Role of intestinal flora in the development of nonalcoholic fatty liver disease in children.

In China, 45% of adolescents with obesity develop fatty liver disease, a condition that increases the long-term risk of developing cirrhosis and liver cancer. Although the factors triggering nonalcoholic fatty liver disease (NAFLD) vary in children, the composition of intestinal microflora has been found to play an increasingly important role. However, evidence is limited on the prevalence of nonalcoholic fatty liver (NAFL) and nonalcoholic steatohepatitis (NASH) in Chinese children. Therefore, this study aimed to evaluate the fecal microbiome of Chinese children with NAFLD and further analyze the potential of flora in regulating NAFLD-related symptoms and metabolic functions. Specifically, the study applied a 16S rRNA and metagenomic sequencing to the fecal samples of pediatric patients with NAFLD, NASH, and NAFL, as well as healthy controls, to explore the correlation among NAFLD-related indexes, metabolic pathways, and gut flora. The findings showed that some fecal microbiota had a negative correlation with body mass index, and various NAFLD-related bacteria, including Lachnoclostridium, Escherichia-Shigella, and Faecalibacterium prausnitzii, were detected. Consequently, the study concluded that the variation in gut microbiota might be more important in improving NAFLD/NASH compared with single species, providing a microbiota diagnostic profile of NAFLD/NASH.IMPORTANCEThis study aims to characterize the gut microbiota in Chinese children with nonalcoholic fatty liver disease (NAFLD) through 16S rRNA and metagenomic sequencing. The results highlight the association between fecal microbiota and NAFLD in Chinese children, demonstrating distinct characteristics compared to adults and children from other countries. Based on the sequencing data from our cohort's fecal samples, we propose a microbiota model with a high area under the curve for distinguishing between NAFLD and healthy individuals. Furthermore, our follow-up study reveals that changes in the relative abundance of microbial biomarkers in this model are consistent with variations in patients' body mass index. These findings suggest the potential utility of the microbiota model and microbial biomarkers for diagnosing and treating NAFLD in children.

BMI at different childhood age periods associated with cardiometabolic disorders in young adulthood.

OBJECTIVE: The aim of this study was to investigate when the association between childhood obesity and adult cardiometabolic disorders starts to be operative. METHODS: The study cohort included 811 participants who had data on blood pressure, lipid profile, fasting blood glucose, fasting insulin, or alanine aminotransferase in adulthood and had at least one measurement of BMI in childhood. RESULTS: Childhood BMI z score was significantly associated with increased risks of ≥1 cardiometabolic disorders, ≥2 cardiometabolic disorders, and elevated blood pressure in adulthood for age groups of 6 to 9 years, 10 to 12 years, 13 to 15 years, and 16 to 18 years, after adjustment for covariates. For low high-density lipoprotein cholesterol, significant associations were observed for age groups of 10 to 12 years, 13 to 15 years, and 16 to 18 years. For elevated triglyceride and elevated alanine aminotransferase, significant associations were observed for age groups of 13 to 15 years and 16 to 18 years. For insulin resistance, significant associations were observed for age groups of 10 to 12 years and 16 to 18 years. For elevated total cholesterol, elevated low-density lipoprotein cholesterol, or elevated fasting blood glucose, no association was observed in any age group. CONCLUSIONS: The association between childhood BMI and adult cardiometabolic disorders begins to be operative from early life. These results support universal screening of childhood obesity starting at an early age.

Evaluating Differentiation Status of Mesenchymal Stem Cells by Label-Free Microscopy System and Machine Learning.

Mesenchymal stem cells (MSCs) play a crucial role in tissue engineering, as their differentiation status directly affects the quality of the final cultured tissue, which is critical to the success of transplantation therapy. Furthermore, the precise control of MSC differentiation is essential for stem cell therapy in clinical settings, as low-purity stem cells can lead to tumorigenic problems. Therefore, to address the heterogeneity of MSCs during their differentiation into adipogenic or osteogenic lineages, numerous label-free microscopic images were acquired using fluorescence lifetime imaging microscopy (FLIM) and stimulated Raman scattering (SRS), and an automated evaluation model for the differentiation status of MSCs was built based on the K-means machine learning algorithm. The model is capable of highly sensitive analysis of individual cell differentiation status, so it has great potential for stem cell differentiation research.

