Anti-Inflammatory Effect of Chamaecyparis obtusa (Siebold & Zucc.) Endl. Leaf Essential Oil.

Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) belongs to the Cupressaceae family and is native to East Asian regions. Essential oils extracted from the leaves, bark, branches, and roots of C. obtusa have both aesthetic and medicinal properties and are thus widely used. However, detailed analyses of the active ingredients of C. obtusa extract are lacking. In this study, the sabinene content in the hydro-distillation of C. obtusa leaf essential oil (COD) was analyzed using GC-MS, and the anti-inflammatory effect of COD was compared with that of pure sabinene. Cell viability was evaluated by MTT assay, and nitric oxide (NO) production was measured using Griess reagent. Relative mRNA and protein levels were analyzed using RT-qPCR and western blot, and secreted cytokines were analyzed using a cytokine array kit. The results showed that both COD and sabinene inhibited the expression of inducible nitric oxide synthase (iNOS) and the phosphorylation of c-Jun N-terminal kinase (JNK) and p38 in lipopolysaccharide (LPS)-induced RAW 264.7 cells. COD and sabinene also reduced the production of pro-inflammatory cytokines interleukin (IL)-1ß, IL-6, IL-27, IL-1 receptor antagonist (IL-1ra), and granulocyte-macrophage colony-stimulating factor (GM-CSF). The anti-inflammatory mechanisms of COD and sabinene partially overlap, as COD was shown to inhibit MAPKs and the JAK/STAT axis, and sabinene inhibited MAPKs, thereby preventing LPS-induced macrophage activation.

The acidic tumor microenvironment enhances PD-L1 expression via activation of STAT3 in MDA-MB-231 breast cancer cells.

Tumor acidosis, a common phenomenon in solid cancers such as breast cancer, is caused by the abnormal metabolism of cancer cells. The low pH affects cells surrounding the cancer, and tumor acidosis has been shown to inhibit the activity of immune cells. Despite many previous studies, the immune surveillance mechanisms are not fully understood. We found that the expression of PD-L1 was significantly increased under conditions of extracellular acidosis in MDA-MB-231 cells. We also confirmed that the increased expression of PD-L1 mediated by extracellular acidosis was decreased when the pH was raised to the normal range. Gene set enrichment analysis (GSEA) of public breast cancer patient databases showed that PD-L1 expression was also highly correlated with IL-6/JAK/STAT3 signaling. Surprisingly, the expression of both phospho-tyrosine STAT3 and PD-L1 was significantly increased under conditions of extracellular acidosis, and inhibition of STAT3 did not increase the expression of PD-L1 even under acidic conditions in MDA-MB-231 cells. Based on these results, we suggest that the expression of PD-L1 is increased by tumor acidosis via activation of STAT3 in MDA-MB-231 cells.

Development and validation of a scale to assess the attitude of healthcare professionals toward persons with disability.

OBJECTIVE: This study aimed to develop and validate an instrument that measures the attitudes of healthcare professionals toward persons with disabilities. DESIGN: The Delphi survey consensus method was used to compose a preliminary questionnaire for survey. Then, healthcare professionals responded to the questionnaire online, and the results were used for psychometric analysis. SETTING: Three tertiary hospitals. SUBJECTS: A total of 993 healthcare professionals. INTERVENTIONS: Not applicable. MAIN MEASURES: Attitudes of healthcare professionals toward persons with disabilities. RESULTS: A 28-item preliminary questionnaire was initially developed after a two-round Delphi survey with 16 experts. Through an online survey, the final 20 items were categorized as follows: behaviors in clinical situation, knowledge and skills, emotional response, and responsibility of healthcare professionals. The Cronbach's alpha coefficient for the total score was 0.890 and varied between 0.856 and 0.892 in the four subdomains. The newly developed tool showed an acceptable model fit. Healthcare professionals who participated in an education module showed higher scores in behavior and knowledge/skills subdomains than those who did not, whereas participants who had clinical experience with persons with disabilities demonstrated higher scores in emotional response and knowledge/skills subdomains. CONCLUSIONS: We developed a scale to measure the attitudes toward persons with disabilities of healthcare professionals. The scale demonstrated a sensitivity to measure a difference by participation in an education module or clinical experience with persons with disabilities.

