METTL14 contributes to the progression of nasopharyngeal carcinoma through regulating the stability of AOC1 mRNA.

BACKGROUND: Nasopharyngeal carcinoma (NPC) is a malignant epithelial tumor of the nasopharyngeal mucosa with a high incidence rate all over the world. Methyltransferase-like 14 (METTL14) is a major RNA N6-adenosine methyltransferase implicated in tumor progression by regulating RNA function. This study is designed to explore the biological function and mechanism of METTL14 in NPC. METHODS: METTL14 and Amine oxidase copper containing 1 (AOC1) expression were detected by real-time quantitative polymerase chain reaction (RT-qPCR). The protein levels of METTL14, AOC1, Cyclin D1, B-cell lymphoma-2 (Bcl-2), and N-cadherin were measured using western blot. Cell proliferation, cycle progression, apoptosis, migration, and invasion were assessed using 5-ethynyl-2'-deoxyuridine (EdU), Colony formation, flow cytometry, wound scratch, and transwell assays. The interaction between METTL14 and AOC1 was verified using RNA immunoprecipitation (RIP), methylated RNA immunoprecipitation (MeRIP), and dual-luciferase reporter assays. The biological role of METTL14 on NPC tumor growth was examined by the xenograft tumor model in vivo. RESULTS: METTL14 and AOC1 were highly expressed in NPC tissues and cells. Moreover, METTL14 knockdown might block NPC cell proliferation, migration, invasion, and induce cell apoptosis in vitro. In mechanism, METTL14 might enhance the stability of AOC1 mRNA via m6A methylation. METTL14 silencing might repress NPC tumor growth in vivo. CONCLUSION: METTL14 might boosted the development of NPC cells partly by regulating the stability of AOC1 mRNA, which provided a promising therapeutic target for NPC treatment.

Network Pharmacology along with Molecular Docking to Explore the Mechanism of Danshen Injection against Anthracycline-induced Cardiotoxicity and Transcriptome Validation.

INTRODUCTION: Although anthracyclines have demonstrated efficacy in cancer therapy, their utilization is constrained by cardiotoxicity. In contrast, Danshen injection (DSI), derived from Salvia miltiorrhiza, has a longstanding tradition of being employed to ameliorate cardiovascular ailments, including anthracycline- induced cardiotoxicity (AIC). Nonetheless, there is a notable dearth of comprehensive systematic investigation into the molecular mechanisms underlying DSI's effects on AIC. Consequently, this study was undertaken to explore the underlying mechanism by which DSI acted against AIC. METHODS: Employing network pharmacology approach, the current investigation undertook a comprehensive analysis of the impact of DSI on AIC, which was further validated by transcriptome sequencing with in vitro AIC model. Additionally, molecular docking was conducted to evaluate the binding of active ingredients to core targets. A total of 3,404 AIC-related targets and 12 active ingredients in DSI, including chrysophanol, luteolin, tanshinone IIA, isoimperatorin, among others, were collected by differentially expressed analysis and database search, respectively. RESULTS: The network pharmacology and enrichment analysis suggested 102 potential targets and 29 signaling pathways associated with the protective effect of DSI on AIC. Three core targets (CA12, NOS3, and POLH) and calcium signaling pathways were further validated by transcriptomic analysis of the in-vitro model. The high affinity of the active ingredients binding to corresponding targets was confirmed by molecular docking. CONCLUSION: The present study suggested that DSI might exert a cardioprotective effect on AIC via the inhibition of CA12, NOS3, and POLH, as well as the modulation of calcium signaling. Further experiments are warranted to verify the findings.

Effect of intraoperative noise on postoperative pain in surgery patients under general anesthesia: evidence from a prospective study and mouse model.

