Targeting Aging and Longevity with Exogenous Nucleotides (TALENTs): Rationale, Design, and Baseline Characteristics from a Randomized Controlled Trial in Older Adults.

Nucleotides (NTs), important biomolecules involved in numerous cellular processes, have been proposed as potential candidates for anti-aging interventions. However, whether nucleotides can act as an anti-aging supplement in older adults remains unclear. TALENTs is a randomized, double-blinded, placebo-controlled trial that evaluates the efficacy and safety of NTs as an anti-aging supplement in older adults by exploring the effects of NTs on multiple dimensions of aging in a rigorous scientific setting. Eligible community-dwelling adults aged 60-70 years were randomly assigned equally to two groups: nucleotides intervention group and placebo control group. Comprehensive geriatric health assessments were performed at baseline, 2-months, and 4-months of the intervention. Biological specimens were collected and stored for age-related biomarker testing and multi-omics sequencing. The primary outcome was the change from baseline to 4 months on leukocyte telomere length and DNA methylation age. The secondary aims were the changes in possible mechanisms underlying aging processes (immunity, inflammatory profile, oxidative stress, gene stability, endocrine, metabolism, and cardiovascular function). Other outcomes were changes in physical function, body composition and geriatric health assessment (including sleep quality, cognitive function, fatigue, frailty, and psychology). In the RCT, 301 participants were assessed for eligibility and 122 were enrolled. Participants averaged 65.65 years of age, and were predominately female (67.21%). All baseline characteristics were well-balanced between groups, as expected due to randomization. The majority of participants were pre-frailty and had at least one chronic condition. The mean scores for physical activity, psychological, fatigue and quality of life were within the normal range. However, nearly half of the participants still had room for improvement in cognitive level and sleep quality. This TALENTs trial will represent one of the most comprehensive experimental clinical trials in which supplements are administered to elderly participants. The findings of this study will contribute to our understanding of the anti-aging effects of NTs and provide insights into their potential applications in geriatric healthcare.

Treatment of malignant airway obstruction with Y-shape sigma stent loaded with I125 seeds installed via rigid bronchoscopy.

PURPOSE: To summarize and analyze the safety and efficacy of a Y-shape Sigma stent loaded with I125 in patients with inoperable malignant main airway obstruction. METHODS: This study was approved by the Institutional Ethics Committee, and a written informed consent was obtained from each participant. A Y-shape Sigma stent loaded with I125 was placed under vision from rigid bronchoscopy. The primary endpoint was alleviation of symptoms and improvement of Karnofsky Performance Status (KPS) score, and the secondary endpoint was complications and technical success. RESULTS: From November 2018 through June 2023, total 33 patients with malignant airway obstruction were palliatively treated by installing Y-shape Sigma stents loaded with I125. The airway lumen was immediately restored and the average airway opening significantly increased to 70 ± 9.4% after the procedure from baseline 30.2 ± 10.5% (p < 0.05). Average KPS score was improved from baseline 30.0 ± 10.0 to 70.0 ± 10.0 (p < 0.05) as well as PaO2 from baseline 50.1 ± 15.4 mmHg to 89.3 ± 8.6 mmHg (p < 0.05). The technical success rate of placing the stent in this study was 73%, and adverse events or complications including bleeding, I125 loss, and airway infection occurred during or after the procedure. CONCLUSION: Placement of Y-shape Sigma stents under vision from rigid bronchoscopy in the patients with malignant airway obstruction is feasible and it immediately alleviates dyspnea and significantly improves quality of life.

The Validity and Feasibility of Utilizing the Photo-Assisted Dietary Intake Assessment among College Students and Elderly Individuals in China.

