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The current standard for the pharmacological management of lumbar disk herniation (LDH), involving non-steroidal anti-inflammatory drugs, muscle relaxants, and opioid analgesics, often carries a risk of adverse events. The search for alternative therapeutic options remains a vital objective, given the high prevalence of LDH and the critical impact on the quality of life. Shinbaro 2 is a clinically effective herbal acupuncture against inflammation and various musculoskeletal disorders. Therefore, we explored whether Shinbaro 2 exerts protective effects in an LDH rat model. The results showed that Shinbaro 2 suppressed pro-inflammatory cytokines, interleukin-1 beta, tumor necrosis factor-alpha, disk degeneration-related factors, matrix metalloproteinase-1,−3,−9, and ADAMTS-5 in LDH rats. Shinbaro 2 administration reinstated a behavioral activity to a normal level in the windmill test. The results indicated that Shinbaro 2 administration restored spinal cord morphology and functions in the LDH model. Therefore, Shinbaro 2 exerted a protective effect in LDH via actions on inflammatory responses and disk degeneration, indicating that future research is warranted to assess the action mechanisms further and validate its effects.

Despite the epidemiological association between intrahepatic cholangiocarcinoma (iCCA) and HBV infection, little is known about the relevant oncogenic effects. We sought to identify the landscape and mechanism of HBV integration, along with the genomic architecture of HBV-infected iCCA (HBV-iCCA) tumors.We profiled a cohort of 108 HBV-iCCAs using whole-genome sequencing, deep sequencing, and RNA sequencing, together with preconstructed data sets of HBV-infected HCC (HBV-HCC; n = 167) and combined hepatocellular cholangiocarcinoma (HBV-cHCC/CCA; n = 59), and conventional (n = 154) and fluke-related iCCAs (n = 16). Platforms based on primary iCCA cell lines to evaluate the functional effects of chimeric transcripts were also used. We found that HBV had inserted at multiple sites in the iCCA genomes in 45 (41.7%) of the tumors. Recurrent viral integration breakpoints were found at nine different sites. The most common insertional hotspot (7 tumors) was in the TERT (telomerase reverse transcriptase) promoter, where insertions and mutations (11 tumors) were mutually exclusive, and were accompanied by promoter hyperactivity. Recurrent HBV integration events (5 tumors) were also detected in FAT2 (FAT atypical cadherin 2), and were associated with enrichment of epithelial-mesenchymal transition-related genes. A distinctive intergenic insertion (chr9p21.3), between DMRTA1 (DMRT like family A1) and LINC01239 (long intergenic non-protein coding RNA 1239), had oncogenic effects through activation of the mammalian target of rapamycin (mTOR)/4EBP/S6K pathway. Regarding the mutational profiles of primary liver cancers, the overall landscape of HBV-iCCA was closer to that of nonviral conventional iCCA, than to HBV-HCC and HBV-cHCC/CCA.Our findings provide insight into the behavior of iCCAs driven by various pathogenic mechanisms involving HBV integration events and associated genomic aberrations. This knowledge should be of use in managing HBV carriers.

The androgen receptor (AR)-signaling pathways are essential for prostate tumorigenesis. Although significant effort has been devoted to directly targeting AR-expressing tumor cells, these therapies failed in most prostate cancer patients. Here, we demonstrate that loss of AR in stromal sonic-hedgehog Gli1-lineage cells diminishes prostate epithelial oncogenesis and tumor development using in vivo assays and mouse models. Single-cell RNA sequencing and other analyses identified a robust increase of insulin-like growth factor (IGF) binding protein 3 expression in AR-deficient stroma through attenuation of AR suppression on Sp1-regulated transcription, which further inhibits IGF1-induced Wnt/β-catenin activation in adjacent basal epithelial cells and represses their oncogenic growth and tumor development. Epithelial organoids from stromal AR-deficient mice can regain IGF1-induced oncogenic growth. Loss of human prostate tumor basal cell signatures reveals in basal cells of stromal AR-deficient mice. These data demonstrate a distinct mechanism for prostate tumorigenesis and implicate co-targeting stromal and epithelial AR-signaling for prostate cancer.