BACKGROUND: Most patients are in a noisy environment during abdominal surgery under general anesthesia. This study included patients who underwent abdominal surgery under general anesthesia and established an animal model to determine whether intraoperative noise affects postoperative pain. MATERIALS AND METHODS: This prospective study included 200 patients who underwent abdominal surgery under general anesthesia. Intraoperative noise and electroencephalograms were continuously recorded, and the mean level and time proportion of noise intensity of greater than 70 dB were calculated. Maximum postoperative pain was assessed using a numerical rating scale at 0-12 h and 12-24 h after surgery, and postoperative analgesia consumption in patients receiving patient-controlled intravenous analgesia was recorded. Postoperative pain intensity and electroencephalogram amplitude were compared between patients with high-noise exposure (time proportion of noise intensity greater than 70 dB ≥40%) and low-noise exposure (<40%). Mechanical pain sensitivity was tested in two groups of mice with plantar incisions exposed to 40 dB or 70-100 dB. RESULTS: The time proportion of noise intensity greater than 70 dB was identified as an independent risk factor for postoperative pain intensity ( P <0.001). P ain numerical rating scale 0-12 h (4.5±1.5 vs. 3.7±1.3, P =0.001) and 12-24 h (3.9±1.5 vs. 3.2±1.1, P =0.004) after surgery in patients with high-noise exposure was significantly higher than in patients with low-noise exposure. The electroencephalogram amplitude of patients with high-noise exposure was significantly lower than that of patients with low-noise exposure ( P <0.05). In the mouse model, mechanical hyperalgesia in the 70-100 dB group was significantly greater than that in the 40 dB group ( P <0.001). CONCLUSION: High-level intraoperative noise exposure aggravates the degree of postoperative pain and analgesic needs of patients undergoing abdominal surgery, which may be related to the impact of noise on the neurophysiological activity of the brain and postoperative hyperalgesia.

Effects of single-use alfentanil versus propofol on cognitive functions after colonoscopy: A randomized controlled trial.

Purpose: Colonoscopy is often accompanied by short-term postoperative cognitive decline. We aimed to explore whether single-use alfentanil for patients undergoing elective colonoscopy could reduce cognitive impairment at discharge compared with propofol. Patients and methods: 172 adult patients undergoing elective colonoscopy were randomized to receive intravenous propofol at 2 mg/kg (group P) or alfentanil at 10 µg/kg (group A); 40 healthy volunteers were included in the blank group. Cognitive function was considered the primary outcome and was measured using five neuropsychological tests before sedation and discharge. The z-score method was used to determine cognitive dysfunction according to z-score >1.96 in two types of neuropsychological tests. Other outcomes included discharge time, vital signs, associated adverse events during colonoscopy, and the satisfaction level of patients and endoscopic physicians. Results: 164 patients (78 in group A and 86 in group P) completed the study protocol. At discharge, the incidence of cognitive dysfunction in group P was 23% and was significantly lower in the alfentanil group (2.5%), with a relative risk of 0.11 (95% confidence interval: 0.03-0.46, P < 0.001). The incidence of hypotension in group A was lower than that in group P (3.8% vs 22.1%, relative risk = 0.17 [95% confidence interval: 0.05-0.46, P = 0.001]), and the discharge time in group A was shorter than that in group P (5 [(Rutter and et al., 2016; Zhang and et al., 2013; Hirsh and et al., 2006; Zhou and et al., 2021; Singh and et al., 2008; Ko and et al., 2010; Sargin et al., 2019) 3-93-9 vs 13 [(Ekmekci and et al., 2017; Eberl and et al., 2012; Eberl and et al., 2014; N'Kaoua and et al., 2002; Chung et al., 1995; Berger and et al., 2019; Quan and et al., 2019; Deng and et al., 2021; Gualtieri and Johnson, 2006) 10-1810-18 min, P < 0.001). Conclusion: For patients undergoing colonoscopy, single-use alfentanil causes less damage to postoperative cognitive function, less risk of hypotension, and shorter discharge time than propofol.

Tracheal Extubation Under Deep Anesthesia Using Transnasal Humidified Rapid Insufflation Ventilatory Exchange vs. Awake Extubation: An Open-Labeled Randomized Controlled Trial.

Background: Tracheal extubation can be associated with several complications, including desaturation, agitation, hypertension, and tachycardia. We hypothesize that the use of transnasal humidified rapid insufflation ventilator exchange (THRIVE) immediately after extubation under deep anesthesia reduces the incidence of these adverse events. Methods: One hundred patients who underwent elective abdominal surgery under general anesthesia were randomly assigned to undergo tracheal extubation under deep anesthesia employing THRIVE (THRIVE group) or awake extubation (CONTROL group). The primary outcome was the incidence of experiencing desaturation (SpO2 < 90%) at any time during emergence from anesthesia. Secondary outcomes included variations in heart rate and blood pressure, comfort level, bucking, and agitation. Results: The THRIVE group showed a lower incidence of desaturation than the CONTROL group (12 vs. 54%, OR = 0.22 [95% CI, 0.10-0.49], P < 0.001). Less patients in the THRIVE group experienced a 20% (or more) increase in mean arterial pressure (4 vs. 26%, OR = 0.15 [95% CI, 0.04-0.65], P = 0.002). THRIVE patients did not suffer from agitation or bucking, while in the CONTROL group agitation and bucking occurred in 22 and 58% of the patients, respectively. Additionally, the THRIVE group showed a lower incidence of uncomfortable experience than the CONTROL group (8 vs. 36%, OR = 0.22 [95% CI, 0.08-0.61], P = 0.001). Conclusion: Tracheal extubation under deep anesthesia using THRIVE decreases the incidence of desaturation and adverse haemodynamic events and increases patient satisfaction. Extubation under deep anesthesia using THRIVE might be an alternative strategy in selected patient populations.