Patient-Centeredness during In-Depth Consultation in the Outpatient Clinic of a Tertiary Hospital in Korea: Paradigm Shift from Disease to Patient.

BACKGROUND: Patient-centered care (PCC) and integrative care approach are widely advocated. However, their implementation usually requires an extended consultation time. Despite significant advances in medical diagnosis and treatment, no studies have examined consultation time and patient centeredness in Korea. METHODS: We conducted a "15-Minute Consultation" for first-time patients in outpatient clinics of 13 departments. A control group was selected from the same physicians' first-time patients, adjusting for age and gender. A total of 275 patients were selected for receiving in-depth consultation and 141 control patients were selected for regular consultation. Data were collected from patients using a questionnaire comprising a patient-centeredness scale and items on potential predictors such as socio-demographic and clinical factors. We also investigated the participating physician's professionalism. RESULTS: As compared to the control group, the in-depth consultation group scored higher on 5 variables associated with PCC, including (patients' perception of) medical professionals, wait and consultation times, treatment, patient advocacy, and patient satisfaction. While 92.4% of patients in the in-depth consultation group reported that the consultation time was sufficient, only 69.0% of those in the control group reported the same (P < 0.01). In the in-depth consultation group, scores on satisfaction level were the highest for the department of internal medicine, followed by departments of surgery and pediatrics. Participating physicians' improved satisfaction following the intervention proved that in-depth consultation facilitated building a rapport with patients. CONCLUSION: This study illustrated that the provision of sufficiently long consultation for serious and rare diseases could improve PCC and physicians' professionalism. Health authorities should reshuffle the healthcare delivery system and provide sufficient consultation time to ensure PCC and medical professionalism.

Prevalence of Possible Depression and Post-traumatic Stress Disorder among Community Dwelling Adult Refugees and Refugee Applicants in South Korea.

BACKGROUND: Because there has been a recent increase in refugee applications in Korea, the mental health of these refugees merits greater study. METHODS: We surveyed 129 refugees (including those in process of refugee application) and 121 migrant workers living in urban communities, using: the Patient Health Questionnaire-9 for depressive symptoms, the Impact Event Scale-Revised for post-traumatic stress disorder (PTSD) symptoms, and the health questionnaires used in 2016 Korean National Health and Nutrition Examination Survey (KNHANES). The majority of refugee subjects were from sub-Saharan Africa and Middle East. We compared the prevalence of possible depression and possible PTSD between refugees and migrant workers and refugees and age-gender matched samples from the KNHANES 2016. RESULTS: Frequency of suicidal planning during the last year was higher in the refugee group than Korean nationals, but frequency of suicidal attempt was not. High risk drinking was found in 0.8% of refugees, 6.6% of migrant workers and 27.2% of Korean nationals. Possible depression was present in 42.9% of refugee subjects, 33.3% of migrant workers, and 4.2% of Korean controls. Possible PTSD was present in 38.9% of refugees compared to 12.5% of migrant workers. Only major risk factor for depression among refugees was a traumatic event before entering Korea. CONCLUSION: Possible depression and PTSD are significantly more prevalent in refugees, compared to both migrant workers and Korean nationals. Prevalence rates are commensurate with refugee studies worldwide. Appropriate early screening and intervention schemes need to be developed for refugees entering Korea.

Successful management of proteinuria and systemic hypertension in a dog with renal cell carcinoma with surgery, telmisartan, and amlodipine.

An 11-year-old neutered male Yorkshire terrier dog was presented with a 3-week history of hematuria and anorexia. A unilateral renal mass was detected and surgically removed. The renal mass was diagnosed on histopathologic examination as a renal carcinoma. Supportive medical therapy was carried out and persistent systemic hypertension was managed using telmisartan.

