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Differentiation of preadipocytes into functional adipocytes could be a major target for repressing obesity-induced insulin resistance (IR). However, the molecular mechanisms involved in adipogenesis and the development of IR are unclear. We report, for the first time, that miR-574-5p, a novel miRNA, promotes adipogenesis to suppress IR. An increase in the level of miR-574-5p significantly induced the differentiation of preadipocytes into mature adipocytes. Conversely, reduction of miR-574-5p levels blocked the differentiation of preadipocytes in vitro. In a dual-luciferase reporter assay, it was shown that homeobox A5 (HOXA5) promoted the transcription of miR-574-5p to induce the differentiation of preadipocytes. Hdac9, a direct downstream target of miR-574-5p, was involved in the regulation of adipocyte differentiation. The overexpression of miR-574-5p also promoted adipogenesis in subcutaneous fat to alleviate IR in high-fat-diet-fed mice. Additionally, miR-574-5p expression was significantly higher in the subcutaneous adipose tissue of obese patients without type 2 diabetes than in those with type 2 diabetes. There was an increase in HOXA5 expression and a decrease in histone deacetylase 9 (HDAC9) expression in the subcutaneous fat of obese patients without type 2 diabetes. These results suggest that miR-574-5p may be a potential therapeutic target for combating obesity-related IR., Graphical abstract, This is the first study to report that the upregulation of the expression of HOXA5-miR-574-5p-HDAC9 axis components can promote adipogenesis and maintenance of subcutaneous adipose expansion to suppress insulin resistance; this could be a potential therapeutic target for combating obesity-related insulin resistance.

In recent years, high-fat diet (HFD) has been widely applied in aquaculture, which reduces the intestinal health of cultured fish. The current study evaluated the protective effects of nano-selenium (nano-Se) on intestinal health of juvenile grass carp (Ctenopharyngodon idella) fed with HFD. A total of 135 experimental fish were fed with a regular diet (Con), a HFD (HFD) and a HFD containing nano-Se at 0.6 mg/kg (HSe) for 10 weeks. The results showed that dietary nano-Se significantly improved the survival rate and feed efficiency which were reduced by HFD in juvenile grass carp (P

The urokinase plasminogen activator (uPA) plays a critical role in tumor cell invasion and migration and is a promising anti-metastasis target. 6-Substituted analogs of 5-N,N-(hexamethylene)amiloride (HMA) are potent and selective uPA inhibitors that lack the diuretic and anti-kaliuretic properties of the parent drug amiloride. However, the compounds display pronounced selectivity for human over mouse uPA, thus confounding interpretation of data from human xenografted mouse models of cancer. Here, computational and experimental findings reveal that residue 99 is a key contributor to the observed species selectivity, whereby enthalpically unfavorable expulsion of a water molecule by the 5-N,N-hexamethylene ring occurs when residue 99 is Tyr (as in mouse uPA). Analog 7 lacking the 5-N,N-hexamethylene ring maintained similar water networks when bound to human and mouse uPA and displayed reduced selectivity, thus supporting this conclusion. The study will guide further optimization of dual-potent human/mouse uPA inhibitors from the amiloride class as anti-metastasis drugs.

Background The aim of this study was to investigate the prognostic value of arginase-1 (Arg-1) and glypican-3 (GPC-3) in patients with intrahepatic cholangiocarcinoma (ICC). Methods Two hundred and thirty-seven patients with ICC were included in this study. All patients had undergone radical surgery and had complete clinical information. Immunohistochemistry was used to assess the levels of Arg-1 and GPC-3 in ICC tissues. Univariate and multivariate analyses were conducted to identify independent risk factors in ICC. The relationship between Arg-1 and GPC-3 levels and patient survival was determined using the Kaplan-Meier method. Results High Arg-1 and GPC-3 expression levels were associated with poor prognosis in patients with ICC, and they could be as new prognostic biomarkers in ICC. Conclusion Arg-1 and GPC-3 can serve as independent prognostic biomarkers in ICC.