Kupffer cells depletion alters cytokine expression and delays liver regeneration after Radio-frequency-assisted Liver Partition with Portal Vein Ligation.

Radio-frequency-assisted Liver Partition with Portal Vein Ligation (RALPP) induces comparable hypertrophy of the liver remnant compared to Associating Liver Partition and Portal vein ligation for Staged hepatectomy (ALPPS) in humans. However, whether it is significantly improved compared to ALPPS is unclear, and the underlying mechanisms of liver regeneration after RALPP need to further investigate. The present study was to develop an animal model mimicking RALPP and explore mechanisms of liver regeneration. The mice in RALPP group received liver radiofrequency ablation and 90% portal vein ligation (PVL), followed by resection of the targeted liver within two days after the first surgery. The mice in ALPPS group underwent 90% PVL combined with parenchyma transection. Controls received liver radiofrequency ablation (RAF group) or PVL (PVL group) or small left lateral lobe (LLL group) resection alone. Liver regeneration was assessed by liver weight and proliferation-associated molecules. The role of Kupffer cells (KCs) in liver regeneration was investigated after RALPP. The results showed that RALPP induced comparable liver regeneration compared to ALPPS, but with less liver injury and mortality in mice. RALPP led to over-expression of TNF-α and IL-6 in the circulating plasma compared with PVL. KCs infiltrating in liver tissues was a characteristic of mice in the RALPP group. KCs depletion markedly depressed cytokine expression and delayed liver regeneration after RALPP. These results suggested that RALPP in mice induced accelerated liver regeneration similar to ALPPS, but safer than ALPPS. KCs depletion altered cytokine expression and delayed liver regeneration after RALPP.

The Potential Effect of Na v 1.8 in Autism Spectrum Disorder: Evidence From a Congenital Case With Compound Heterozygous SCN10A Mutations.

Apart from the most prominent symptoms in Autism spectrum disorder (ASD), namely deficits in social interaction, communication and repetitive behavior, patients often show abnormal sensory reactivity to environmental stimuli. Especially potentially painful stimuli are reported to be experienced in a different way compared to healthy persons. In our present study, we identified an ASD patient carrying compound heterozygous mutations in the voltage-gated sodium channel (VGSC) Na v 1.8, which is preferentially expressed in sensory neurons. We expressed both mutations, p.I1511M and p.R512∗, in a heterologous expression system and investigated their biophysical properties using patch-clamp recordings. The results of these experiments reveal that the p.R512∗ mutation renders the channel non-functional, while the p.I1511M mutation showed only minor effects on the channel's function. Behavioral experiments in a Na v 1.8 loss-of-function mouse model additionally revealed that Na v 1.8 may play a role in autism-like symptomatology. Our results present Na v 1.8 as a protein potentially involved in ASD pathophysiology and may therefore offer new insights into the genetic basis of this disease.

A High Power-Conversion-Efficiency Voltage Boost Converter with MPPT for Wireless Sensor Nodes.

A high power-conversion-efficiency voltage boost converter with MPPT for wireless sensor nodes (WSNs) is proposed in this paper. Since tiny wireless sensor nodes are all over complex environments, an efficient power management system (PMS) must be equipped to achieve long-term self-power supply and maintain regular operation. It is common to use Photovoltaic cells (PV) to harvest sunlight in the environment. However, most existing interface boost integrated circuits for the PV cell have low efficiency. This paper presents a voltage boost converter (VBC) with high power conversion efficiency (PCE) for WSNs. The integrated circuit (IC) designed in this paper includes a novel four-phase high-efficiency charge pump module, an ultra-low-power perturbation observation (P&O) MPPT control circuit module, a feedback control module, a nano-ampere current reference, etc. Manufactured in a standard 0.35 um complementary metal-oxide-semiconductor (CMOS) technology, the chip area is 3.15 mm × 2.43 mm. Test results demonstrate that when the output voltage of the PV cell is more than 0.5 V, VBC can improve the voltage to 3Vin, and the calculated voltage conversion efficiency can reach 99.4%. P&O MPPT algorithm makes output power improving 8.53%. Furthermore, when the output load current is 297uA, the output PCE achieves 85.1%.