Dietary assessments hold significant importance within the field of public health. However, the current methods employed for dietary assessments face certain limitations and challenges that necessitate improvement. The aim of our study was to develop a reliable and practical dietary assessment tool known as photo-assisted dietary intake assessment (PAD). In order to evaluate its validity, we conducted an analysis on a sample of 71 college students' dinners at a buffet in a canteen. We compared estimates of food weights obtained through the 24-h recall (24 HR) or PAD method with those obtained through the weighing method; we also evaluated the feasibility of PAD for recording dinner intakes among a sample of college students (n = 76) and elderly individuals (n = 121). In addition, we successfully identified the dietary factors that have a significant impact on the bias observed in weight estimation. The findings of the study indicated that the PAD method exhibited a higher level of consistency with the weighing method compared to the 24 HR method. The discrepancy in D% values between cereals (14.28% vs. 40.59%, P < 0.05), vegetables (17.67% vs. 44.44%, P < 0.05), and meats (14.29% vs. 33.33%, P < 0.05) was clearly apparent. Moreover, a significant proportion of the food mass value acquired through the PAD method fell within the limits of agreement (LOAs), in closer proximity to the central horizontal line. Furthermore, vegetables, cereals, eggs, and meats, for which the primary importance lies in accuracy, exhibited a considerably higher bias with the 24 HR method compared to the PAD method (P < 0.05), implying that the PAD method has the potential to mitigate the quality bias associated with these food items in the 24 HR method. Additionally, the PAD method was well received and easily implemented by the college students and elderly individuals. In conclusion, the PAD method demonstrates a considerable level of accuracy and feasibility as a dietary assessment method that can be effectively employed across diverse populations.

Bioactive Alkaloids as Secondary Metabolites from Plant Endophytic Aspergillus Genus.

Alkaloids represent a large family of natural products with diverse structures and bioactivities. These compounds and their derivatives have been widely used in clinics to treat various diseases. The endophytic Aspergillus is a filamentous fungus renowned for its extraordinary ability to produce active natural products of high therapeutic value and economic importance. This review is the first to focus on Aspergillus-derived alkaloids. Through an extensive literature review and data analysis, 263 alkaloids are categorized according to their structural features into those containing cytochalasans, diketopiperazine alkaloids, quinazoline alkaloids, quinoline alkaloids, indole alkaloids, pyrrolidine alkaloids, and others. These metabolites exhibited diverse biological activities, such as antibacterial activity, cytotoxicity, anti-inflammatory activity, and α-glucosidase, ACE, and DPPH inhibitory activities. The bioactivity, structural diversity, and occurrence of these alkaloids are reviewed in detail.

Anti-Alzheimer's Natural Products Derived from Plant Endophytic Fungi.

Alzheimer's is the most common cause of dementia worldwide and seriously affects patients' daily tasks. Plant endophytic fungi are known for providing novel and unique secondary metabolites with diverse activities. This review focuses primarily on the published research regarding anti-Alzheimer's natural products derived from endophytic fungi between 2002 and 2022. Following a thorough review of the literature, 468 compounds with anti-Alzheimer's-related activities are reviewed and classified based on their structural skeletons, primarily including alkaloids, peptides, polyketides, terpenoids, and sterides. The classification, occurrences, and bioactivities of these natural products from endophytic fungi are summarized in detail. Our results provide a reference on endophytic fungi natural products that may assist in the development of new anti-Alzheimer's compounds.

Perioperative safety assessment of patients undergoing unilateral or bilateral hip replacements.

Introduction: Due to the aging of the world population and the increase of obesity rate, it is expected that the number of joint replacement surgery will continue to increase in the next few years. This study evaluated the safety differences between unilateral and bilateral hip replacement surgeries. Methods: The data for patients who underwent hip arthroplasty in 2021 and 2022 were examined. The data set included 68 patients who were grouped according to the type of hip replacement needed, sex, age, and body mass index. Total blood loss and operative time were the safety-related indicators used to compare the groups. Results: Regardless of whether the unilateral replacement group was compared with the overall bilateral replacement group or separately with the staged and simultaneous bilateral replacement groups, simultaneous bilateral replacement surgeries were equally safe as the other types of hip replacements. The total blood loss and operative time for the simultaneous bilateral replacement group were not significantly different from those in the unilateral and staged bilateral replacement groups. For overweight patients, the operative time for simultaneous bilateral replacements was significantly shorter than that for unilateral replacements. Conclusions: These findings suggest that for patients requiring bilateral hip replacements, the blood loss risk for patients undergoing simultaneous bilateral hip replacements was similar to that for patients undergoing either unilateral or staged bilateral hip replacements. Thus, simultaneous bilateral total hip replacement (THR) are safe and should be considered for candidate patients.