Effect of Insulin Resistance on Recurrence after Radiofrequency Catheter Ablation in Patients with Atrial Fibrillation.

BACKGROUND: Whether there are many risk factors for recurrence of atrial fibrillation (AF) after ablation is unclear. The aim of this study was to investigate the relationship between insulin resistance (IR) and AF recurrence in patients without diabetes who underwent catheter ablation. METHODS: This retrospective study included patients who underwent AF ablation between 2018 and 2019 at the First Affiliated Hospital of Zhengzhou University. Homeostasis model assessment of insulin resistance (HOMA-IR) was calculated, and a value of ≥2.69 was defined as IR. The patients were divided into two groups (group 1 HOMA-IR < 2.69, n = 163; group 2 HOMA-IR ≥ 2.69, n = 69). AF recurrence was defined as the occurrence of atrial arrhythmias of more than 30 s after the first 3 months. Univariate and multivariable Cox regression models were used to analyse the risk of AF recurrence. RESULTS: Overall, 232 patients were enrolled (mean age, 59.9 ± 10.2 years old; female, 37.5%; paroxysmal AF, 71.6%). We found that dyslipidaemia, antiarrhythmic drug use, fasting blood glucose and fasting insulin were significantly higher in the IR group (P < 0.05). During the follow-up 1 year after ablation, 62 (26.7%) patients experienced AF recurrence. After adjusting for traditional risk factors, multivariable analysis showed that the HOMA-IR value (HR 1.259, 95% CI 1.086-1.460, P = 0.002) and left atrial diameter (LAD; HR 1.043, 95% CI 1.005-1.083, P = 0.026) were independently associated with AF recurrence. CONCLUSIONS: The present results provide evidence that IR patients are more likely to experience AF recurrence. Improving IR status may be a potential target for reducing the postoperative recurrence rate.

Long-Term Adiposity and Midlife Carotid Intima-Media Thickness Are Linked Partly Through Intermediate Risk Factors.

OBJECTIVE: This study aims to determine quantitatively the mediation effects of multiple cardiovascular risk factors on the associations of childhood body mass index (BMI) and its cumulative burden with adult carotid intima-media thickness (cIMT). METHODS: The longitudinal cohort consisted of 1391 adults who had been examined for BMI 4-15 times over 35.0 years on average since childhood and had data on adult cIMT, systolic blood pressure, low-density lipoprotein cholesterol, atherogenic index of plasma, and serum glucose. The area under the curve was used as a measure of cumulative burden of BMI. RESULTS: After adjusting for covariates, the total effects (standardized regression coefficient) of childhood BMI (0.138), adult BMI (0.111), and area under the curve of BMI (0.150) on cIMT were all significant (P<0.001) without mediators included in the model. The mediation effects of adult systolic blood pressure, glucose, atherogenic index of plasma and low-density lipoprotein cholesterol were 8.0%, 4.3%, 3.6%, and 0.0%, respectively, in the model with childhood BMI as the predictor, 23.4%, 15.3%, 12.6%, and 7.2%, respectively, with adult BMI as the predictor, and 14.7%, 8.7%, 6.0%, and 2.0%, respectively, with area under the curve of BMI as the predictor. The direct effects on cIMT were 0.117 (P<0.001) for childhood BMI, 0.046 (P=0.224) for adult BMI, and 0.103 (P<0.001) for area under the curve of BMI after removing the mediation effects. CONCLUSIONS: The long-term deleterious impact of adiposity on subclinical changes in vascular structure begins early in life and is accumulated over lifetime. Excess adiposity and higher cIMT are linked partly through other cardiovascular risk factors in later life, especially elevated blood pressure and glucose.

Early Detection of Cervical Cancer by Fluorescence Lifetime Imaging Microscopy Combined with Unsupervised Machine Learning.