Effect of Team Health Education on Radiodermatitis in Patients with Head and Neck Tumor Radiotherapy Under the Joint Committee International Standards.

OBJECTIVES: This study aimed to investigate the impacts of team health education on radiodermatitis in patients undergoing head and neck radiotherapy. DATA SOURCES: A total of 118 patients undergoing radiotherapy at the Oncology Department of Qingdao Municipal Hospital under the Joint Committee International (JCI) accreditation standards of medical and health institutions were divided into two groups according to the order of the admission: the intervention group (n = 66) and the control group (n = 52). The patients in the control group were given routine nursing, while those in the intervention group received team health education based on the control group. The incidence and satisfaction of radiodermatitis were observed and compared between the groups, and the EORTC QLQ-C30 and General Quality of Life Inventory-74 were used to evaluate the patients' quality of life when after radiotherapy and when after 6 months post the radiotherapy, respectively. CONCLUSION: The incidence of radiodermatitis was 100% in both groups, and the difference in the grade of radiodermatitis and quality of life was significant (P < .05) between them. IMPLICATIONS FOR NURSING PRACTICE: Team health education under the JCI standards team can reduce the degree of skin injury due to radiodermatitis and improve the quality of life.

CX3CR1 deficiency aggravates brain white matter injury and affects expression of the CD36/15LO/NR4A1 signal.

OBJECTIVE: To study the effects of CX3CR1 on white matter injury, neurofunction, recognition, and expression of the CD36/15LO/NR4A1 signal in mice with traumatic brain injury (TBI). METHODS: CX3CR1GFP/GFP, CX3CR1GFP/+ and C57BL/6 male mice were randomly divided into 3 groups. We used a controlled cortical impact (CCI) to establish a TBI model and T2wt MRI to detect the TBI lesion. FA and DTI allowed for quantitative evaluation of the structural integrity of white matter tracts. Several behavior tests were used to investigate nerve function; a computer-based tracing system was used to trace and analyze dendrites and cell bodies of microglia and astrocytes in the peri-lesional brain areas. We also used RT-PCR and western blot to detect the effect of CX3CL1/CX3CR1 axis on CD36/15LO/NR4A1 signal. RESULTS: The fractional anisotropy (FA) at the corpus callosum area of brain was decreased at 3 days post TBI, the average lesion volume CX3CR1GFP/GFP group was increased, and the neurologic deficit scores of mice of Cx3Cr1GFP/+ and wild-type groups were significantly increased compared to Cx3Cr1GFP/GFP group mice. In the Corner turn test, TBI induced impairments in forelimb function that were more severe than Cx3Cr11GFP/+ and wild-type TBI mice. We operated the Y-maze at 3 days post-TBI and the NOR test at 28 days after TBI. There was a significant TBI effect induced in decreased percentage entries into the novel arm in Cx3Cr1GFP/+ and wild-type TBI mice, compared with Cx3Cr1GFP/GFP; Cx3Cr1GFP/+. Wild-type mice showed decreased exploration time in new objects compared with Cx3Cr1GFP/GFP. Those two behavior tests demonstrated that Cx3Cr1 knock-out increased the damage caused by TBI to memory. In the tail suspension and force swimming tests, there was no significant difference between those three groups. CD36 increased in Cx3Cr1GFP/GFP compared with the other three groups at 3 days after TBI. TBI inhibited the expression of NR4A1 at 3 d after damage. Cx3Cr1 deficiency can induce high expression of 15LO, this was unaffected by TBI. CONCLUSION: CX3CR1 deletion can enhance white matter injury. It increased the expression of CD36 and 15LO and increased expression of NR4A1. The lack of CX3CR1 can affect the recovery of nerve function.

Effect of RFA and TACE combined with postoperative cytokine-induced killer cell immunotherapy in primary hepatocellular carcinoma.