Design of an Injectable Magnetic Hydrogel Based on the Tumor Microenvironment for Multimodal Synergistic Cancer Therapy.

Conventional tumor chemotherapy is limited by its low therapeutic efficacy and side effects, which severely hold back its further application. Drug delivery systems (DDSs) based on nanomaterials have attracted wide interest in cancer treatment; especially, the system can realize efficient synergistic therapies. Here, we designed a smart hydrogel drug delivery system with multiple responses to enhance the tumor treatment effect. By cross-linking oxidized hydroxypropyl cellulose with carboxymethyl chitosan, an injectable hydrogel was obtained, into which artesunate (ART), ferroferric oxide (Fe3O4) nanoparticles, and black phosphorus nanosheets (BPs) were preloaded. This DDS has multiple functions including magnetic targeting, pH sensitivity, chemodynamic therapy, and photothermal response. This nanoparticle-composited hydrogel not only preserved excellent rheological properties but also allowed for an accurate stable drug release at tumor sites and synergistic effects of multiple therapies. The in vitro and in vivo experiments revealed that this DDS could efficiently eliminate the HepG2 tumor with good biocompatibility. Taken together, this study clarifies the possible antitumor mechanism of this ART-loaded nanoparticle-composited hydrogel and provides a new strategy for synergistic photothermal-chemo-chemodynamic therapy.

Chinese medicine Fufang Zhenzhu Tiaozhi capsule ameliorates coronary atherosclerosis in diabetes mellitus-related coronary heart disease minipigs.

BACKGROUND: Diabetes mellitus-related coronary heart disease (DM-CHD) is the most common cause of death in diabetic patients. Various studies have shown that Chinese medicine Fufang-Zhenzhu-Tiaozhi capsule (FTZ) has therapeutic effects on cardiovascular diseases. More research is required to determine the mechanism of FTZ protection against coronary atherosclerosis. OBJECTIVE: To investigate the unique mechanism of FTZ in treatment of DM-CHD minipigs with coronary atherosclerosis. METHODS: High-fat/high-sucrose/high-cholesterol diet combined with streptozotocin and coronary balloon injury were used to induce DM-CHD minipig model, which was then randomly divided into: DM-CHD model, DM-CHD treated with FTZ or positive drug (Metformin + Atorvastatin, M+A). After twenty-two weeks, ultrasonography, electrocardiography, and image detection were employed to detect cardiac functions and assess coronary artery stenosis and plaque. Human umbilical vein endothelial cells (HUVECs) were treated high glucose or/and FTZ. Pigs tissues and treated-cells were collected for further testing. RESULTS: In DM-CHD minipigs, FTZ treatment significantly reduced disordered glycolipid metabolism similar as M+A administration. FTZ and M+A also alleviated coronary stenosis and myocardial injury. In addition, IκB and NF-κB phosphorylation levels, as well as the protein levels of IL-1ß, Bax, cleave-Caspase 3, Bcl-2, and α-SMA were dramatically increased in the DM-CHD coronary artery, whereas CD31 and VE-cadherin expressions were decreased. Similar to M+A, FTZ reversed these protein levels in the DM-CHD coronary artery. Furthermore, FTZ ameliorated the damage and high migration activity of HUVECs induced by high glucose. CONCLUSIONS: FTZ improves coronary atherosclerosis through modulating inflammation, alleviating apoptosis, and inhibiting EndMT of coronary artery to protects against DM-CHD.

Biochar combined with organic and inorganic fertilizers promoted the rapeseed nutrient uptake and improved the purple soil quality.

