Establishment of Patient-Derived Organoids Using Ascitic or Pleural Fluid from Cancer Patients / Journal of the Korean Cancer Association, 대한암학회지

Purpose@#Patient-derived tumor cells can be a powerful resource for studying pathophysiological mechanisms and developing robust strategies for precision medicine. However, establishing organoids from patient-derived cells is challenging because of limited access to tissue specimens. Therefore, we aimed to establish organoids from malignant ascites and pleural effusions. @*Materials and Methods@#Ascitic or pleural fluid from pancreatic, gastric, and breast cancer patients was collected and concentrated to culture tumor cells ex vivo. Organoids were considered to be successfully cultured when maintained for five or more passages. Immunohistochemical staining was performed to compare the molecular features, and drug sensitivity was assayed to analyze the clinical responses of original patients. @*Results@#We collected 70 fluid samples from 58 patients (pancreatic cancer, n=39; gastric cancer, n=21; and breast cancer, n=10). The overall success rate was 40%; however, it differed with types of malignancy, with pancreatic, gastric, and breast cancers showing 48.7%, 33.3%, and 20%, respectively. Cytopathological results significantly differed between successful and failed cases (p=0.014). Immunohistochemical staining of breast cancer organoids showed molecular features identical to those of tumor tissues. In drug sensitivity assays, pancreatic cancer organoids recapitulated the clinical responses of the original patients. @*Conclusion@#Tumor organoids established from malignant ascites or pleural effusion of pancreatic, gastric, and breast cancers reflect the molecular characteristics and drug sensitivity profiles. Our organoid platform could be used as a testbed for patients with pleural and peritoneal metastases to guide precision oncology and drug discovery.

Regulation of gastrointestinal hormones during laxative activity of gallotannin-enriched extract isolated from Galla Rhois in loperamide-induced constipation of SD rats / 한국실험동물학회지

Regulation of gastrointestinal hormones have been reported in animal models for constipation undergoing laxative therapy when administered herbal products. We undertook to investigate whether the laxative activity of gallotannin-enriched extracts isolated from Galla Rhois (GEGR) affects the regulation of gastrointestinal hormones, by examining the concentration of four hormones and the activation of their receptors in the loperamide (Lop)-induced constipation model. Stool parameters, including number, weight and water content, were significantly recovered in the Lop+GEGR treated group, relative to the Lop+vehicle treated group; however, food intake and water consumption were maintained at a constant level. Also, a similar recovery was detected for thickness of mucosa, muscle and flat luminal surface in the Lop+GEGR treated group. Furthermore, concentration of the four gastrointestinal hormones evaluated, namely, cholecystokinin (CCK), gastrin (GAS), somatostatin (SS) and motilin (MTL), were lower in the Lop+vehicle treated group than the No treated group, but were remarkably enhanced in the Lop+GEGR treated group. Moreover, the downstream signaling pathway of MTL and SS receptors were recovered after GEGR administration. Results of the present study therefore indicate that the laxative effects of GEGR treatment may be tightly related with the regulation of gastrointestinal hormones in the Lop-induced constipation model.

Inhibition of endoplasmic reticulum stress in high-fat-diet-induced obese C57BL/6 mice: Efficacy of a novel extract from mulberry (Morus alba) leaves fermented with Cordyceps militaris / 한국실험동물학회지