PURPOSE: To evaluate the efficacy and safety of radiofrequency ablation (RFA) and transcatheter arterial chemoembolization (TACE) combined with postoperative cytokine-induced killer (CIK) cell immunotherapy in the treatment of primary hepatocellular carcinoma (HCC). METHODS: The clinical data of 116 patients with primary HCC treated in our hospital from March 2016 to January 2018 were collected. 58 patients were treated with RFA+TACE (RFA+TACE group), and the other 58 patients underwent RFA+TACE+CIK cell immunotherapy (RFA+TACE+CIK group). Before and after treatment, the proportions of cluster of differentiation 3+ (CD3+), CD3+CD4+, and CD3+CD8+ T cells, regulatory T cells (Tregs) and natural killer (NK) cells and the CD4+/CD8+ ratio were detected via flow cytometry, and the levels of serum interferon-γ (IFN-γ), interleukin-2 (IL-2) and IL-6 were detected via enzyme-linked immunosorbent assay (ELISA). The incidence of adverse reactions and the quality of life score of patients after treatment were compared between the two groups, and the patient's survival status was recorded through follow-up. RESULTS: After treatment, the levels of CD3+ T cells, CD3+CD4+ T cells, CD4+/CD8+ ratio, Tregs and NK cells were significantly higher, while the level of CD3+CD8+ T cells was significantly lower in RFA+TACE+CIK group than those in RFA+TACE group. After treatment, the level of alpha fetoprotein (AFP) obviously declined in both groups compared with that before treatment, and it was significantly lower in RFA+TACE+CIK group than that in RFA+TACE group. After treatment, the scores of the QLQ-C30 questionnaire were all significantly higher in RFA+TACE+CIK group than those in RFA+TACE group. After treatment, the general functioning score rose from (58.55±11.82) and (59.39±10.97) points to (74.74±15.58) and (68.42±14.85) points, respectively, in RFA+TACE+CIK group and RFA+TACE group, and it was significantly higher in RFA+TACE+CIK group than that in RFA+TACE group. According to the follow-up results, the mean overall survival (OS) of patients was (42.1±5.6) months and (37.8±4.8) months, and the 5-year OS rate was 29.3% (17/58) and 13.8% (8/58), respectively, in RFA+TACE+CIK group and RFA+TACE group. The results of log-rank test showed that the OS in RFA+TACE+CIK group was significantly superior to that in RFA+TACE group. CONCLUSIONS: RFA and TACE combined with postoperative autologous CIK cell reinfusion have significant efficacy in the treatment of primary HCC, which can enhance the immune function, improve the postoperative quality of life and raise the survival rate of patients, with tolerable adverse reactions.

[Splenic tyrosine kinase promotes pulmonary angiogenesis in rats with hepatopulmonary syndrome].

The present paper was aimed to study the role of spleen tyrosine kinase (Syk) in angiogenesis in hepatopulmonary syndrome (HPS) and the underlying mechanism. Sprague Dawley (SD) rats were randomly divided into three groups: sham operation group (sham group), common bile duct ligation (CBDL) 5-week group (5W group) and R788 intervention group (R788 group). HPS model was established by CBDL. Rats in R788 group were intraperitoneally injected with R788 (20 mg/kg) once daily to week 5 after CBDL operation. The protein expression levels and distribution of Syk, p-Erk1/2, and p-Akt in lung tissue were detected by Western blot and immunohistochemistry. Immunofluorescence staining was used to observe the location of Syk expression and the number of angiogenesis in lung tissue. The results showed that, compared with sham group, 5W group exhibited up-regulated protein expression level of Syk, increased phosphorylation levels of Erk1/2 and Akt, and increased number of pulmonary microvessels. Compared with 5W group, R788 group exhibited down-regulated protein expression level of Syk, decreased phosphorylation levels of Erk1/2 and Akt, and decreased number of pulmonary microvessels. These results suggest that Syk may promote pulmonary angiogenesis in HPS model rats by activating downstream Erk1/2 and Akt signaling pathways, which provides a theoretical basis and potential drug therapeutic targets for the clinical treatment of HPS.

Effect of Intraoperative Dexmedetomidine Dose on Postoperative First Night Sleep Quality in Elderly Surgery Patients: A Retrospective Study With Propensity Score-Matched Analysis.