Gestion réussie de la protéinurie et de l'hypertension systémique chez un chien atteint d'un carcinome rénal à l'aide d'une chirurgie, de telmisartan et d'amlodipine. Un chien Yorkshire terrier mâle stérilisé âgé de 11 ans a été présenté avec une anamnèse de 3 semaines d'hématurie et d'anorexie. Une masse rénale unilatérale a été détectée et excisée par chirurgie. La masse rénale a été diagnostiquée à l'examen histopathologique comme étant un carcinome rénal. Une thérapie médicale de soutien a été réalisée et l'hypertension systémique persistante a été gérée à l'aide de telmisartan.(Traduit par Isabelle Vallières).

Differential expressions of NOD-like receptors and their associations with inflammatory responses in rheumatoid arthritis.

OBJECTIVES: To determine the differential expressions of nucleotide oligomerisation domain (NOD)-like receptors (NLRs) and to investigate their association with inflammatory responses in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS). METHODS: Gene expression and protein levels of various NLRs, including NOD1, NOD2, NLRP1, NLRP3, NLRP12, NLRX1, and NLRC3, were determined in FLS and synovial tissues from patients with RA and patients with osteoarthritis (OA) using quantitative real-time PCR and immunohistochemistry. After transfection of NOD2 RNAi plasmids or a pcDNA3.1-NLRX1 vector, gene expression levels of pro-inflammatory cytokines in RA FLS and the protein levels of these cytokines in culture supernatants were determined using quantitative real-time PCR and enzyme-linked immunosorbent assays. The effects of NLR gene regulation on NF-κB and caspase-1 were evaluated using Western blot analysis. RESULTS: Gene expression levels of NOD1, NLRP1, NLRP3, NLRP12, and NLRC3 were not different between RA and OA samples. NOD2 gene expression and protein levels were significantly increased in RA samples, whereas the levels of NLRX1 were significantly decreased. Downregulation of NOD2 gene expression by transfection with NOD2 RNAi plasmid significantly reduced pro-inflammatory cytokine levels in RA FLS, while transfection with adenoviral vectors encoding NLRX1 had no effect on pro-inflammatory cytokine levels. Downregulation of NOD2 gene expression significantly decreased NF-κB, TRAF6, and IKK levels, but not caspase-1 levels, in RA FLS. CONCLUSIONS: NOD2 is upregulated in RA FLS; moreover, downregulation of NOD2 gene expression reduces pro-inflammatory cytokine and NF-κB levels in RA FLS. These findings provide evidence that NOD2 exerts pro-inflammatory effects in RA.

Comparison of short- and long-term outcomes of an early experience with robotic and laparoscopic-assisted resection for rectal cancer.

BACKGROUND/AIMS: Robotic surgery is increasingly used for rectal cancer. We compared the short- and long-term outcomes between robotic- and laparoscopic-assisted resection for rectal cancer. METHODOLOGY: A retrospective chart review was performed between 2006 and 2010. RESULTS: Seventeen robotic and 61 laparoscopic surgeries were performed consecutively. Median follow-up time was 58.2 months. No operation was converted to open surgery. No difference was observed between the groups for types of operations, diverting ileostomy rate, operation time, blood loss, and postoperative hospital stay, tumor diameter, distal margin, circumferential margin, tumor stage, differentiation, lymphovascular, or perineural invasion. However, the number of harvested lymph nodes was higher in the robot than that in the laparoscopy group (p = 0.017). Overall morbidity and reoperation rates were similar between the groups. The 5-yr overall and disease-free survival rates of all patients were 82.5% and 81.3%, respectively. The 5-yr overall and disease-free survival rates of the robotic and the laparoscopy groups were 94.1% and 79.7% (p = 0.241), and 94.1% and 77.9% (p = 0.159), respectively. CONCLUSIONS: Robot-assisted resection for rectal cancer resulted in harvesting more lymph nodes without increasing morbidity and showed a comparable survival rate, compared with those of laparoscopy.

Secreted frizzled-related protein 5 suppresses inflammatory response in rheumatoid arthritis fibroblast-like synoviocytes through down-regulation of c-Jun N-terminal kinase.