A few clues about correlation between endoplasmic reticulum (ER) stress and mulberry (Morus alba) leaves were investigated in only the experimental autoimmune myocarditis and streptozotocin-induced diabetes. To investigate whether a novel extract of mulberry leaves fermented with Cordyceps militaris (EMfC) could suppress ER in fatty liver, alterations in the key parameters for ER stress response were measured in high fat diet (HFD)-induced obese C57L/6 mice treated with EMfC for 12 weeks. The area of adipocytes in the liver section were significantly decreased in the HFD+EMfC treated group as compared to the HFD+Vehicle treated group, while their level was higher in HFD+Vehicle treated group than No treated group. The level of the eukaryotic initiation factor 2 alpha (eIF2α) and inositol-requiring enzyme 1 beta (IRE1α) phosphorylation and CCAAT-enhancer-binding protein homologous protein (CHOP) expression were remarkably enhanced in the HFD+Vehicle treated group. However, their levels were restored in the HFD+EMfC treated group, although some differences were detected in the decrease rate. Similar recovery was observed on the ER stress-induced apoptosis. The level of Caspase-3, Bcl-2 and Bax were decreased in the HFD+EMfC and HFD+orlistat (OT) treated group compared to the HFD+Vehicle treated group. The results of the present study therefore provide first evidence that EMfC with the anti-obesity effects can be suppressed ER stress and ER stress-induced apoptosis in the hepatic steatosis of HFD-induced obesity model.

Dose dependence and durability of the therapeutic effects of Asparagus cochinchinensis fermented extract in an ovalbumin-challenged asthma model / 한국실험동물학회지

The butanol extract of Asparagus cochinchinensis roots fermented with Weissella cibaria (BAfW) significantly suppressed the inflammatory response induced by lipopolysaccharide (LPS) treatment in RAW264.7 cells. To investigate the dose dependence and durability of BAfW on the anti-asthma effects, alterations in key parameters were measured in ovalbumin (OVA)-challenged Balb/c mice treated with the different doses of BAfW at three different time points. The number of immune cells, OVA-specific IgE level, thickness of respiratory epithelium and mucus score decreased significantly in a dose-dependent manner in response to treatment with 125 to 500 mg/kg BAfW (P < 0.05), although the highest level was detected in the 500 mg/kg treated group. Moreover, the decrease in these parameters was maintained from 24 to 48 h in the 500 mg/kg of BAfW treated group. At 72 h, the effects of BAfW on the number of immune cells, OVA-specific IgE level and thickness of respiratory epithelium partially disappeared. Overall, this study provides the first evidence that the anti-asthma effect of BAfW may reach the maximum level in OVA-challenged Balb/c mice treated with 500 mg/kg and that these effects can last for 48 h.

Successful development of squamous cell carcinoma and hyperplasia in RGEN-mediated p27 KO mice after the treatment of DMBA and TPA / 한국실험동물학회지

To evaluate the carcinogenicity of p27 knockout (KO) mice with RNA-guided endonuclease (RGENs)-mediated p27 mutant exon I gene (IΔ), alterations in the carcinogenic phenotypes including tumor spectrum, tumor suppressor proteins, apoptotic proteins and cell cycle regulators were observed in p27 (IΔ) KO mice after treatment with 7,12-Dimethylbenz[a]anthracene (DMBA) and 12-O-tetradecanoylphorbol-13-acetate (TPA)(DT) for 5 months. The target region (544~571 nt) in exon I of the p27 gene was successfully disrupted in p27 (IΔ) KO mice using the RGEN-induced non-homologous end joining (NHEJ) technique. After DT exposure for 5 months, a few solid tumors (identified as squamous cell carcinoma) developed on the surface of back skin of DT-treated p27 (IΔ) KO mice. Also, squamous cell hyperplasia with chronic inflammation was detected in the skin dermis of DT-treated p27 (IΔ) KO mice, while the Vehicle+p27 (IΔ) KO mice and WT mice maintained their normal histological skin structure. A significant increase was observed in the expression levels of tumor suppressor protein (p53), apoptotic proteins (Bax, Bcl-2 and Caspase-3) and cell-cycle regulator proteins (Cyclin D1, CDK2 and CDK4) in the skin of DT-treated p27 (IΔ) KO mice, although their enhancement ratio was varied. Taken together, the results of the present study suggest that squamous cell carcinoma and hyperplasia of skin tissue can be successfully developed in new p27 (IΔ) KO mice produced by RGEN-induced NHEJ technique following DT exposure for 5 months.

