Structural characterization of a novel polysaccharide from Tremella fuciformis and its interaction with gut microbiota.

BACKGROUND: Because of their diverse biological activities, polysaccharides derived from Tremella fuciformis have received growing attention. This study aimed to investigate the structural characterization of a purified polysaccharide (designated as PTP-3a) derived from T. fuciformis and explore its interaction with gut microbiota in vitro. RESULTS: The findings revealed that PTP-3a had a molecular weight of 1.22 × 103 kDa and consisted of fucose, glucose, xylose, mannose and glucuronic acid in a molar ratio of 0.271:0.016:0.275:0.400:0.038. The primary linkage types identified in PTP-3a were 1,3-linked-manp, 1,4-linked-xylp and 1,2,3-linked-fucp, with corresponding ratios of 0.215:0.161:0.15. In addition, PTP-3a demonstrated notable thermal stability and exhibited a triple-helical structure. Moreover, following in vitro fermentation for 48 h, PTP-3a was efficiently utilized, resulting in a reduction in carbohydrate levels, the production of short-chain fatty acids (SCFAs) and pH adjustment. Furthermore, during in vitro fecal microbial fermentation, PTP-3a decreased the relative abundance of Firmicutes while increasing the proportions of Bacteroidetes and Proteobacteria, resulting in a significantly reduced Firmicutes/Bacteroidetes ratio. Additionally, PTP-3a stimulated the growth of beneficial bacteria such as Parabacteroides merdae, Gordonibacter pamelaeae, Bifidobacterium pseudolongum and Parabacteroides distasonis. Importantly, a strong correlation was observed between the production of SCFAs and specific microorganisms. CONCLUSION: These findings suggested that PTP-3a has potential as a prebiotic for modulating the gut microbiota. © 2024 Society of Chemical Industry.

Crude Tieguanyin oolong tea polysaccharides regulate intestinal immune and gut microflora in dextran sulfate sodium-induced mice colitis.

BACKGROUND: The global incidence and prevalence of inflammatory bowel diseases (IBDs) have been increasing. Epidemiological studies, clinical trials, and animal experiments have indicated a negative association between the consumption of tea and IBD. This study aims to investigate the protective effects of crude Tieguanyin oolong tea polysaccharides (CTPS) on experimental colitis, while also exploring the underlying mechanisms. RESULTS: The administration of CTPS significantly alleviated IBD in the mouse model, and was found to regulate T-cell mediated immune responses in the colon by modulating cytokine production associated with T cells. Furthermore, CTPS demonstrated a positive impact on the gut microbiota, reversing the increase in pathogenic Helicobacter and enhancing the relative abundances of beneficial bacteria such as Akkermansia, Lachnospiraceae, and Odoribacter. Oral administration of CTPS also led to an improvement in intestinal metabolism, specifically by increasing the levels of short-chain fatty acids. CONCLUSION: This study provides the first in vivo evidence of the protective effects of CTPS on colitis in mice. The effects are likely facilitated through the regulation of T cell-mediated responses and modulation of the gut microbiota, suggesting that CTPS may be a potential preventive and therapeutic approach for IBD. © 2023 Society of Chemical Industry.

Sirtuin 3 ameliorates inflammatory bowel disease via inhibiting intestinal inflammation and oxidative stress.

Sirtuin 3 involved in development of various diseases, but its role in inflammatory bowel disease is still unknown. We used inflammatory bowel disease biopsies, colitis animal model, and vitro cells RAW264.7 to study the role of Sirtuin 3 in the pathophysiology of inflammatory bowel disease. Sirtuin 3 negatively correlated with intestinal TNF-α. Sirt3 was less pronounced in pediatric and adult inflammatory bowel disease patients compared with corresponding control group. Sirtuin 3 activator Honokiol suppressed dextran sulfate sodium induced colonic manifestations, while Sirt3 inhibitor caused opposite results. Honokiol inhibited colonic oxidative stress by and reduced intestinal permeability. Honokiol repressed inflammatory response by reducing macrophage infiltration, pro-inflammatory cytokines TNF-α, IL-1ß, and IL-6 levels, and inhibiting activation of NF-κB p65 in the colitis mice. However, Sirt3 inhibitor amplified colonic oxidative stress and inflammatory response. In vitro study, Sirt3 inhibitor or siRNA Sirtuin 3 activated NF-κB p65 and enhanced TNF-α, IL-1ß, and IL-6 secretion from LPS stimulated RAW264.7, while Honokiol remarkably attenuated these pro-inflammatory cytokines secretion. Finally, knockdown of Sirt3 in Caco-2 cells enhanced TNF-α induced intestinal barrier integrity injury. Sirtuin 3 negatively regulates inflammatory bowel disease progression via reducing colonic inflammation and oxidative stress. Sirtuin 3 is a promising therapeutic target in clinical application for inflammatory bowel disease therapy.

