inclusion at low levels decreased the whole‐body lipid content and increased the antioxidant activity of juvenile mirror carp.

Currently, the mechanisms underlying the peritoneal metastasis of gastric cancer cells and the function of mesothelial cells during this process are unclear, primarily due to the absence of an effective in vitro peritoneal model. In this study, we constructed a biomimetic peritoneal model using a digital micromirror device-based optical projection lithography system. This model enabled the simulation of a damaged peritoneum, which allowed for a comparison of the characteristics of an undamaged peritoneum, such as porosity, mechanical properties, and surface morphology, with those of a damaged peritoneum. Biological inertness and removability of the polyethylene glycol dimethacrylate hydrogel were exploited to fabricate an arrayed heterogeneous interface that imitated a damaged human peritoneum. The porous structure of the peritoneum was achieved by adjusting the ratio of collagen I to gelatin methacryloyl; this structure of the peritoneum might contribute to its shock absorption property. Atomic force microscopy characterization showed that the outermost layers of the model peritoneum and real peritoneum were similar in surface morphology and mechanical properties. Furthermore, we reproduced the process of peritoneal metastasis in vitro. The numbers of gastric cancer cells that adhered to the heterogeneous interface were different, and mesothelial cells played an essential role in peritoneal metastasis. Our findings indicate that this model can be utilized in preclinical drug screening and personalized therapy.

Endometrial carcinoma (EC) is the most common malignant tumors in female derived from the endometrial epithelium. Several previous studies have described estrogen receptors (ER), progesterone Receptor (PR) and phosphatase and tensin homolog (PTEN) are associated with clinicopathological factors and prognosis in EC patients. However, during EC patients follow-up, we found that some EC patients with down-regulation of PTEN, but up-regulation of ER or PR , and some EC patients with down-regulation of ER or PR, but up-regulation of PTEN also had a poor prognosis. Therefore, to reveal the prognosis of EC patients with different phenotypes based on PTEN, ER and PR expression, 120 cases formalin-fixed paraffin-embedded EC tissues and 543 cases uterine corpus endometrial carcinoma (UCEC) patients from the cancer genome atlas (TCGA) UCEC datasets were analyzed. Results showed that EC tissues can be classified to PTENLERLPRL, PTENHERLPRL, PTENHERHPRH, PTENLERHPRH, PTENHERHPRL, PTENHERLPRH, and PTENLERHPRL phenotypes basing on IHC analysis. Additionally, EC patients with PTENLERLPRL showed high malignancy, while patients with PTENHERHPRH showed low malignancy. Therefore, combined detection of PTEN, ER, PR may help identify a small subset of EC with more aggressive behavior and may aid in risk stratification.

Hemodynamic forces have an important role in venous intimal hyperplasia, which is the main cause of arteriovenous fistula dysfunction. Endothelial cells (ECs) constantly exposed to the shear stress of blood flow, converted the mechanical stimuli into intracellular signals, and interacted with the underlying vascular smooth muscle cells (VSMCs). Caveolin-1 is one of the important mechanoreceptors on cytomembrane, which is related to vascular abnormalities. Extracellular signal-regulated kinase1/2 (ERK1/2) pathway is involved in the process of VSMCs proliferation and migration. In the present study, we explore the effects of Caveolin-1-ERK1/2 pathway and uremia toxins on the endothelial cells and VSMCs following shear stress application. Different shear stress was simulated with a ECs/VSMCs cocultured parallel-plate flow chamber system. Low shear stress and oscillating shear stress up-regulated the expression of fibroblast growth factor-4, platelet-derived growth factor-BB, vascular endothelial growth factor-A, ERK1/2 phosphorylation in endothelial cells, and proliferation and migration of VSMCs but down-regulated the Caveolin-1 expression in endothelial cells. Uremia toxin induces the proliferation and migration of VSMCs but not in a Caveolin-1-dependent manner in the static environment. Low shear stress-induced proliferation and migration of VSMCs is inhibited by Caveolin-1 overexpression and ERK1/2 suppression. Shear stress-regulated VSMC proliferation and migration is an endothelial cells-dependent process. Low shear stress and oscillating shear stress exert atherosclerotic influences on endothelial cells and VSMCs. Low shear stress modulated proliferation and migration of VSMCs through Caveolin-1-ERK1/2 pathway, which suggested that Caveolin-1 and ERK1/2 can be used as a new therapeutic target for the treatment of arteriovenous fistula dysfunction.Impact statementVenous intimal hyperplasia is the leading cause of arteriovenous fistula (AVF) dysfunction. This article reports that shear stress-regulated vascular smooth muscle cells (VSMCs) proliferation and migration is an endothelial cell (EC)-dependent process. Low shear stress (LSS) and oscillating shear stress (OSS) exert atherosclerotic influences on the ECs and VSMCs. LSS-induced proliferation and migration of VSMCs is inhibited by Caveolin-1 overexpression and extracellular signal-regulated kinase1/2 (ERK1/2) suppression, which suggested that Caveolin-1 and ERK1/2 can be used as a new therapeutic target for the treatment of AVF dysfunction.

To evaluate the effects of dietary nano‐selenium (Nano‐Se) on antioxidant capacity and hypoxia tolerance of grass carp fed with high‐fat diet, experimental fishes were fed Nano‐Se supplemented diets at doses of 0 (Control), 0.3, 0.6, 0.9 and 1.2 mg/kg for 10 weeks. After feeding trial, a part of the fishes were exposed to hypoxia stress. Results showed that the survival ratio of grass carp significantly increased in 0.6 and 0.9 mg/kg Nano‐Se group, and the content of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) significantly decreased in 0.6–1.2 mg/kg Nano‐Se groups compared with the control group. In addition, dietary Nano‐Se significantly enhanced glutathione peroxidase (GPX) activity and reduced the malondialdehyde (MDA) content in fishes fed diets with 0.3 and 0.6 mg/kg Nano‐Se. Dietary Nano‐Se significantly elevated mRNA expression of GPX1 and catalase (CAT) by promoting the mRNA expression of NF‐E2‐related nuclear factor 2 (Nrf2) in the hepatopancreas. After hypoxia stress, the GPX and superoxide dismutase (SOD) activities were significantly enhanced, and the MDA content and mortality rate consequently decreased in fishes fed diets with 0.3 and 0.6 mg/kg Nano‐Se. In summary, these results suggested that optimal Nano‐Se in diet enhanced the antioxidant capacity and hypoxia tolerance of grass carp.

