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SCOPE: Osteopontin (OPN) is a multifunctional protein naturally present in mammals' milk, associated with immune homeostasis and intestinal maturation. This study aims to investigate the protein digestion pattern and the cellular bioactivity of bovine milk OPN digesta in vitro. METHODS AND RESULTS: A modified INFOGEST static in vitro infant digestion protocol and a Caco-2/HT-29 co-culture cell model are employed to evaluate the digestion properties and the anti-inflammatory effects of OPN. OPN is resistant to gastric hydrolysis but degraded into large peptides during intestinal digestion. Its 10 kDa digesta permeate with predicted extensive bioactivities protects the co-culture cell model from the inflammation-induced dysfunction by dose-dependently recovering the expression of occludin, claudin-3, and ZO-1. Low dosage of OPN significantly decreases the production of IL-8 and IL-6, and downregulates the mRNA and protein expression of MyD88, NF-κB p65, and IκB-α, whereas a high dose evokes a mild pro-inflammatory response. Interestingly, anti-inflammatory effect of OPN digesta is stronger than lactoferrin and whey protein concentrate counterparts. CONCLUSION: The findings demonstrate that the bioactive peptides released from in vitro infant gastrointestinal digestion of bovine milk OPN alleviates intestinal epithelial cell inflammation by inhibiting NF-κB pathway activation and potentiates the barrier function of the intestinal epithelium.
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Leche , FN-kappa B , Humanos , Lactante , Animales , Leche/química , Células CACO-2 , Osteopontina/genética , Osteopontina/metabolismo , Inflamación , Biomarcadores/análisis , Antiinflamatorios , Mamíferos/metabolismoRESUMEN
Purpose: The underlying mechanism of congenital cataracts caused by deficiency or mutation of junctional adhesion molecule C (JAM-C) gene remains unclear. Our study aims to elucidate the abnormal developmental process in Jamc-/- lenses and reveal the genes related to lens development that JAM-C may regulate. Methods: Jamc knockout (Jamc-/-) mouse embryos and pups were generated for in vivo studies. Four key developmental stages from embryonic day (E) 12.5 to postnatal day (P) 0.5 were selected for the following experiments. Hematoxylin and eosin staining was used for histological analysis. The 5-bromo-2'-deoxyuridine (BrdU) incorporation assay and TUNEL staining were performed to label lens epithelial cell (LEC) proliferation and apoptosis, respectively. Immunofluorescence and Western blot were used to analyze the markers of lens epithelium, cell cycle exit, and lens fiber differentiation. Results: JAM-C was expressed throughout the process of lens development. Deletion of Jamc resulted in decreased lens size and disorganized lens fibers, which arose from E16.5 and aggravated gradually. The LECs of Jamc-/- lenses showed decreased quantity and proliferation, accompanied with reduction of key transcription factor, FOXE3. The fibers in Jamc-/- lenses were disorganized. Moreover, Jamc-deficient lens fibers showed significantly altered distribution patterns of Cx46 and Cx50. The marker of fiber homeostasis, γ-crystallin, was also decreased in the inner cortex and core fibers of Jamc-/- lenses. Conclusions: Deletion of JAM-C exhibits malfunction of LEC proliferation and fiber maturation during murine lens development, which may be related to the downregulation of FOXE3 expression and abnormal localization patterns of Cx46 and Cx50.
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Molécula C de Adhesión de Unión , Cristalino , Animales , Ratones , Diferenciación Celular/fisiología , Proliferación Celular , Células Epiteliales/metabolismo , Epitelio , Molécula C de Adhesión de Unión/metabolismo , Cristalino/metabolismo , Ratones NoqueadosRESUMEN
Cataract, the leading cause of blindness worldwide, is caused by crystallin protein aggregation within the protected lens environment. Phase separation has been implicated as an important mechanism of protein aggregation diseases, such as neurodegeneration. Similarly, cataract has been proposed to be a protein condensation disease in the last century. However, whether crystallin proteins aggregate via a phase separation mechanism and which crystallin protein initiates the aggregation remain unclear. Here, we showed that all types of crystallin-GFP proteins remain soluble under physiological conditions, including protein concentrations, ion strength, and crowding environments. However, in age or disease-induced aberrant conditions, α-crystallin-GFP, including αA- and αB-crystallin-GFP, but not other crystallin-GFP proteins, undergo phase separation in vivo and in vitro. We found that aging-related changes, including higher crystallin concentrations, increased Na+, and decreased K+ concentrations, induced the aggregation of α-crystallin-GFP. Furthermore, H2O2, glucose, and sorbitol, the well-known risk factors for cataract, significantly enhanced the aggregation of αB-crystallin-GFP. Taken together, our results revealed that α-crystallin-GFP forms aggregates via a phase transition process, which may play roles in cataract disease. Opposite to the previously reported function of enhancing the solubility of other crystallin, α-crystallin may be the major aggregated crystallin in the lens of cataract patients.