Biochar is a kind of organic matter that can be added into soil to improve soil quality. To study the effect of biochar combined with organic and inorganic fertilizers on rapeseed growth and purple soil fertility and microbial community, a completely randomized block design was designed with three levels of biochar (B0: no biochar, B1: low-rate biochar, B2: high-rate biochar); two levels of inorganic fertilizers (F1: low-rate inorganic fertilizer; F2: high-rate inorganic fertilizer); and two levels of organic fertilizers (M1: no organic fertilizer; M2: with organic fertilizer). All combinations were repeated three times. The combined application of biochar and organic and inorganic fertilizers could improve soil pH, soil fertility and soil microbial community richness: The pH of B1F2M1 increased 0.41 compared with the control, the nitrogen, phosphorus and potassium content increased by 103.95, 117.88, and 99.05%. Meanwhile, soil microbial community richness was also improved. Our research showed that biochar could promote the Nutrient Uptake of rapeseed, and the combined application of biochar with organic and inorganic fertilizers could improve soil fertility and increase microbial diversity. Low-rate biochar combined with organic fertilizer and low-rate inorganic fertilizer was the most suitable application mode in rapeseed production in purple soil area of Southwest China.

Short-term Effectiveness of a Modified Osteotomy for Total Elbow Joint Replacement in Patients of Hemophilic Elbow Arthritis with Severe Flexion Contracture Deformity: A Three-Cases Series Report.

OBJECTIVE: To report the modified osteotomy and the short-term effectiveness of the total elbow joint replacement in patients of hemophilic elbow arthritis with severe flexion contracture deformity. METHODS: This study introduced the therapeutic approach of the total elbow joint replacement in patients of hemophilic elbow arthritis with severe flexion contracture deformity, and assessed the short-term effectiveness in three cases (three elbows) of end-stage hemophilic elbow arthritis admitted from October 2020 to December 2020. The included patients were all diagnosed with hemophilia A (factor VII deficiency), accompanied by severe bilateral elbow joint flexion contracture, which seriously affects daily life and requires surgical intervention. Clinical data and follow-up results were analyzed before total elbow arthroplasty and 1, 3, and 6 months postoperatively. Pre- and postoperative range of motion, pain score, and function score were compared, and intraoperative and postoperative complications are reported. RESULTS: All three patients were male, with an average age of 31 years. The main clinical manifestations were bilateral elbow arthritis with flexion contracture. Two of the patients underwent right elbow replacement, and one patient underwent left elbow replacement. All cases were followed up for 6 months postoperatively. No incision infection or ulnar nerve injury occurred. Postoperative triceps brachii muscle strength was slightly weakened compared with preoperative muscle strength. Average elbow flexion and extension range of motion was 60° (30°-100°) preoperatively and increased to 127° (110°-140°) postoperatively; rotational range of motion of the affected forearm was 47° (10°-85°) preoperatively and increased to 117° postoperatively. The mean visual analogue scale (VAS) was 6 (5-8) preoperatively and decreased to 3 (2-4) postoperatively. The mean MEPS score was 62 (55-75) and increased to 87 (80-95) postoperatively. During the follow-up, anteroposterior and lateral radiographs showed no signs of prosthesis loosening in the elbow. CONCLUSIONS: For severe hemophilic elbow arthritis patients, the short-term treatment effect of total elbow replacement is good, following the strict adherence to the surgical indications and proper preparation for the perioperative period. The modified osteotomy can fully expose the visual field and reduce complications of ulnar nerve injury. The long-term effects need to be study future.

Traditional Chinese Medicine Fufang-Zhenzhu-Tiaozhi capsule prevents renal injury in diabetic minipigs with coronary heart disease.