Background This study investigates the characteristics of a special type of cancer of unknown primary site (CUP, type 2), which is a metastasis of a definite pathological diagnosis without a detectable primary site. Patients and methods Patients diagnosed between 2004 and 2014 were identified from the Surveillance Epidemiology and End Results (SEER) database. The characteristics of type 2 CUP from different sources were analyzed. For each source of type 2 CUP, tumors of the corresponding TnN0‐XM1 stage were used as controls. Results A total of 8505 patients with type 2 CUP were included in this analysis. Type 2 CUP shows an increasing trend, while type 1 shows the opposite. Type 2 CUPs have significant differences with stage IV of the same pathological primary lesion. Many characteristics influenced the prognosis of type 2 CUP patients, including marital status, age, race, sex, registration time, lymph node metastasis, surgery, chemotherapy, and radiation. Conclusion Our study suggests that identifying the source of metastasis is the key to the selection of treatment and the determination of the prognosis for CUP., The number of type 2 cancer of unknown primary site (CUP) (metastatic cancer that the tissue or organ source can be determined but not the primary tumor, T0N0‐XM1) is increasing. Type 2 CUP (T0N0‐XM1) is a special cancer status, whose clinical characteristics and prognosis are significantly different from the classical CUP (type 1 CUP) and corresponding TnN0‐XM1. Identifying the source of metastatic focus is the key to the selection of treatment and prognosis for CUP.

BackgroundThe intensity of electrical acupoint stimulation such as electroacupuncture (EA) and transcutaneous electrical nerve stimulation (TENS) is regulated by the observation of skin shivering or the participant’s comfort response. However, the specific intensity and spatial scope following EA or TENS stimulation are unclear.ObjectiveThis study aimed to test the stimulatory current intensities of lower and upper sensation thresholds in TENS- and EA-based treatment of Bell’s palsy patients. Also, the spatial scope of the stimulation at these current intensities was simulated and measured quantitatively.MethodsA total of 19 Bell’s palsy patients were recruited. Six acupoints on the affected side of the face were stimulated by TENS and EA successively at 30-min intervals. During the stimulation, the current intensity was regulated gradually from 0 to 20 mA, and we simultaneously measured the lower (sensory) and upper (tolerability) sensations. After the treatment by TENS and EA, the modified Chinese version of the Massachusetts General Hospital Acupuncture Sensation Scales (C-MMASS) was applied to survey the de-qi sensations during stimulation. Additionally, we analyzed the correlation between current intensities and C-MMASS and comfort scores. Finite element models were established to depict the spatial distribution of electric field gradients at the lower and upper thresholds.ResultsThe mean sensory and tolerability thresholds of TENS were 3.91–4.37 mA and 12.33–16.35 mA, respectively. The median sensory and tolerability thresholds of EA were 0.2 mA and 2.0–3.2 mA, respectively. We found a significant correlation between total C-MMASS scores and the current intensities at the tolerability threshold of TENS. The finite element model showed that the activated depths of TENS and EA at the lower threshold were 3.8 and 7 mm, respectively, whereas those at the upper threshold were both 13.8 mm. The cross-sectional diameter of the activated area during TENS was 2.5–4 times larger than that during EA.ConclusionThis pilot study provided a method for exploring the current intensity at which the de-qi sensations can be elicited by TENS or EA. The finite element analysis potentially revealed the spatial scope of the electrical stimulation at a specific current intensity.

Microlenses have become an indispensable optical element in many optical systems. The advancement of technology has led to a wider variety of microlenses fabrication methods, but these methods suffer from, more or less, some limitations. In this article, we review the manufacturing technology of microlenses from the direct and indirect perspectives. First, we present several fabrication methods and their advantages and disadvantages are discussed. Then, we discuss the commonly used materials for fabricating microlenses and the applications of microlenses in various fields. Finally, we point out the prospects for the future development of microlenses and their fabrication methods.

During orthodontic treatment, mechanical force is applied to the teeth, and following a series of complex metabolism changes, the position of the teeth in the alveolar bone change. This process is closely associated with primitive bone mesenchymal stem cells (BMSCs), which may differentiate into osteoblasts precursor cell. A hypoxic microenvironment may be caused by orthodontic mechanical forces between the alveolar bone and the root. Hypoxia‑inducible factor 1α (HIF‑1α) is a specific receptor that adapts to a hypoxic environment. The present study was designed to investigate whether HIF‑1α was involved in the osteoblastic differentiation of BMSCs induced by cyclic tensile stress. During this process, HIF‑1α mRNA and protein expression were detected using a reverse transcription‑quantitative polymerase chain reaction and western blotting. It was revealed that alkaline phosphatase activity increased in a time‑dependent manner in three different stretching strength groups, which indicates that cyclic stretch promotes the osteogenic differentiation of BMSCs. The optimal force stage of osteogenesis was an unexpected discovery, which will provide theoretical guidance for selecting the most suitable orthodontic force for tooth movement in clinical orthodontic treatment. Most importantly, all experiments revealed that HIF‑1α mRNA and protein were significantly increased following stretching treatment in BMSCs. It was therefore concluded that HIF‑1α may be involved in BMSCs modulating osteogenic metabolism during exposure to cyclic stretch and a hypoxic microenvironment, which may prove useful for the reconstruction of a jaw during orthodontic treatment.

Seven indole alkaloid glycosides containing a 1′-(4″-hydroxy-3″,5″-dimethoxyphenyl)ethyl unit (1–7) were isolated from an aqueous extract of Isatis indigotica leaves (da qing ye). Their structures were determined by spectroscopic data analysis combined with enzymatic hydrolysis as well as comparison of their experimental CD (circular dichroism) and calculated ECD (electrostatic circular dichroism) spectra. Based on analysis of [α]D20 and/or Cotton effect (CE) data of 1–7, two simple roles to assign location and/or configuration of β-glycopyranosyloxy and 1′-(phenyl)ethyl units in the indole alkaloid glycosides are proposed. Stereoselectivity in plausible biosynthetic pathways of 1–7 is discussed. Compounds 3 and 4 and their mixture in a 3:2 ratio showed activity against KCNQ2 in CHO cells. The mixture of 5 and 6 (3:2) exhibited antiviral activity against influenza virus H1N1 PR8 with IC50 64.7 μmol/L (ribavirin, IC50 54.3 μmol/L), however, the individual 5 or 6 was inactive. Preliminary structure–activity relationships were observed., Graphical abstract Image 1Seven indole alkaloid glycosides (1–7) were isolated from an aqueous extract of Isatis indigotica leaves (da qing ye), representing the first isomers biogenetically originated from indole glycosides and sinapinic acid in differential stereoselective manners, respectively. The mixture of 5 and 6 (3:2) exhibited antiviral activity against influenza virus H1N1 PR8.

