Tumor Necrosis Factor-α Protein Research Articles

Zoledronic Acid Inhibits Lipopolysaccharide-Induced Osteoclastogenesis by Suppressing Macrophage NLRP3-Mediated Autophagy Pathway.

Inflammatory factors leading to bone loss significantly increase the risk of tooth loosening or implantation failure. Zoledronic acid (ZOL) is a widely used medication for effectively inhibiting excessive bone destruction, but its effect on alleviating inflammatory bone loss remains to be elucidated. In this study, we investigated whether ZOL alleviates inflammatory bone resorption through immunomodulatory effect. The viability of the cells was evaluated by Cell Counting Kit 8 (CCK8) assay. Osteoclast (OC) differentiation and function were determined by tartrate-resistant acid phosphatase (TRAP) staining and bone resorption pits assays, respectively. Autophagosomes and actin ring structures of OC were observed using transmission electron microscopy (TEM) and F-actin ring staining, respectively. The microstructure in mice maxillary alveolar bone model was observed by micro computed tomography (Miro-CT). Reverse transcription-quantitative PCR (RT-qPCR) to detect the mRNA expression of osteoclast-related genes and Western blot (WB) analysis to evaluate the protein expression levels of autophagy-related proteins and the NOD-like receptor family pyrin domain-containing protein 3 (NLRP3)-related proteins in pre-OCs. The findings indicated that ZOL hindered lipopolysaccharide (LPS)-mediated OC differentiation, formation, bone resorption activity and autophagosome levels. Furthermore, ZOL diminished the expression of genes associated with OC. And the expression of proteins ATG7, LC3II, Beclin1, NLRP3-related proteins and tumor necrosis factor-α (TNF-α) protein were markedly decreased while P62 was increased, especially in the 1 μM ZOL group or MCC950 + ZOL group. ZOL has a certain immunomodulatory effect that exhibits anti-inflammatory properties at lower concentrations, which can weaken LPS-induced OCs differentiation and function, and NLRP3-mediated autophagy pathway may participate in this process.

Dipeptidyl peptidase-4 disturbs adipocyte differentiation via the negative regulation of the glucagon-like peptide-1/adiponectin-cathepsin K axis in mice under chronic stress conditions.

Exposure to chronic psychosocial stress is a risk factor for metabolic disorders. Because dipeptidyl peptidase-4 (DPP4) and cysteinyl cathepsin K (CTSK) play important roles in human pathobiology, we investigated the role(s) of DPP4 in stress-related adipocyte differentiation, with a focus on the glucagon-like peptide-1 (GLP-1)/adiponectin-CTSK axis invivo and invitro. Plasma and inguinal adipose tissue from non-stress wild-type (DPP4+/+), DPP4-knockout (DPP4-/-) and CTSK-knockout (CTSK-/-) mice, and stressed DPP4+/+, DPP4-/-, CTSK-/-, and DPP4+/+ mice underwent stress exposure plus GLP-1 receptor agonist exenatide loading for 2 weeks and then were analyzed for stress-related biological and/or morphological alterations. On day 14 under chronic stress, stress decreased the weights of adipose tissue and resulted in harmful changes in the plasma levels of DPP4, GLP-1, CTSK, adiponectin, and tumor necrosis factor-α proteins and the adipose tissue levels of CTSK, preadipocyte factor-1, fatty acid binding protein-4, CCAAT/enhancer binding protein-α, GLP-1 receptor, peroxisome proliferator-activated receptor-γ, perilipin2, secreted frizzled-related protein-4, Wnt5α, Wnt11 and β-catenin proteins and/or mRNAs as well as macrophage infiltration in adipose tissue; these changes were rectified by DPP4 deletion. GLP-1 receptor activation and CTSK deletion mimic the adipose benefits of DPP4 deficiency. Invitro, CTSK silencing and overexpression respectively prevented and facilitated stress serum and oxidative stress-induced adipocyte differentiation accompanied with changes in the levels of pref-1, C/EBP-α, and PPAR-γ in 3T3-L1 cells. Thus, these findings indicated that increased DPP4 plays an essential role in stress-related adipocyte differentiation, possibly through a negative regulation of GLP-1/adiponectin-CTSK axis activation in mice under chronic stress conditions.

RNA Activators of Stress Kinase PKR within Human Genes That Control Splicing or Translation Create Novel Targets for Hereditary Diseases.