Implantable metaverse with retinal prostheses and bionic vision processing.

We present an implantable metaverse featuring retinal prostheses in association with bionic vision processing. Unlike conventional retinal prostheses, whose electrodes are spaced equidistantly, our solution is to rearrange the electrodes to match the distribution of ganglion cells. To naturally imitate the human vision, a scheme of bionic vision processing is developed. On top of a three-dimensional eye model, our bionic vision processing is able to visualize the monocular image, binocular image fusion, and parallax-induced depth map.

Yellow tea: more than turning green leaves to yellow.

Yellow tea (YT), a slightly-fermented tea originated from Ming Dynasty with distinctive "Three yellows," mild-sweet smell, and mellow taste attributed to the unique yellowing process. Based on current literature and our previous work, we aim to comprehensively illustrate the key processing procedures, characteristic chemical compounds, health benefits and applications, as well as the interlocking relationships among them. Yellowing is the most vital procedure anchored on the organoleptic quality, characteristic chemical components, and bioactivities of YT, which is influenced by temperature, moisture content, duration, and ventilation conditions. Pheophorbides, carotenoids, thearubigins and theabrownins are the major pigments contributing to the "three yellows" appearance. Alcohols, such as terpinol and nerol, are attributed to the refreshing and sweet aroma of bud and small-leaf YT, while heterocyclics and aromatics forming during roasting result in the crispy rice-like large-leaf YT. Hygrothermal effects and enzymatic reactions during yellowing result in the decline of astringent substances. Meanwhile, multiple bioactive compounds such as catechins, ellagitannins, and vitexin, endow YT with antioxidant, anti-metabolic syndrome, anti-cancer, gut microbiota regulation, and organ injury protection effects. Future studies focusing on the standard yellowing process technology, quality evaluation system, and functional factors and mechanisms, possible orientations, and perspectives are guaranteed.

Transdural circumferential decompression for thoracic spinal stenosis caused by beak-type ossification of the posterior longitudinal ligament: a technical note.

PURPOSE: Thoracic myelopathy caused by ossification of the posterior longitudinal ligament (OPLL) in the thoracic spine is usually progressive and responds poorly to conservative therapy, making surgery the only effective treatment option. A variety of surgical procedures have been developed to treat thoracic OPLL. However, the optimal surgical approach for removal of thoracic OPLL remains unclear. In the present study, we described a newly modified posterior approach for the removal of OPLL: circular decompression via dural approach, and complete removal of OPLL can be achieved under direct vision and without neurological deficit. MATERIALS AND METHODS: Three patients with beak-type thoracic OPLL presented with progressive thoracic myelopathy and leg weakness. Magnetic resonance imaging showed the spinal cord severely compressed. The surgical management of the three patients involved the 'cave-in' circular decompression and transdural resection of OPLL. RESULTS: Transdural circumferential decompression was successfully performed in all three patients. Clinical outcome measures, including pre- and postoperative radiographic parameters, were assessed. All of the patients were followed up for an average of 12 months (ranging from 10 to 15 months), and no surgery-related complications occurred. Weakness relief and neural function recovery were satisfactorily achieved in all patients by the final follow-up. CONCLUSIONS: Transdural circumferential decompression was an effective method for thoracic spinal stenosis caused by concurrent beak-type OPLL, by which OPLL could be safely removed. It is especially useful when there is a severe adhesion between the dura OPLL.

A systemic review on Liubao tea: A time-honored dark tea with distinctive raw materials, process techniques, chemical profiles, and biological activities.