Cervical cancer has high morbidity and mortality rates, affecting hundreds of thousands of women worldwide and requiring more accurate screening for early intervention and follow-up treatment. Cytology is the current dominant clinical screening approach, and though it has been used for decades, it has unsatisfactory sensitivity and specificity. In this work, fluorescence lifetime imaging microscopy (FLIM) was used for the imaging of exfoliated cervical cells in which an endogenous coenzyme involved in metabolism, namely, reduced nicotinamide adenine dinucleotide (phosphate) [NAD(P)H], was detected to evaluate the metabolic status of cells. FLIM images from 71 participants were analyzed by the unsupervised machine learning method to build a prediction model for cervical cancer risk. The FLIM method combined with unsupervised machine learning (FLIM-ML) had a sensitivity and specificity of 90.9% and 100%, respectively, significantly higher than those of the cytology approach. One cancer recurrence case was predicted as high-risk several months earlier using this method as compared to using current clinical methods, implying that FLIM-ML may be very helpful for follow-up cancer care. This study illustrates the clinical applicability of FLIM-ML as a detection method for cervical cancer screening and a convenient tool for follow-up cancer care.

Application of Dual-Enhanced Surface-Enhanced Raman Scattering Probe Technology in the Diagnosis of Tumor Cells in Vitro.

With the development of precision medicine, antigen/antibody-targeted therapy has brought great hope to tumor patients; however, the migration of tumor cells, especially a small number of cells flowing into blood or other tissues, remains a clinical challenge. In particular, it is difficult to use functional gold nanomaterials for targeted clinical tumor diagnosis while simultaneously obtaining stable and highly sensitive Raman signals. Therefore, we developed a detection method for functional Au Nanostars (AuNSs) with dual signal enhancement that can specifically track location and obtain high-intensity surface-enhanced Raman scattering (SERS) signals. First, AuNSs with specific optical properties were synthesized and functionalized. The Raman dye 4-mercapto-hydroxybenzoic acid and polyethylene glycol were coupled with the tumor marker, epidermal growth factor receptor, to obtain the targeted SERS probes. In addition, a detection chip was prepared for Raman detection with physical enhancement, exhibiting a 40-times higher signal intensity than that of quartz glass. This study combines physical enhancement and SERS enhancement technologies to achieve dual enhancement, enabling the detection of a highly sensitive and stable Raman signal; this has potential clinical value for antigen/antibody-targeted tumor diagnosis and treatment.

Associations between body mass index in different childhood age periods and hyperuricemia in young adulthood: the China Health and Nutrition Survey cohort study.

BACKGROUND: Few studies have evaluated the specific age period in childhood when the association of body mass index with adult hyperuricemia begins to be operative. This study aimed to examine the associations between body mass index in different childhood age periods and the risk of adult hyperuricemia in China. METHODS: The study cohort from the China Health and Nutrition Survey included 676 participants who were aged ≥ 18 years and had data on uric acid in 2009 with at least one measurement of body mass index in childhood surveys before 2009. There were 357, 365, 358, 427, and 432 observations in childhood age groups of ≤ 5 years, 6-9 years, 10-12 years, 13-15 years, and 16-18 years, respectively. Body mass index Z score was calculated based on 2000 Center for Disease Control and Prevention growth charts for the United States. RESULTS: Childhood body mass index Z scores measured at age ≤ 5 years, 6-9 years, 10-12 years, and 13-15 years had no statistical association with adult uric acid. In comparison, childhood body mass index Z scores measured at age 16-18 years were significantly associated with adult uric acid (ß = 11.539, P = 0.007), and the strength of association was stronger in girls (ß = 18.565, P = 0.002) than in boys (ß = 9.209, P = 0.087). In addition, childhood body mass index Z scores measured at age 16-18 years were significantly associated with an increased risk of adult hyperuricemia (odds ratio = 1.323, 95% confidence interval = 1.003-1.746, P = 0.048), but not for other age groups. CONCLUSION: The association between childhood body mass index and young adulthood hyperuricemia was influenced by childhood age.

Single cell sorting of young yeast based on label-free fluorescence lifetime imaging microscopy.