Background: Extensive research has shown that long noncoding RNA (lncRNA) is involved in tumorigenesis, including hepatocellular carcinoma (HCC). The lncRNA LINC00339 was reported to regulate the development of lung cancer or breast cancer. However, whether LINC00339 participates in HCC progression remains unclear. Here, our results showed that LINC00339 was upregulated in HCC. Methods: qRT-PCR and in situ hybridization (ISH) was used to analyze LINC00339 expression in tumor tissues and cell lines. CCK8 and colony formation assays were used to analyze cell proliferation. Transwell assay was used to analyze cell migration and invasion. Xenograft experiment was used to test tumor growth in vivo. Results: LINC00339 overexpression was correlated with an advanced stage, metastasis, and bad prognosis in HCC patients. Functional investigation showed that LINC00339 knockdown significantly suppressed HCC cell proliferation, migration, and invasion. Moreover, decreased LINC00339 expression inhibited HCC growth in vivo. Mechanistically, LINC00339 could interact with miR-1182 to promote SKA1 expression. We also demonstrated that SKA1 acted as an oncogene and SKA1 upregulation reversed the effect of LINC00339 silencing. Conclusion: Our results illustrated that the LINC00339/miR-1182/SKA1 axis plays an essential role in HCC progression.

The dysregulation of microRNA (miRNA) expression has been highlighted in a variety of human malignant conditions with reports implicating a critical role in the process of tumor growth. The role of miR-539 in pancreatic cancer (PC) is yet to be fully elucidated, hence the aim of the current study was to investigate the effect of miR-539 expression in relation to a cohort of 52 PC specimens. The application of a real-time quantitative polymerase chain reaction (qRT-PCR) revealed a significantly down-regulated miR-539 level, which was accompanied by an increased TWIST1 expression in PC when compared with the controls. The in vitro experiment results demonstrated that the endogenic mimic of miR-539 significantly suppressed the growth of the xenograft tumors in PANC-1 cells, when compared to the delivery of the control miRNA and blank control. Meanwhile, the key epithelial-mesenchymal transition (EMT) inducer, TWIST1 was verified as a direct target gene of miR-539 through the application of a luciferase reporter assay. In conclusion, the results of the current study present evidence emphasizing the significance of the interactions between miR-539 and TWIST1 in the development of and progression of PC, highlighting its potential as a therapeutic target in the treatment of PC patients.

Aim: Mechanical strain plays a crucial role in bone formation and remodeling. Hypoxia-inducible factor (HIF)-1α and TWIST are upstream of master regulators of osteogenesis, including runt-related transcription factor 2 (RUNX2) and bone morphogenetic proteins (BMPs). This study investigated the effect of the HIF-1α-TWIST pathway on cyclic mechanical stretch-induced osteogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs) and the underlying mechanism. Materials and Methods: BMSCs were isolated from bone marrow derived from the femurs and humeri of Sprague-Dawley rats. Osteogenic differentiation of BMSCs was induced by applying cyclic mechanical stretch using the Flexcell Tension System. HIF-1α and TWIST were knocked down using recombinant lentiviral vectors. Osteogenic differentiation was evaluated by real-time qPCR, western blotting, and the alkaline phosphatase (ALP) activity assay. Results: Cyclic mechanical stretch increased ALP activity and expression of HIF-1α and TWIST in BMSCs. Knockdown of HIF-1α decreased TWIST expression in stretched BMSCs. Moreover, knockdown of HIF-1α or TWIST enhanced cyclic mechanical stretch-induced osteogenic differentiation of BMSCs. In addition, knockdown of TWIST increased expression of RUNX2 and BMP2 in stretched BMSCs. Conclusions: The HIF-1α-TWIST signaling pathway inhibits cyclic mechanical stretch-induced osteogenic differentiation of BMSCs. This finding may facilitate cell and tissue engineering for clinical applications.

Fatty acid metabolism is crucial for maintaining energy homeostasis in aquatic vertebrates experiencing environmental stress. Both sterol regulatory element-binding protein 1 (SREBP-1) and peroxisome proliferator-activated receptor α (PPARα) are the key regulators of fatty acid metabolism. In this study, the coding sequences (CDS) of SREBP-1 and PPARα were firstly identified and characterized from Onychostoma macrolepis, encoding peptides of 1136 and 470 amino acids, respectively. The functional domains in O. macrolepis SREBP-1 and PPARα proteins retained the high similarity with those of other animals, at 74.69% and 77.29%, respectively. The mRNA encoding SREBP-1 was primarily expressed in the muscle and PPARα was highly expressed in the liver and intestine. Under thermal exposure, the content of non-esterified fatty acid (NEFA) decreased gradually after 1 h in the liver and muscle of O. macrolepis, which might be due to that the organism meet more energy expenditure via fatty acid β-oxidation. Furthermore, the mRNA expression level of SREBP-1 decreased, while the mRNA expression level of PPARα increased from 0 h to 6 h in the liver. And we found that the mRNA expression levels of both SREBP-1 and PPARα decreased significantly at 48 h (P < .05) in the muscle, which was in accordance with the significant decrease of target gene FAS and CPT1A mRNA expression levels, respectively. It might be the physiological adjustment that the fish adapted to thermal exposure at the end of experiment. These results illustrate that O. macrolepis SREBP-1 and PPARα-mediated fatty acid metabolism is a fundamental requirement for thermal adaptation.

BACKGROUND Intrahepatic and distant metastases could be the major cause of treatment failure in hepatocellular carcinoma (HCC). The deep mechanism of HCC metastasis is closely related to the interaction between integrins and extracellular matrix (ECM) in tumor microenvironment. MATERIAL AND METHODS In vitro cell adhesion assay was performed to determine the capability of adhering to ECM elements of HCC cells. To modulate the expression status of ADAR1 p110 in tumor cells, lentivirus system was applied. Meanwhile, patients' HCC samples and orthotopic xenograft mouse model were used for verifying our in vitro data. RESULTS ADAR1 p110 could strongly enhance the adhesion of HCC tumor cells to ECM, which was usually regarded as the initiation of tumor invasion. Such phenotype was caused due to up-regulation of ITGA2 both in mRNA and protein level. Moreover, specimen collected from HCC patients revealed a positive correlation between ADAR1 and ITGA2. Finally, ADAR1 p110 promoted HCC metastasis was verified when we applied orthotopic xenograft mouse model. CONCLUSIONS ADAR1 could enhance HCC metastasis by promoting tumor cells adhering to ECM via increasing ITGA2 expression. This phenomenon could provide novel information to better understanding the mechanism of HCC metastasis procedure.