Liubao tea (LBT) is a unique microbial-fermented tea that boasts a long consumption history spanning 1500 years. Through a specific post-fermentation process, LBT crafted from local tea cultivars in Liubao town Guangxi acquires four distinct traits, namely, vibrant redness, thickness, aging aroma, and purity. The intricate transformations that occur during post-fermentation involve oxidation, degradation, methylation, glycosylation, and so forth, laying the substance foundation for the distinctive sensory traits. Additionally, LBT contains multitudinous bioactive compounds, such as ellagic acid, catechins, polysaccharides, and theabrownins, which contributes to the diverse modulation abilities on oxidative stress, metabolic syndromes, organic damage, and microbiota flora. However, research on LBT is currently scattered, and there is an urgent need for a systematical recapitulation of the manufacturing process, the dominant microorganisms during fermentation, the dynamic chemical alterations, the sensory traits, and the underlying health benefits. In this review, current research progresses on the peculiar tea varieties, the traditional and modern process technologies, the substance basis of sensory traits, and the latent bioactivities of LBT were comprehensively summarized. Furthermore, the present challenges and deficiencies that hinder the development of LBT, and the possible orientations and future perspectives were thoroughly discussed. By far, the productivity and quality of LBT remain restricted due to the reliance on labor and experience, as well as the incomplete understanding of the intricate interactions and underlying mechanisms involved in processing, organoleptic quality, and bioactivities. Consequently, further research is urgently warranted to address these gaps.

Near-eye display with a triple-channel waveguide for metaverse.

We present a near-eye display featuring a triple-channel waveguide with chiral liquid crystal gratings. Our triple-channel waveguide is capable of dividing one field of view into three through both the polarization orthogonality and angular separation. To illustrate its principle, a k-space diagram, which takes into account the aspect ratio of field of view, is depicted. Our results demonstrate that its diagonal field of view reaches 90°, eye relief is 10 mm, exit pupil is 4.9 × 4.9 mm2, transmittance is 4.9%, and uniformity is 89%.

Radix Tetrastigma Inhibits the Non-Small Cell Lung Cancer via Bax/Bcl-2/Caspase-9/Caspase-3 Pathway.

Lung cancer with high mortality is regarded as a challenging global problem with unsatisfied curative effects. Clinically, the chemotherapy drugs are often faced with side-effects and tumor resistance. Radix Tetrastigma (RT) is a traditional Chinese herb and now regarded as a kind of functional food. In this study, A549-bearing nude mice control was adopted to evaluate the anti-tumor capacity of RT. Results demonstrated that RT showed excellent anti-tumor ability with no side-effect on mice compared to chemotherapy drug (5-Fu).Further studies proved that RT down-regulated the proliferation-related proteins (PCNA, Ki67) and vascular endothelial growth factor (VEGF). Additionally, RT up-regulated the ratio of Bax/Bcl-2, which caused the over-expression of Caspase-9, leading to the activation of downstream protein caspase-3, eventually resulting in apoptosis of A549 in solid tumor. These results together suggest that RT inhibits the non-small cell lung cancer (NSCLC) via Bax/Bcl2/Caspase-9/Caspase-3 pathway. Furthermore, the anti-A549 abilities of the main flavonoid components from RT were compared, and kaempferol-3-O-rutinoside would play a role in RT's outstanding anti-NSCLC ability.

Pomegranate peel anthocyanins prevent diet-induced obesity and insulin resistance in association with modulation of the gut microbiota in mice.