Specific sequences within RNA encoded by human genes essential for survival possess the ability to activate the RNA-dependent stress kinase PKR, resulting in phosphorylation of its substrate, eukaryotic translation initiation factor-2α (eIF2α), either to curb their mRNA translation or to enhance mRNA splicing. Thus, interferon-γ (IFNG) mRNA activates PKR through a 5'-terminal 203-nucleotide pseudoknot structure, thereby strongly downregulating its own translation and preventing a harmful hyper-inflammatory response. Tumor necrosis factor-α (TNF) pre-mRNA encodes within the 3'-untranslated region (3'-UTR) a 104-nucleotide RNA pseudoknot that activates PKR to enhance its splicing by an order of magnitude while leaving mRNA translation intact, thereby promoting effective TNF protein expression. Adult and fetal globin genes encode pre-mRNA structures that strongly activate PKR, leading to eIF2α phosphorylation that greatly enhances spliceosome assembly and splicing, yet also structures that silence PKR activation upon splicing to allow for unabated globin mRNA translation essential for life. Regulatory circuits resulting in each case from PKR activation were reviewed previously. Here, we analyze mutations within these genes created to delineate the RNA structures that activate PKR and to deconvolute their folding. Given the critical role of intragenic RNA activators of PKR in gene regulation, such mutations reveal novel potential RNA targets for human disease.

Phenolic-rich fraction of green tea attenuates histamine-mediated cardiopulmonary toxicity by inhibiting Cox-2/NF-κB signaling pathway and regulating oxidant/antioxidant balance

BackgroundHistamine (HIS) has a substantial impact on the development of numerous allergic disorders including asthma. Antihistamines mostly target histamine receptor-1 alone, so it is not entirely effective in the treatment of allergic diseases. In the current investigation, we examine the growing evidence for novel therapeutic strategies that aim to treat histamine-mediated cardiopulmonary toxicity with the phenolic-rich fraction of green tea (PRFGT).ResultsOur findings demonstrated that weekly ingestion of HIS to rats induced oxidant/antioxidant imbalance in both lung and heart homogenates. The histopathological examination demonstrated extensive interstitial pneumonia with progressive alveolar and bronchial damage in HIS receiving groups. Heart sections showed severe myocardial necrosis and hemorrhage. All lesions were confirmed by the immunohistochemical staining that demonstrated strong caspase-3, cyclooxygenase-2 (Cox-2), and tumor necrosis factor-α (TNF-α) protein expressions along with upregulation of the pulmonary m-RNA expression of TNF-α, nuclear factor kappa-B (NF-κB), and interleukin-1β (IL-1β) genes and cardiac levels of many apoptotic genes. Otherwise, the pretreatment of rats with PRFGT had the ability to alleviate all the aforementioned toxicological parameters and return the microscopic picture of both lung and heart sections to normal histology.ConclusionsWe concluded that PRFGT’s powerful antioxidant, anti-inflammatory, and anti-apoptotic properties can reduce cardiopulmonary toxicity caused by HIS. We recommended daily intake of green tea as a beverage or adding it to foods containing elevated levels of HIS to prevent its possible toxicity.

Freshwater Clam Extract Attenuates Indomethacin-Induced Gastric Damage In Vitro and In Vivo.

Contemporary pharmacological studies have reported that freshwater clam (Corbicula fluminea) can provide a broad spectrum of bioactivities, including antioxidant, anticancer, antihypertensive, hepatoprotective, and hypocholesterolemic effects. The aim of this study was to evaluate the gastroprotective effects of water extract of freshwater clam (WEC) on indomethacin (IND)-induced gastric mucosal cell damage in vitro and gastric ulcer in vivo. The cell viability of rat gastric mucosa RGM-1 cells was markedly decreased by 0.8 mM of IND treatment, and pre-treated with various concentration of WEC significantly restored IND-induced cell damage in a dose-dependent manner. WEC also significantly attenuated the elevated reactive oxygen species (ROS) levels, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression, and nuclear factor-κB (NF-κB) p65 nuclear translocation induced by IND. In the in vivo study, IND caused severe gastric ulcer in Wistar rats, while WEC pretreatment effectively reduced the ulcer area and edema in the submucosa. We found that WEC significantly restored glutathione (GSH) content in gastric mucosa in a dose-dependent manner (p < 0.05). The reduction of prostaglandin E2 (PGE2) caused by IND was also improved with higher doses of WEC administration. Moreover, the overexpression of COX-2, iNOS, and tumor necrosis factor-α (TNF-α) proteins in gastric mucosa was downregulated by administration of WEC. Consequently, WEC can be used as a potential nutritional supplement to improve NSAIDs-caused gastric mucosal lesions.