BACKGROUND: Renal injury is one of the common microvascular complications of diabetes, known as diabetic kidney disease (DKD) seriously threatening human health. Previous research has reported that the Chinese Medicine Fufang-Zhenzhu-Tiaozhi (FTZ) capsule protected myocardia from injury in diabetic minipigs with coronary heart disease (DM-CHD). And we found significant renal injury in the minipigs. Therefore, we further investigated whether FTZ prevents renal injury of DM-CHD minipig and H2O2-induced oxidative injury of HK-2 cells. METHODS: DM-CHD model was established by streptozotocin injection, high fat/high-sucrose/high-cholesterol diet combined with balloon injury in the coronary artery. Blood lipid profile, fasting blood glucose (FBG), and SOD were measured with kits. The levels of blood urea nitrogen (BUN), serum creatinine (Scr), urine trace albumin (UALB), urine creatinine (UCR) (calculate UACR), cystatin (Cys-C), and ß-microglobulin (ß-MG) were measured by ELISA kits to evaluate renal function. TUNEL assay was performed to observe the apoptosis. qPCR was used to detect the mRNA expression levels of HO-1, NQO1, and SOD in kidney tissue. The protein expressions of Nrf2, HO-1, NQO1, Bax, Bcl-2, and Caspase 3 in the kidney tissue and HK-2 cells were detected by western blot. Meanwhile, HK-2 cells were induced by H2O2 to establish an oxidative stress injury model to verify the protective effect and mechanisms of FTZ. RESULTS: In DM-CHD minipigs, blood lipid profile and FBG were elevated significantly, and the renal function was decreased with the increase of BUN, Scr, UACR, Cys-c, and ß-MG. A large number of inflammatory and apoptotic cells in the kidney were observed accompanied with lower levels of SOD, Bcl-2, Nrf2, HO-1, and NQO1, but high levels of Bax and Cleaved-caspase 3. FTZ alleviated glucose-lipid metabolic disorders and the pathological morphology of the kidney. The renal function was improved and the apoptotic cells were reduced by FTZ administration. FTZ could also enhance the levels of SOD, Nrf2, HO-1, and NQO1 proteins to promote antioxidant effect, down-regulate the expression of Bax and Caspase3, as well as up-regulate the expression of Bcl-2 to inhibit cell apoptosis in the kidney tissue and HK-2 cells. CONCLUSIONS: We concluded that FTZ prevents renal injury of DM-CHD through activating anti-oxidative capacity to reduce apoptosis and inhibiting inflammation, which may be a new candidate for DKD treatment.

Perioperative blood loss during joint replacement: comparison between patients with and without hemophilia.

BACKGROUND: Although arthroplasty provides satisfactory pain relief, functional improvement, and reduced flexion contracture in patients with hemophilia arthropathy, bleeding remains the primary problem associated with hemophilic arthropathy joint replacement. We aimed to explore the differences in perioperative blood loss (PBL) associated with joint replacement surgery in patients with and without hemophilia. METHODS: This study retrospectively analyzed 61 cases of PBL in patients undergoing joint replacement surgery, including 37 patients with hemophilia and 24 patients without hemophilia. All patients demonstrated severe joint flexion contractures that seriously affected their daily lives and required surgical intervention. PBL was compared between the two groups. RESULTS: In univariate analysis, the overall (p < 0.001) and hidden (p < 0.001) blood losses were significantly higher for patients with hemophilia than those for patients without hemophilia. However, after adjustment for multiple factors, there was no significant difference in overall blood loss between the two groups (p = 0.731). In addition, sex, age group, and surgical site did not affect blood loss in patients with hemophilia. CONCLUSION: Overt bleeding did not increase significantly in patients with hemophilia, compared with that in patients without hemophilia. In terms of blood loss, joint replacement surgery for patients with hemophilia is relatively safe. The results of this study must be verified by a prospective follow-up study with larger sample size. Trial registration Retrospectively registered.

Human epididymis protein 4: a novel predictor of ischemic cardiomyopathy.

BACKGROUND: The prognostic value of human epididymis protein 4 (HE4) in patients with ischemic cardiomyopathy (ICM) is unknown. METHODS: A total of 103 patients with ICM were prospectively enrolled in this study from Hunan Provincial People's Hospital between February 2019 and June 2019. All patients were tested for HE4 levels at baseline and follow-up. Endpoints of the study included cardiovascular death and heart failure-related hospitalization. RESULTS: A total of 96 patients with ICM were included for analysis. After a mean follow-up period of 263 (153-313) days, cardiovascular events were observed in 45 patients. Serum HE4 levels in patients with events were significantly higher than those in patients without events [188.70 (113.35-326.82) pmol/L versus 92.90 (61.50-123.20) pmol/L, P < 0.001]. Multivariate Cox regression analysis revealed that HE4 [χ2: 9.602, hazard ratio (HR): 1.003, 95% confidence interval (CI): 1.001-1.005, P = 0.002] and age [χ2: 4.55, HR: 1.044, 95% CI: 1.003-1.085, P = 0.033] were independent predictors of events. After adjusting for age and sex, the risk of events in patients with HE4 > 100.2 pmol/L was higher than that in patients with HE4 ≤ 100.2 pmol/L [HR: 3.372, 95% CI: 1.409-8.065, P < 0.001]. CONCLUSION: HE4 is an independent predictor of cardiovascular death and heart failure-related rehospitalization in patients with ICM.