PURPOSE: Pomegranate peels are rich in anthocyanins. The present study aimed to explore the beneficial effects of pomegranate peel anthocyanins (PPA) on obesity and gut microbiota in mice with high-fat diet (HFD)-induced obesity. METHODS: Specific pathogen-free (SPF) male C57BL/6 J mice were randomly divided into three groups and fed with low-fat diet (LFD, 10% fat energy), HFD (45% fat energy), or HFD supplemented with PPA by intragastric administration for 15 weeks. Body weight and food intake were monitored weekly. The obesity-related biochemical indexes and hepatic gene expression levels were determined. The compositions of the gut microbiota were analyzed by 16S rRNA sequencing, and the association between the gut microbiota and obesity-related indicators was investigated by Spearman correlation analysis. RESULTS: The results showed that the body weight gain, steatosis scores and insulin resistance index in the PPA group decreased by 27.46%, 56.25%, and 46.07%, respectively, compared to the HFD group. Gene expression analysis indicated that PPA supplement improved the genes expression profiles involved in glucose and lipid metabolism compared with the mice fed HFD alone. Meanwhile, PPA significantly changed the composition of the gut microbiota, which were closely correlated with the obesity-related biomarkers. CONCLUSION: This study suggested that PPA could be a beneficial treatment option for alleviating HFD-induced obesity and related metabolic disorders by targeting microbiota and lipid metabolism.

Weakened fertilization impact of anthropogenic aerosols on marine phytoplankton-A comparative analysis of dust and haze particles.

Although increases in air pollutants are changing chemical compositions of atmosphere, the resultant impacts on marine biogeochemistry remains elusive. We performed a collective analysis of 12 microcosm experimental data concerning treatments of dust particles (DPs, typically mineral aerosols), haze particles (HPs, typically anthropogenic aerosols), and various nutrients in varying trophic seawaters of the Northwest Pacific Ocean. The addition of DPs and HPs generally stimulated phytoplankton growth, as indicated by total chlorophyll a (Chl a), and shifted the phytoplankton size structure towards larger cells (> 2 µm in cell size), as indicated by size-fractionated Chl a. We further found that DP/HP-derived Chl a increase relative to the control (RCChl a) was proportional to the proportion of nitrogen (N) supplied by DPs/HPs relative to the baseline N concentration in seawater (PSN) and was higher than that in the N alone treatment when the PSN exceeded ~480%. The enhanced utilization of dissolved organic P potentially contributed to the stimulation of DPs/HPs. The slope of fitted line based on RCChl a and PSN in the DP treatments (0.14) was higher than that in the HP treatments (0.11). When the particle loading was extremely high (2 mg L-1), the addition of HPs exhibited an obvious inhibition impact on phytoplankton and was adverse to the shift of the size structure towards larger cells. These results suggest that the impact of HPs on phytoplankton is a composite result of stimulation by nutrients and inhibition by toxic matter, which may affect carbon sequestration efficiency in the ocean by regulating phytoplankton biomass and size structure.

Sensory-Guided Identification and Characterization of Kokumi-Tasting Compounds in Green Tea (Camellia sinensis L.).

The chemical substances responsible for the kokumi taste of green tea infusion are still unclear. Here, we isolated the kokumi compound-containing fractions from green tea infusion through ultrafiltration, and the major kokumi compounds were characterized as γ-Glu-Gln and γ-Glu-Cys-Gly (GSH) through ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS). The results indicated that peptides and amino acids were essential compounds in the kokumi-enriched fractions for conducting the sense of kokumi. L-theanine had an enhancing effect on the kokumi taste of green tea infusion, which was confirmed in the sensory reconstitution study. Thus, peptides, especially γ-Glu-Gln and GSH, are the major kokumi compounds in green tea infusion, which has the potential of improving the flavor of tea beverages.

Pomegranate fruit pulp polyphenols reduce diet-induced obesity with modulation of gut microbiota in mice.

BACKGROUND: Pomegranate is a rich source of polyphenols and has been used as a dietary supplement and pharmaceutical ingredient. This study aimed to investigate the pomegranate fruit pulp polyphenols (PFP) with regard to their anti-obesity activity and gut microbiota-modulating effect in mice. Thirty-six 4-week-old specific pathogen-free C57BL/6J mice (weight: 17.7-20.8 g) were randomly divided into three groups and fed with low-fat diet (10% fat energy), high-fat diet (HFD) (45% fat energy), or HFD supplemented with PFP by intragastric administration for 14 weeks. The obesity-related clinical indicators were investigated, and the composition of fecal microbiota was analyzed by 16S rRNA sequencing. RESULTS: Our results showed that PFP treatment reduced HFD-induced body weight gain by 35.23% (P < 0.05), steatosis scores by 50% (P < 0.05) and insulin resistance by 56.84% (P < 0.05), compared with the mice fed HFD alone. Moreover, compared with the mice in the HFD group, PFP supplement changed the composition of the gut microbiota, and enriched Akkermansia muciniphila, Parabacteroides distasonis, Bacteroides acidifaciens, Mucispirillum schaedleri and Lachnospiraceae bacterium 28-4, which were negatively correlated with physical biomarkers, including body weight, glucose, triglycerides and total cholesterol. CONCLUSION: PFP alleviated HFD-induced obesity, insulin resistance and hepatic steatosis in mice, and the changes in the gut microbiota might be one of the potential mechanisms through which PFP improved obesity and obesity-related disorders, eventually benefiting the recipient. © 2021 Society of Chemical Industry.