Lipolytic effect of novel extracts from mulberry (Morus alba) leaves fermented with Cordyceps militaris in the primary adipocytes derived from SD rats / 한국실험동물학회지

Mulberry (Morus alba) leaves are known to have therapeutic effects on lipid metabolism including lipogenesis, lipolysis and hyperlipidemia. However, novel compounds with strong lipolytic ability among 27 extracts of the mulberry leaves fermented with Cordyceps militaris (EMfCs) have not yet been identified. Therefore, the cAMP concentration and cell viability were measured in the primary adipocytes of SD (Sprague Dawley) rats and 3T3-L1 cells after treatment of 27 EMfCs. Briefly, mulberry leaves powders amended with three different concentrations (0, 25 and 50%) of silkworm pupae (SWP) powder were fermented with 10% C. militaris (v/w) during three different periods (3, 4 and 6 weeks). A total of 27 extracts were obtained from the fermented mulberry leaves powders using three different solvents (dH2O, 50% EtOH and 95% EtOH). Among the 27 EMfCs treated groups, a significant increase in the concentration of cAMP was detected in primary adipocytes treated with 10 extracts when compared with the Vehicle treated group. However, their cAMP concentration did not agree completely with the non-toxicity, although most extracts showed non-toxicity. Furthermore, the concentration of cAMP and level of free glycerol gradually increased in a dose dependent manner (100, 200 and 400 µg/mL) of 4M3-95 contained cordycepin without any significant toxicity. Overall, the results of this study provide strong evidence that 4M3-95 extract derived from EMfCs can stimulate the lipolysis of primary adipocytes at an appropriate concentration and therefore have the potential for use as lipolytic agents to treat obesity.

Correlation between laxative effects of uridine and suppression of ER stress in loperamide induced constipated SD rats / 한국실험동물학회지

A correlation between endoplasmic reticulum (ER) stress and laxative effects was first reported in a constipation model treated with an aqueous extract of Liriope platyphylla (AEtLP) roots. To investigate the correlation between the laxative effect of uridine (Urd) and ER stress response, alterations in the key parameters for ER stress were measured in loperamide (Lop) induced constipation Sprague Dawley (SD) rats treated with Urd. The efficacy of the laxative effect of Urd was notable on the symptoms of chronic constipation, including alteration of stool parameters and structure of the transverse colon, in Lop induced constipated SD rats. In the PERK/eIF2-ATF4 pathway of ER stress response, the levels of eukaryotic initiation factor 2 alpha (eIF2α) phosphorylation and DNA damage-inducible protein (GADD34) transcripts were significantly enhanced in the Lop+Vehicle treated group. However, the levels were restored in the Lop+Urd treated group, although few differences were detected in the decrease rate. Similar changes were observed for levels of inositol-requiring enzyme 1 beta (IRE1β) phosphorylation and X-box binding protein 1 (XBP-1) transcript in the IRE1α/XBP pathway. Furthermore, the number of ER stress-induced apoptotic cells and Bax and Bcl-2 expression were recovered in the Lop+Urd treated group compared to the Lop+Vehicle treated group. The results of the present study therefore provide first evidence that the laxative effects of Urd may be tightly correlated with the recovery of ER stress response in constipation models.

Erratum: Comparative study of fertilization rates of C57BL/6NKorl and C57BL/6N mice obtained from two other sources / 한국실험동물학회지

One of the authors' names was misprinted.

Comparative study of fertilization rates of C57BL/6NKorl and C57BL/6N mice obtained from two other sources / 한국실험동물학회지