OBJECTIVE: This study was performed to investigate the effect of secreted frizzled-related protein 5 (Sfrp5), a novel anti-inflammatory adipokine that competes with the frizzled proteins for Wnt binding, on inflammatory response and the c-Jun N-terminal kinase (JNK) signalling pathway in RA. METHODS: Expression of Sfrp5 mRNA in peripheral blood mononuclear cells (PBMCs) and fibroblast-like synoviocytes (FLSs) from patients with RA and OA was determined using real-time quantitative PCR (qPCR). Sfrp5 RNA interference (RNAi) plasmids were transfected to abrogate Sfrp5 expression in RA FLSs, and adenovirus containing the Sfrp5 transcript was delivered into RA FLSs to strengthen Sfrp5 expression. Levels of pro-inflammatory genes and their protein products were determined using real-time qPCR and ELISA in RA FLSs. Production of mitogen-activated protein kinase kinase 7 (MKK-7), JNK and c-Jun were assessed by Western blot analysis. RESULTS: Expression of Sfrp5 mRNA was decreased in PMBCs and FLSs from patients with RA compared with patients with OA. Gene expression and production of IL-1ß, IL-6, chemokine ligand 2 (CCL-2), CCL-7, cyclooxygenase 2 and MMP-9 were markedly increased in Sfrp5 RNAi plasmid-transfected RA FLSs, while transfection with adenoviral vectors encoding Sfrp5 induced reductions in those levels. Phosphorylated forms of MKK-7, JNK and c-Jun were increased by Sfrp5 RNAi plasmids and were decreased by adenoviral vectors encoding Sfrp5. CONCLUSION: Sfrp5 suppressed the inflammatory response and down-regulated JNK signalling in RA FLSs. These findings provide evidence for the anti-inflammatory effect of Sfrp5 in RA.

Is T-stage the only significant factor determining the extent of surgery for gallbladder cancer?

BACKGROUND/AIMS: T-stage is currently the only factor determining the extent of surgery of gallbladder cancer (GBCA). We hypothesized that perineural invasion could be another predictive factor determining the extent of surgery because it is very powerful prognostic factor for GBCA. METHODOLOGY: A retrospective analysis was carried out of patients who underwent operation for gallbladder cancer between February 1991 and November 2011. The data were retrospectively analyzed and reviewed and the microscopic findings were checked by a pathologist. RESULT: Simple and extended cholecystectomy was performed in 82 patients during the study period. In univariate analysis, CA 19-9 level, gross type, depth of invasion, lymph node metastasis, distant metastasis, perineural invasion, lymphatic invasion and vascular invasion were associated with survival (P < 0.05). In multivariate analysis, perineural invasion and vascular invasion had an impact on survival (P < 0.05). CONCLUSIONS: T and N stage are powerful prognostic factor for GBCA, but perineural invasion and vascular invasion are also significant prognostic factors. To improve survival radical resection should be considered in early GBCA with perineural and vascular invasion.

Active tuberculosis risk with tumor necrosis factor inhibitors after treating latent tuberculosis.

BACKGROUND: Active tuberculosis (TB) risk increases during anti-tumor necrosis factor (TNF) therapy; therefore, latent TB infection (LTBI) screening is recommended in potential TNF inhibitor users. It is unclear whether anti-TNF therapy increases the risk of active TB infection even after standard LTBI treatment. OBJECTIVE: The objective of this study was to compare the risk of active TB development in LTBI-positive versus LBTI-negative TNF inhibitor users following the current national LTBI treatment guidelines for LTBI. METHODS: We retrospectively studied 949 TNF inhibitor users with immune-mediated inflammatory diseases from 2005 to 2012 at the Yonsei University Health System. We compared the incidence of active TB among LTBI-positive TNF inhibitor users treated according to national guidelines (n = 256) and LTBI-negative TNF inhibitor users (n = 521), using Poisson regression. RESULTS: The active TB incidence was 1107 per 100,000 patient-years in LTBI-positive TNF inhibitor users who received standard LTBI treatment and 490 per 100,000 patient-years in LTBI-negative TNF inhibitor users. Analysis showed that despite this numerical trend active TB risk was not statistically significantly elevated in LTBI-positive versus LTBI-negative TNF inhibitor users (incidence risk ratio, 2.15; P = 0.24; 95% confidence interval, 0.6-7.7). CONCLUSIONS: This study demonstrated no statistically significantly increased risk of active TB in LTBI-positive TNF inhibitor users who received standard LTBI treatment compared with LTBI-negative TNF inhibitor users.