Fe3O4@Pt nanoparticles to enable combinational electrodynamic/chemodynamic therapy.

Electrodynamic therapy (EDT) has recently emerged as a potential external field responsive approach for tumor treatment. While it presents a number of clear superiorities, EDT inherits the intrinsic challenges of current reactive oxygen species (ROS) based therapeutic treatments owing to the complex tumor microenvironment, including glutathione (GSH) overexpression, acidity and others. Herein for the first time, iron oxide nanoparticles are decorated using platinum nanocrystals (Fe3O4@Pt NPs) to integrate the current EDT with chemodynamic phenomenon and GSH depletion. Fe3O4@Pt NPs can effectively induce ROS generation based on the catalytic reaction on the surface of Pt nanoparticles triggered by electric field (E), and meanwhile it may catalyze intracellular H2O2 into ROS via Fenton reaction. In addition, Fe3+ ions released from Fe3O4@Pt NPs under the acidic condition in tumor cells consume GSH in a rapid fashion, inhibiting ROS clearance to enhance its antitumor efficacy. As a result, considerable in vitro and in vivo tumor inhibition phenomena are observed. This study has demonstrated an alternative concept of combinational therapeutic modality with superior efficacy.

Catalytic core-shell nanoparticles with self-supplied calcium and H2O2 to enable combinational tumor inhibition.

Nanoparticles, presenting catalytic activity to induce intracellular oxidative species, have been extensively explored for tumor treatment, but suffer daunting challenges in the limited intracellular H2O2 and thus suppressed therapeutic efficacy. Here in this study, a type of composite nanoparticles, consisting CaO2 core and Co-ferrocene shell, is designed and synthesized for combinational tumor treatment. The findings indicate that CaO2 core can be hydrolyzed to produce large amounts of H2O2 and calcium ions at the acidic tumor sites. Meanwhile, Co-ferrocene shell acts as an excellent Fenton catalyst, inducing considerable ROS generation following its reaction with H2O2. Excessive cellular oxidative stress triggers agitated calcium accumulation in addition to the calcium ions released from the particles. The combined effect of intracellular ROS and calcium overload causes significant tumor inhibition both in vitro and in vivo.

Application of open-door laminoplasty with ARCH plate fixation in cervical intraspinal tumors.

BACKGROUND: The open-door laminoplasty is an effective procedure for the treatment of cervical spondylotic myelopathy. However, little information is available about the surgical results of open-door laminoplasty in the treatment of intraspinal tumors. In the present study, we aimed to investigate the clinical effect of open-door laminoplasty with ARCH plate fixation in the treatment of cervical intraspinal tumors. METHODS: This was a retrospective study. From January 2013 to May 2018, 38 patients (13 males and 25 females, the average age of 44 ± 17 years) with cervical intraspinal tumors underwent open-door laminoplasty with ARCH plate fixation in our hospital. The operation time, blood loss, pre- and postoperative visual analog scale (VAS), and Japanese Orthopedic Association (JOA) scores were determined. To determine the radiographic outcomes, cervical X-ray film and magnetic resonance imaging (MRI) were performed before and after the operation, and cervical X-ray sagittal film was used to measure Cobb angle. The clinical data before and after the operation were compared by t-test. RESULTS: A total of 38 patients underwent a successful operation and demonstrated primary healing. The average operation time was 113 ± 12 min. The average blood loss was 120 ± 19 mL. All patients were followed up for 26.1 ± 2.8 months, and the final follow-up time was more than 24 months. VAS scores were much better at 24 months after operation compared with those before the operation, which were decreased from 6.1 ± 1.1 to 1.4 ± 0.7 (t = 32.63, P < 0.01). The JOA score was improved from 9.9 ± 1.5 to 15.5 ± 0.6 (t = - 18.36, P < 0.01), and the mean JOA recovery rate was 79% ± 11% at 24 months after the operation. There was no significant difference in Cobb angle between pre-operation and 24 months after the operation, which was 9.8 ± 2.6 and 10.3 ± 3.1 respectively (t = - 0.61, P > 0.05). Neither spinal malalignment on the coronal plane nor displacement of the laminoplasty flap was observed on postoperative cervical X-ray and MRI examinations at the final follow-up. CONCLUSIONS: Open-door laminoplasty with ARCH plate fixation was a safe and effective surgical approach for the treatment of cervical intraspinal tumors.