C57BL/6N is the most widely used inbred mouse strain applied in a wide variety of research areas including cancer, cardiovascular biology, developmental biology, diabetes and obesity, genetics, immunology, neurobiology, and sensorineural research. To compare the fertilization rates of C57BL/6NKorl mice with two commercial C57BL/6N stocks, differences in reproductive organ structures, sperm and egg numbers, fertilization rates, and embryo development rates among C57BL/6NKorl (Korea FDA source), C57BL/6NA (USA source), and C57BL/6NB (Japan source) mice were determined. Among the stocks, no significant differences were detected in organ weight and histological structure of male and female reproductive organs, although body weight was higher in C57BL/6NKorl mice than that in the other groups. The concentration and morphology of sperm and eggs in C57BL/6NKorl mice were similar to those of C57BL/6NA and C57BL/6NB mice. Furthermore, the three stocks had similar in vitro fertilization and embryo development rates, although these rates tended to be higher in C57BL/6NB mice. Pup body weight was higher in C57BL/6NKorl and C57BL/6NB mice than that in C57BL/6NA mice. The results of the present study suggest that C57BL/6NKorl, C57BL/6NA, and C57BL/6NB mice obtained from three different sources have similar fertilization and embryo development rates, although there were slight differences in the magnitude of their responses rates.

Comparison of therapeutic responses to an anticancer drug in three stocks of ICR mice derived from three different sources / 한국실험동물학회지

Korl:ICR mice, established by the Korean National Institute of Food and Drug Safety Evaluation (NIFDS), are characterized based on their genetic variation, response to gastric injury, and response to constipation inducers. To compare the inhibitory responses of ICR stocks obtained from three different sources to the anticancer drug cisplatin (Cis), alterations in tumor volume, histopathological structure, and toxicity were examined in Sarcoma 180 tumor-bearing Korl:ICR, A:ICR (USA source), and B:ICR (Japan source) mice treated with low and high concentrations of Cis (L-Cis and H-Cis, respectively). Tumor size and volume were lower in H-Cis-treated mice than in L-Cis-treated mice in all three ICR stocks with no significant differences among stocks. There was a significant enhancement of the necrotizing areas in the histological structures in the L-Cis- and H-Cis-treated groups relative to that in the untreated group. The necrotizing area changes were similar in the Sarcoma 180 tumor-bearing Korl:ICR, A:ICR, and B:ICR mice. However, there were stock-bases differences in the serum biomarkers for liver and kidney toxic effects. In particular, the levels of AST, ALT and BUN increased differently in the three H-Cis-treated ICR stocks, whereas the levels of ALP and CRE were constant. Taken together, the results of the present study indicate that Korl:ICR, A:ICR, and B:ICR mice have similar overall inhibitory responses following Cis treatment of Sarcoma 180-derived solid tumors, although there were some differences in the magnitude of the toxic effects in the three ICR stocks.

Continuous Abstinence Rates from Smoking Over 12 Months according to the Frequency of Participation in a Hospital-based Smoking Cessation Program among Patients Discharged after Acute Myocardial Infarction

BACKGROUND: This study evaluated the continuous abstinence rates from smoking at 12-month after a hospital-based smoking cessation program was applied for smokers hospitalized for acute myocardial infarction. METHODS: Among those who are hospitalized for acute myocardial infarction from January 2012 to December 2013, ninety-eight smokers agreed to quit smoking were eligible for follow up to 12 months. Each of them underwent six consecutive sessions (first during admission, the other 5 sessions after discharge) of behavioral modification, counseling for withdrawal symptoms, and anti-smoking advices by a trained nurse. Exhaled carbon monoxide measurements less than 6 ppm were used to confirm the smoking cessation status of each participant. RESULTS: Mean age of all participants was 55.2±10.8 years old, and their continuous abstinence rates at 1, 3, 6, 12 months were 63.3%, 49.0%, 43.9%, and 37.8% for each. The continuous abstinence rate from smoking after 12 months was 69.7% and significantly higher in those who completed the 6 sessions than 21.5% in those who completed 5 sessions or less (P<0.001). After adjustment for general and smoking-related characteristics, multivariate logistic regression analysis revealed that full participation relative to 5 or less participation was significantly associated with higher continuous abstinence rate from smoking at 12 months (odds ratio: 7.96; 95% confidence interval: 2.07-30.55). CONCLUSIONS: The consistency of participating in a hospital-based smoking cessation program, described herein, significantly improved success rates of smoking cessation in patients discharged after acute myocardial infarction. Hospital-based smoking cessation program based on education and counseling should be included as an important part of patient management for acute myocardial infarction.