Anti-Inflammatory Effects of Chamaecyparis obtusa (Siebold & Zucc.) Endl. Leaf Extract Fermented by Ganoderma applanatum Mycelia.

Corticosteroids are commonly used anti-inflammatory agents. However, their prolonged use can lead to side effects. Therefore, the development of natural compounds with minimal side effects is necessary. This study was performed to investigate the anti-inflammatory effects and mechanisms of action of Chamaecyparis obtusa (Siebold & Zucc.) Endl. leaf (COL), bioconverted using Ganoderma applanatum (G. applanatum) in lipopolysaccharide (LPS)-induced RAW264.7 cells. The COL 70% EtOH extract fermented by G. applanatum (70COLGA) improved the high cytotoxicity of 70% EtOH extracts (70COL). When RAW264.7 cells were pre-treated with 100 and 200 µg/mL of 70COLGA for 2 h and then treated with LPS for 16 h, LPS induced the production of nitric oxide (NO), and the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) were significantly inhibited. When RAW264.7 cells were pre-treated with 100 and 200 µg/mL of 70COLGA for 2 h and then treated with LPS for 4 h, the phosphorylation of signal transducers and activators of transcription (STAT) was markedly decreased. In addition, 70COLGA markedly suppressed the production of the inflammatory cytokines interleukin (IL)-1ß and IL-6 in LPS-induced RAW264.7 cells. Analysis of pro-inflammatory molecules using cytokine arrays showed that macrophage inflammatory protein (MIP)-2, granulocyte-macrophage colony-stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) and IL-27 expressions were also suppressed by 200 µg/mL of 70COLGA in LPS-induced RAW264.7 cells. These results demonstrate that 70COLGA significantly prevented inflammatory responses by inhibiting the secretion of pro-inflammatory molecules in LPS-induced RAW264.7 cells. When RAW264.7 cells were pre-treated with 100 and 200 µg/mL of 70COLGA for 2 h and then treated with LPS-conditioned medium (LPS-CM) for 30 min, 70COLGA directly inhibited STAT activation. In summary, our findings suggest that 70COLGA has therapeutic potential for the treatment of inflammatory diseases.

Protective Effects of Repetitive Transcranial Magnetic Stimulation Against Streptozotocin-Induced Alzheimer's Disease.

Repetitive transcranial magnetic stimulation (rTMS) is a non-invasive brain stimulation under investigation for treatment of a wide range of neurological disorders. In particular, the therapeutic application of rTMS for neurodegenerative diseases such as Alzheimer's disease (AD) is attracting attention. However, the mechanisms underlying the therapeutic efficacy of rTMS have not yet been elucidated, and few studies have systematically analyzed the stimulation parameters. In this study, we found that treatment with rTMS contributed to restoration of memory deficits by activating genes involved in synaptic plasticity and long-term memory. We evaluated changes in several intracellular signaling pathways in response to rTMS stimulation; rTMS treatment activated STAT, MAPK, Akt/p70S6K, and CREB signaling. We also systematically investigated the influence of rTMS parameters. We found an effective range of applications for rTMS and determined the optimal combination to achieve the highest efficiency. Moreover, application of rTMS inhibited the increase in cell death induced by hydrogen peroxide. These results suggest that rTMS treatment exerts a neuroprotective effect on cellular damage induced by oxidative stress, which plays an important role in the pathogenesis of neurological disorders. rTMS treatment attenuated streptozotocin (STZ)-mediated cell death and AD-like pathology in neuronal cells. In an animal model of sporadic AD caused by intracerebroventricular STZ injection, rTMS application improved cognitive decline and showed neuroprotective effects on hippocampal histology. Overall, this study will help in the design of stimulation protocols for rTMS application and presents a novel mechanism that may explain the therapeutic effects of rTMS in neurodegenerative diseases, including AD.

Activation of liver X receptors suppresses inflammatory gene expressions and transcriptional corepressor clearance in rheumatoid arthritis fibroblast like synoviocytes.