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Catarata , Cristalinas , Cristalino , Cadena A de alfa-Cristalina , alfa-Cristalinas , Humanos , alfa-Cristalinas/metabolismo , Cristalinas/genética , Cristalinas/metabolismo , Agregado de Proteínas , Peróxido de Hidrógeno/metabolismo , Catarata/metabolismo , Cristalino/metabolismoRESUMEN
AIMS: Inflammatory bowel disease is a complex, refractory disorder characterised by chronic gastrointestinal inflammation. Studies have reported that Lactobacillus reuteri alleviates gastrointestinal inflammation and strengthens the intestinal barrier. However, further biochemical and genetic studies are required to correctly understand the therapeutic potential of L. reuteri. MATERIALS AND METHODS: This study sought to further understand the anti-colitis effect of L. reuteri isolated from faecal samples of healthy locals by focusing on biochemical (immunological, mechanical, chemical and biological barriers) and genetic studies. KEY FINDINGS: In this study, we assessed and compared the benefits and efficacy of L. reuteri FYNDL13 and FCQHC8L in the treatment of colitis and found strain FYNDL13 to be superior to FCQHC8L in this regard. Compared with FCQHC8L, FYNDL13 was associated with more diverse and powerful regulatory pathways. Meanwhile, it encouraged butyric acid formation, upregulated antimicrobial peptide-coding gene transcription and prevented hyperimmune reactions on the intestinal periphery and within the intestine. Moreover, it enhanced the abundance of beneficial bacteria (Bifidobacterium, Akkermansia, Blautia and Oscillospira), thereby limiting the relative abundance of harmful bacteria (Bacteroides and Sutterella). Furthermore, the advantage might be attributed to metabolism- and defence system-related genomic characteristics. SIGNIFICANCE: Taken together, our study compares and summarizes a pathway paradigm of these two L. reuteri strains in reinforcing the intestinal barrier against colitis and identifies candidate genes responsible for microbiota-immune axis balance.
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Colitis , Microbioma Gastrointestinal , Limosilactobacillus reuteri , Probióticos , Ratones , Animales , Probióticos/uso terapéutico , Colitis/inducido químicamente , Colitis/terapia , Colitis/microbiología , Inflamación , Sulfato de Dextran/toxicidad , Ratones Endogámicos C57BLRESUMEN
Glutamate receptors (GLR) are widely present in animals and plants, playing essential roles in regulating plant growth, development and stress response. At present, most studies of GLRs in plants are focused on Arabidopsis thaliana, while there have been few studies on rice. In this study, we identified 26 OsGLR genes in rice (Oryza sativa L.). Then, we analyzed the chromosomal location, physical and chemical properties, subcellular location, transmembrane (TM) helices, signal peptides, three-dimensional (3D) structure, cis-acting elements, evolution, chromatin accessibility, population variation, gene-coding sequence haplotype (gcHap) and gene expression under multiple abiotic stress and hormone treatments. The results showed that out of the 26 OsGLR genes, ten genes had the TM domain, signal peptides and similar 3D structures. Most OsGLRs exhibited high tissue specificity in expression under drought stress. In addition, several OsGLR genes were specifically responsive to certain hormones. The favorable gcHap of many OsGLR genes in modern varieties showed obvious differentiation between Xian/indica and Geng/japonica subspecies. This study, for the first time, comprehensively analyzes the OsGLR genes in rice, and provides an important reference for further research on their molecular function.