FTZ attenuates liver steatosis and fibrosis in the minipigs with type 2 diabetes by regulating the AMPK signaling pathway.

Fufang Zhenzhu Tiaozhi formula (FTZ), a preparation of Chinese herbal medicine, has various pharmacological properties, such as hypoglycemic, hypolipidemic, anticoagulant, and anti-inflammatory activities. Hepatocyte apoptosis is a marker of nonalcoholic steatohepatitis (NASH) and contributes to liver injury, fibrosis, and inflammation. Given the multiple effects of FTZ, we investigated whether FTZ can be a therapeutic agent for NASH and its mechanism. In the present study, we observed that FTZ treatment had an obviously favorable influence on hepatic steatosis and fibrosis in the histopathologic features of type 2 diabetes mellitus (T2DM) and coronary heart disease (CHD) with NASH minipigs. In addition, immunohistochemical analysis showed increased expression of the fibrotic marker α-smooth muscle actin (α-SMA), and a TUNEL assay revealed increased apoptotic positive hepatic cells in the liver tissues of the model group. Furthermore, FTZ administration reduced the increased expression of α-SMA, and FTZ inhibited apoptosis by affecting Bcl-2/Bax and cleaved caspase-3 expression. Mechanistically, our data suggested that FTZ treatment attenuated hepatic steatosis and fibrosis via the adenosine monophosphate-activated protein kinase (AMPK) pathway. In vitro studies showed that FTZ also attenuated intracellular lipid accumulation in HepG2 cells exposed to palmitic acid (PA) and oleic acid (OA). FTZ upregulated the expression levels of P-AMPK and BCL-2 and downregulated BAX. The changes induced by FTZ were reversed by Compound C, an inhibitor of AMPK. In conclusion, FTZ attenuated NASH by ameliorating steatosis and hepatocyte apoptosis, which is attributable to the regulation of the AMPK pathway.

[Design and Clinical Application of Split Memory Alloy Sternum Plate].

OBJECTIVE: The design and development of split memory alloy sternum bone plate are discussed, and the effect of split memory alloy sternum bone plate internal fixation in the treatment of sternal fractures are analysed. METHODS: The structure of the product is designed according to the anatomy and physiological characteristics of human bones, and the cross section shape of the product is designed according to the cross section shape of human bones. Internal fixation is effective in the treatment of sternal fracture. RESULTS: The split memory alloy sternal plate was successfully designed and developed, and all the patients with sternal fractures treated by internal fixation were clinically healed, the hospitalization and fracture healing time were significantly shortened, and no obvious complications occurred. CONCLUSIONS: The application of split memory alloy sternal plate internal fixation in the treatment of sternal fracture has the advantages of small trauma, simple operation, safety, reliable fixation, good histocompatibility and less complications, and is conducive to promoting fracture healing and respiratory function improvement.

The traditional Chinese medicine formula Fufang-Zhenzhu-Tiaozhi protects myocardia from injury in diabetic minipigs with coronary heart disease.

