Seo, Min-gyu;Kang, Yun-Mi;Chung, Kyung-Sook;Cheon, Se-Yun;Park, Jong Hyuk;Lee, Young-Cheol;An, Hyo-Jin
The Korea Journal of Herbology
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v.32
no.2
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pp.17-24
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2017
Objective : This study was designed to evaluate candidate materials as anti-inflammation agent from extracts of various Korean Compositae herbs in Hwaak mountain. Among Korea medicinal herbs, Ainsliaea acerifolia (AA) belongs to the Compositae family, has been used for the treatment of rheumatic arthritis. However, AA has not been previously reported to have an anti-inflammatory effect. Therefore, we investigated the anti-inflammatory effects of AA and its underlying molecular mechanisms in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Methods : Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in RAW 264.7 macrophages. Nitric oxide (NO) was measured with Griess reagent and pro-inflammatory cytokines were detected by enzyme immunoassay (EIA) kits in LPS-stimulated RAW 264.7 macrophages. Protein expressions of inducible nitric oxide synthase, and cyclooxygenase-2 (COX-2) and p65 subunit of nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) were determined by Western blot analysis. Results : Among 8 extracts of Korean Compositae herbs tested, AA showed the inhibition of NO production without cytotoxicity. Consistent with the observation, AA reduced the expression levels of iNOS and COX-2 proteins in LPS-simulated RAW 264.7 macrophages in dose-dependent manner. In addition, AA inhibited the productions of $TNF-{\alpha}$ and IL-6 in LPS-simulated RAW 264.7 macrophages. However, AA did not inhibit activation of p65 $NF-{\kappa}B$ in LPS-simulated RAW 264.7 macrophages. Conclusion : These results suggest that down-regulation of iNOS, COX-2 protein expression and $TNF-{\alpha}$ and IL-6 production by AA are responsible for its anti-inflammatory effects.
Objective : This study is conducted to investigate the effect of Astragali Radix on Post Traumatic Stress Disorder(PTSD) induced by the Single Proposed Stress(SPS). Methods : The experiment was conducted with five groups; SAL groups with only saline treatment, SPS group, SPS + ARX25 group, SPS + ARX50 group, and SPS + ARX100 group. After applying SPS, saline and ARX were administered for 14 days to identify the change of body weight, sucrose intake amount, and behavioral changes through Open Field Test(OFT) and Forced Swimming Test(FST). After the behavioral experiment, plasma corticosterone levels, serotonin, norepinephrine and dopamine concentrations were measured by enzyme-linked immunoassay in medical prefrontal cortex, hippocampus, and amygdala. Brain-derived neurotrophic factor(BDNF) in the hippocampus was measured using Reverse Transcription Polymerase Chain Reaction. Results : Weight change has significantly decreased in the SPS group compared to the SAL group(p<0.05). On day 14, the sucrose intake of rats has significantly increased in the SPS + ARX100 group compared to the SPS group(p<0.05). In OFT, the number of staying in the central space has significantly increased in the SPS + ARX100 group(p<0.01). In FST, immobility has significantly decreased in SPS + ARX50 group and SPS + ARX100 group(p<0.05). The concentration of serotonine, dopamine and BDNF expression has increased significantly in SPS + ARX100 group compared to SPS group(p<0.05) Conclusions : In the SPS-induced PTSD experiment, ARX increased sucrose intake and the numbers of crossing in the central zone space in OFT, decreased immobility time in FST, and increased concentration of serotonin, dopamine, and BDNF. It can be postulated that the ARX could be effective for the treatment of PTSD.