Background and purpose KV 1.3 potassium channels play a predominant role in regulating calcium signalling that is essential for the activation and proliferation of effector memory T (TEM ) cells. This ion channel has been recognized as a promising therapeutic target against various autoimmune diseases. Experimental approach In a high-throughput screening programme, WP1066 was identified as a KV 1.3 channel inhibitor. Using molecular biology and electrophysiological methods, the mechanism(s) underlying WP1066 blockade of Kv1.3 channels was investigated. Using TEM cell proliferation assay and mouse delayed-type hypersensitivity (DTH) model, the effects of WP1066 were examined. Key results WP1066 blocked KV 1.3 channels in a dose-dependent manner with an IC50 of 3.2 μM and induced a hyperpolarizing shift of the steady-state inactivation curve. This blockade was use-dependent, as WP1066 interacted preferentially with channels in their open state, rather than the closed state or inactivated state. When the residues located in the S6 domain scaffolding the inner vestibule, were sequentially mutated, the potency of WP1066 was significantly impaired, especially by mutations A413C and I420C, indicating a higher affinity of interacting sites for WP1066. Moreover, WP1066 effectively suppressed mouse TEM cell proliferation in vitro and mouse DTH reaction in vivo. Conclusions and implications The results presented here have identified WP1066 as a KV 1.3 channel blocker with an open-state-dependent property, providing fundamental evidence for the application of WP1066 in further immunomodulatory studies targeting KV 1.3 channels.

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.

Aims To establish a simple criterion for determining a failed His-bundle pacing (HBP). This criterion states that if stimulus to QRS end interval is longer than His-bundle potential to QRS end interval (‘S-QRSend > H-QRSend’) then a failed HBP can be determined. Methods and results We performed retrospective analysis on 737 pacing tests around His-bundle in 241 patients and prospective analysis on 400 tests in 123 patients. A successful HBP is defined as that whole His-bundle is captured with or without capture of adjacent ventricular myocardium, otherwise, a failed HBP was considered. The output criteria and effective refractory period criteria were used as the gold standards for determining a successful HBP. The gold standards are that if decreasing the pacing output or pacing cycle length to a certain level results in duration or morphology changes of QRS, then a successful HBP is ascertained. In retrospective analysis of patients with normal His-Purkinje conduction, a failed HBP was determined in 31% (154/492) of pacing tests according to ‘S-QRSend > H-QRSend’; all of them were validated by the gold standards (specificity = 100%). In prospective study, a failed HBP was confirmed according to the simple criterion with 100% accuracy in 33% (79/241) pacing tests. This simple criterion was also suitable for patients with His-Purkinje conduction disease although cases with ‘S-QRSend > H-QRSend’ rarely occurred. Conclusion A failed HBP can be easily and reliably determined solely by ‘S-QRSend > H-QRSend’ in more than 30% pacing tests.

Objective: We conducted this study to assess the predictive value of tumor-infiltrating lymphocytes (TILs) for progression-free survival (PFS) and overall survival (OS) in intrahepatic cholangiocarcinoma (ICC) patients after complete resection. Methods: Sixty-eight patients with resectable ICC were included in this study. We studied stromal TIL density and scored it by staining sections from surgically resected ICC patients with hematoxylin and eosin (HE). The clinical data and prognosis of ICC patients were obtained by searching clinical and follow-up records. Results: A stromal TIL negative status was an independent predictor of poor OS (HR=2.84, 95% CI 1.48- 5.46, P

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.

Introduction The safety and feasibility of laparoscopic splenectomy plus selective esophagogastric devascularization (LSSD) via the spleen bed for cirrhotic portal hypertension have not been well studied. Aim To assess the safety and feasibility of LSSD via the spleen bed for patients with cirrhotic portal hypertension. Material and methods From June 2012 to December 2017, 423 patients suffering from portal hypertension and hypersplenism with liver cirrhosis underwent surgery in our department. One hundred and sixty-seven of these patients received totally LSSD, and the others received open splenectomy and esophagogastric devascularization (OSD). The characteristics, intraoperative and postoperative details and complications of the two groups were compared. Results The operations were successfully performed in all patients. Intraoperative blood loss volume and blood transfusion were similar between the two groups (all p-values > 0.05). Postoperative length of hospital stay and time to oral intake were significantly shorter, but operation time was longer in the LSSD group compared with the OSD group (all p < 0.05). However, postoperative portal vein diameter was significantly smaller in the LSSD group (p < 0.001). The postoperative grade of varices was significantly lower in the LSSD group (p = 0.030). No significant differences were detected between the two groups regarding postoperative liver function, but the incidences of pancreatic leakage, pleural effusion, and wound infections were higher in the OSD group (all p < 0.05). Conclusions LSSD via the spleen bed is safe and feasible for liver cirrhosis and portal hypertension.