OBJECTIVES: Liver X receptors (LXR) are nuclear receptors that play important roles in lipid metabolism and transport. LXR also suppress inflammatory responses in macrophages through a unique mechanism of transrepression. This study was performed to investigate whether the synthetic LXR agonist GW3965 can modulate the inflammatory status of fibroblast-like synoviocytes (FLS) from patients with rheumatoid arthritis (RA) and to identify the mechanism for their effect. METHODS: RA FLS were treated with 0.1 and 1 µM of GW3965, a synthetic LXR agonist. The mRNA expressions of pro-inflammatory mediators were measured using quantitative real-time PCR. Apoptotic cell death of RA FLS was assessed using TUNEL assay and determination of caspase-3 activity by a colorimetric assay. The levels of transcriptional corepressors including NCoR and SMRT were determined using western blot analyses. RESULTS: Treatment of RA FLS with GW3965 induced dose-dependent reductions in mRNA expression of pro-inflammatory mediators (IL-1ß, IL-6, MMP-9, CCL-2, CCL-7, and COX-2). However, treatment with GW3965 at the concentration selected for this study had no effect on apoptosis of RA FLS. Decreased productions of NCoR and SMRT by LPS stimulation was attenuated by GW3965 treatment. CONCLUSIONS: GW3965 treatment suppressed mRNA expressions of pro-inflammatory mediators from RA FLS and inhibited the clearance of transcriptional corepressors. These data suggest that LXR activation can be used as a therapeutic approach to reduce the synovial inflammation in RA.

What is the optimal time for laparoscopic cholecystectomy in gallbladder empyema?

PURPOSE: With the accumulating experience in laparoscopic surgery, early laparoscopic cholecystectomy (LC) is increasingly offered for acute cholecystitis. However, early LC without percutaneous transhepatic gallbladder drainage (PTGBD) for gallbladder empyema is still believed to be unsafe. The purpose of this study was to determine the optimal time for LC in gallbladder empyema. METHODS: A retrospective analysis was carried out of patients who underwent LC without PTGBD for gallbladder empyema between August 2007 and December 2010. All cases were confirmed by biopsy. The patients were divided into two groups on the basis of a cutoff of 72 h. RESULTS: LC for gallbladder empyema was performed without PTGBD in 61 patients during the study period. The overall conversion rate was 6.6 %. Based on the 72 h cutoff, there were 33 patients in the early group and 28 in the delayed group. There were no significant differences between early and late patients with respect to operation duration (75.5 vs. 71.4 min, p = 0.537), postoperative hospital stay (4.2 vs. 3.3 days, p = 0.109), conversion rate (12.1 vs. 0 %, p = 0.118), and complication rate (12.1 vs. 3.6 %, p = 0.363). However, the early group had a significantly shorter total hospital stay (5.3 vs. 8.7 days, p = 0.001). CONCLUSIONS: Early LC without PTGBD is safe and feasible for gallbladder empyema and is associated with a low conversion rate. Delayed LC for gallbladder empyema has no advantages and results in longer total hospital stays. LC should be performed as soon as possible within 72 h after admission to decrease length of hospital stay.

One layer end-to-side pancreaticojejunostomy using reinforcing suture on the pancreatic stump.

BACKGROUND/AIMS: Pancreaticoduodenectomy is associated with a high incidence of postoperative morbidity, and pancreatic fistulas are the most important complication. We therefore designed a modified method of pancreaticojejunostomy using reinforcing suture on the pancreatic stump. METHODOLOGY: The procedure is performed as follow: first, insert a soft rubber catheter into the pancreatic duct, then make reinforcing sutures with 3-0 vicryl. These sutures are parallel to the cut edge of the pancreatic stump, and 5mm from the cut edge. The whole layer of jejunum is sewed to the pancreas including the reinforcing sutures. RESULTS: Between July 2002 and November 2011, 52 consecutive cases of pancreaticoduodenectomy were performed using this method. We used The International Study Group for Pancreatic Surgery (ISGPS) severity grading for postoperative pancreatic fistula (POPF) and delayed gastric emptying (DGE). There was 1 grade A fistula (1.9%), 16 grade B fistulas (30.8%), and 2 grade C fistulas (3.8%). CONCLUSIONS: This method of one layer anastomosis is straightforward and practicable; hence it saves operative time, and is not much affected by lack of familiarity with the surgical technique. Even if it does not actually prevent pancreatic fistula formation, it does prevent the life-threatening complete breakdown of the pancreaticojejunostomy.