Saccharomyces cerevisiae is an attractive organism used in the fermentation industry and is an important model organism for virus research. The ability to sort yeast cells is important for diverse applications. Replicative aging of Saccharomyces Cerevisiae is accompanied by metabolic changes that are related to an essential coenzyme, reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H). Here, a single cell sorting method based on fluorescence lifetime imaging microscopy (FLIM) and laser-induced forward transfer (LIFT) was implemented for the first time. The aging level of yeast was determined based on the FLIM by NAD(P)H, which was a label-free and noninvasive method for studying individual cells. Then, young and active yeast cells were sorted by the LIFT system at the single cell level. During the entire experiment, a sterile and humid environment was maintained to ensure the activity of cells. The high viability of sorted cells was achieved by the LIFT combining with FLIM.

Reversing the Polarity of MoS2 with PTFE.

Pristine monolayer molybdenum disulfide (MoS2) demonstrates predominant and persistent n-type semiconducting polarity due to the natural sulfur vacancy, which hinders its electronic and optoelectronic applications in the rich bipolarity area of semiconductors. Current doping strategies in single-layer MoS2 are either too mild to reverse the heavily n-doped polarity or too volatile to create a robust electronic device meeting the requirements of both a long lifetime and compatibility for mass production. Herein, we demonstrate that MoS2 can be transferred onto polytetrafluoroethylene (PTFE), one of the most electronegative substrates. After transfer, the MoS2 photoluminescence exhibits an obvious blueshift from 1.83 to 1.89 eV and a prolonged lifetime, from 0.13 to 3.19 ns. The Fermi level of MoS2 experiences a remarkable 510 meV decrease, transforming its electronic structure into that of a hole-rich p-type semiconductor. Our work reveals a strong p-doping effect and charge transfer between MoS2 and PTFE, shedding light on a new nonvolatile strategy to fabricate p-type MoS2 devices.

Metabolic Changes in Maternal and Cord Blood in One Case of Pregnancy-Associated Breast Cancer Seen by Fluorescence Lifetime Imaging Microscopy.

Pregnancy-associated breast cancer (PABC) is a rare disease, which is frequently diagnosed at an advanced stage due to limitations in current diagnostic methods. In this study, fluorescence lifetime imaging microscopy (FLIM) was used to study the metabolic changes by measuring maternal blood and umbilical cord blood via the autofluorescence of coenzymes, reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), and flavin adenine dinucleotide (FAD). The NAD(P)H data showed that a PABC case had significant differences compared with normal cases, which may indicate increased glycolysis. The FAD data showed that both maternal and cord blood of PABC had shorter mean lifetimes and higher bound-FAD ratios. The significant differences suggested that FLIM testing of blood samples may be a potential method to assist in PABC non-radiative screening.

Label-free and noninvasive method for assessing the metabolic status in type 2 diabetic rats with myocardium diastolic dysfunction.

This study assesses the metabolic status of rat diabetic cardiomyopathy (DCM) models. Echocardiography is used to detect the diastolic dysfunction in type 2 diabetic rats, and a lower threshold for inducible atrial fibrillation is found in type 2 diabetic rats with diastolic dysfunction compared to the control. Metabolic abnormalities are detected by status changes of reduced nicotinamide adenine dinucleotide (phosphate) (NAD(P)H), which is an essential coenzyme in cells or tissues. Fluorescence lifetime imaging microscopy (FLIM) is used to monitor changes in NAD(P)H in both myocardial tissues and blood. FLIM reveals that the protein-bound proportion of NAD(P)H in rat myocardium in the DCM group is smaller than the control group, which indicates the oxidative phosphorylation rate of the DCM group decreased. Similar results are found for blood plasma of DCM rats by the FLIM study. FLIM exhibits high potential for screening DCM as a label-free, sensitive, and noninvasive method.

Intermediate Effects of Body Mass Index and C-Reactive Protein on the Serum Cotinine- Leukocyte Telomere Length Association.