Background: Postoperative sleep disorder is common in elderly surgery patients, and it often worsens their recovery after surgery. This study aimed to explore the effect of intraoperative dexmedetomidine dose on postoperative sleep quality. Methods: Based on information regarding dexmedetomidine use during surgery from an electronic medical record system, 4,349 elderly surgery patients were divided into three groups: 1,374 without intraoperative use of dexmedetomidine (Non-DEX), 917 with dexmedetomidine 0.1-0.2 µg/kg/h (Low-DEX), and 2,058 with dexmedetomidine >0.2 µg/kg/h (High-DEX). The numerical rating scale (NRS) for sleep disturbance during the first night after surgery was recorded, and the incidence of NRS ≥ 6 was considered the primary outcome. Results: NRS (P < 0.001) and incidence of severe sleep disturbance (P < 0.001) were lower in patients receiving intraoperative dexmedetomidine than in those without the intraoperative use of dexmedetomidine. Patients in the Low-DEX group had the lowest incidence, followed by those in the High-DEX and Non-DEX groups (6.7% vs. 13.7% vs. 19.5%). After propensity score matching, 906 pairs of elderly surgery patients were included in the Low-DEX and High-DEX groups, and the Low-DEX group had lower NRS (2.7 ± 2.1 vs. 3.1 ± 2.4, P < 0.001) than the High-DEX group. The incidence of severe sleep disturbance was lower in the Low-DEX group than in the High-DEX group (6.6% vs. 12.8%) with an odds rate of 0.48 (95% confidence interval, 0.35 to 0.67). Conclusions: For elderly patients, intraoperative dexmedetomidine use can significantly improve the quality of the first night sleep after surgery. Low-dose (0.1-0.2 µg/kg/h) dexmedetomidine can have an improvement effect on sleep quality, and it is recommended to improve the quality of postoperative sleep.

MiR-155 contributes to intestinal barrier dysfunction in DSS-induced mice colitis via targeting HIF-1α/TFF-3 axis.

Intestinal barrier dysfunction is a hallmark of inflammatory bowel disease (IBD). MiR-155 is increased in colitis and downregulates expression of hypoxia-inducible factor 1α (HIF-1α). Here, we investigated the effects of miR-155 on intestinal barrier dysfunction in dextran sulfate sodium (DSS)-induced colitis. We found that miR-155 antagomir treatment relieved weight loss and intestinal damage in IBD mouse models (P < 0.05). Furthermore, electron microscopy and immunofluorescence imaging showed that miR-155 increased intestinal barrier dysfunction and downregulated the expression of tight junction proteins in DSS-induced colitis. FG-4497, which upregulates HIF-1α expression, elicited protective effects on the intestinal barrier in DSS-induced colitis. Dual luciferase reporter assays also confirmed that miR-155 downregulated expression of HIF-1α. Finally, we discovered that HIF-1α levels were elevated by miR-155 antagomir treatment (P < 0.05) and that TFF-3 expression correlated positively with HIF-1α expression. These results suggest that miR-155 contributes to DSS-induced colitis by promoting intestinal barrier dysfunction and inhibiting the HIF-1α/TFF-3 axis.

miR-155 antagomir protect against DSS-induced colitis in mice through regulating Th17/Treg cell balance by Jarid2/Wnt/ß-catenin.

BACKGROUND: Th subsets particularly T helper 17 and regulatory T cells play a critical role in immune balance in colonic mucosa of Inflammatory Bowel Disease. Recent studies have indicated miR-155 is overexpressed in the colonic mucosa in IBD patients. Thus, whether and how miR-155 influences Th17/Treg cell balance in IBD patients is worthy of researching. METHODS: We divided mice into four groups: the mice oral administration of 3.0 % DSS in fresh drinking water for 7 days except normal group. In this period, starting from the fifth day, the miR-155 and NC antagomir group were carried out by intraperitoneal injection of miR-155 antagomirs and corresponding negative controls. In vitro, we isolated naïve CD4+T cells and divided into two groups: the cells were transfected with mmu-miR-155-5p inhibitor or corresponding negative controls and then induced differentiation. RESULTS: We found miR-155 antagomir can reach colon tissues in DSS-induced colitis and indeed ameliorated DSS-induced experimental colitis. Subsequently, we proved the levels of Th17 cells in spleens and Mesenteric lymph nodes and its associated IL-6, IL-17A and RORγt in colonic tissues were dramatically decreased and TGF-ß1 raised in DSS + miR-155 antagomir group. However, miR-155 antagomir significantly increased the expression of Tregs. In vitro, we found miR-155 inhibitor could improve the Tregs but decrease Th17 cells. Finally, we dig out that Jarid2 was apparently improved by miR-155 antagomir, Wnt/ß-catenin and its associated T cell factor-4 (TCF-4) and Cyclin D1 expression were positively correlated with Jarid2. CONCLUSION: Silencing of miR-155 attenuates DSS-induced colitis by regulating Th17/Treg cell balance and Jarid2/Wnt/ß-catenin participated in the process.