Acetaminophen (paracetamol, APAP) is a major component of Tylenol, Penzal Q, and Panpyrin, and is the most commonly used antipyretic analgesic in children. The conventional oral drug delivery systems of APAP are pills and tablets. However alternative drug delivery methods are desirable in case of pediatric or geriatric patients, especially for drugs like APAP that must be taken in large doses at once. Another requirement for a good drug delivery system is the rapid dissolution to ensure a rapid therapeutic action as pain reliever. In this study Acetaminophen (paracetamol, APAP) was encapsulated in a water-soluble polymer. After the preparation of the solid dispersion by encapsulating acetaminophen in polyvinylpyrrolidone, the resultant granules were used in three formulations: tablets, chewable tablets, and oral dissolving films (ODF). Solid dispersions and prepared formulations were evaluated by Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), X-ray diffractometer (XRD), and universal tensile machine (UTM), and the release behavior was compared. As a result, it was confirmed that the oral dissolving films can be taken easily because it has the advantages of both tablet and liquid dosage form accurate dosage, easy administration, easy swallowing, and fast bioavailability. Furthermore, the drug absorption rate can be effectively increased by changing the formulation.

Redox status is a main indicator of cancer cell adaptation to chemotherapy. Nuclear factor erythroid 2-related factor 2 (Nrf2) is one of the crucial regulators of redox condition and can affected to drug resistance through redox imbalance. The incidence and mortality of pancreatic cancer (PC) have been gradually increasing globally. Although the first line treatment with anticancer agents improves the disease status, acquired drug resistance is major huddles in the treatment failure. Targeting Nrf2 has been suggested as a promising therapeutic approach for the overcoming drug resistance. In the present study, Nrf2 was found to be upregulated in tumor tissues and gemcitabine-resistant PC cells. In addition, Nrf2 knockdown exhibited to recover gemcitabine sensitivity in the gemcitabine-resistant PC cells. To further validate the role of Nrf2 in gemcitabine-resistant cells, the effects of Nrf2 inhibitor, periplocin, were employed. Periplocin exerted the antiproliferative activities by regulating Nrf2 suppression which leads to the increase of reactive oxygen species generation. Furthermore, the downregulation of Nrf2 by periplocin was correlated to Keap1-mediated protein degradation. Docking simulation also confirmed the binding interaction of Nrf2 and Keap1 by periplocin. These findings suggest that targeting the Nrf2 might be a therapeutic strategy for overcoming acquired gemcitabine PC. Keywords: Pancreatic cancer cell, Gemcitabine resistance, Nuclear factor erythroid-2-related factor 2 (Nrf2), Kelch Like ECH Associated Protein 1 (Keap1), protein degradation Acknowledgements: This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2022R1I1A1A01063346). Citation Format: Sang Kook Lee, Eun Seo Bae, Woong Sub Byun, Chae Won Ock, Won Kyung Kim, Hyen Joo Park. Targeting Nrf2 signaling pathway by Keap1-mediated degradation in acquired gemcitabine resistant pancreatic cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2787.