We aimed to examine the association between serum cotinine and leukocyte telomere length (LTL) and the intermediate effects of body mass index (BMI) and C-reactive protein (CRP) on modulating the association. This study included 4,047 adults from the 1999 to 2002 National Health and Nutrition Examination Survey. In the combined sample, after adjusting for age, race, sex, physical activity, and alcohol use, the total effect of serum cotinine on LTL was significant (standardized regression coefficient, ß = -0.049, p = 0.001) without BMI and CRP included in the model. With inclusion of BMI but without CRP in the model, the direct effect of cotinine on LTL in its absolute value increased to ß = -0.053 (p < 0.001), and the suppression effect of BMI was estimated at 8.8%. With inclusion of CRP but without BMI in the model, the direct effect of cotinine on LTL in its absolute value decreased to ß = -0.040 (p = 0.008), and the mediation effect of CRP was estimated at 16.9%. With inclusion of both BMI and CRP in the model, BMI and CRP still had significant suppression and mediation effects, respectively, on the cotinine-LTL association. These findings suggest that weight and inflammation have different roles in the inverse association between serum cotinine and LTL.

Moderate Mechanical Stimulation Protects Rats against Osteoarthritis through the Regulation of TRAIL via the NF-κB/NLRP3 Pathway.

The aim of this study was to examine exercise-related genes in articular cartilage identified through bioinformatics analysis to dissect the potential signaling pathway involved in mechanical stimulation in osteoarthritis (OA). To this end, we evaluated the GSE74898 dataset from the Gene Expression Omnibus database for exercise-related differentially expressed miRNAs (DE-miRNAs) using the R software package and predicted potential target genes for these miRNAs using miRTarBase. Functional annotation and pathway enrichment analysis were performed for these potential DE-miRNA targets. The effects of mechanical stimulation on the tumor necrosis factor-related apoptosis-induced ligand (TRAIL)/nuclear factor-kappa B (NF-κB)/nucleotide-binding and oligomerization domain-like receptor containing protein 3 (NLRP3) signaling pathway were evaluated in articular cartilage and chondrocytes. A total of 394 DE-miRNAs were identified (103 upregulated miRNAs; 291 downregulated miRNAs) in the cartilage of rats following treadmill exercise compared to the cartilage of unexercised control rats. Thus, mechanical stimulation could modulate the TRAIL/NF-κB/NLRP3 signaling pathway on OA. Histological and protein analysis demonstrated that moderate-intensity treadmill exercise could ameliorate OA through the downregulation of TRAIL. Furthermore, moderate cyclic tensile strain (CTS) could rescue chondrocytes from the effects of TRAIL via the inhibition of the nuclear translocation of NF-κB p65 and formation of NLRP3. Our findings indicate that moderate mechanical stimulation could ameliorate the degeneration of cartilage and chondrocyte damage through the inhibition of the TRAIL/NF-κB/NLRP3 pathway.

A cathelicidin-related antimicrobial peptide suppresses cardiac hypertrophy induced by pressure overload by regulating IGFR1/PI3K/AKT and TLR9/AMPKα.

Cathelicidin-related antimicrobial peptide (CRAMP), an antimicrobial peptide, was reported to protect against myocardial ischemia/reperfusion injury. However, the effect of CRAMP on pressure overload-induced cardiac hypertrophy was unknown. This study explored the role of CRAMP on cardiac hypertrophy. A cardiac hypertrophy mouse model was induced by aortic banding surgery. Seven days after surgery, mice were given mCRAMP by intraperitoneal injection (8 mg/kg/d) for 7 weeks. Cardiac hypertrophy was evaluated by the hypertrophic response and fibrosis level as well as cardiac function. Mice were also injected with AAV9-shCRAMP to knockdown CRAMP in the mouse heart. CRAMP levels first increased and then reduced in the remodeling heart, as well as in angiotensin II-stimulated endothelial cells but not in cardiomyocytes and fibroblasts. mCRAMP protected against the pressure overload-induced cardiac remodeling process, while CRAMP knockdown accelerated this process. mCRAMP reduced the inflammatory response and oxidative stress in the hypertrophic heart, while mCRAMP deficiency deteriorated the pressure overload-induced inflammatory response and oxidative stress. mCRAMP inhibited the angiotensin II-stimulated hypertrophic response and oxidative stress in neonatal rat cardiomyocytes, but mCRAMP did not help the angiotensin II-induced inflammatory response and oxidative stress in endothelial cells. Mechanistically, we found that mCRAMP suppressed the cardiac hypertrophic response by activating the IGFR1/PI3K/AKT pathway via directly binding to IGFR1. AKT knockout mice completely reversed the anti-hypertrophic effect of mCRAMP but not its anti-oxidative effect. We also found that mCRAMP ameliorated cardiac oxidative stress by activating the TLR9/AMPKa pathway. This was confirmed by a TLR9 knockout mouse experiment, in which a TLR9 knockout partly reversed the anti-hypertrophic effect of mCRAMP and completely counteracted the anti-oxidative effect of mCRAMP. In summary, mCRAMP protected against pressure overload-induced cardiac hypertrophy by activating both the IGFR1/PI3K/AKT and TLR9/AMPKa pathways in cardiomyocytes.