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.

Establishing a physiological microenvironment in vitro that is suitable for cell and tissue growth is essential for medical research. Microfluidic chips are widely used in the construction of a microenvironment and the analysis of cell behavior in vitro ; however, the design and manufacture of microfluidic chips for the long-term culture of a tumor model tends to be highly complex and time-consuming. In this paper, we propose a method for the rapid fabrication of a microfluidic chip for multi-dimensional cell co-culture. A major advantage of this method is that the microfluidic chip can be divided into several sections by micro-pillar arrays to form different functional regions to grow two- and three-dimensional cell culture on the same matrix. At the micro-scale, the surface tension between the gelatin methacryloyl-encapsulated cells and micro-pillars prevents the leakage of the hydrogel, and the hydrogel provides a three-dimensional microenvironment for cell growth. Our results of long-term cell culture and preclinical drug screening showed that cells cultured in a two-dimensional monolayer differ from three-dimensional cultured cells in terms of morphology, area, survival rate, proliferation, and drug resistance. This method shows potential for use in the study of cell behavior, drug screening, and tissue engineering.

Tumor cell clusters of varying sizes and densities have different metastatic potentials. Three-dimensional (3D) patterned structures with rational topographical and mechanical properties are capable of guiding the 3D clustering of tumor cells. In this study, single femtosecond laser pulses were used to fabricate individual high-aspect-ratio micropillars via two-photon polymerization (TPP). By combining this approach with capillary-force self-assembly, complex 3D microstructure patterns were constructed with a high efficiency. The microstructures were able to regulate the formation of cell clusters at different cell seeding densities and direct self-guided 3D assembly of cell clusters of various sizes and densities. Localization of cell clusters was achieved using grid-indexed samples to address individual cell clusters, which holds great promise for in situ cell cluster culture and monitoring and for applications such as RNA sequencing of cell clusters.

Objective: To investigate the association between susceptibility to type 1 diabetes mellitus (T1DM) and polymorphisms (rs1143627 and rs1143643) in the interleukin 1 beta (IL1B) gene in the Chinese Han population.Methods: The Meso Scale Discovery (MSD) method was used to detect the concentration of IL-1β in 24 T1DM patients and 27 healthy controls. MassARRAY was used to analyze the polymorphisms in the IL1B gene in 510 patients with classic T1DM and 531 healthy controls. The general data of the T1DM patients and healthy controls were compared by the chi-square test and Mann-Whitney U test. The chi-square test and logistic regression were used to analyze the frequency distributions of alleles and genotypes of polymorphisms in the IL1B gene. The Kruskal-Wallis H test and chi-square test were used for the genotype-phenotype analysis of rs1143627 and rs1143643 in the IL1B gene.Results: ① The concentration of IL-1β in T1DM patients was significantly higher than that in healthy controls. ② rs1143627 and rs1143643 in the IL1B gene were significantly correlated with the positivity rates for IA-2A and ZnT8A; genotype GG at rs1143627 and genotype CC at rs1143643 in the case group showed lower positivity rates for IA-2A and ZnT8A. ③ There was no significant difference in the genotypes or allele frequencies at rs1143627 (GG/GA/AA) or rs1143643 (CC/CT/TT) between the case group and control group (p > 0.05). ④ rs1143627 and rs1143643 were not found to be linked to T1DM susceptibility under different genetic models.Conclusion: rs1143627 and rs1143643 in the IL1B gene correlate with the positivity rate of IA-2A and ZnT8A in Chinese Han individuals with T1DM.