Background In China, Jin's three-needle acupuncture therapy has been widely used for stroke treatment. However, its mechanism remains unclear. The aim of this study was to investigate the effects of Jin's three-needle acupuncture therapy on stroke patients using multi-channel EEGs. Materials and methods Fifty stroke patients participated and their EEG signals were recorded before, during, and after acupuncture. Results Comparing with baseline before acupuncture, the alpha rhythm was significantly increased during the acupuncture needle retention stage and became stronger after removing the needles. The increase of alpha rhythm occurred when inserting and removing the needles, with its high amplitude remaining during retention and after removal of the needles. As the alpha rhythm was significantly correlated with patients' movement and daily-living abilities, the alteration of alpha rhythm during and after acupuncture suggests a possible electrophysiological mechanism of the curative effect of acupuncture for stroke rehabilitation. Conclusion From the electrophysiological study, Jin's three-needle acupuncture therapy is efficient in stroke patients.

Objectives: To summarize development processes and research hotspots of MRI research on acupuncture and to provide new insights for researchers in future studies.Methods: Publications regarding MRI on acupuncture from inception to 2020 were downloaded from the Web of Science Core Collection. VOSviewer 1.6.15 and CiteSpace V software were used for bibliometric analyses. The main analyses include collaboration analyses between countries/institutions/authors, co-occurrence analysis between keywords, as well as analyses on keyword bursts, citation references, and clusters of references.Results: A total of 829 papers were obtained with a continually increased trend over time. The most productive country and institution in this field were the People's Republic of China (475) and KyungHee University (70), respectively. Evidence-based Complementary and Alternative Medicine (83) was the most productive journal, and Neuroimage (454) was the most co-cited journal. Dhond's et al. (2008) article (co-citation counts: 58) and Napadow's et al. (2005) article (centrality: 0.21) were the most representative and symbolic references, with the highest co-citation number and centrality, respectively. Jie Tian had the highest number of publications (35) and Kathleen K S Hui was the most influential author (280 co-citations). The four hot topics in MRI on acupuncture were acupuncture, fMRI, pain, and stimulation. The three frontier topics were connectivity, modulation, and fMRI. Based on the clustering of co-cited documents, chronic low back pain, sham electro-acupuncture treatment, and clinical research were the main research directions.Conclusion: This study provides an in-depth perspective for MRI research on acupuncture and provides researchers with valuable information to determine the current status, hot spots, and frontier trends of MRI research on acupuncture.

Facial muscle activities are essential for the appearance and communication of human beings. Therefore, exploring the activation patterns of facial muscles can help understand facial neuromuscular disorders such as Bell’s palsy. Given the irregular shape of the facial muscles as well as their different locations, it should be difficult to detect the activities of whole facial muscles with a few electrodes. In this study, a high-density surface electromyogram (HD sEMG) system with 90 electrodes was used to record EMG signals of facial muscles in both healthy and Bell’s palsy subjects when they did different facial movements. The electrodes were arranged in rectangular arrays covering the forehead and cheek regions of the face. The muscle activation patterns were shown on maps, which were constructed from the Root Mean Square (RMS) values of all the 90-channel EMG recordings. The experimental results showed that the activation patterns of facial muscles were distinct during doing different facial movements and the activated muscle regions could be clearly observed. Moreover, two features of the activation patterns, 2D correlation coefficient (corr2) and Centre of Gravity (CG) were extracted to quantify the spatial symmetry and the location of activated muscle regions respectively. Furthermore, the deviation of activated muscle regions on the paralyzed side of a face compared to the healthy side was quantified by calculating the distance between two sides of CGs. The results revealed that corr2 of the activated facial muscle region (classified into forehead region and cheek region) in Bell’s palsy subjects was significantly (p < 0.05) lower than that in healthy subjects, while CG distance of activated facial region in Bell’s palsy subjects was significantly (p < 0.05) higher than that in healthy subjects. The correlation between corr2 of these regions and Bell’s palsy [assessed by the Facial Nerve Grading Scale (FNGS) 2.0] was also significant (p < 0.05) in Bell’s palsy subjects. The spatial information on activated muscle regions may be useful in the diagnosis and treatment of Bell’s palsy in the future.

Introduction and objective: The purpose of this study was to investigate the expression levels and prognostic roles of α-fetoprotein (AFP), carcinoembryonic antigen (CEA), and Ki67 in tumor tissues of intrahepatic cholangiocarcinoma (ICC) patients. Patients or materials and methods: The study involved ninety-two ICC patients with complete clinicopathological data and follow-up information, who had previously undergone radical surgery. AFP, CEA, CD10, CD34, and Ki67 were detected in tumor tissues using immunohistochemistry. Statistical tests were used to identify independent risk factors and their associations with overall survival (OS) and disease-free survival (DFS). Results: AFP, CEA and Ki67 were strongly correlated with prognosis. Univariate analysis indicated that higher AFP (P = 0.002), CEA (P

Three-dimensional multicellular spheroids (MCSs) have received extensive attention in the field of biomedicine due to their ability to simulate the structure and function of tissues in vivo more accurately than traditional in vitro two-dimensional models and to simulate cell–cell and cell extracellular matrix (ECM) interactions. It has become an important in vitro three-dimensional model for tumor research, high-throughput drug screening, tissue engineering, and basic biology research. In the review, we first summarize methods for MCSs generation and their respective advantages and disadvantages and highlight the advances of hydrogel and microfluidic systems in the generation of spheroids. Then, we look at the application of MCSs in cancer research and other aspects. Finally, we discuss the development direction and prospects of MCSs

Angiotensin-converting enzyme-2 (ACE2) has been recognized as the binding receptor for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Flow cytometry demonstrated that there was little to no expression of ACE2 on most of the human peripheral blood-derived immune cells including CD4+ T, CD8+ T, activated CD4+ /CD8+ T, Tregs, Th17, NKT, B, NK cells, monocytes, dendritic cells, and granulocytes. There was no ACE2 expression on platelets and very low level of ACE2 protein expression on the surface of human primary pulmonary alveolar epithelial cells. The ACE2 expression was markedly upregulated on the activated type 1 macrophages (M1). Immunohistochemistry demonstrated high expressions of ACE2 on human tissue macrophages, such as alveolar macrophages, Kupffer cells within livers, and microglial cells in brain at steady state. The data suggest that alveolar macrophages, as the frontline immune cells, may be directly targeted by the SARS-CoV-2 infection and therefore need to be considered for the prevention and treatment of COVID-19.