Carboxyl hydrogel particle film as a local pH buffer for voltammetric determination of luteolin and baicalein.

Electrochemical determination of luteolin and baicalein always needs acidic supporting electrolyte to guarantee good sensitivity. Therefore, most of the reported electrochemical sensors of luteolin and baicalein are unsuitable for detection of neutral actual samples. It is necessary to design a highly sensitive sensor for direct determination of them in neutral conditions. In this study, poly(N-isopropylacrylamide-acrylic acid) hydrogel particles (NIPA/AA) and multiwall carbon nanotubes (MWCNTs) composite modified glassy carbon electrode (GCE) was fabricated by a simple casting method. The voltammetric results showed that the NIPA/AA particle film provided acidic environment for proton-electron coupled reaction in neutral mediums. The near-surface pH of the electrode was related on the loaded amount of the NIPA/AA particles in pH range from 4.2 to 5.9. The voltammetric behaviors of luteolin and baicalein at the NIPA/AA-MWCNTs-GCE were studied by cyclic voltammetry. The peak separations between cathodic and anodic peaks were decreased and peak currents were increased because of decrease in pH and increase in ion conductivity at the local electrode surface. The sensitivity of the electrode was investigated by differential pulse voltammetry. Even under neutral conditions, the plots of the oxidation currents of luteolin and baicalein were dependent linearly on their concentration with detection limit of 14.5 pM and 44.4 pM, respectively. Moreover, the proposed NIPA/AA-MWCNTs-GCE was also successfully applied for determination of luteolin and baicalein in peanut shell, Huang-qin and tomato samples.

Mechanical stress protects against osteoarthritis via regulation of the AMPK/NF-κB signaling pathway.

Mechanical stress plays a key role in regulating cartilage degradation in osteoarthritis (OA). The aim of this study was to evaluate the effects and mechanisms of mechanical stress on articular cartilage. A total of 80 male Sprague-Dawley rats were randomly divided into eight groups (n = 10 for each group): control group (CG), OA group (OAG), and CG or OAG subjected to low-, moderate-, or high-intensity treadmill exercise (CL, CM, CH, OAL, OAM, and OAH, respectively). Chondrocytes were obtained from the knee joints of rats; they were cultured on Bioflex 6-well culture plates and subjected to different durations of cyclic tensile strain (CTS) with or without exposure to interleukin-1ß (IL-1ß). The results of the histological score, immunohistochemistry, enzyme-linked immunosorbent assay, and western-blot analyses indicated that there were no differences between CM and CG, but OAM showed therapeutic effects compared with OAG. However, CH and OAH experienced more cartilage damage than CG and OAG, respectively. CTS had no therapeutic effects on collagen II of normal chondrocytes, which is consistent with findings after treadmill exercise. However, CTS for 4 hr could alleviate the chondrocyte damage induced by IL-1ß by activating AMP-activated protein kinase (AMPK) phosphorylation and suppressing nuclear translocation of nuclear factor (NF)-κB p65. Our findings indicate that mechanical stress had no therapeutic effects on normal articular cartilage and chondrocytes; mechanical stress only caused damage with excessive stimulation. Still, moderate biomechanical stress could reduce sensitization to the inflammatory response of articular cartilage and chondrocytes through the AMPK/NF-κB signaling pathway.