BackgroundWe developed the Stem Cell Educator therapy among multiple clinical trials based on the immune modulations of multipotent cord blood-derived stem cells (CB-SC) on different compartments of immune cells such as T cells and monocytes/macrophages in diabetes and other autoimmune diseases. However, the effects of CB-SC on the B cells remained unclear. To better understand the molecular mechanisms underlying the immune education of CB-SC, we explored the modulations of CB-SC on human B cells.MethodsCB-SC were isolated from human cord blood units and confirmed by flow cytometry with different markers for their purity. B cells were purified by using anti-CD19 immunomagnetic beads from human peripheral blood mononuclear cells (PBMC). Next, the activated B cells were treated in the presence or absence of coculture with CB-SC for 7 days before undergoing flow cytometry analysis of phenotypic change with different markers. RT-PCR was utilized to evaluate the levels of galectin expressions with or without treatment of activated B cells in order to find the key galectin contributing to the B-cell modulation.ResultsFlow cytometry demonstrated that the proliferation of activated B cells was markedly suppressed in the presence of CB-SC, leading to the down-regulation of immunoglobulin productions from the activated B cells. Phenotypic analysis revealed that treatment with CB-SC increased the percentage of IgD+CD27-naïve B cells, but decreased the percentage of IgD-CD27+switched B cells. Transwell assay showed that the immune suppression of CB-SC on B cells was dependent on the manner of cell-cell contact via Gal-9 molecule, as confirmed by the blocking experiment with the anti-Gal-9 monoclonal antibody. Mechanistic studies demonstrated that both calcium levels of cytoplasm and mitochondria were down-regulated after the treatment with CB-SC, causing the decline of mitochondrial membrane potential in the activated B cells. Western blot exhibited that the levels of phosphorylated Akt and Erk1/2 signaling proteins in the activated B cells were also markedly reduced in the presence of CB-SC.ConclusionsCB-SC displayed multiple immune modulations on B cells through the Gal-9-mediated cell-cell contact mechanism and calcium flux/Akt/Erk1/2 signaling pathways. The data advances current understanding about the molecular mechanisms underlying the Stem Cell Educator therapy to treat autoimmune diseases in clinics.

Art therapy, as a non-pharmacological medical complementary and alternative therapy, has been used as one of medical interventions with good clinical effects on mental disorders. However, systematically reviewed in detail in clinical situations is lacking. Here, we searched on PubMed for art therapy in an attempt to explore its theoretical basis, clinical applications, and future perspectives to summary its global pictures. Since drawings and paintings have been historically recognized as a useful part of therapeutic processes in art therapy, we focused on studies of art therapy which mainly includes painting and drawing as media. As a result, a total of 413 literature were identified. After carefully reading full articles, we found that art therapy has been gradually and successfully used for patients with mental disorders with positive outcomes, mainly reducing suffering from mental symptoms. These disorders mainly include depression disorders and anxiety, cognitive impairment and dementias, Alzheimer’s disease, schizophrenia, and autism. These findings suggest that art therapy can not only be served as an useful therapeutic method to assist patients to open up and share their feelings, views, and experiences, but also as an auxiliary treatment for diagnosing diseases to help medical specialists obtain complementary information different from conventional tests. We humbly believe that art therapy has great potential in clinical applications on mental disorders to be further explored.

Here we report the vibrational spectra of deprotonated serine calculated from the classical molecular dynamics (MD) simulations and thermostated ring-polymer molecular dynamics (TRPMD) simulation with third-order density-functional tight-binding. In our earlier study [Inakollu and Yu, “A systematic benchmarking of computational vibrational spectroscopy with DFTB3: Normal mode analysis and fast Fourier transform dipole autocorrelation function,” J. Comput. Chem. 39, 2067 (2018)] of deprotonated serine, we observed a significant difference in the vibrational spectra with the classical MD simulations compared to the infrared multiple photon dissociation spectra. It was postulated that this is due to neglecting the nuclear quantum effects (NQEs). In this work, NQEs are considered in spectral calculation using the TRPMD simulations. With the help of potential of mean force calculations, the conformational space of deprotonated serine is analyzed and used to understand the difference in the spectra of classical MD and TRPMD simulations at 298.15 and 100 K. The high-frequency vibrational bands in the spectra are characterized using Fourier transform localized vibrational mode (FT-νNAC) and interatomic distance histograms. At room temperature, the quantum effects are less significant, and the free energy profiles in the classical MD and the TRPMD simulations are very similar. However, the hydrogen bond between the hydroxyl–carboxyl bond is slightly stronger in TRPMD simulations. At 100 K, the quantum effects are more prominent, especially in the 2600–3600 cm−1, and the free energy profile slightly differs between the classical MD and TRPMD simulations. Using the FT-νNAC and the interatomic distance histograms, the high-frequency vibrational bands are discussed in detail.