MiR-143 Targeting TAK1 Attenuates Pancreatic Ductal Adenocarcinoma Progression via MAPK and NF-κB Pathway In Vitro.

BACKGROUND: Transforming growth factor (TGF)-ß-activated kinase 1 (TAK1) is one of the major regulators of inflammation-induced cancer cell growth and progression. MiR-143 dysregulation is a common event in a variety of human diseases including pancreatic ductal adenocarcinoma (PDA). AIMS: To identify the interaction between TAK1 and miR-143 in PDA. METHODS: Data mining of TAK1 expression in PDA patient gene profiling was conducted. QRT-PCR and western blot were performed to detect the expression of TAK1 in PDA tissues and cell lines. Ectopic miR-143 and TAK1 were introduced to PDA cells. Cell growth, apoptosis and migration were examined. Xenograft models were used to examine the function of TAK1 in vivo. Western blot and luciferase assay were carried out to investigate the direct target of miR-143. RESULTS: PDA patient gene profiling data (GSE15471 and GSE16515) showed that TAK1 mRNA was aberrantly up-regulated in PDA tissues. TAK1 protein levels were overexpressed in PDA tissues and cell lines. Overexpression of TAK1 was strongly associated with positive lymph node metastasis. Inhibition of TAK1 suppressed cell growth, migration, and induced cell apoptosis in vitro and in vivo. Further studies demonstrated that TAK1 was a direct target gene of miR-143. MiR-143 also inhibited PDA cells proliferation and migration, induced apoptosis and G1/S arrest. Moreover, TAK1 depletion inactivated MAPK and NF-κB pathway, mimicking the function of miR-143. CONCLUSIONS: The study highlights that miR-143 acts as a tumor suppressor in PDA through directly targeting TAK1, and their functional regulation may provide potential therapeutic strategies in clinics.

Certain Dietary Habits Contribute to the Functional Dyspepsia in South China Rural Area.

BACKGROUND Functional dyspepsia (FD) refers to a group of upper gastrointestinal syndromes, subdivided into two types: postprandial distress syndrome (PDS) and epigastric pain syndrome (EPS). The etiology of FD remains unclear; however, unhealthy dietary habit is one potential underlying cause. We aim to explore the association of poor dietary habits with FD and its subtypes. MATERIAL AND METHODS A validated epidemiological questionnaire was designed to investigate dietary habits and gastrointestinal syndromes. Citizens in the Baotun community of Dongguan were invited to complete the study questionnaire. All participants were asked to undergo a physical examination and a blinded physician interview. The study was conducted from June 2015 to June 2016. FD was diagnosed using ROME III criteria. The association between investigated dietary habits and dyspeptic symptoms were explored. RESULTS There were 1,304 adult residents recruited for the study survey; 165 residents had existing organic dyspepsia (OD), 203 residents were diagnosed with FD, and the other 936 participants, who were without dyspeptic symptoms or functional gastrointestinal diseases, were regarded as the control group. Subtype diagnosis indicated 61 participants had EPS, 66 participants had PDS, and 76 participants had coexisting EPS and PDS. Unhealthy dietary habits were more prevalent in the FD group than in the control groups (75.86% versus 37.50%; p<0.001). FD was found to be associated with irregular mealtime, dining out, fatty food, sweet food, and coffee (p<0.05). The impact of each dietary factor varied with FD subtypes. CONCLUSIONS Certain types of dietary habits were positively correlated with the prevalence of FD. FD subtypes showed relatively different associations with dietary factors.