The therapeutic effects of lipoxin A4 during treadmill exercise on monosodium iodoacetate-induced osteoarthritis in rats.

Lipoxin A4 (LXA4), a kind of adipokines, is a potent stop signal of inflammation. Our preliminary study found that LXA4 of serum and intra-articular lavage fluid (IALF) was rapidly elevated in 2 h and rapidly reduced to normal level at 4 h after moderate-intensity treadmill exercise. The aim was to confirm the therapeutic effects of LXA4 during treadmill exercise on rat model of monosodium iodoacetate (MIA)-induced OA and the detailed mechanism of LXA4 on OA. One hundred and twenty-four male Sprague-Dawley rats were submitted to two different protocols. A single session of treadmill exercise: sixty-four rats were randomly divided into treadmill exercise of different intensities for 60 min only once (n = 4). Formal treadmill exercise: sixty rats were randomly divided into six groups (n = 10): control group (CG), knee OA group (OAG), OA with treadmill exercise of different intensities (OAL, OAM and OAH), and OAM + BOC-2 (an antagonist of LXA4 receptor). The rats were evaluated by ELISA, histology, immunohistochemistry and western blotting. Fibroblast-like synoviocytes (FLSs) were obtained from knee joint of rats. The effects of LXA4 on interleukin (IL)-1ß induced FLSs were evaluated by western blotting and immunofluorescence. The results of ELISA, histological evaluation, western blotting and immunohistochemistry indicated that OAM had a better treatment which could be suppressed by BOC-2. Moreover, LXA4 could attenuate the expression of matrix metalloproteinase (MMP)-3 and MMP-13 and suppress the expression of nuclear factor-kappa B (NF-κB) p65 induced by IL-1ß in FLSs. The therapeutic effects of LXA4 during treadmill exercise on MIA-induced OA via inhibiting NF-κB signaling pathway.

Carnosine Prevents Type 2 Diabetes-Induced Osteoarthritis Through the ROS/NF-κB Pathway.

Background: The anti-inflammatory and antioxidant capacity of carnosine (CAR) has been investigated in autoimmune diseases. The aim of this study was to evaluate the potential protective effects of oral CAR supplements to ameliorate type 2 diabetes mellitus (T2DM)-induced osteoarthritis (OA) in rats and its mechanism. Methods: Seventy male Sprague-Dawley rats were randomly divided into the control group (CG, n = 10) and the T2DM group (n = 60). A rat model of T2DM was established using a high fat diet and streptozotocin (30 mg/kg, i.p.). The 41 rats that developed T2DM were chosen and randomly divided into four groups: T2DM-induced OA group (OAG, n = 11), and the T2DM-induced OA with low, moderate, and high-doses of CAR for 8 weeks group (CAR-L, CAR-M, and CAR-H, n = 10). After 13 weeks, all rats were evaluated by enzyme-linked immunosorbent assay (ELISA), histology, immunohistochemistry, and western blotting. Fibroblast-like synoviocytes (FLSs) were obtained from the knee joints of all rats. The effects of CAR on the inflammatory response in interleukin (IL)-1ß-stimulated FLSs under a high glucose environment were evaluated by real-time quantitative polymerase chain reaction, western blotting, flow cytometry, and immunofluorescence. Results: The results of ELISA (IL-1ß and tumor necrosis factor-α), the histological evaluation (Mankin and OARSI score), western blotting [COL2A1, matrix metalloproteinase (MMP)-3, MMP-13, IL-1ß, and nuclear factor-kappaB (NF-κB) p65], and immunohistochemistry (COL2A1, MMP-3, and MMP-13) indicated that oral CAR attenuated the development of T2DM-induced OA and suppressed the inflammatory response. Moreover, CAR alleviated MMP-3 and MMP-13 expression levels by decreasing reactive oxygen species content and suppressing nuclear translocation of NF-κB p65 on IL-1ß-induced FLSs in a high glucose environment. Conclusion: These findings indicate that oral CAR had chondroprotective effects on T2DM-induced OA through the reactive oxygen species (ROS)/NF-κB pathway.