Applied research of a four-dimensional CT localization technique in radiotherapy and treatment planning for early lung cancer.

BACKGROUND: To study the feasibility of target contouring and the potential benefits to radiotherapy of four-dimensional computed tomography (4D-CT) for early stage lung cancer. METHODS: We applied Brilliance CT to scan 24 lung cancer patients for 4D localization. Treatment plannings based on different breath phase CT images were designed in the Monaco 5.2 treatment planning system. Different planning was compared to find the difference of target volume, center, and dose. RESULTS: Target volume composed of 10-breath phases (Sum) was considered as the reference volume, which was slightly larger than the volume contouring obtained by the maximum intensity projection (Mip) and extreme phase images (Exs). Target centers for Sum, Mip and Exs of CT images showed little deviation in the X, Y and Z directions. The V20 and V5 for the 4D-CT-based treatment plan showed a markedly lower dose delivered to normal tissues than those based on 3D-CT. CONCLUSIONS: For target contouring, a target contouring by 10-breath phases was superior to Mip and Exs. As the gross target volume (GTV) contouring obtained by the maximum intensity projection and extreme phase were similar to that of the reference target, they can support the contouring target and reduce the time and work. Treatment plan comparisons indicated that normal lung tissues received a remarkably lower dose when a 4D-CT-based plan was used.

SIRT4 silencing in tumor-associated macrophages promotes HCC development via PPARδ signalling-mediated alternative activation of macrophages.

BACKGROUND: The activation of tumour-associated macrophages (TAMs) contributes to the progression of hepatocellular carcinoma (HCC). SIRT4 acts as a tumour suppressor of tumour growth by regulating cell metabolism, inflammation, and anti-tumourigenesis. However, the involvement of SIRT4 in the activation of TAMs is unknown. Based on previous findings, the expression of SIRT4 in distinct groups of TAMs as well as the effect of SIRT4 silencing on macrophage polarization was investigated. METHODS: The expression of SIRT4 in HCC tissues and peritumour tissues was tested by qRT-PCR, western blotting and histological analysis. A Kaplan-Meier survival curve was generated based on the expression of SIRT4 in the HCC samples. Next, immunofluorescence staining was used to evaluate distinct groups of TAMs in human HCC samples, and the expression of SIRT4 in M1 and M2 TAMs was examined by flow cytometry. A homograft mouse model was used to assess the effect of SIRT4 silencing in TAMs on the development of HCC cells. RESULTS: SIRT4 was significantly downregulated in HCC tumour tissues, and the expression of SIRT4 in peritumour tissues was positively associated with survival in patients. We further found that downregulation of SIRT4 was associated with increased macrophage infiltration and a high ratio of M2/M1 macrophages in HCC peritumour tissues. Using gene interference, we found that SIRT4 silencing in TAMs significantly modulated the alternative activation of macrophages and promoted in vitro and in vivo HCC cell growth. Mechanistically, we revealed that HCM restricted the expression of SIRT4 in macrophages and promoted alternative activation of macrophages via the FAO-PPARδ-STAT3 axis. Furthermore, we also revealed that elevated MCP-1 expression induced by SIRT4 downregulation was responsible for increased TAM infiltration in peritumour tissues. CONCLUSIONS: Overall, our results demonstrate that downregulation of SIRT4 in TAMs modulates the alternative activation of macrophages and promotes HCC development via the FAO-PPARδ-STAT3 axis. These results could provide a new therapeutic target for the treatment of HCC.

Uncovering the Anticancer Mechanism of Compound Sophorae Decoction against Ulcerative Colitis-Related Colorectal Cancer in Mice.