Insomnia as an Unmet Need in Patients With Chronic Hematological Cancer: Protocol for a Randomized Controlled Trial Evaluating a Consumer-Based Meditation App for Treatment of Sleep Disturbance.

BackgroundTo address the need for long-term, accessible, nonpharmacologic interventions targeting sleep in patients with chronic hematological cancer, we propose the first randomized controlled trial to determine the effects of a consumer-based mobile meditation app, Calm, on sleep disturbance in this population.ObjectiveThis study aims to test the efficacy of daily meditation delivered via Calm compared with a health education podcast control group in improving the primary outcome of self-reported sleep disturbance, as well as secondary sleep outcomes, including sleep impairment and sleep efficiency; test the efficacy of daily meditation delivered via Calm compared with a health education podcast control group on inflammatory markers, fatigue, and emotional distress; and explore free-living use during a 12-week follow-up period and the sustained effects of Calm in patients with chronic hematological cancer.MethodsIn a double-blinded randomized controlled trial, we will recruit 276 patients with chronic hematological cancer to an 8-week app-based wellness intervention—the active, daily, app-based meditation intervention or the health education podcast app control group, followed by a 12-week follow-up period. Participants will be asked to use their assigned app for at least 10 minutes per day during the 8-week intervention period; complete web-based surveys assessing self-reported sleep disturbance, fatigue, and emotional distress at baseline, 8 weeks, and 20 weeks; complete sleep diaries and wear an actigraphy device during the 8-week intervention period and at 20 weeks; and complete blood draws to assess inflammatory markers (tumor necrosis factor-α, interleukin-6, interleukin-8, and C-reactive protein) at baseline, 8 weeks, and 20 weeks.ResultsThis project was funded by the National Institutes of Health National Cancer Institute (R01CA262041). The projects began in April 2022, and study recruitment is scheduled to begin in October 2022, with a total project duration of 5 years. We anticipate that we will be able to achieve our enrollment goal of 276 patients with chronic hematological cancers within the allotted project time frame.ConclusionsThis research will contribute to broader public health efforts by providing researchers and clinicians with an evidence-based commercial product to improve sleep in the long term in an underserved and understudied cancer population with a high incidence of sleep disturbance.Trial RegistrationClinicalTrials.gov NCT05294991; https://clinicaltrials.gov/ct2/show/NCT05294991International Registered Report Identifier (IRRID)PRR1-10.2196/39007

Effect of Warming Yang, Tonifying Kidney, and Removing Arthralgia Therapy on Cold-Dampness Arthralgia Type Ankylosing Spondylitis and Its Influence on the Levels of Humoral Factor in Human Serum

Objective This study is aimed at exploring the effect of warming Yang, tonifying kidney, and removing arthralgia therapy in the treatment of cold-dampness arthralgia type ankylosing spondylitis (AS) and the effects on the levels of humoral factor in human serum. Method. A total of 72 patients with cold-dampness arthralgia type AS treated in our hospital from May 2020 to June 2021 were selected and divided into the observation group (n = 36) and control group (n = 36) according to the random number table method. The clinical efficacy of the two groups was observed. The traditional Chinese medicine (TCM) syndrome scores and clinical signs of the two groups were compared, and the pain situation of the two groups was evaluated by visual analog scale (VAS). Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Bath Ankylosing Spondylitis Functional Index (BASFI) were used to evaluate the spinal function and activity of the two groups, and the levels of CXC-type chemokine ligand 16 (CXCL16), Dickkopf-1(DKK-1), interleukin-17 (IL-17), tumor necrosis factor-α (TNF-α), sclerostin (SOST) ,and bone morphogenetic protein 2(BMP-2) in serum of the two groups were measured. Results The total effective rate of the observation group (91.67%) was significantly higher than that of the control group (66.67%) (P < 0.05). After treatment, the degree of improvement in TCM syndrome score, clinical signs, VAS score, BASDAI, BASFI, and the levels of CXCL16, TNF-α, IL-17, DKK-1, SOST, and BMP-2 in the observation group were significantly higher than that in the control group (all P < 0.05). Conclusion. Warming Yang, tonifying kidney, and removing arthralgia therapy had a good effect on the treatment of cold-dampness arthralgia type AS, and it could effectively improve the clinical symptoms and signs, relieve pain, improve spinal motion, and relieve inflammation of patients.