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PURPOSE: This study aimed to investigate the regulation of heme oxygenase-1 (HO-1) by paired box gene 6 (Pax6) and their roles in hydrogen peroxide (H2O2)-induced oxidative stress and apoptosis in lens epithelial cells (LECs) (SRA01/04, HLE-B3). METHODS: Lens anterior capsule membranes of mice of different ages were obtained to compare differences in the expression of Pax6 and HO-1 using Western blotting. Pax6-overexpressing plasmid and small interfering RNA were designed to overexpress and silence Pax6, respectively. Cobalt protoporphyrin (CoPP) was used to promote the expression of HO-1. Oxidative damage in LECs was induced by treatment with H2O2 (400 µM) for 24 h. Cell viability was measured using the Cell Counting Kit-8 assay. Intracellular reactive oxygen species (ROS) were detected using flow cytometry and immunofluorescence. Superoxide dismutase (SOD) level was measured using SOD Assay Kit and apoptotic cells were quantified using annexin V-fluorescein isothiocyanate/propidium iodide staining. RESULTS: Pax6 and HO-1 expression levels showed an age-dependent decrease in LECs of mouse. Overexpressing Pax6 upregulated HO-1 expression level. Silencing Pax6 downregulated the HO-1 expression level, resulting in increased generation of ROS, reduced SOD activity, decreased cell viability, and increased apoptotic cells of LECs under H2O2-induced oxidative stress. Overexpressing Pax6 and CoPP both mitigates H2O2-induced oxidative stress by increasing the expression of HO-1 of LECs. CONCLUSION: Pax6 and HO-1 expression levels showed an age-dependent decrease in LECs in mouse anterior capsules. Pax6 could regulate the expression of HO-1 in LECs. The decrease of Pax6 weakened the antioxidant ability of LECs under H2O2-induced oxidative stress by downregulating HO-1, which may be a potential mechanism for the formation of age-related cataract.
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Peróxido de Hidrógeno , Cristalino , Animales , Anexina A5/metabolismo , Antioxidantes/metabolismo , Apoptosis , Cápsulas/metabolismo , Células Epiteliales/metabolismo , Fluoresceínas/metabolismo , Hemo-Oxigenasa 1/genética , Hemo-Oxigenasa 1/metabolismo , Peróxido de Hidrógeno/metabolismo , Isotiocianatos , Cristalino/metabolismo , Proteínas de la Membrana , Ratones , Estrés Oxidativo , Factor de Transcripción PAX6 , Propidio/metabolismo , ARN Interferente Pequeño/genética , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismoRESUMEN
SCOPE: Human milk oligosaccharides (HMOs), multifunctional glycans naturally present in human milk, are known to contribute to the infant's microbiota and immune system development. However, the molecular specificity of HMOs on microbiota and associated fermentation is not yet fully understood, and is important for the development of infant formula optimum functionality. METHODS AND RESULTS: In vitro fermentation is carried out on structurally different HMOs with infant fecal inocula dominated by Bifidobacterium longum, Bifidobacterium breve, and Bacteroides. The gas, metabolite (SCFA, lactate, and succinate) profiles, and microbiota responses differ between individual microbiota inocula patterns regardless of HMO structure. In terms of HMO pairs with same sugar composition but different glycosidic bonds, gas and metabolite profiles are similar with the B. longum- and B. breve-dominated inocula. However, large individual variations are observed with the Bacteroides-dominated inocula. The microbial communities at the end of fermentation are closely related to the initial microbiota composition. CONCLUSION: The findings demonstrate that short-term in vitro fermentation outcomes largely depend on the initial gut microbiota composition more than the impact of HMO molecular specificity. These results advance the current understanding for the design of personalized infant nutritional solutions and therapies in future.