BACKGROUND AND PURPOSE: Diabetes mellitus (DM) is a major risk factor for coronary heart disease (CHD). Previous research has reported that the Fufang-Zhenzhu-Tiaozhi (FTZ) formula has obvious effects on the treatment of dyslipidemia and hyperglycemia. In the present study, we intended to establish a convenient DM-CHD model in minipigs and investigated the protective effect of FTZ against myocardial injury and its mechanism. METHODS: The DM-CHD model was established by a high-fat/high-sucrose/high-cholesterol diet (HFSCD) combined with balloon injury in the coronary artery. Subsequently, sixteen Wuzhishan minipigs were assigned to three groups: control group, model group, and FTZ group. The model group and FTZ group were given a HFSCD, while the control group was given a normal diet (ND). FTZ was given with meals in the FTZ group. During this time, biochemical parameters, such as total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein (HDL-C), and fasting blood glucose (FBG), were measured by using testing kits. Insulin (INS) was determined by ELISA, and the homeostasis model assessment index of insulin resistance (HOMA-IR) was calculated to evaluate insulin resistance levels. After FTZ administration, the plasma levels of lactate dehydrogenase (LDH), creatine kinase isoenzyme MB (CK-MB), and cardiac troponin I (cTnI) were measured by using ELISA kits to evaluate myocardial injury. Coronary artery stenosis was analyzed by angiographic and HE staining. Myocardial ischemia was assayed with electrocardiogram (ECG). Moreover, cytokines, including interleukin-6 (IL-6), hypersensitive C-reactive protein (hs-CRP), and tumor necrosis factor-alpha (TNF-α), were measured by ELISA kits to assess inflammation. The myocardial tissue was collected, and the pathological morphology was observed by transmission electron microscopy (TEM), HE staining, and Masson staining. Western blots were used to detect the expression of PI3K, AKT, p-AKT, p-NF-κB, and NF-κB. RESULTS: A DM-CHD model in minipigs with glucose-lipid metabolism disorder, coronary artery incrassation and myocardial damage was successfully established through balloon injury in the coronary artery combined with HFSCD. FTZ effectively inhibited coronary artery incrassation and protected the myocardium against injury in DM-CHD minipigs. FTZ decreased proinflammatory cytokine levels and upregulated the protein expression of the PI3K/Akt pathway in the myocardium. CONCLUSIONS: A novel DM-CHD model in minipigs was successfully established through balloon injury in the coronary artery combined with HFSCD. FTZ has a protective effect against myocardial injury in DM-CHD by inhibiting inflammation and activating the PI3K/AKT signaling pathway.

Molecular mechanism of Fufang Zhenzhu Tiaozhi capsule in the treatment of type 2 diabetes mellitus with nonalcoholic fatty liver disease based on network pharmacology and validation in minipigs.

ETHNOPHARMACOLOGICAL RELEVANCE: Fufang Zhenzhu Tiaozhi formula (FTZ) of which a patented preparation of Chinese herbal medicine has been well documented with significant clinical curative effect for hyperglycemia and hyperlipidemia. Because of the complexity of the chemical constituents of Chinese herbal formulas, the holistic pharmacological mechanism of FTZ acting on type 2 diabetes mellitus (T2DM) and nonalcoholic fatty liver disease (NAFLD) remains unclear. AIM OF THE STUDY: To investigate the pharmacological efficacy and mechanism of FTZ in the treatment of T2DM accompanied by NAFLD. MATERIALS AND METHODS: Network pharmacology and validation in minipigs were used in this study. First, potential bioactive compounds of FTZ were identified by the traditional Chinese medicine system pharmacology technology platform (TCMSP). Then, targets of compounds were gathered using DrugBank, SwissTargetPrediction and TCMSP, while targets for T2DM and NAFLD were collected from CTD (compounds-targets-diseases network) and GeneCards. Common targets were defined as direct therapeutic targets acting on T2DM with NAFLD. In addition, crucial targets were chosen by the protein-protein interaction (PPI) network and contribution to compound-therapeutic targets in T2DM with the NAFLD network. Furthermore, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze the metabolism-related signaling pathways affected by FTZ. Candidate patterns selected by network pharmacology were tested in the minipigs model of T2DM with NAFLD. Measurements of triglycerides (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), fasting insulin (FINS) and fasting blood glucose (FBG) in the blood and the expression levels of proteins, including PI3K-AKT and HIF-1α, in the livers of the minipigs were followed by the administration of FTZ. RESULTS: A total of 116 active compounds and 82 potential targets related to T2DM and NAFLD were found. Pathway and functional enrichment analysis showed that FTZ mainly regulates metabolism-related pathways, including PI3K-AKT, HIF-1α, TNFα and MAPK. Animal experiments showed that FTZ treatment significantly reduced the serum levels of TG, TC, LDL-C and FBG, increased serum levels of HDL-C, ameliorated systemic insulin resistance (IR), and attenuated liver damage in minipigs with T2DM and NAFLD. FTZ treatment has an obviously favorable influence on hepatic steatosis and liver lipid accumulation in the histopathologic features of HE, Oil red O staining, and electron microscopy. Mechanistically, FTZ improved liver metabolism by increasing the phosphorylation of PI3K-AKT and decreasing the expression of HIF-1α. CONCLUSION: Network pharmacology was supported by experimental studies, which indicated that FTZ has demonstrated therapeutic benefits in T2DM and NAFLD by regulating the PI3K-AKT and HIF-1α signaling pathways.