MO682: Serum Total Indoxyl Sulfate is Associated with Intraperitoneal Inflammation and High Peritonitis Episodes in Peritoneal Dialysis Patients

Abstract BACKGROUND AND AIMS The accumulating evidence presented thus far supports the idea that indoxyl sulfate (IS) is a trigger of chronic inflammation in end-stage kidney disease patients. However, although serum IS is one of the most extensively studied uremic toxins, no single study exists which has investigated the association between serum IS and intraperitoneal inflammation in peritoneal dialysis (PD) patients. The present study was undertaken to evaluate the association between serum total IS (tIS) concentration and the pro-inflammatory markers in peritoneal dialysis effluent (PDE) and peritonitis episodes in PD patients. METHOD In this observational cross-sectional study, we analysed serum tIS concentration and 24-h PDE levels of interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein -1 (MCP-1) in 74 PD patients with an average age of 55 (38–64) years and dialysis vintage of 36 (30–58) months. All patients had been undergoing continuous ambulatory PD and had no PD-associated infectious complications for more than 3 months. Serum concentrations of tIS were determined using the spectrophotometry method. The concentrations of IL-6, TNF-α and MCP-1 in PDE were analysed using ELISA. For the statistical analysis, we stratified this PD patient cohort into two groups depending on experienced peritonitis: the peritonitis group (n = 48) and the peritonitis-free group (n = 26). The data were presented as the median and the interquartile ranges [Me (Q25–Q75)] and compared using the Kruskal–Wallis test. The Spearmen correlation test was performed to assess the association between IS and the pro-inflammatory markers. RESULTS Significant differences were found in serum concentrations of tIS and PDE levels of IL-6 between the peritonitis and the peritonitis-free PD patients (Table 1). In addition, serum tIS was significantly associated with number of experienced peritonitis episodes (r = 0.26, P = 0.022). Although PDE levels of TNF-a and MCP-1 did not differ between the peritonitis and the peritonitis-free groups in our cohort, they had a direct association with serum tIS similar to those of IL-6 (Fig. 1, 2). Moreover, serum tIS was statistically higher in the anuric PD patients compared with the non-anuric patients [33.6 (13.9–74) versus 20.2 (9.3–46) μg/mL, P = 0.043], had a negative association with residual renal function (r = −0.39, P = 0.0017) and, accordingly, total weekly Kt/V (r = −0.27, P = 0.026). CONCLUSION Serum tIS concentration has a significant direct association with PDE levels of the pro-inflammatory markers, high peritonitis episodes and their outcomes in PD patients. Further well-designed studies are required to establish the effect of serum tIS concentration on intraperitoneal inflammation and clinical outcomes in PD patients.

The Clinical Efficacy of Tirofiban Combined with Ticagrelor and Aspirin in Treating Acute Myocardial Infarction by Percutaneous Coronary Intervention and Its Effect on Patients' Cardiac Function.

Objective To explore the clinical efficacy of tirofiban combined with ticagrelor and aspirin in acute myocardial infarction treatment by percutaneous coronary intervention and its effect on patients' cardiac function. Methods We selected 102 patients with acute myocardial infarction who came to The First Hospital of LanZhou University for treatment from July 2018 to May 2021. On the basis of conventional treatment, patients were separated into a joint group (tirofiban combined with ticagrelor and aspirin) comprising 55 cases and a control group (conventional ticagrelor and aspirin dual treatment) involving 47 cases. Blood flow classification of the two groups of patients was immediately recorded and compared after the myocardial infarction thrombolysis test (TIMI). Left ventricular function-related indicators, platelet-related parameters, neutrophil/lymphocyte ratio (NLR), red blood cell distribution width (RDW), and platelet/lymphocyte ratio (PLR) before treatment and 7 days after PCI were evaluated and compared between the groups before treatment and 3 months after treatment. ELISA was utilized to detect the serum levels of inflammatory factors, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and hypersensitive C-reactive protein (hs-CRP) before and after treatment. Incidence of major adverse cardiovascular events (MACEs) and adverse reaction incidence was put into comparison between the two groups in the course of the 3-month follow-up period. Compared with the control group, the joint group accounted for more patients with TIMI blood flow classification level 3 (P < 0.05) and showed more drastic improvement on the left ventricular function, platelet-related parameters, and serum inflammatory factors (P < 0.05). Moreover, patients of the joint group suffered less fluctuation from RDW, NLR, and PLR (P < 0.05), and their incidence of MACE was drastically lower in contrast with the control group (P < 0.05). No notable changes were presented in terms of incidence of adverse reaction (P > 0.05). For patients who suffered from acute myocardial infarction and treated with percutaneous coronary intervention, the application of tirofiban combined with ticagrelor and aspirin could effectively reduce the incidence of no reflow or slow blood flow, improve myocardial perfusion function, and have marked curative effects. It is worthy of clinical promotion and application.