Factors Related to Pre-hospital Delay in Korean Patients with ST-segment Elevation Myocardial Infarction: A Data from the Province of Jeonbuk Regional Cardiovascular Center

OBJECTIVE: Despite successful efforts to shorten the door-to-balloon time in patients with ST-segment elevation myocardial infarction (STEMI), pre-hospital delayremains a problem. We evaluated the factors related to pre-hospital delay using the Jeonbuk regional cardiovascular center database. METHODS: From 2010 to 2013, a total of 384 STEMI patients were enrolled. We analyzed the onset time, door time, and balloon time, and the patients were grouped according to pre-hospital delay (120 minutes). Clinical and socio-demographic variables were compared. RESULTS: 53.2% of patients had prolonged onset-to-door time (median 130, interquartile range [IQR] 66~242 minutes), and 68.5% of patients did not achieve 60 years (OR 1.8, 95% CI 1.1-3.0, p=0.031) and hypertension (OR 1.9, 95% CI 1.2-2.9, p=0.047) were independent predictors of pre-hospital delay. CONCLUSIONS: The present study demonstrated a significant pre-hospital delay in the treatment of STEMI patients in the Province of Jeonbuk. Public campaigns and education are needed to raise the public awareness of STEMI and the use of 119.

Emergency Medical Service Use among Patients with Acute ST-segment Elevation Myocardial Infarction in Jeonbuk Province / 대한내과학회지

BACKGROUND/AIMS: The use of emergency medical services (EMSs) at the onset of pain, and the relationship between transport type and the treatment delay for acute ST-segment elevation myocardial infarction (STEMI) were evaluated using the Jeonbuk Regional Cardiovascular Center database. METHODS: In total, 527 STEMI patients who underwent primary percutaneous coronary intervention (PCI) were enrolled in this study. Basic characteristics, socioeconomic variables, and delay factors were compared between patients that contacted an EMS as first medical contact (FMC) and patients that used other forms of FMC. RESULTS: Only 28.8% of patients used EMS as their FMC. The patients that used EMS showed significantly shorter onset-to-balloon time than those who did not (250.7 ± 366.6 min vs. 405.9 ± 649.8 min, p = 0.001). However, 36.2% of patients that used EMS as FMC were transported to non-PCI-capable centers, which led to significantly prolonged onset-to-balloon time. Multivariate analysis revealed that transfer via another hospital (odds ratio [OR] 2.0, p 65 years (OR 1.9, p = 0.003), and previous history of PCI (OR 0.4, p = 0.033) were independent predictors of pre-hospital delay. CONCLUSIONS: EMS used as FMC at the onset of chest pain was an important factor for decreasing treatment delay in patients with STEMI. However, a small number of patients used EMS as FMC, and some patients that used EMS were transported to non-PCI-capable centers. Public campaigns and education are needed to raise the public awareness of STEMI and the use of EMSs.

The Effects of Mental Fitness, Social Support and Psychological Distress on Suicidal Idea of People with Schizophrenia Using Community Mental Health Services / 신경정신의학

OBJECTIVES: The objective of this study was to analyze the relationships of psychological distress, social support, mental fitness and suicidal idea among patients of chronic schizophrenia using community mental health services. METHODS: This study was conducted in patients who visited the mental health services in Daejeon from October to December 2011. In total, 395 chronic schizophrenic patients were evaluated using Mental Fitness Scale (MFS), Kessler Psychological Distress Scale (K10), and Multidimensional Scale of Perceived Social Support (MSPSS). RESULTS: Results showed that 37% (n=146) of patients had suicidal ideas and 63% (n=249) of patients did not have suicidal ideas. The group that had suicidal ideas showed significantly lower scores on MFS, K10, and MSPSS than the group with no suicidal ideas (p=0.000). Logistic regression analysis showed a significantly lower score on MFS, K10 for the suicidal ideation group than the group with no suicidal ideas. CONCLUSION: These results suggest that health care providers should consider mental fitness and psychological distress, in order to prevent suicidal idea of schizophrenic patients.