Prostate cancer is the second most common cancer in men worldwide. The current treatment targeting androgen receptor (AR) expressing cells, androgen deprivation therapy (ADT), eventually fails in most prostate cancer patients who consequently develop castration resistant prostate cancer, an incurable disease. Although significant effort has been devoted to understanding the promotional role of AR action in prostate tumor cells, there is an urgent need to define the specific role of stromal AR in prostate tumorigenesis and needs to be evaluated for designing effective therapy.We directly examined AR in stromal Shh responsive Gli1 lineage cells in prostate oncogenesis and tumor development utilizing in vivo tissue recombination and novel genetically engineered mouse models. AR loss in stromal Gli1 lineage cells reduces prostate epithelial oncogenesis in xenograft models, combining transformed UGE by expression of stabilized β catenin with AR deficient or control UGM. A Myc induced prostatic epithelial tumor mouse model was combined with conditional deletion of AR in Gli1 lineage cells during prepubescent and adult age which showed significant impairment of prostate tumor development. Single cell RNA sequencing revealed a robust increased expression of insulin like growth factor binding protein 3 (IGFBP3) in AR deleted stromal Gli1 lineage cells. Interestingly although no gene manipulation occurred in the epithelium, basal epithelial cells showed reduced IGF1 induced Wnt signaling, due to the increased IGFBP3 expression. Through chromatin immunoprecipitation qPCR, we examined how deletion of AR in Gli1 lineage cells increases IGFBP3 expression. Stromal AR deletion directly alleviates the suppression of Sp1 regulating transcription of IGFBP3, leading to inhibition of oncogenic growth of tumor epithelium in a paracrine manner. To uncover the mechanism for stromal AR in Gli1 lineage as a tumor niche, we used organoid culture systems. In prostatic epithelial organoid cultures, addition of IGFBP3 or conditional media from AR deleted stromal cells blocked tumor cell growth. Overall, the data implicate the novel and specific role of stromal AR in Gli1 lineage cells as a tumor niche to support prostate epithelial tumorigenesis. Dysregulation of stromal AR on IGFBP3 IGF1 signaling suggest an underlying mechanism for hormone refractoriness through the IGF axes. Cotargeting reciprocal interactions of AR and IGF1 pathways between epithelial tumor cells and surrounding tumor niches may improve clinical outcomes for treating advanced prostate cancer. Therefore, given the importance of sex hormone, hedgehog, and IGF1 signaling pathways in human development and tumorigenesis raises new and relevant questions underlying androgen, Shh, and IGF regulated tumor cell niches in tumor initiation and progression. Citation Format: Alex Hiroto, Won Kyung Kim, Christian H. Nenninger, Alyssa J. Buckley, Zijie Sun. Stromal androgen signaling acts as tumor niches to drive prostatic basal epithelial progenitor initiated oncogenesis through IGFBP3 mediated IGF1 induced Wnt activation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2371.

Although hepatocellular carcinomas (HCCs) often recur in patients undergoing hepatectomy, there are no reliable biomarkers of this undesirable event. Recent RNA-based efforts have developed valuable genetic indices prognostic of cancer outcomes. We aimed to identify molecular predictors of early recurrence after resection of HCC, and reveal the genomolecular structure of the resected tumors. Based on the transcriptomic and genomic datasets of 206 HCC samples surgically resected in the Asan Medical Center (AMC), we performed a differential gene expression analysis to identify quantitative markers associated with early recurrence and used the unsupervised clustering method to classify genomolecular subtypes. Differential gene expression profiling revealed that S100P was the highest-ranked overexpressed gene in HCCs that recurred within 2 years of surgery. This trend was reproduced in immunohistochemical studies of the original cohort and an independent AMC cohort. S100P expression also independently predicted HCC-specific mortality post-resection (adjusted hazard ratio 1.09, 95% confidence interval 1.01–1.19; p = 0.042). Validation in a Chinese cohort and in in vitro experiments confirmed the prognostic value of S100P in HCC. We further identified five discrete molecular subtypes of HCC; a subtype with stem cell features (‘AMC-C4’) was associated with the worst prognosis, both in our series and another two Asian datasets, and S100P was most strongly upregulated in that subtype. We identified a promising prognostic biomolecule, S100P, associated with early recurrence after HCC resection, and established the genomolecular architecture of tumors affecting clinical outcomes, particularly in Asian patients. These new insights into molecular mediators should contribute to effective care for affected patients.

Patients at risk for hypertension with comorbidities such as diabetes and metabolic syndrome often require two or more antihypertensive drugs to lower their blood pressure. Telmisartan and amlodipine are widely known drugs to treat hypertension. However, telmisartan suffer of poor solubility in water that makes necessary to increase its dosage to reach a sufficient therapeutic concentration. In this study, a solid dispersion containing a water-soluble polymer was prepared to make the absorption rate of telmisartan similar to that of amlodipine, a water soluble drug. As water-soluble polymers, polyvinylpyrrolidone K30, polyethylene glycol 6000, and poloxamer 407 were used. The encapsulation of the solid dispersion was analyzed by differential scanning calorimetry, Xray diffraction, and high-performance liquid chromatography. The reduced flowability of the solid dispersion was improved by a wet granulation. Scanning Electron Microscopy was used to study the morphology and the flowability was verified by measuring the angle of repose, bulk density, and tap density, and expressed by Hausner ratio and Carr index. The release behavior was confirmed by dissolution test and high-performance liquid chromatography. As result we proved that telmisartan a higher release rate when encapsulated with PVP K30. Finally, we designed a drug formulation in which amlodipine and telmisartan could be absorbed at a similar rate.