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Microbiota , Leche Humana , Bacteroides , Fermentación , Humanos , Lactante , Lactatos , Leche Humana/química , Oligosacáridos/metabolismo , Succinatos , AzúcaresRESUMEN
BACKGROUND: Diabetic retinopathy (DR) is the common complication of diabetes, accounting for most blindness cases worldwide. MicroRNAs (miRs) are small non-coding RNAs and participate in the pathogenesis and develop-ment of various diseases, including DR. The present study aimed to investigate miR-335-3p and vascular endothelial growth factor (EGFR) roles in DR diagnosis and development. METHODS: A total of 104 healthy volunteers, 96 type 2 diabetes mellitus (T2DM) patients, and 102 DR cases were enrolled in this study. The clinicopathological information of all subjects were collected and analyzed using chisquared test. After collecting plasma from each participant, a ROC assay was conducted to determine the dis-criminative value of miR-335-3p and EGFR in DR diagnosis. The targeted relationship between miR-335-3p and EGFR was examined by dual-luciferase reporter assay and correlation analysis. After exposing APRE-19 cells to different concentrations of high glucose (HG), the DR in vitro cell model was constructed. The expression levels of miR-335-3p and EGFR were detected using RT-qPCR. The effects of miR-335-3p and EGFR on HG-treated APRE-19 cell viability were determined by CCK-8 assay. RESULTS: Clinicopathological information presented that BMI index, fasting blood glucose (FBP), 2h-BG, HbA1c, miR-335-3p, and EGFR levels were strongly associated with DR pathogenesis. MiR-335-3p was significantly decreased while EGFR was increased in DR patients and HG-treated APRE-19 cells. MiR-335-3p and EGFR presented high accuracy, sensitivity, and specificity in differentiating DR from healthy cases and T2DM patients; moreover, miR-335-3p and EGFR could also discriminate proliferative DR (PDR) cases from healthy controls. After confirming that miR-335-3p was negatively correlated with its target EGFR, we found miR-335-3p could increase the viability in HG-treated APRE-19 cells while silencing of EGFR could also reverse the inhibitory effects of HG conditions on APRE-19 cell viability. CONCLUSIONS: We concluded that plasma miR-335-3p and EGFR may be utilized as non-invasive biomarkers for screening DR cases, contributing to DR diagnosis and treatment.
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Diabetes Mellitus Tipo 2 , Retinopatía Diabética , MicroARNs , Proliferación Celular , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/diagnóstico , Diabetes Mellitus Tipo 2/genética , Retinopatía Diabética/diagnóstico , Retinopatía Diabética/genética , Factores de Crecimiento Endotelial/farmacología , Receptores ErbB/genética , Humanos , Factor A de Crecimiento Endotelial VascularRESUMEN
PURPOSE: Autophagy plays a crucial role in intracellular quality control of crystalline lens and AMPK has regulatory effect on autophagy. However, whether AMPK regulated autophagy is involved in diabetic cataract (DC) progression remains unknown. This study aims to investigate the AMPK-FOXO3 and AMPK-TFEB induced autophagy activity in DC patients. MATERIALS AND METHODS: First, anterior capsule specimens from DC and age-related cataract (ARC) patients were obtained to compare the expression difference of autophagy-related genes. The phosphorylation levels of AMPK, AKT, and mTOR and the expression of FOXO3 and TFEB were measured. Then, human lens epithelial cells (LECs, SRA 01/04) were cultured with 30 mM or 5.5 mM glucose, and AMPK activator (AICAR) and inhibitor (Compound C) were applied to further investigate the regulatory role of AMPK on autophagy. RESULTS: Compared with ARC patients, the expression of autophagy-related genes ATG5, FYCO1, ATG8, ATG12, Beclin1, and ULK1 in anterior capsules LECs of DC patients were significantly down-regulated. Meanwhile, AMPK and AMPK-dependent transcription factors, FOXO3 and TFEB were also inhibited. Similar results were found in high glucose (HG) treated SRA 01/04 model. Notably, this down-regulation of autophagy activity was rescued by AICAR in vitro, which was manifested by inhibition of AKT and mTOR phosphorylation and up-regulation of FOXO3, TFEB, Beclin1 and LC3B-II expression. CONCLUSIONS: Down-regulation of AMPK-FOXO3 and AMPK-TFEB induced autophagy activity was found in both LECs of anterior capsule from DC patients and SRA 01/04 cells under HG condition, which may be the underlying mechanism of DC formation. Thus, targeting AMPK-induced autophagy may be a potential therapeutic approach for diabetic cataract.