Purpose: The cow's milk-related-symptom-score (CoMiSS) tool was developed as an awareness tool for the assessment of cow's milk-related symptoms in infants or children. Fecal calprotectin (FC) is a noninvasive biomarker of gut inflammation that can be measured in serum and stool. This study aimed to investigate the relationship between FC levels and CoMiSS scores in infants with cow's milk protein allergy. Methods: Infants (aged 6-12 months) who were allergic to cow's milk protein were enrolled prospectively. Following completion of the CoMiSS scoring, the infants were divided into group 1 (positive CoMiSS scores ≥12) and group 2 (negative CoMiSS scores <12). FC was measured using immunoassay. Results: Of the 120 infants enrolled in this study, 60 (50.0%) had positive CoMiSS scores (group 1), while 60 (50.0%) had negative scores (group 2). The mean FC level was higher in the infants in group 1 than those in group 2 (2,934.57 ㎍/g vs. 955.13 ㎍/g; p<0.001). In addition, there was a positive correlation between FC and CoMiSS scores (R=0.168, p<0.0001). A FC level of 1,700 ㎍/g provided a sensitivity of 98.3%, specificity of 93.3%, and accuracy of 95.8% for the diagnosis of cow's milk protein allergy (CMPA). Conclusion: FC measurement may have a role in the assessing infants with CMPA.
Journal of Radiopharmaceuticals and Molecular Probes
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v.8
no.2
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pp.95-101
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2022
Microfluidic radioimmunoassay (RIA) platform called µ-RIA spends less reagent and shorter reaction time for the analysis compared to the conventional tube-based radioimmunoassay. This study reported the design of µ-RIA chips optimized for the gamma counter which could measure the small samples of radioactive materials automatically. Compared with the previous study, the µ-RIA chips developed in this study were designed to be compatible with conventional RIA test tubes. And, the automatic gamma counter could detect radioactivity from the 125I labeled anti-PSA attached to the chips. Effects of the multi-layer microchannels and two-phase flow in the µ-RIA chips were investigated in this study. The measured radioactivity from the 125I labeled anti-PSA was linearly proportional to the number of stacked chips, representing that the radioactivity in µ-RIA platform could be amplified by designing the chips with multi-layers. In addition, we designed µ-RIA chip to generate liquid-gas plug flow inside the microfluidic channel. The plug flow can promote binding of the biomolecules onto the microfluidic channel surface with recirculation in the liquid phase. The ratio of liquid slug and air slug length was 1 : 1 when the 125I labeled anti-PSA and the air were injected at 1 and 35 µL/min, respectively, exhibiting 1.6 times higher biomolecule attachment compared to the microfluidic chip without the air injection. This experimental result indicated that the biomolecular reaction was improved by generating liquid-gas slugs inside the microfluidic channel. In this study, we presented a novel µ-RIA chips that is compatible with the conventional gamma counter with automated sampler. Therefore, high-throughput radioimmunoassay can be carried out by the automatic measurement of radioactivity with reduced radiowaste generation. We expect the µ-RIA platform can successfully replace conventional tube-based radioimmunoassay in the future.
Purpose: With an increase in the aging population, the number of patients with degenerative spinal diseases undergoing surgery has risen, as has the incidence of postoperative delirium. This study aimed to investigate the risk factors affecting postoperative delirium in older adults who had undergone spine surgery and to identify the associated biomarkers. Methods: This study is a prospective study. Data of 100 patients aged ≥ 70 years who underwent spinal surgery were analyzed. Demographic data, medical history, clinical characteristics, cognitive function, depression symptoms, functional status, frailty, and nutritional status were investigated to identify the risk factors for delirium. The Confusion Assessment Method, Delirium Rating Scale-R-98, and Nursing Delirium Scale were also used for diagnosing delirium. To discover the biomarkers, urine extracellular vesicles (EVs) were analyzed for tau, ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), neurofilament light, and glial fibrillary acidic protein using digital immunoassay technology. Results: Nine patients were excluded, and data obtained from the remaining 91 were analyzed. Among them, 18 (19.8%) developed delirium. Differences were observed between participants with and without delirium in the contexts of a history of mental disorder and use of benzodiazepines (p = .005 and p = .026, respectively). Tau and UCH-L1-concentrations of urine EVs-were comparatively higher in participants with severe delirium than that in participants without delirium (p = .002 and p = .001, respectively). Conclusion: These findings can assist clinicians in accurately identifying the risk factors before surgery, classifying high-risk patients, and predicting and detecting delirium in older patients. Moreover, urine EV analysis revealed that postoperative delirium following spinal surgery is most likely associated with brain damage.