Compound sophorae decoction (CSD), a traditional Chinese medicine (TCM) formula, has been voluminously used in China to deal with ulcerative colitis and gained significant therapeutic effect. Tremendous explorations have unraveled a contributory role of inflammatory bowel disease (IBD) like ulcerative colitis (UC) and Crohn's disease (CD) at the onset of colorectal cancer, scilicet, and colitis-related cancer (CRC). In light of the anti-inflammatory properties of CSD in UC, we appraised its chemoprevention capacity and underlying mechanism in ulcerative colitis-related colorectal cancer (UCRCC), employing a model of azoxymethane (AOM) plus dextran sulfate sodium- (DSS-) induced colorectal cancer (CRC) in C57BL/6 mice. Rapturously, our results illuminated the ameliorative effect of CSD against UCRCC in mice portrayed by lesser polyps or adenomas, attenuated colonic xenograft tumor growth in company with the preferable well-being of mice in contrast to the Model Group. We examined significant downregulation of proinflammatory cytokines such as TNF-α, NF-κB, IL-6, STAT3, and IL-17 after exposure to CSD, with the concomitant repression of inflammation-associated proteins, including COX-2 and iNOS. Independent of this, treatment with CSD declined the proportion of T helper 17 cells (Th17) and protein level of matrix metallopeptidase 9 (MMP-9). Moreover, transmission electron microscopy (TEM) detected observably suppressed mitophagy in mice administered with CSD and that was paralleled by the pro-apoptotic effect as indicated by upregulating caspase-3 together with caspase-9 and deregulating B-cell lymphoma 2 (Bcl-2). In closing, these findings suggest CSD executes the UCRCC-inhibitory activity through counteracting inflammatory responses and rescuing detuning of apoptosis as well as neutralizing overactive mitophagy, concurring to build up an oncosuppressive microenvironment.

USP25 promotes endotoxin tolerance via suppressing K48-linked ubiquitination and degradation of TRAF3 in Kupffer cells.

The inhibition of tumor necrosis factor receptor-associated factor 3 (TRAF3) degradation induces endotoxin tolerance (ET) in macrophages. However, the mechanisms leading to TRAF3 inhibition by ET are largely unknown. Here, we found that ubiquitin-specific peptidase 25 (USP25), a deubiquitinating enzyme (DUB), interacted with TRAF3 and stabilized ET in Kupffer cells (KCs). Lentiviral knockdown of USP25 activated K48-linked ubiquitination of TRAF3 and the cytoplasmic translocation of the Myd88-associated multiprotein complex in tolerized KCs. This outcome led to a subsequent activation of Myd88-dependent c-Jun N-terminal kinase (JNK) and p38-mediated downregulation of inflammatory cytokines. The overexpression of TRAF3 attenuated the proinflammatory effects of USP25 knockdown in tolerized KCs. Thus, our findings reveal a novel mechanism of endotoxin-mediated TRAF3 degradation in KCs.

MiR-155 inhibition ameliorates 2, 4, 6-Trinitrobenzenesulfonic acid (TNBS)-induced experimental colitis in rat via influencing the differentiation of Th17 cells by Jarid2.

Th17 cells play an important role in the immune imbalance and inflammatory state in colonic mucosa of Inflammatory Bowel Disease (IBD) and to clarify the mechanism that affect the differentiation of Th17 cells will help us find a new target for the treatment of IBD. MiR-155 which is reported to have an important role in regulating immune system function is also detected to be significantly up-regulated in colonic tissues of IBD patients. However, whether and how miR-155 affects the differentiation of Th17 cells in the colon of IBD patients is still worth studying. Here, we investigated the role of miR-155 in TNBS-induced rat colitis. Firstly, we found that the disease activity index (DAI) and Colon pathological changes were significantly reduced (P < 0.05) by using miR-155 inhibition sequences delivered by lentiviral vector, which revealed that miR-155 inhibition ameliorated TNBS-Induced experimental colitis. Then, we carried out flow cytometry, ELISA, qRT-PCR, and found that in TNBS+miR-155 inhibition group, the proportion of Th17 cells in spleens and mesenteric lymph nodes (MLNs) and the level of the Th17 cell-associated cytokines IL-6, IL-17A, IL-17F and IL-21 in colon tissues were significantly reduced (P < 0.05), which revealed that miR-155 inhibition regulated the differentiation and function of Th17 cells. Finally, we discovered that Jarid2 was significantly elevated (P < 0.05) by miR-155 inhibition and notch1 expression was inversely correlated with Jarid2 by using Immunohistochemistry and western blot. This study suggests that miR-155 inhibition ameliorates TNBS-induced colitis by regulating the Th17 cells differentiation and function and Jarid2/notch1 is closely related with the process.