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Catarata , Diabetes Mellitus , Proteínas Quinasas Activadas por AMP/metabolismo , Autofagia , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/genética , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/metabolismo , Factores de Transcripción Básicos con Cremalleras de Leucinas y Motivos Hélice-Asa-Hélice/farmacología , Beclina-1/metabolismo , Regulación hacia Abajo , Proteína Forkhead Box O3/genética , Proteína Forkhead Box O3/metabolismo , Glucosa/farmacología , Humanos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Serina-Treonina Quinasas TOR/genética , Serina-Treonina Quinasas TOR/metabolismo , Serina-Treonina Quinasas TOR/farmacologíaRESUMEN
BACKGROUND Online blended learning, also known as "smart classes", has benefits when compared with traditional teaching methods that use books and lectures. This study aimed to compare the use of the Smart Class teaching module with traditional teaching on the topic of psychosocial dysfunction during the training of undergraduate occupational therapy (OT) students in China. MATERIAL AND METHODS We recruited Grade 2017 OT students as the Smart Class teaching module group and Grade 2016 OT students as the Traditional Class teaching module group to participate in the study. The objective evaluation (assignment score, practical exam score, written exam score, and final score) and subjective evaluation (data from student questionnaires and information from interviews with the lead teacher and assistant teachers) were performed in both groups. RESULTS No significant difference was found in the final scores (P=0.874) and students' questionnaire results between the 2 groups. However, data from the student questionnaires and teacher interviews indicated a preference for combining the Smart Class teaching module and the Traditional Class teaching module. CONCLUSIONS The advantage of the Smart Class teaching module is that it can effectively integrate excellent teaching resources across geographical restrictions and it is conducive to promoting independent learning for students and all-around supervision for teaching. The Smart Class teaching module was comparable to traditional teaching methods for the training of undergraduate OT students in China, but was preferred by the students.
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Educación a Distancia/métodos , Adolescente , Adulto , China , Curriculum , Femenino , Humanos , Masculino , Estudiantes de Medicina , Encuestas y Cuestionarios , Adulto JovenRESUMEN
OBJECTIVE: To investigate the protective efficacies and potent mechanisms of combination therapy with semaglutide and rosiglitazone (RSG) on the high-glucose incubated human ARPE-19 cells and diabetic retinopathy (DR) model rats. MAIN METHODS: The CCK-8 methods were used to evaluate the protective effects of semaglutide and RSG alone or combination on the cell viability of high-glucose treated ARPE-19 cells. After the DR rat model was established, the effects of combined treatment on general indexes, retinal morphological changes, retinal Müller cells as well as PI3K/Akt/MTOR related factors of DR model rats were investigated. RESULTS: The CCK-8 assay showed obviously enhanced protective efficacies of combination therapy with semaglutide and RSG on the ARPE-19 with oxidative stress induced by high-glucose with combination index all below 1.5 demonstrating obvious synergistic effects. Combined incubation could also effectively decrease the expression of inflammatory factors, including TNF-α, IL-1ß, IL-6, and the increase of ROS content in ARPE cell culture supernatant induced by high-glucose. Combined use of the antioxidant, PI3K/Akt and mTOR inhibitors, we further demonstrated that combined incubation of semaglutide and RSG could effectively by reduce high glucose-induced inflammatory injury inhibiting ROS/PI3K/Akt/mTOR signaling. Furthermore, chronic combination treatment effectively improved the histopathological characteristics and down-regulated the GFAP expression in Müller cells as well as PI3K/Akt/MTOR signaling pathway-related factors in retina which was better than any monomer treatment group. CONCLUSIONS: Combined semaglutide with RSG exhibited synergistically protective efficacies on retinal cells by decreasing the GFAP expression, inhibiting oxidative stress and PI3K/Akt/MTOR signaling-transduction in DR model rats.