The phosphoenolpyruvate-dependent glucose phosphotransferase system (PTSGlc) is the major uptake system responsible for transporting glucose, and is involved in glucose translocation and phosphorylation in Corynebacterium glutamicum. For the longest time, the PTSGlc was considered as the only uptake system for glucose. However, some PTS-independent glucose uptake systems (non-PTSGlc) were discovered in recent years, such as the coupling system of inositol permeases and glucokinases (IPGS) and the coupling system of β-glucoside-PTS permease and glucokinases (GPGS). The products (e.g. lysine, phenylalanine and leucine) will be increased because of the increasing intracellular level of phosphoenolpyruvate (PEP), while some by-products (e.g. lactic acid, alanine and acetic acid) will be reduced when this system become the main uptake pathway for glucose. In this review, we survey the uptake systems for glucose in C. glutamicum and their composition. Furthermore, we summarize the latest research of the regulatory mechanisms among these glucose uptake systems. Detailed strategies to manipulate glucose uptake system are addressed based on this knowledge.

In addition to cognitive impairments, depression symptoms were reported in subcortical vascular mild cognitive impairment. Although hippocampal alterations were associated with cognitive decline in subcortical vascular mild cognitive impairment, the neural mechanism underlying depression symptoms remains unclear. Thus, a cohort of 18 patients with depression symptoms, 17 patients without depression symptoms, and 23 normal controls was used. Functionally, significantly altered resting-state functional connectivity between hippocampal emotional sub-region and right posterior cingulate cortex, between hippocampal cognitive sub-region and right inferior parietal gyrus and between hippocampal perceptual sub-region and left inferior temporal gyrus were identified among three groups. Structurally, significantly altered structural associations between hippocampal emotional sub-region and 6 frontal regions/right pole part of superior temporal gyrus/right inferior occipital gyrus, between hippocampal cognitive sub-region and right orbital part of inferior frontal gyrus /right anterior cingulate cortex, and between hippocampal perceptual and right orbital part of inferior frontal gyrus / left inferior temporal gyrus / left thalamus were identified among the three groups. Further analyses also showed correlations between functional connectivity and depression symptoms and/or cognitive impairments of patients. Together, these results showed different patterns of functional and structural alterations of the hippocampal sub-regions in the subcortical vascular mild cognitive impairment with and without depression, which might be specially associated with the depression symptoms and cognitive impairments in these patients.

Background Hepatic adrenal ectopia is a common clinical diagnosis, whereas adrenal tumors developed from hepatic adrenal ectopia are rare. Hepatic adrenal tumors are easily misdiagnosed as hepatic carcinoma and frequently treated by unnecessary operations. Case presentation A 50-year-old female patient was hospitalized due to B-ultrasonic detection of “right focal liver lesions.” After hospitalization, enhanced CT examination was performed. A 2.2 cm × 1.8 cm tumor was found in the seventh section of the right liver, as indicated by obvious enhancement of the arterial phase and low density during the portal vein and delay stages. Enhanced MRI examination detected a 2.0 cm × 1.8 cm tumor on the right liver, which was considered a “primary hepatic carcinoma”. The patient was treated by open hepatectomy and recovered well after the operation. The postoperative pathological diagnosis was hepatic adrenal adenoma. No relapse was observed through the 1-year follow-up visit. Conclusions According to imaging manifestations, pathological immunohistochemical treatment, alpha fetoprotein (AFP) and clinical features, hepatic adrenal tumors should be considered in the diagnosis of hepatic carcinoma to prevent misdiagnosis. Hepatic adrenal tumors should be ruled out during the diagnosis to avoid unnecessary operation.

Lysine is widely used in food, medical and feed industries. The biosynthesis of l-lysine is closely related to NADPH level, but the regulation mechanism between the biosynthesis of l-lysine in C. glutamicum and the cofactor NADPH is still not clear. Here, a high intracellular NADPH level strain C. glutamicum XQ-5Δpgi::(zwf-gnd) was constructed by blocking the glycolytic pathway and overexpressing the pentose phosphate pathway in the lysine-producing strain C. glutamicum XQ-5, and the intracellular NADPH level in strain XQ-5Δpgi::(zwf-gnd) was increased from 3.57 × 10–5 nmol/(104 cells) to 1.8 × 10–4 nmol/(104 cell). Transcriptome analyses pointed to Cgl2680 as an important regulator of NADPH levels and l-lysine biosynthesis in C. glutamicum. By knocking out the gene Cgl2680, the intracellular NADPH level of the recombinant C. glutamicum lysCfbr ΔCgl2680 was raised from 7.95 × 10–5 nmol/(104 cells) to 2.04 × 10–4 nmol/(104 cells), consequently leading to a 2.3-fold increase in the NADPH/NADP+ ratio. These results indicated that the regulator Cgl2680 showed the negative regulation for NADPH regeneration. In addition, Cgl2680-deficient strain C. glutamicum lysCfbr ΔCgl2680 showed the increase of yield of both l-lysine and l-leucine as well as the increase of H2O2 tolerance. Collectively, our data demonstrated that Cgl2680 plays an important role in negatively regulating NADPH regeneration, and these results provides new insights for breeding l-lysine or l-leucine high-yielding strain.

Traditionally, the visual motion area (MT) is considered as a brain region specialized for visual motion perception. However, accumulating evidence showed that MT is also related to various functions, suggesting that it is a complex functional area and different functional subregions might exist in this area. To delineate functional subregions of this area, left and right masks of MT were defined using meta-analysis in the BrainMap database, and coactivation-based parcellation was then performed on these two masks. Two dorsal subregions (Cl1 and Cl2) and one ventral subregion (Cl3) of left MT, as well as two dorsal-anterior subregions (Cl1 and Cl2), one ventral-anterior subregion (Cl3), and an additional posterior subregion (Cl4) of right MT were identified. In addition to vision motion, distinct and specific functions were identified in different subregions characterized by task-dependent functional connectivity mapping and forward/reverse inference on associated functions. These results not only were in accordance with the previous findings of a hemispheric asymmetry of MT, but also strongly confirmed the existence of subregions in this region with distinct and specific functions. Furthermore, our results extend the special role of visual motion perception on this area and might facilitate future cognitive study.