Mosaicisms caused by postzygotic mutational events are of increasing interest because of their potential association with various human diseases. Postzygotic somatic mutations have not been well characterized however in terms of their developmental lineage in humans. We conducted whole-genome sequencing (WGS) and targeted deep sequencing in 15 organs across three developmental lineages from a single male fetus with polycystic kidney disease (PKD) of 21 weeks gestational age. This fetus had no detectable neurological abnormalities at autopsy but germline mutations in the PKHD1 gene were identified that may have been associated with the PKD. Eight early embryonic mosaic variants with no alteration of protein function were detected. These variants were thought to have occurred at the two or four cell stages after fertilization with a mutational pattern involving frequent C>T and T>C transitions. In our current analyses, no tendency toward organ-specific mutation occurrences was found as the eight variants were detected in all 15 organs. However different allele fractions of these variants were found in different organs, suggesting a tissue-specific asymmetric growth of cells that reflected the developmental germ layer of each organ. This indicated that somatic mutation occurrences, even in early embryogenesis, can affect specific organ development or disease. Our current analyses demonstrate that multi-organ analysis is helpful for understanding genomic mosaicism. Our results also provide insights into the biological role of mosaicism in embryonic development and disease.

Colorectal cancer (CRC) is a common and intractable malignancy with a high mortality risk. Conventional chemotherapeutics are effective for patients with early stage CRC, but the majority of deaths of CRC patients are linked to acquired drug resistance or metastasis occurrence. Asperphenin B (1), a lipopeptidyl benzophenone isolated from a marine-derived Aspergillus sp. fungus, reportedly possesses antiproliferative activity against cancer cells. However, its antitumor activity and the underlying molecular mechanisms remain unexplored. In this study, 1 induced G2/M phase cell cycle arrest and subsequent apoptotic cell death and inhibited tumor growth in a xenograft model. The 1-induced G2/M phase arrest was associated with the regulation of checkpoint proteins, including Chk1/2 and Cdc25c. The 1-induced apoptosis was correlated with an upregulation of p53 and cleaved caspases and a downregulation of survivin. Further experiments revealed that 1-mediated suppression of migration and invasion of metastatic HCT116 cells was partially associated with the downregulation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression. The antimetastatic potential of 1 was also confirmed by E-cadherin upregulation and N-cadherin and Snail downregulation, which were in turn associated with the GAPDH regulation. These findings highlight the potential use of 1 as a novel candidate for treating metastatic CRC with the modulation of GAPDH function.

Early diagnosis of hepatocellular carcinoma (HCC) increases the chances of curative treatment and long-term survival.1 2 We read with interest the analytical study by Zhang et al presenting nomogram models predicting recurrence-free survival and guiding therapeutic decisions in a patient-tailored approach to treatment of early-stage HCC within Milan criteria.3 The refined Barcelona Clinic Liver Cancer (BCLC) guidelines no longer limit the size of solitary nodules in the definition of stage A HCC. However, there is continuing debate about the need to restrict the maximum diameter of single HCCs to inside the Milan limit (≤5 cm).4–6 In a survival-based confirmatory investigation, we primarily used a Korean nationwide cancer registry database of 10 510 new HCC cases. These cases were randomly sampled and registered in 54 hospitals, and investigated by the Korea Liver Cancer Study Group and the governmental organisation of the Korea Central Cancer Registry in 2008–2014. We included in the final analysis 1610 asymptomatic HCC subjects with BCLC stage A or stage B who had preserved liver function levels referred to as Child-Pugh class A without ascites and who initially received standard treatments as per BCLC stage (online supplemental figure S1).4 The population was divided into four groups: group 1, stage A disease fulfilling Milan criteria undergoing liver resection or transplantation, or local ablation; group 2, potential stage A with single nodules >5 cm undergoing resection; group 3, potential stage B with single nodules >5 cm undergoing transarterial chemoembolisation (TACE); and group 4, stage B undergoing TACE. Five-year overall survival (OS) rates were significantly different across the four groups (p