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Retinopatía Diabética/tratamiento farmacológico , Péptidos Similares al Glucagón/uso terapéutico , Rosiglitazona/uso terapéutico , Animales , Antioxidantes/metabolismo , Línea Celular , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Quimioterapia Combinada , Células Ependimogliales/efectos de los fármacos , Células Ependimogliales/metabolismo , Células Ependimogliales/patología , Proteína Ácida Fibrilar de la Glía/metabolismo , Péptidos Similares al Glucagón/farmacología , Humanos , Inflamación/patología , Masculino , Fosfatidilinositol 3-Quinasas/metabolismo , Sustancias Protectoras/farmacología , Sustancias Protectoras/uso terapéutico , Proteínas Proto-Oncogénicas c-akt/metabolismo , Ratas Sprague-Dawley , Rosiglitazona/farmacología , Transducción de Señal/efectos de los fármacos , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
The leucine-rich repeat (LRR) proteins play important roles in the recognition of corresponding ligands and signal transduction networks in plant defence responses. Herein, a novel LRR protein from Capsicum annuum, CaLRR51, was identified and characterized. It was localized to the plasma membrane and transcriptionally up-regulated by Ralstonia solanacearum infection (RSI), as well as the exogenous application of salicylic acid (SA), jasmonic acid (JA) and ethephon (ETH). Virus-induced gene silencing of CaLRR51 significantly increased the susceptibility of pepper to RSI. By contrast, transient overexpression of CaLRR51 in pepper plants activated hypersensitive response (HR)-like cell death, and up-regulated the defence-related marker genes, including PO2, HIR1, PR1, DEF1 and ACO1. Moreover, ectopic overexpression of CaLRR51 in transgenic tobacco plants significantly enhanced the resistance to RSI. Transcriptional expression of the corresponding defence-related marker genes in transgenic tobacco plants was also found to be enhanced by the overexpression of CaLRR51, which was potentiated by RSI. These loss- and gain-of-function assays suggest that CaLRR51 acts as a positive regulator in the response of pepper to RSI. In addition, the putative signal peptide and transmembrane region were found to be required for plasma membrane targeting of CaLRR51, which is indispensable for the role of CaLRR51 in plant immunity.
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Capsicum/microbiología , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/metabolismo , Proteínas/metabolismo , Ralstonia solanacearum/patogenicidad , Secuencia de Aminoácidos , Capsicum/efectos de los fármacos , Capsicum/genética , Capsicum/inmunología , Muerte Celular/efectos de los fármacos , Clonación Molecular , Resistencia a la Enfermedad/genética , Eliminación de Gen , Regulación de la Expresión Génica de las Plantas , Silenciador del Gen/efectos de los fármacos , Proteínas Repetidas Ricas en Leucina , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/inmunología , Reguladores del Crecimiento de las Plantas/farmacología , Virus de Plantas/fisiología , Plantas Modificadas Genéticamente , Dominios Proteicos , Proteínas/química , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ralstonia solanacearum/efectos de los fármacos , Análisis de Secuencia de Proteína , Fracciones Subcelulares/metabolismo , Regulación hacia ArribaRESUMEN
The blood-nourishing and hard-softening (BNHS) capsule is a traditional Chinese formula used in the symptomatic treatment of inflammation and pain. We conducted this randomized controlled trial to compare the efficacy of BNHS with other commonly prescribed drugs. We recruited 120 patients from two teaching hospitals; 30 patients in each hospital were randomly assigned to receive BNHS. In one hospital, the 30 controls were given another traditional Chinese drug; whereas a Western medicine (chondroprotection drug/Viartril-s) was used as the control in the other hospital. Intervention was carried out over a period of 4 weeks. Primary outcome measures included self-reported pain level, and changes in stiffness and functional ability as measured by the Western Ontario McMaster Universities Osteoarthritis (WOMAC) index. Mixed models were used for statistical analysis. Substantial improvements in disease-specific symptoms were observed, after 4 weeks of treatment, in patients taking BNHS capsules. As assessed by the WOMAC index, pain level of the BNHS group decreased by 57% [95% confidence interval (CI) = 50, 63], stiffness by 63% (95% CI = 55, 71) and functional ability increased by 56% (95% CI = 50, 63). No significant differences were found in any of the outcome measures between the BNHS group and either of the comparison groups. No severe adverse effects were reported. However, this study lacked a placebo group; therefore, we conclude that BNHS appears to be as effective as commonly prescribed medicines for the relief of pain and dysfunction in knee osteoarthritis patients, but costs a lot less than other Western and herbal drugs in the study.