TNF-α-Mediated RIPK1 Pathway Participates in the Development of Trigeminal Neuropathic Pain in Rats.

Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) participates in the regulation of cellular stress and inflammatory responses, but its function in neuropathic pain remains poorly understood. This study evaluated the role of RIPK1 in neuropathic pain following inferior alveolar nerve injury. We developed a model using malpositioned dental implants in male Sprague Dawley rats. This model resulted in significant mechanical allodynia and upregulated RIPK1 expression in the trigeminal subnucleus caudalis (TSC). The intracisternal administration of Necrosatin-1 (Nec-1), an RIPK1 inhibitor, blocked the mechanical allodynia produced by inferior alveolar nerve injury The intracisternal administration of recombinant rat tumor necrosis factor-α (rrTNF-α) protein in naive rats produced mechanical allodynia and upregulated RIPK1 expression in the TSC. Moreover, an intracisternal pretreatment with Nec-1 inhibited the mechanical allodynia produced by rrTNF-α protein. Nerve injury caused elevated TNF-α concentration in the TSC and a TNF-α block had anti-allodynic effects, thereby attenuating RIPK1 expression in the TSC. Finally, double immunofluorescence analyses revealed the colocalization of TNF receptor and RIPK1 with astrocytes. Hence, we have identified that astroglial RIPK1, activated by the TNF-α pathway, is a central driver of neuropathic pain and that the TNF-α-mediated RIPK1 pathway is a potential therapeutic target for reducing neuropathic pain following nerve injury.

Cinnamon(Cinnamomum japonicum) subcritical water extract suppresses gut damage induced by dextran sodium sulfate in mouse colitis model

• Clinical features and symptoms were improved in CSE-treated mice. • CSE inhibited MPO activity and reduced colonic inflammation. • CSE reduced pro-inflammatory reaction by inhibiting NF-κB inflammatory pathway. • CSE altered the gut microbiome and increases alpha-diversity in mice. Inflammatory bowel disease involves the breaking down of tight junctions and triggers leaky gut syndrome. In this study, we evaluated the effect of cinnamon subcritical water extract (CSE) on gut health improvement using a mouse colitis model induced by 5 % dextran sodium sulfate. The mice were treated with varying concentrations of CSE for 21 days and assessed. We examined histopathological changes, mRNA expression of cytokines and tight junction proteins, and alteration of the gut microbiome. CSE treatment improved clinical symptoms. Moreover, myeloperoxidase activity, interleukin (IL)-1β, IL-6, tumor necrosis factor-α protein, and mRNA expression were reduced in the CSE-treated groups. However, CSE increased the mRNA expression of tight junction proteins, such as ZO-1, occludin, mucin-1, mucin-2, and E-cadherin. In addition, CSE altered the microbiome to reduce inflammation. These results suggested that CSE could prevent dextran sodium sulfate sodium-induced colitis and disruption of the intestinal environment.

TLR4 and AT1R mediate blood-brain barrier disruption, neuroinflammation, and autonomic dysfunction in spontaneously hypertensive rats