Anti-inflammatory effects of Allium cepa L. peel extracts via inhibition of JAK-STAT pathway in LPS-stimulated RAW264.7 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Allium cepa L. (A. cepa) is one of the oldest cultivated plants in the world. A. cepa has been used in traditional folk medicine to treat inflammatory disease in several regions, such as Palestine and Serbia. A. cepa peel has a higher content of flavonoids, such as quercetin, than the edible parts. These flavonoids alleviate inflammatory diseases. However, the anti-inflammatory effects of A. cepa peel extract-obtained using various extraction methods-and their underlying mechanisms require further investigation. AIM OF THE STUDY: Although research to find safe anti-inflammatory substances in various natural products has been actively conducted for many years, it is important to continue identifying potential anti-inflammatory effects in natural materials. The purpose of this study was to investigate the ethnopharmacological properties of the A. cepa peel extract, whose efficacy when obtained through different extraction methods and underlying action mechanisms is not well known. The present study specifically aimed to observe the anti-inflammatory effects of the A. cepa peel extracts obtained using various extraction methods and the related detailed mechanisms of A. cepa peel extracts in lipopolysaccharide (LPS)-induced RAW264.7 cells. MATERIALS AND METHODS: The total flavonoid content of the A. cepa peel extracts was determined the diethylene glycol colorimetric method and measured using a calibration curve prepared using quercetin as a standard solution. The antioxidant activity was evaluated using the ABTS assay, and cytotoxicity was measured using the MTT assay. NO production was measured using Griess reagent. Protein levels were measured by western blotting, and mRNA expression was measured by RT-qPCR. Secreted cytokines were analyzed using ELISA or cytokine arrays. In the GSE160086 dataset, we calculated Z-scores for individual genes of interest and displayed using a heat map. RESULTS: Of the three A. cepa peel extracts obtained using different extraction methods, the A. cepa peel 50% EtOH extract (AP50E) was the most effective at inhibiting LPS-induced nitric oxide (NO) and inducible nitric oxide synthase (iNOS). Furthermore, AP50E significantly reduced the levels of pro-inflammation cytokines interleukin (IL)-1α, IL-1ß, IL-6, and IL-27. Additionally, AP50E directly inhibited the Janus kinase-signaling transducer and activator of transcription (JAK-STAT) pathway. CONCLUSIONS: These results showed that AP50E exhibited an anti-inflammatory effect in LPS-induced RAW264.7 mouse macrophages by directly inhibiting JAK-STAT signaling. Based on these findings, we propose AP50E as a potential candidate for the development of preventive or therapeutic agents against inflammatory diseases.