Acute lung injury is the main causative factor in paraquat dichloride (PQ)-induced mortality. The innate immune system-triggered detrimental inflammatory cascade plays a vital role in PQ-induced acute lung injury. However, the role of natural killer (NK) cells, which are essential for innate response, in PQ-induced acute lung injury remains largely unknown. Here, we found that in an acute PQ poisoning model, depletion of NK cells attenuated PQ-induced lung injury by inhibiting macrophage polarization towards the M1 type. Specifically, the percentages of NK cells were reduced in the lung, spleen, and peripheral blood in a murine model of acute PQ poisoning. NK cells were aberrantly activated, evidenced by upregulation of the activating markers CD69, CD107a, and NKG2D and downregulation of the inhibitive marker KLRG1. Further, NK-specific depletion in mice greatly prolonged the survival time and ameliorated reactive oxygen species-induced damage following PQ treatment compared with the control group. Importantly, NK cell depletion alleviated macrophage and neutrophil infiltration in the lung and reversed PQ induced-macrophage polarization towards the pro-inflammatory M1 type. Our study demonstrates a crucial role of NK cells and NK cell-to-macrophage interaction in PQ-induced acute lung injury.

The main pathogenesis of type 1 diabetes mellitus (T1DM) is autoimmune-mediated apoptosis of pancreatic islet β cells. We sought to characterize the function of microRNA-203a (miR-203a) on pancreatic islet β cell proliferation and apoptosis. In situ hybridization was used to detect the expression of miR-203a in islet β cells in normal and hyperglycaemic non-obese diabetic (NOD) mice. Cell proliferation was measured by cell counting kit eight and cell apoptosis was detected using flow cytometry. Insulin receptor substrate 2 (IRS2/Irs2) was determined to be a direct target of miR-203a by Luciferase reporter assay. We detected the effects of miR-203a overexpression or inhibition on proliferation and apoptosis of IRS2-overexpressing or IRS2-knockdown MIN6 cells respectively, and preliminarily explored the downstream targets of the IRS2 pathway. NOD mice model was used to detect miR-203a inhibitor treatment for diabetes. Our experiment showed miR-203a was upregulated in pancreatic β cells of hyperglycaemic NOD mice. Elevated miR-203a expression inhibited the proliferation and promoted the apoptosis of MIN6 cells. IRS2/Irs2 is a novel target gene directly regulated by miR-203a and miR-203a overexpression downregulated the expression of IRS2. Irs2 silencing reduced cell proliferation and increased apoptosis. Irs2 overexpression could abolish the pro-apoptotic and anti-proliferative effects of miR-203a on MIN6 cells. Hyperglycemia in newly hyperglycemic NOD mice was under control after treatment with miR-203a inhibitor. Our study suggests that miR-203a regulates pancreatic β cell proliferation and apoptosis by targeting IRS2, treatment with miR-203a inhibitors and IRS2 might provide a new therapeutic strategy for T1DM.

Background. Clinical and experimental data suggest that ultrasound stimulation (US) at acupoints can produce similar effective treatment compared to manual acupuncture (MA). Although the brain activation to MA at acupoints is investigated by numerous studies, the brain activation to US at acupoints remains unclear. Methods. In the present work, we employed task state functional magnetic resonance imaging (fMRI) to explore the human brain’s activation to US and MA at ST 36 (Zusanli) which is one of the most commonly used acupoints in acupuncture-related studies. 16 healthy subjects underwent US and MA procedures in an interval of more than one week. On-off block design stimulation was used for the recording of fMRI-related brain patterns. Results. Both US and MA at ST 36 produced activations in somatosensory and limbic/paralimbic regions (postcentral gyrus, insula, middle prefrontal cortex, and anterior cingulate cortex). Only US at ST 36 produced a significant signal increase in the inferior parietal lobule and decrease in the posterior cingulate cortex, whereas MA at ST 36 produced a significant signal increase in the lentiform nucleus and cerebellum. Conclusions. Our results indicate that US may be a possible noninvasive alternative method to MA due to its similar activation patterns.

Background. Acupuncture has been found to be an effective treatment for functional dyspepsia (FD). Currently, several types of acupuncture have been developed but it is not clear which type is suitable for FD. Currently, doctors often rely on experience to decide which form of acupuncture to apply. Herein, we employed network meta-analysis (NMA) to compare the effectiveness of various methods of acupuncture in the treatment of functional dyspepsia. Methods. We searched for randomized controlled trials (RCTs) of acupuncture treatments for functional dyspepsia in seven databases; PubMed, the Cochrane Library, Embase, Wanfang database, China National Knowledge Infrastructure (CNKI) database, Chinese Science and Technique Journals (CQVIP), and Chinese Biomedical Database (CBM) from the date of database inception to October 10, 2019. Cochrane risk of bias tool was used to analyze the risk of bias of the included RCTs. Pairwise meta-analyses were performed with RevMan 5.3 and the network meta-analysis of the included RCTs was performed using the frequentist framework. Results. A total of 35 studies involving 3301 patients and 10 interventions were eligible for this study. NMA results showed that five types of acupuncture (manual acupuncture, acupoint application, moxibustion, acupoint catgut embedding, and warm acupuncture alone) all were superior to prokinetics (itopride, mosapride, and domperidone) and sham acupuncture in terms of improving the symptoms of functional dyspepsia. Specifically, manual acupuncture and electroacupuncture were more effective in improving the MOS 36 Item Short-Form Health Survey (SF-36) compared to itopride and sham acupuncture, and electroacupuncture was the best among the three acupuncture therapies (acupuncture, electroacupuncture, and acupoint catgut embedding). Moxibustion and manual acupuncture were more effective in improving Nepean Dyspepsia Life Quality Index (NDLQI) compared to itopride, domperidone, and sham acupuncture; moxibustion ranks first among the three acupuncture therapies (acupuncture, electroacupuncture, moxibustion). Conclusions. These results showed that manual acupuncture alone was the most effective therapy for FD. It should, therefore, be considered as an alternative treatment for FD patients who are unresponsive to prokinetics or intolerant to the adverse effects of prokinetics. We recommend further multiple centers and high-quality RCT studies to confirm the present findings.