Objectives : In this study, the effects of ethylacetate extract of Ostericum koreanum on inflammation in RAW264.7 cells were investigated. Methods : Dried roots of Ostericum koreanum was extracted with 80% methanol for 24 h, and then fractionated with n-butanol, n-hexan and ethylacetate. RAW264.7 cells, a mouse macrophage line were incubated with different concentrations of the extract for 30 min and then stimulated with LPS at indicated times. Cell toxicity was determined by MTT assay. The concentrations of nitric oxide (NO) and prostaglandin E2 ($PGE_2$) were measured by Griess assay and enzyme immunoassay (EIA), respectively. The expressions of inducible nitric oxide synthease (iNOS) and cyclooxyganase (COX) -2 mRNA and protein were determined by RT-PCR and Western blot. Results : The methanol extract of Ostericum koreanuman and its fractions were significantly inhibited the NO and PGE2 productions in LPS-stimulated RAW264.7 cells. Among the fractions of Ostericum koreanuman the ethylacetate fraction was more strongly inhibited NO and $PGE_2$ productions compared with other fractions. The ethylacetate fraction was also suppressed LPS-induced mRNA expressions of iNOS and its protein levels in RAW264.7 cells. Conclusions : This study suggests that the ethylacetate fraction of Ostericum koreanum may have an anti-inflammatory property through suppressing inflammatory mediator productions in activated macrophages, suggesting have a therapeutic potential for the treatment of various inflammatory diseases.
Journal of The Korean Society of Integrative Medicine
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v.11
no.2
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pp.119-128
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2023
Purpose : The fruit of Actinidia polygama has been used in oriental medicine for the treatment of gout, rheumatoid arthritis, and inflammation. Though A. polygama exhibited anti-inflammatory activity in RAW 264.7 cells and carrageenan-induced rat paw edema, the exact mechanism for anti-inflammation was not evaluated yet. In this study, the anti-inflammatory mechanisms of A. polygama ethanol extract (APEE) in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Methods : WST-1 assay was applied to analyze the cytotoxic effect of APEE in RAW 264.7 cells. The productions of nitric oxide (NO) and prostaglandin (PG) E2 were analyzed by the Griess reaction and enzyme immunoassay (EIA) assay, respectively. In addition, protein expressions for inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 were measured by Western blot analysis. The activated status of an inflammatory transcription factor, NF-κ B, and its upstream signaling molecules, mitogen-activated protein kinases (MAPKs), was also evaluated by Western blot analysis. Results : As a result, APEE treatment did not exhibit any cytotoxicity until the concentration of 200 ㎍/㎖. APEE treatment significantly inhibited NO and PGE2 productions as well as their enzymes, iNOS and COX-2 in a dose-dependent manner. The inflammatory transcription factor, NF-κ B, was also attenuated by APEE treatment. In addition, the phosphorylated status of MAPKs such as extracellular regulated kinase (ERK), c-jun NH2 kinase (JNK), and p38, were significantly diminished by APEE treatment in LPS stimulated RAW 264.7 cells. Conclusion : Consequently, APEE treatment significantly attenuated the production of inflammatory mediators and their enzyme expressions in LPS-stimulated RAW 264.7 cells. The inflammatory transcription factor, NF-κ B, and upstream signaling molecules, MAPKs, were also significantly attenuated by APEE treatment in LPS-activated RAW 264.7 cells. These results indicate that APEE might be a candidate to be utilized as a promising candidate for the treatment of inflammatory disorders.