Angiotensin II (AngII) is implicated in neuroinflammation, blood-brain barrier (BBB) disruption, and autonomic dysfunction in hypertension. We have previously shown that exogenous AngII stimulates Toll-like receptor 4 (TLR4) via AngII type 1 receptor (AT1R), inducing activation of hypothalamic microglia ex vivo, and that AngII-AT1R signaling is necessary for the loss of BBB integrity in spontaneously hypertensive rats (SHRs). Herein, we hypothesized that microglial TLR4 and AT1R signaling interactions represent a crucial mechanistic link between AngII-mediated neuroinflammation and BBB disruption, thereby contributing to sympathoexcitation in SHRs. Male SHRs were treated with TAK-242 (TLR4 inhibitor; 2 weeks), Losartan (AT1R inhibitor; 4 weeks), or vehicle, and age-matched to control Wistar Kyoto rats (WKYs). TLR4 and AT1R inhibitions normalized increased TLR4, interleukin-6, and tumor necrosis factor-α protein densities in SHR cardioregulatory nuclei (hypothalamic paraventricular nucleus [PVN], rostral ventrolateral medulla [RVLM], and nucleus tractus solitarius [NTS]), and abolished enhanced microglial activation. PVN, RVLM, and NTS BBB permeability analyses revealed complete restoration after TAK-242 treatment, whereas SHRs presented with elevated dye leakage. Mean arterial pressure was normalized in Losartan-treated SHRs, and attenuated with TLR4 inhibition. In conscious assessments, TLR4 blockade rescued SHR baroreflex sensitivity to vasoactive drugs, and reduced the SHR pressor response to ganglionic blockade to normal levels. These data suggest that TLR4 activation plays a substantial role in mediating a feed-forward pro-hypertensive cycle involving BBB disruption, neuroinflammation, and autonomic dysfunction, and that TLR4-specific therapeutic interventions may represent viable alternatives in the treatment of hypertension.

Fullerene C60 nanoparticle attenuates pain and tumor necrosis factor-α protein expression in hippocampus following diabetic neuropathy in rats

Fullerene C60 nanoparticle attenuates pain and tumor necrosis factor-α protein expression in hippocampus following diabetic neuropathy in rats

Fast and sensitive immuno-PCR assisted by plasmonic magnetic nanoparticles

Immunoassay for quantitative protein detection is an important tool in biomedical and clinical researches. Various types of detection platforms have emerged from typical ELISA using chromogenic reporter to more advanced approaches including electric, electrochemiluminescent and fluorogenic reporters to create quantitative analysis. Among them, fluorescence-based immuno-polymerase chain reaction (iPCR) is regarded as one of the simplest and most sensitive strategy that combines the specificity of immunological recognition technology with the amplification power of PCR. However, most PCR-based immunoassays to date have relied on the costly and time-consuming Peltier block heating technology. Here, we introduce a plasmonic photothermal (PPT)-iPCR assay method based on plasmonic magnetic nanoparticles to reduce the required thermal cycling time of amplification without compromising the superior sensitivity of iPCR. PPT-iPCR is capable of sensitive and quantitative detection of tumor necrosis factor-α (TNF-α) proteins ranging from 0.1 pg/mL to 1000 pg/mL within 8 min. PPT-iPCR uses fast, energy-efficient and inexpensive LED-based devices and can be used with commercial kits without modification while achieving excellent sensitivity. Our strategy can be easily combined with mobile phones or micro-optics, thus laying the foundation for the development of inexpensive, highly mobile detection devices that can quickly and accurately analyze biomarkers in a variety of diagnostic applications.

Samsum ant venom protects against carbon tetrachloride-induced acute spleen toxicity in vivo.

Many active molecules used in the development of new drugs are produced by ants. Present study assessed antioxidant and anti-inflammatory properties of Samsum ant venom (SAV) extract in carbon tetrachloride (CCL4)-induced spleen toxicity. Toxicity and oxidative stress were measured in four experimental groups: a negative control group without any treatment, a positive control group (CCl4-treated rats; a single dose of 1ml/kg CCL4), an experimental group of CCl4-treated rats co-treated daily with SAV (100μl), and a group to determine safe use with rats treated only with SAV (100μl) daily for 3weeks. CCl4-treatment led to an elevation in toxicity and oxidative stress. CCl4 significantly elevated malondialdehyde (MDA) levels, as well as expression of inhibitor of κB (IκB) and tumor necrosis factor-α (TNF-α) proteins. On the other hand, a decrease in glutathione (GSH) and catalase (CAT) levels were detected in CCl4-treated rats. Co-treatment with SAV was found to reduce these inflammatory and oxidative parameters. SAV elucidated a significant recovery of MDA concentration as well as a significant restoration in GSH levels compared to CCl4-treated rats; however, SAV increased CAT levels compared to normal rats. Hence, SAV was found to restore splenomegaly induced in CCl4-treated rats. Histopathological analysis also favored the biochemical analysis showing improvement in splenic architecture in CCl4 and SAV co-treated rats. The antioxidant properties of SAV may potentially enhance anti-inflammatory actions and improve spleen structure and function in CCl4-challenged rats.