Background. Heart failure (HF) is the terminal stage of all cardiovascular events. Although implantable cardioverter defibrillator (ICD) therapies have reduced mortality among the high-risk HF population, it is necessary to determine whether certain factors can predict mortality even after cardiac device implantation. Growth stimulation expressed gene 2 (ST2) is an emerging biomarker for HF patient stratification in different clinical settings. Aims. This study aimed to investigate the relationship between baseline soluble ST2 (sST2) levels in serum and the clinical outcomes of high-risk HF patients with device implantation. Methods. Between January 2017 and August 2018, we prospectively recruited consecutive patients implanted with an ICD for heart failure, with LVEF ≤35% as recommended, and analyzed the basic characteristics, baseline serum sST2, and NT-proBNP levels, with at least 1-year follow-up. All-cause mortality was the primary endpoint. Results. During a 643-day follow-up, all-cause mortality occurred in 16 of 150 patients (10.67%). Incidence of all-cause mortality increased significantly in patients with sST2 levels above 34.98846 ng/ml (16.00% vs. 5.33%, P = 0.034 ). After adjusting the model (age, gender, device implantation, prevention of sudden death, LVEDD, LVEF, WBC and CLBBB, hsTNT, etiology, and eGFR) and the model combined with NT-proBNP, the risk of all-cause death was increased by 2.5% and 1.9%, respectively, per ng/ml of sST2. The best sST2 cutoff for predicting all-cause death was 43.42671 ng/ml (area under the curve: 0.72, sensitive: 0.69, and specificity: 0.69). Compared to patients with sST2 levels below 43.42671 ng/ml, the risk of all-cause mortality was higher in those with values above the threshold (5.1% vs. 21.2%, P = 0.002 ). ST2 level ≥43.42671 ng/ml was an independent predictor of all-cause mortality (HR: 3.30 [95% CI 1.02–10.67]). Age (HR: 1.06 [95% CI: 1.01–1.12]) and increased NT-proBNP per 100 (HR: 1.02 [95% CI: 1.01–1.03]) were also associated with all-cause mortality in ICD patients. Conclusions. sST2 level was associated with risk of all-cause mortality, and a threshold of 43.43 ng/ml showed good distinguishing performance to predict all-cause mortality in patients with severe heart failure, recommended for ICD implantation. Patients with sST2 levels more than 43.42671 ng/ml even after ICD implantation should therefore be monitored carefully.

Adenosine monophosphate-activated protein kinase (AMPK) is the main energy sensor in mammals, but limited information is available regarding its role as an energy sensor in nutrient-restricted fish particularly in period of overwinter starvation. The present study aimed to investigate the role of AMPK in the grass carp Ctenopharyngodon idellus through characterization of AMPK full-length cDNAs and the measurement of transcriptional activity in response to overwinter starvation. AMPK is a heterotrimeric serine/threonine kinase that consists of a catalytic alpha (α) subunit complexed with two regulatory subunits, beta (β) and gamma (γ). In our study, we identified nine isoforms of the AMPK family in grass carp and obtained their complete coding sequences (CDS). In the grass carp, the α subunit is encoded by two isoforms (α1 and α2). The β and γ subunits are encoded by three (β1a, β1b, β2) and four isoforms (γ1, γ2a, γ2b, γ3), respectively. AMPK isoforms in grass carp possess structural features similar to mammalian AMPK and exhibit a high degree of homology with other fish and vertebrate AMPK sequences. The mRNA of nine grass carp AMPK isoforms were found to be expressed in a wide range of tissues in vivo, but the abundance of each AMPK mRNA demonstrated a tissue-dependent expression pattern, indicating that they might be key complexes playing the role of energy metabolism sensors during overwinter starvation conditions. Compared to expression levels in control fish (week 0), the expression of various AMPK isoforms significantly increased in the hepatopancreas of fish exposed to 1 week or more of overwinter starvation conditions as follows: week 1 (AMPK α1 and AMPK α2), week8 (AMPK β1b and AMPK γ2b), week 12 (AMPK β2 and AMPK γ1), and week 16 (AMPK β1a, AMPK γ2a, and AMPK γ3). Additionally, compared to expression levels in control fish (week 0), the expression of various AMPK isoforms significantly increased in the adipose tissue of fish exposed to 1 week or more of overwinter starvation conditions as follows: week 1(AMPK β1a and AMPK β1b), week 4 (AMPK α1, AMPK α2, AMPK γ1, AMPK γ2b and AMPK γ3), and week 8 (AMPK β2 and AMPK γ2a). Further in vitro analysis revealed that the mRNA levels of AMPK isoforms in hepatocytes (AMPK α1, AMPK α2, AMPK β1a, AMPK β1b, AMPK β2, AMPK γ2b and AMPK γ3) and adipocytes (AMPK γ2a, AMPK γ2b and AMPK γ3) changed significantly with in the first 24 h of exposure to the overwinter starvation conditions. These findings confirm that nine AMPK subunits are present in grass carp and that all encode proteins with conserved functional domains. The nine AMPK subunits are all regulated at the transcriptional levels to manage excess energy expenditure during overwinter starvation stress.

Temperature is a key environmental factor, and understanding how its fluctuations affect physiological and metabolic processes is critical for fish. The present study characterizes the energy response and fatty acid metabolism in Onychostoma macrolepis exposed to low temperature (10 °C). The results demonstrated that cold stress remarkably disrupted the energy homeostasis of O. macrolepis, then the AMP-activated protein kinase (AMPK) could strategically mobilize carbohydrates and lipids. In particular, when the O. macrolepis were faced with cold stress, the lipolysis was stimulated along with the enhanced fatty acid β-oxidation for energy, while the fatty acid synthesis was supressed in the early stage. Additionally, the fatty acid composition analysis suggested that saturated fatty acid (SFA) might accumulate while monounsaturated fatty acid (MUFA) and polyunsaturated fatty acid (PUFA) in storage lipids (mainly containing non-polar lipid, NPL) could be utilized to supply energy during cold acclimation. Altogether, this study may provide some meritorious for understanding the cold-tolerant mechanism of fish in the viewpoint of energy balance combined with fatty acid metabolism, and thus to contribute to this species rearing in fish farms in the future.

The quality of biomolecular simulations critically depends on the accuracy of the force field used to calculate the potential energy of the molecular configurations. Currently, most simulations employ non-polarisable force fields, which describe electrostatic interactions as the sum of Coulombic interactions between fixed atomic charges. Polarization of these charge distributions is incorporated only in a mean-field manner. In the past decade, extensive efforts have been devoted to developing simple, efficient, and yet generally applicable polarisable force fields for biomolecular simulations. In this review, we summarise the latest developments in accounting for key biomolecular interactions with polarisable force fields and applications to address challenging biological questions. In the end, we provide an outlook for future development in polarisable force fields., Comment: 25 pages, 3 figures

With the development of biomedical technology, personalized diagnosis and treatment at the single-cell level are becoming more important in the medical field. As one of the most powerful tools, microfluidic chips have shown significant potential for various applications related to cell separation, cell proliferation, and cell behavior analysis. However, fabricating microfluidic devices requires complicated procedures and high-cost equipment. In this study, an optofluidic maskless lithography method was proposed for rapid fabrication of microfluidic devices integrated with microwells. Through the use of this approach, microwells can be on-line designed and the exposure patterns can be modulated. Single or multi polystyrene microspheres were successfully trapped by using the designed microwells. The capture of MCF-7 cells and cell arrays indicated that the microfluidic devices fabricated in the present study can be applied for cell research.