Accumulating studies had been performed using magnetic resonance imaging (MRI) to understand the neural mechanism of acupuncture therapy for depression. However, inconsistencies remain due to differences in research designs and MRI analytical methods. Therefore, we aim to summarize the current MRI research and provide useful information for further research by identifying papers published in English and Chinese about MRI studies on acupuncture for depression up to November 2020. A total of 22 studies met the inclusion criteria, including 810 depression patients and 416 health controls (HCs). The applied designs of these studies are mainly random control trial and pre–post designs. The MRI analytical methods are mainly (fractional) amplitude of low-frequency fluctuation (fALFF/ALFF) and functional connectivity (FC), whereas a small subset of studies used voxel-based morphometry (VBM) and diffusion tensor imaging (DTI). The most consistent functional MRI (fMRI) results showed increased N-acetylaspartate/creatine (NAA/Cr) ratios, increased ALFF in the right precuneus, decreased ALFF in the inferior frontal gyrus (IFG), and increased FC of the anterior cingulate cortex (ACC). In contrast, no significant neurological changes were identified in any of the DTI or VBM studies. However, clear, reliable conclusions cannot be drawn due to the use of different designs, analytical methods, seed points selected, types of depression, acupuncture points, and so on. Improved report specifications, well-designed studies, consistent analytical methods, and larger sample sizes will enable the field to better elucidate the underlying mechanisms of acupuncture in depressed patients.

The neural mechanisms of acupuncture are not well-understood. Over the past decades, an increasing number of studies have used MRI to investigate the response of the brain to acupuncture. The current review aims to provide an update on acupuncture therapy in disease. The PubMed, Embase, Web of Science, and Cochrane Library databases were searched from inception to January 31, 2021. Article selection and data extraction were conducted by two review authors. A total of 107 publications about MRI in acupuncture were included, the collective findings of which were as follows: (1) stroke and GB34 (Yanglingquan) are the most studied disease and acupoint. Related studies suggested that the mechanism of acupuncture treatment for stroke may associate with structural and functional plasticity, left and right hemispheres balance, and activation of brain areas related to movement and cognition. GB34 is mainly used in stroke and Parkinson's disease, which mainly activates brain response in the premotor cortex, the supplementary motor area, and the supramarginal gyrus; (2) resting-state functional MRI (rs-fMRI) and functional connectivity (FC) analysis are the most frequently used approaches; (3) estimates of efficacy and brain response to acupuncture depend on the type of sham acupuncture (SA) used for comparison. Brain processing after acupuncture differs between patients and health controls (HC) and occurs mainly in disorder-related areas. Factors that influence the effect of acupuncture include depth of needling, number and locations of acupoints, and deqi and expectation effect, each contributing to the brain response. While studies using MRI have increased understanding of the mechanism underlying the effects of acupuncture, there is scope for development in this field. Due to the small sample sizes, heterogeneous study designs, and analytical methods, the results were inconsistent. Further studies with larger sample sizes, careful experimental design, multimodal neuroimaging techniques, and standardized methods should be conducted to better explain the efficacy and specificity of acupuncture, and to prepare for accurate efficacy prediction in the future.