Sinueretone A, a Diterpenoid with Unprecedented Tricyclo[12.1.0.05,9]pentadecane Carbon Scaffold from the South China Sea Soft Coral Sinularia erecta.

A novel diterpenoid, sinueretone A (1), featuring an unprecedented tricyclo[12.1.0.05,9]pentadecane carbon framework, along with two new (2 and 3) and one known (4) casbane diterpenoids were isolated from the South China Sea soft coral Sinularia erecta. The structures of the new compounds, especially their absolute stereochemistry, were established by extensive spectroscopic analysis, various quantum chemical calculations, and/or X-ray diffraction analyses. A plausible biogenetic relationship of 1-4 was proposed, which gave an insight for future biomimetic synthesis of the novel compounds. In a bioassay, compounds 1 and 2 displayed interesting anti-inflammatory activity by the inhibition of lipopolysaccharide-induced tumor necrosis factor-α protein release.

Anticancer and cytotoxic effects of troxerutin on HeLa cell line: an in-vitro model of cervical cancer.

Cervical cancer is one of the grave uterine tumors which leads to death in women worldwide. Troxerutin (TRX) as a bioflavonoid compound has many pharmacological effects such as anti-neoplastic, radioprotective, and anti-cancer. The present study was designed to examine the cytotoxic effect of TRX on human HeLa tumor cells. Human HeLa cells were cultured and treated with different doses of TRX (20-640 mg/ml) to evaluate the effective half-maximal inhibitory concentration (IC50) after 24 h. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test was used for cell proliferation assay. Also, the Bax, Bcl-2, cleaved caspase-3, and tumor necrosis factor-α (TNF-α) protein expression levels were detected with immunoblotting analysis. The malondialdehyde (MDA) concentration, glutathione peroxidase (GPx) and superoxide dismutase (SOD) activity levels were measured via their commercial kits. Data were analyzed using one-way ANOVA. The result showed that TRX at 320 mg/ml concentration (IC50) has a growth inhibitory effect against HeLa cells at 24 h treatment (P ˂ 0.01). Moreover, it increased the MDA concentration and also decreased the GPx and SOD activity levels at 320 mg/ml concentration versus control (P < 0.001). Also, TRX significantly up-regulated the Bax, cleaved caspase-3 and TNF-α proteins expression levels (P < 0.01) and down-regulated the Bcl-2 protein expression in HeLa tumor cells at 320 mg/ml concentration compared to control (P < 0.05). Our study showed that 24 h of treatment with TRX (320 mg/ml) has apoptotic and growth inhibitory effects against HeLa cells. It can induce inflammation (at least via up-regulating the TNF-α protein expression) and oxidative stress in human HeLa cells.

Effect of electroacupuncture on recognition memory and levels of Aβ, inflammatory factor proteins and aquaporin 4 in hippocampus of APP/PS1 double transgenic mice