Red pitaya betacyanins protects from diet-induced obesity, liver steatosis and insulin resistance in association with modulation of gut microbiota in mice.

BACKGROUND AND AIM: Growing evidence indicates that gut microbiota contributes to obesity and its related metabolic disorders. Betacyanins possess free radical scavenging and antioxidant activities, suggesting its potential beneficial effects on metabolic diseases. The present study aimed to investigate the metabolic effect of red pitaya (Hylocereus polyrhizus) fruit betacyanins (HPBN) on high-fat diet-fed mice and determine whether the beneficial effects of HPBN are associated with the modulation of gut microbiota. METHODS: Thirty-six male C57BL/6J mice were divided into three groups and fed low-fat diet (LFD), high-fat diet (HFD), or high-fat diet plus HPBN of 200 mg/kg for 14 weeks. Sixteen seconds rRNA sequencing was used to analyze the composition of gut microbiota. RESULTS: Our results indicated that administration of HPBN reduced HFD-induced body weight gain and visceral obesity and improved hepatic steatosis, adipose hypertrophy, and insulin resistance in mice. Sixteen seconds rRNA sequencing performed on the MiSeq Illumina platform (Illumina, Inc., San Diego, CA, USA) showed that HPBN supplement not only decreased the proportion of Firmicutes and increased the proportion of Bacteroidetes at the phylum level but also induced a dramatic increase in the relative abundance of Akkermansia at the genus level. CONCLUSIONS: Red pitaya betacyanins protect from diet-induced obesity and its related metabolic disorders, which is associated with improved inflammatory status and modulation of gut microbiota, especially its ability to decrease the ratio of Firmicutes and Bacteroidetes and increase the relative abundance of Akkermansia. The study suggested a clinical implication of HPBN in the management of obesity, non-alcoholic fatty liver disease, and type 2 diabetes.

Lentivirus-mediated short hairpin RNA interference targeting TNF-alpha in macrophages inhibits particle-induced inflammation and osteolysis in vitro and in vivo.

BACKGROUND: Aseptic loosening is a significant impediment to joint implant longevity. Prosthetic wear particles are postulated to play a central role in the onset and progression of periprosthetic osteolysis, leading to aseptic loosening of the prosthesis. METHODS: We investigated the inhibitory effects of a lentivirus-mediated short hairpin RNA that targets the TNF-alpha gene on the particle-induced inflammatory and osteolytic changes via macrophages both in vitro and in vivo. An siRNA sequence targeting the mouse TNF-alpha gene from four candidates, transcribed in vitro, was screened and identified. A lentivirus vector expressing short hairpin RNA (shRNA) was then constructed in order to facilitate efficient expression of TNF-alpha-siRNA. Lentivirus-mediated shRNA was transduced into cells of the mouse macrophage line RAW 264.7. Ceramic and titanium particles were introduced 24 h after lentivirus transduction to stimulate cells. TNF-alpha expression, represented by both mRNA and protein levels, was quantified with real-time PCR and ELISA at all time intervals. Lentivirus-mediated shRNA suspension was locally administered into the murine calvarial model, followed by local injection of particles. A multi-slice spiral CT scan was used to evaluate the osteolysis of the calvaria by detecting the width of the cranial sutures. RESULTS: Macrophages developed pseudopods when co-cultured with particles. Lentivirus-mediated shRNA was shown to effectively inhibit the expression of TNF-alpha at both the mRNA and protein levels in RAW 264.7. The multi-slice spiral CT scan showed that the lentivirus-mediated shRNA significantly suppressed osteolysis of mouse calvaria. CONCLUSIONS: Our investigation highlighted the results that lentivirus-mediated shRNA targeting the TNF-alpha gene successfully inhibited particle-induced inflammatory and osteolytic changes both in vitro and in vivo. Therefore, lentivirus-mediated gene therapy may provide a novel therapeutic approach to aseptic joint loosening.