Background: The association between vitamin D and canine keratoconjunctivitis sicca (KCS) has not been investigated in dogs. Objectives: To investigate the association of serum 25-hydroxyvitamin D [25(OH)D] concentrations with Schirmer tear test 1 (STT-1) and tear film breakup time (TFBUT) in dogs. Methods: Sixty-one clinically healthy, client-owned dogs were enrolled. STT-1 and TFBUT were measured in 122 (61dogs) and 82 (41 dogs out of total 61 dogs) eyes, respectively. Serum 25(OH)D concentrations were evaluated by quantitative chemiluminescent immunoassay. The dogs were classified into 6 groups according to the evaluations (STT-1: group 1, normal [≥ 15 mm/min] in both eyes; group 2, normal in one eye and abnormal [< 15 mm/min] in the fellow eye; group 3, abnormal in both eyes; TFBUT: group 4, normal [≥ 20 sec] in both eyes; group 5, normal in one eye and abnormal [< 20 sec] in the fellow eye; group 6, abnormal in both eyes). Results: STT-1 was positively correlated with TFBUT (p < 0.001). Among the STT-1 groups, the mean serum 25(OH)D concentration in group 1 was significantly higher than in groups 2 and 3 with positive correlation (p < 0.001). However, there were no significant differences among the TFBUT groups 4, 5, and 6. Conclusions: In dogs, it was found that serum 25(OH)D concentrations had a greater effect on quantitative KCS than qualitative KCS. Therefore, it is considered that measurement of serum 25(OH)D concentration could be included in the diagnostic tests in canine quantitative KCS patients.
Linda Ratna Wati;Djanggan Sargowo;Tatit Nurseta;Lilik Zuhriyah;Bambang Rahardjo
Journal of Preventive Medicine and Public Health
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v.56
no.5
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pp.422-430
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2023
Objectives: Prolactin is vital for breastfeeding and milk production, and its secretion is influenced by factors related to the mother, infant, and environment. To date, no study has concurrently investigated the correlation of these factors with serum prolactin levels during lactation. Therefore, the objective of this study was to investigate the correlations among maternal and infant factors, lead exposure, and serum prolactin levels during lactation. Methods: A cross-sectional approach was employed in Surabaya, Indonesia, among 110 exclusively lactating mothers. The mothers' daily diets were determined using multiple 24-hour recalls, while blood lead levels were measured with inductively coupled plasma mass spectrometry. Serum prolactin levels were assessed using the electrochemiluminescence immunoassay. For bivariate analysis, we employed the Spearman correlation, Mann-Whitney, and Kruskal-Wallis tests, while for multivariate analysis, we utilized multiple linear regression. Results: The average serum prolactin level of the lactating mothers was 129.19±88.96 ng/mL. Positive correlations were found between serum prolactin levels and breastfeeding frequency (p<0.001), protein intake (p<0.001), and calcium intake (p=0.011) but had negative correlation with blood lead levels (p<0.001) and vitamin B6 intake (p=0.003). Additionally, prolactin levels were not significantly associated with maternal age; parity; intake of calories, vitamin D, vitamin E, zinc, folic acid, magnesium, or iron; infant age; or infant sex. Conclusions: Breastfeeding frequency had a stronger positive relationship with serum prolactin levels than protein and calcium intake. However, lead exposure was associated with reduced serum prolactin levels during lactation. Consequently, specific interventions from policymakers are necessary to manage breastfeeding in mothers exposed to lead.
This study aimed to investigate the immunomodulatory function of Pyropia yezoensis hydrothermal (water) extract (PYWE) in comparison to the group treated only with lipopolysaccharides (LPS) in RAW264.7 cells. LPS is known to be an inflammatory mediator that activates macrophages, leading to the secretion of nitric oxide (NO), inducible nitric oxide synthase (iNOS), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) as defense responses. Through enzyme-linked immunoassay and western blot analyses, it was observed that PYWE increased the expression levels of NO, iNOS, TNF-α, and IL-6 in RAW264.7 cells in a dose-dependent manner, although to a lesser extent compared with the group treated with LPS alone. In addition, the study examined the mitogen-activated protein kinases (MAPKs) pathway, which regulates various cellular activities, including gene expression, mitosis, cell differentiation, transformation, survival, and death. The western blot analysis confirmed that PYWE also regulated the MAPKs pathway. Furthermore, the expression levels of immunomodulatory-related factors increased in the group treated with PYWE compared with the control group. Even though the effects of PYWE were usually less strong than those of LPS, the effects of PYWE increased with increasing doses compared to the control group. This suggests that PYWE could be used to control the immune system.
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