To investigate the effect of electroacupuncture (EA) at "Baihui "(GV20) and "Shenshu "(BL23) on activation of glial cells, expression of inflammatory factor proteins and aquaporin 4 (AQP4)in the hippocampus of amyloid precursor protein/presenilin-1 (APP/PS1) transgenic mice, so as to explore its mechanisms underlying improvement of Alzheimer's disease(AD). Twenty C57/BL6 background male APP695/PS1-dE9(APP/PS1) double transgenic mice (model group) and 20 wild type (WT) C57/BL6 mice (blank group) were respectively randomized into control and EA groups. EA (2 Hz/15 Hz, 1－2 mA) was applied to GV20 and bilateral BL23 for 30 min, once daily, 6 days a week for 4 weeks. The recognition memory ability was detected by novel object recognition tests in a behavior test box. The percentage of time spent in close interaction with novel object (C) relative to the total time was used to generate preference index. The contents of hippocampal β amyloid protein (Aβ)1-40 and Aβ1-42 were assayed using ELISA, and the expression levels of glial fibrillary acidic protein (GFAP), ionic calcium binding receptor molecule-1 (Iba-1), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) proteins in the hippocampus measured by Western blot. The activities of hippocampal astrocytes (GFAP-labelled cells), microglia (Iba-1-labelled cells) and the polarity expression of AQP4 (for removing Aβ) were measured by immunohistochemistry. The preference index was significantly decreased in the model group relatively to the blank control group (P<0.05) and considerably increased in the model＋EA group relatively to the model group (P<0.05), suggesting an improvement of the recognition memory after EA. The contents of Aβ1-40 and Aβ1-42, immunoactivity of GFAP and Iba-1, expression levels of GFAP, Iba-1, IL-1β, IL-6 and TNF-α proteins were significantly higher in the model group than in the blank control group (P<0.01，P<0.05), while the AQP4 immunoactivity was notably lower in the model group than in the blank control group (P<0.05). Compared with the model group, the levels of Aβ1-40 and Aβ1-42, GFAP, Iba-1, IL-1β, IL-6 and TNF-α proteins, and the percentage of Aβ plaque area were significantly decreased in the model＋EA group (P<0.01，P<0.05), and the immunoactivity of AQP4 was significantly increased in the mo-del＋EA group (P<0.05). No significant changes were found in the above-mentioned indexes in the blank＋EA group relevant to the blank control group (P>0.05)．. EA at GV20 and BL23 can reduce inflammatory reaction and Aβ level, suppress activation of astrocytes and microglia, and up-regulate expression of AQP4 in the hippocampus tissue in APP/PS1 transgenic mice, which may contribute to its effect in improving recognition memory ability, suggesting a role of EA intervention in delaying the development of AD via promoting the drainage of Aβ by the glymphatic system in the brain.

Acute changes in the retina and central retinal artery with methamphetamine

Methamphetamine (METH), an addictive stimulant of neurotransmitters, is associated with cardiovascular and neurological diseases. METH-induced ophthalmic complications are also present but have been insufficiently investigated. The purpose of this study is to investigate the retinal effects of METH. C57BL/6 mice were administrated progressively increasing doses of METH (0–6 mg/kg) by repetitive intraperitoneal injections for 5 days (4 times per day). Retinal degeneration was examined by morphological changes and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) assay. Norepinephrine levels were measured by ELISA, protein expression levels were determined by immunoblot and immunostaining, and gelatinase activity was examined by zymography. The thickness of the retina and the number of nuclei in the inner and outer nuclear layers were decreased by METH. Retinal cell death and astrocyte activation by METH treatment were confirmed by TUNEL assay and glial fibrillary acidic protein expression, respectively. Increased tumor necrosis factor-α protein in the retina and elevated norepinephrine levels in plasma were found in METH-treated mice. Platelet endothelial cell adhesion molecule-1 (PECAM-1) protein expression level was decreased in the retina and central retinal artery (CRA) by METH treatment, along with the endothelial proteoglycans glypican-1 and syndecan-1. Moreover, a regulator of the extracellular matrix, matrix metalloproteinase-14 (MMP-14) in the retina, and MMP-2 and MMP-9 in plasma, were increased by METH treatment. In conclusion, METH administration is involved in retinal degeneration with a vascular loss of PECAM-1 and the glycocalyx in the CRA and retina, and an increase of MMPs.

Clinical study of different doses of atorvastatin combined with febuxostat in patients with gout and carotid atherosclerosis

Objective:To evaluate the efficacy of atorvastatin combined with febuxostat in the treatment of gout patients with carotid atherosclerosis and to observe the effects on serum tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and C-reactive protein (CRP) levels, carotid plaques, and the adverse reactions.Methods:Seventy patients with gout and carotid atherosclerosis admitted to Affiliated Hospital of Hebei University from January 2014 to June 2017 were randomly divided into a treatment group and a control group. The treatment group received oral febuxostat 40 mg/day combined with atorvastatin 40 mg/day. The control group was given 40 mg/day febuxostat combined with 20 mg/day atorvastatin for 90 days. The effects of treatment on TNF-α, IL-1β, and CRP levels and carotid plaques of the patients were observed.Results:After 90 days of treatment, serum TNF-α, IL-1β, and CRP levels, as well as HUA and total cholesterol (TC), decreased in both groups after treatment. There were significant differences observed (p < 0.05). The carotid artery plaques in the two groups were significantly smaller after treatment (P<0.05). There was no significant difference in adverse reactions between the two groups (P > 0.05).Conclusion:Double doses of atorvastatin combined with febuxostat can effectively reduce uric acid to improve the inflammatory state in patients and reduce carotid plaques without increasing the incidence of adverse reactions.