Purpose To establish a prediction model for stroke side identification.Methods A total of 168 patients (89 left-sided stroke patients and 79 right-sided stroke patients) were recruited from the Shenzhen Traditional Chinese Medicine Hospital in the study. Retinal characteristics were analyzed using an automated retinal image analysis (ARIA) system. Multivariable logistic regression was used to identify and develop predictive models. Results Each unit increase in the right eye bifurcation coefficient of arterioles increased the risk of right-side stroke by 7.523 times (95% CI, 1.823-31.044). Additionally, an elevated bifurcation coefficient of venules in the right eye also increased the risk of stroke in the right side of the brain, with an odds ratio (OR) of 7.377 (95% CI, 1.771-30.724). A complex retinal composite score was also associated with a higher risk of right-side stroke (OR, 4.955; 95% CI, 3.061-8.022). Conclusion This study demonstrated that retinal image analysis can provide useful information for stroke side identification and the specific retinal characteristics may help in predicting stroke occurrence.

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.

As well as their ion transportation function, the voltage-dependent potassium channels could act as the cell signal inducer in a variety of pathogenic processes. However, their roles in neurogenesis after stroke insults have not been clearly illustrated. In our preliminary study, the expressions of voltage-dependent potassium channels Kv4.2 was significantly decreased after stroke in cortex, striatum and hippocampus by real-time quantitative PCR assay. To underlie the neuroprotection of Kv4.2 in stroke rehabilitation, recombinant plasmids encoding the cDNAs of mouse Kv4.2 was constructed. Behavioral tests showed that the increased Kv4.2 could be beneficial to the recovery of the sensory, the motor functions and the cognitive deficits after stroke. Temozolomide (TMZ), an inhibitor of neurogenesis, could partially abolish the mentioned protections of Kv4.2. The immunocytochemical staining showed that Kv4.2 could promote the proliferations of neural stem cells and induce the neural stem cells to differentiate into neurons in vitro and in vivo. And Kv4.2 could up-regulate the expressions of ERK1/2, p-ERK1/2, p-STAT3, NGF, p-TrkA, and BDNF, CAMKII and the concentration of intracellular Ca2+. Namely, we concluded that Kv4.2 promoted neurogenesis through ERK1/2/STAT3, NGF/TrkA, Ca2+/CAMKII signal pathways and rescued the ischemic impairments. Kv4.2 might be a potential drug target for ischemic stroke intervention.

Acupuncture is widely recognized as a potentially effective treatment for stroke rehabilitation. Researchers in this area are actively investigating its therapeutic mechanisms. Magnetic resonance imaging (MRI), as a noninvasive, high anatomical resolution technique, has been employed to investigate neuroplasticity on acupuncture in stroke patients from a system level. However, there is no review on the mechanism of acupuncture treatment for stroke based on MRI. Therefore, we aim to summarize the current evidence about this aspect and provide useful information for future research. After searching PubMed, Web of Science, and Embase databases, 24 human and five animal studies were identified. This review focuses on the evidence on the possible mechanisms underlying mechanisms of acupuncture therapy in treating stroke by regulating brain plasticity. We found that acupuncture reorganizes not only motor-related network, including primary motor cortex (M1), premotor cortex, supplementary motor area (SMA), frontoparietal network (LFPN and RFPN), and sensorimotor network (SMN), as well as default mode network (aDMN and pDMN), but also language-related brain areas including inferior frontal gyrus frontal, temporal, parietal, and occipital lobes, as well as cognition-related brain regions. In addition, acupuncture therapy can modulate the function and structural plasticity of post-stroke, which may be linked to the mechanism effect of acupuncture.

The regenerative and immunomodulatory properties of mesenchymal stem cells (MSCs) have laid a sound foundation for their clinical application in various diseases. However, the clinical efficiency of MSC treatments varies depending on certain cell characteristics. Among these, the roles of cell aging or senescence cannot be excluded. Despite their stemness, evidence of senescence in MSCs has recently gained attention. Many factors may contribute to the senescence of MSCs, including MSC origin (biological niche), donor conditions (age, obesity, diseases, or unknown factors), and culture conditions in vitro. With the rapidly increasing prevalence of diabetes mellitus (DM) and gestational diabetes mellitus (GDM), the effects of hyperglycemia on the senescence of MSCs should be evaluated to improve the application of autologous MSCs. This review aims to present the available data on the senescence of MSCs, its relationship with hyperglycemia, and the strategies to suppress the senescence of MSCs in a hyperglycemic environment.

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.