Pharmacological anti-tumor effects of natural Chamaecyparis obtusa (siebold & zucc.) endl. Leaf extracts on breast cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: Chamaecyparis obtusa (C. obtusa, cypress species) is a plant that grows mainly in the temperate Northern Hemisphere and has long been used as a traditional anti-inflammatory treatment in East Asia. C. obtusa contains phytoncides, flavonoids, and terpenes, which have excellent anti-cancer effects and have been reported to prevent the progression of various cancers. However, the detailed mechanisms underlying the anti-cancer effects of C. obtusa extracts are unknown. AIM OF THE STUDY: We sought to confirm the anti-cancer effects of C. obtusa leaf extracts and to reveal the mechanism of action, with the possibility of its application in the treatment or prevention of cancer. MATERIAL &METHODS: The cytotoxicity of C. obtusa leaf extracts was confirmed using an MTT assay. Intracellular changes in protein levels were measured by immunoblotting, and mRNA levels were measured with qRT-PCR. Wound healing assay and transwell migration assay were used to evaluate the metastatic potential of breast cancer cells. The extract-induced apoptosis was observed using IncuCyte Annexin V Red staining analysis. A syngeneic breast cancer mouse model was established by injecting 4T1-Luc mouse breast cancer cells into the fat pad of female BALB/c mice, and the extract was administered orally. Luciferin solution was injected intraperitoneally to assess primary tumor development and metastasis by bioluminescence. RESULTS: C. obtusa leaf extracts were extracted with boiling water, 70% EtOH, and 99% EtOH. Among the extracts, the 99% EtOH extract of C. obtusa leaf (CO99EL) most clearly inhibited the tyrosine phosphorylation of Signal Transducer and Activator of Transcription 3 (pY-STAT3) in MDA-MB-231 breast cancer cells at a concentration of 25 and 50 µg/mL. In addition, CO99EL strongly inhibited not only endogenous pY-STAT3 levels but also IL-6-induced STAT3 activation in various types of cancer cells, including breast cancer. CO99EL inhibited metastatic potential by downregulating the expression of N-cadherin, fibronectin, TWIST, MMP2, and MMP9 in MDA-MB-231 breast cancer cells. CO99EL also induced apoptotic cell death by increasing cleaved caspase-3 and decreasing anti-apoptotic proteins Bcl-2 and Bcl-xL. In an in vivo syngeneic breast cancer mouse model, 100 mg/kg CO99EL suppressed tumor growth and induced apoptosis of cancer cells. Moreover, CO99EL significantly inhibited lung metastasis from primary breast cancer. CONCLUSIONS: Our study demonstrated that 100 mg/kg CO99EL has potent anti-tumor effects against breast cancer, thus suggesting that 100 mg/kg CO99EL has potential applications in the treatment and prevention of breast cancer.

Targeted drug release system based on pH-responsive PAA-POSS nanoparticles.

An amphipathic PAA-POSS@DOX drug delivery system that responds sensitively to pH changes in the cancer microenvironment has been developed using a nanoparticle based on inorganic polyhedral oligomeric silsesquioxane (POSS). POSS was introduced to the carboxylic acid group of polyacrylic acid to which doxorubicin anticancer drug was loaded to prepare 480 ± 192 nm self-assembled nanoparticles. PAA-POSS had a high loading efficiency of over 75% and doxorubicin was quickly released to the target area responding sensitively to weakly acidic conditions. The possibility of employing PAA-POSS as a targeted drug delivery system has been confirmed by observing the death of cells of the MDA-MB-231 breast cancer line.

Chamaecyparis obtusa (Siebold & Zucc.) Endl. leaf extracts prevent inflammatory responses via inhibition of the JAK/STAT axis in RAW264.7 cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Chamaecyparis obtusa (Siebold & Zucc.) Endl. (C. obtusa) has been used as folk medicine in East Asia and has been reported to alleviate inflammatory diseases. However, the detailed mechanisms for the anti-inflammatory effects of C. obtusa remain unclear. AIM OF THE STUDY: Although the anti-inflammatory mechanisms of natural products have been studied for decades, it is still important to identify the potential anti-inflammatory effects of natural sources. In this study, we investigated the anti-inflammatory effects and underlying mechanism of C. obtusa leaf extracts. MATERIAL &METHODS: The cell viability was determined by MTT and crystal violet staining. NO production in the supernatant was measured using Griess reagent. The cell lysates were analyzed by immunoblotting and RT-qPCR. Secreted cytokines were analyzed using ELISA kit and cytokine array kit. mRNA expression from the GSE9632 database set. Z-scores were calculated for each gene and visualized by heat map. RESULTS: Among the extracts of C. obtusa obtained with different extraction methods, the 99% ethanol leaf extract (CO99EL) strongly inhibited lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression and Janus kinase/signaling transducer and activator of transcription (JAK/STAT) phosphorylation in RAW264.7 cells. In addition, CO99EL strongly inhibited LPS-induced interleukin (IL)-1ß, IL-6, IL-27, and C-C motif chemokine ligand (CCL)-1 production and directly inhibited LPS-induced JAK/STAT phosphorylation in RAW264.7 cells. CONCLUSIONS: These findings demonstrate that CO99EL significantly prevents LPS-induced macrophage activation by inhibiting the JAK/STAT axis. Therefore, we suggest the use of C. obtusa extracts as therapeutic approach for inflammatory diseases.