Urotoxin (α-KTx 6), a peptide from venom of the Australian scorpion Urodacus yaschenkoi, is the most potent inhibitor of Kv1.2 described to date (IC50 = 160 pM). The native peptide also inhibits Kv1.1, Kv1.3 and KCa3.1 with nanomolar affinity but its low abundance in venom precluded further studies of its actions. Here we produced recombinant Urotoxin (rUro) and characterized the molecular determinants of Kv1 channel inhibition. The 3D structure of rUro determined using NMR spectroscopy revealed a canonical cysteine-stabilised α/β (CSα/β) fold. Functional assessment of rUro using patch-clamp electrophysiology revealed the importance of C-terminal amidation for potency against Kv1.1–1.3 and Kv1.5. Neutralization of the putative pore-blocking K25 residue in rUro by mutation to Ala resulted in a major decrease in rUro potency against all Kv channels tested, without perturbing the toxin’s structure. Reciprocal mutations in the pore of Uro-sensitive Kv1.2 and Uro-resistant Kv1.5 channels revealed a direct interaction between Urotoxin and the Kv channel pore. Our experimental work supports postulating a mechanism of action in which occlusion of the permeation pathway by the K25 residue in Urotoxin is the basis of its Kv1 inhibitory activity. Docking analysis was consistent with occlusion of the pore by K25 and the requirement of a small, non-charged amino acid in the Kv1 channel vestibule to facilitate toxin-channel interactions. Finally, computational studies revealed key interactions between the amidated C-terminus of Urotoxin and a conserved Asp residue in the turret of Kv1 channels, offering a potential rationale for potency differences between native and recombinant Urotoxin.

Sodium channels are important proteins in modulating neuronal membrane excitability. Genetic studies from patients and animals have indicated neuronal sodium channels play key roles in pain sensitization. We identified WB4101 (2-(2,6-Dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane hydrochloride), an antagonist of α1-adrenoceptor, as a Nav1.7 inhibitor from a screen. The present study characterized the effects of WB4101 on sodium channels. We demonstrated that WB4101 inhibited both Nav1.7 and Nav1.8 channels with similar levels of potency. The half-inhibition concentrations (IC50 values) of WB4101 were 11.6 ± 2.07 and 1.0 ± 0.07 µM for the resting and inactivated Nav1.7 channels, respectively, and 8.67 ± 1.31 and 0.91 ± 0.25 µM for the resting and inactivated Nav1.8 channels, respectively. WB4101 induced a hyperpolarizing shift in the voltage-dependent inactivation for both Nav1.7 (15 mV) and Nav1.8 (20 mV) channels. The IC50 values for the open-state sodium channel were 2.50 ± 1.16 µM for Nav1.7 and 1.1 ± 0.2 µM for Nav1.8, as determined by the block of persistent late currents in inactivation-deficient Nav1.7 and Nav1.8 channels, respectively. Consistent with the state-dependent block, the drug also displayed use-dependent inhibitory properties on both wild-type Nav1.7 and Nav1.8 channels, which were removed by the local anesthetic-insensitive mutations but still existed in the inactivation-deficient channels. Further, the state-dependent inhibition on sodium channels induced by WB4101 was demonstrated in dorsal root ganglion neurons. In conclusion, the present study identified WB4101 as a sodium channel blocker with an open-state-dependent property, which may contribute to WB4101's analgesic action.

This study was performed to evaluate the feasibility of the splenic bed laparoscopic splenectomy approach (SBLS) for massive splenomegaly (≥30 cm) in patients with hypersplenism secondary to portal hypertension and liver cirrhosis. Patients who underwent laparoscopic splenectomy (LS) from January 2012 to December 2016 were retrospectively reviewed. We performed LS in 83 patients with massive splenomegaly (≥30 cm) secondary to portal hypertension and liver cirrhosis. Of these patients, 37 underwent the SBLS and 46 underwent anterior LS (ALS). Five patients in the ALS group and none in the SBLS group underwent conversion to open surgery. The operation time, intraoperative blood loss volume, transfusion volume, frequency of transfusion, hemorrhage of short gastric vessels, conversion rate, postoperative hospital stay, and incidence of pancreatic fistula were all significantly lower in the SBLS than ALS group (all P < 0.05). No death or postoperative bleeding occurred in the two groups, and there were no significant differences in age, gender, spleen size, hemoglobin level, platelet count, prothrombin time, Child-Pugh class, hypoproteinemia, or ascites (all P > 0.05). The SBLS is more feasible and effective than ALS in patients with massive splenomegaly (≥30 cm) secondary to portal hypertension and liver cirrhosis.

To compare the safety and efficacy of benzbromarone and febuxostat in hyperuricemia patients with estimated glomerular filtration rate (eGFR) 20–60 mL/min/1.73 m2. This study was a single-centered, parallel-grouped, randomized clinical trial (RCT). We randomly assigned hyperuricemia participants with eGFR 20–60 mL/min/1.73 m2 into benzbromarone and febuxostat treatment group. Drugs were adjusted by titration from small doses. Seventy-three eligible participants enrolled, 66 subjects (33 in each group) were included finally for analysis. When compared to baseline, serum uric acid (SUA) decreased significantly after treatment in both groups, but no differences were detected among all the follow-up points. After 12-month treatment, eGFR did not have significant change in both groups. In the benzbromarone group, kidney stones in one case increased in quantity. In the febuxostat group, kidney stones in one case became smaller in size and in two cases vanished completely. Both drugs did not increase myocardial enzymes significantly after the treatment. In addition, hemoglobin increased significantly in the two groups (p

Cytochrome P450 enzymes (CYP enzymes) catalyze important metabolic reactions of exogenous and endogenous substrates, including fatty acid. In this study, we cloned the complete CDS of the cytochrome P450 2AA (CYP2AA) gene from the grass carp (Ctenopharyngodon idella) for the first time. CYP2AA consisted of 1500 bp, which encoded a predicted protein of 499 amino acids. The identities of CYP2AA between C. idella and zebrafish were 86%. It consists of the conserved heme-binding motif FXXGXXXCXG. Quantitative real-time PCR analysis indicated that CYP2AA mRNA in C. idella was highly expressed in liver and adipose tissue. The effects of fish oil and lard oil in diets on expression of CYP2AA mRNA in vivo were also investigated. The fish oil (FO) group exhibited significantly higher CYP2AA expression in adipose tissue than the lard oil (LO) group (P