PLOD1 Is a Prognostic Biomarker and Mediator of Proliferation and Invasion in Osteosarcoma.

OBJECTIVE: Osteosarcoma is the most common primary bone tumor and most frequently develops during adolescence. PLOD family was mainly involved in lysyl hydroxylation and rarely investigated in cancers, especially in osteosarcoma. The aim of this study was to investigate the expression pattern and oncogenic role of PLODs in osteosarcoma. METHODS: GEO datasets (GSE16088, GSE33382, and GSE16091) and validation cohort were used to analyze the expression pattern of PLODs in osteosarcoma. Kaplan-Meier survival analysis was used to explore the prognostic role of PLODs in patients with osteosarcoma. RNA interference of KRT19 was performed using small interfering RNA (siRNA) in MG-63 and U-2OS cells. The proliferation was detected using CCK8, clone formation assay, and EdU staining. Migration and invasion were determined using the transwell assay. Western blots and luciferase assays for ß-catenin-T-cell factor protein/ß-catenin-lymphoid enhancer factor- (ß-catenin-TCF/LEF-) driven transcriptional activity. RESULTS: PLOD1 was upregulated in osteosarcoma tissues compared with control tissues both in public datasets and in in-house cohort. The expression of PLOD1 in osteosarcoma tissues was significantly associated with the status of distance metastasis and Enneking stage, while PLOD2 and PLOD3 expressed no difference between osteosarcoma and benign tissues and showed no correlation with tumor malignancy. Furthermore, Kaplan-Meier survival analysis revealed that patients with a higher level of PLOD1 had worse prognosis than those with a lower level of PLOD1. Downregulation of PLOD1 dramatically inhibited proliferation, migration, and invasion of MG-63 cells and U-2OS cells in vitro. Mechanistically, PLOD1 regulated ß-catenin signaling pathway in osteosarcoma. CONCLUSION: Our results indicated that PLOD1 promoted proliferation, migration, and invasion of osteosarcoma cells. PLOD1 was a novel prognostic marker, as well as a therapeutic target in osteosarcoma.

Bilobalide reversibly modulates blood-brain barrier permeability through promoting adenosine A1 receptor-mediated phosphorylation of actin-binding proteins.

Bilobalide, one of the key bioactive components of Ginkgo biloba leaves, exerts prominent neuroprotective properties in central nervous system (CNS) disease. However, the effect of bilobalide on blood-brain barrier (BBB) permeability remains unknown. In this study, we investigated the effect of bilobalide on BBB permeability and its potential mechanism involved. Both the in vitro and in vivo results showed that significant enhancement of BBB permeability was found following bilobalide treatment, evidenced by the reduced transendothelial electrical resistance (TEER), the increased fluorescein sodium (Na-F) penetration rate in vitro and the leakage of FITC-dextran in vivo. Transmission electron microscope (TEM) images demonstrated that bilobalide modulated BBB permeability by changing the ultrastructure of tight junctions (TJs). In addition, actin-binding proteins ezrin, radixin and moesin (ERM) and Myosin light chain (MLC) phosphorylation was observed following bilobalide treatment. Moreover, the effect of bilobalide on TEER reduction and ERM/MLC phosphorylation was counteracted by adenosine A1 receptor (A1R) siRNA. The current findings suggested that bilobalide might reversibly modulate BBB permeability by the alteration of TJs ultrastructure through A1R-mediated phosphorylation of actin-binding proteins.