Hyaluronic acid (HA) is one of the most applied biomaterials in a tissue engineering field due to its biocompatibility and its presence in the native extracellular matrix (ECM) of tissues. However, the mechanical property of the HA is weak and requires specific treatment to improve its properties. The application of Agarose (AG) hydrogel is widely studied and used as a support for the three-dimensional culture of cells due to its biocompatibility. Nevertheless, AG itself lacks the biological environment of the matrix which is unsuitable for the growth of the encapsulated cells. In this study, the composite of HA hydrogel and AG hydrogel (HA/AG hydrogel) is proposed to supplement the drawbacks of each hydrogel. HA provided enhanced microenvironment of matrix and AG improved the mechanical properties and assisted the cells. The characterization of the blended hydrogels was carried out with FT-IR, weight loss, swelling ratio, and compressive strength study. The biocompatibility and biological environment of the composite was evaluated by dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide; thiazolyl blue (MTT), live/dead staining, and morphological study. The composite biomaterial exhibited applicability for cartilage tissue engineering and the in vitro study of the cell-laden HA/AG hydrogel displayed potential for cartilage tissue engineering in the future.

Nifedipine is a calcium channel blocker widely used for the treatment of blood pressure related diseases such as angina, hypertension and Raynaud. However, since the drug after the administration reaches the highest blood concentration between 30 min and 2 h, its insolubility may expose the patient to various side effects. In this study, nifedipine was prepared as a solid dispersion using a water-soluble polymer (PVP-K30) in order to increase the releasing time and postpone the time at which the drug reaches its highest concentration, thereby increasing its bioavailability. SEM, XRD, DSC, and FT-IR were used for characterization of the solid dispersion. In order to test the release of the drug, a dissolution test was performed with a serous fluid for 12 h. In this work a drug formulation capable of sustained release of nifedipine using a water-soluble polymer was developed.

Polyglutamine (polyQ) tract polymorphism within the human androgen receptor (AR) shows population heterogeneity. African American men possess short polyQ tracts significantly more frequently than Caucasian American men. The length of polyQ tracts is inversely correlated with the risk of prostate cancer, age of onset, and aggressiveness at diagnosis. Aberrant activation of Wnt signaling also reveals frequently in advanced prostate cancer, and an enrichment of androgen and Wnt signaling activation has been observed in African American patients. Here, we assessed aberrant expression of AR bearing different polyQ tracts and stabilized β-catenin in prostate tumorigenesis using newly generated mouse models. We observed an early onset oncogenic transformation, accelerated tumor cell growth, and aggressive tumor phenotypes in the compound mice bearing short polyQ tract AR and stabilized β-catenin. RNA sequencing analysis showed a robust enrichment of Myc-regulated downstream genes in tumor samples bearing short polyQ AR versus those with longer polyQ tract AR. Upstream regulator analysis further identified Myc as the top candidate of transcriptional regulators in tumor cells from the above mouse samples with short polyQ tract AR and β-catenin. Chromatin immunoprecipitation analyses revealed increased recruitment of β-catenin and AR on the c-Myc gene regulatory locus in the tumor tissues expressing stabilized β-catenin and shorter polyQ tract AR. These data demonstrate a promotional role of aberrant activation of Wnt/β-catenin in combination with short polyQ AR expression in prostate tumorigenesis and suggest a potential mechanism underlying aggressive prostatic tumor development, which has been frequently observed in African American patients.

Ohmyungsamycin A (1), a novel cyclic peptide discovered from a marine Streptomyces sp., was previously reported with antibacterial and anticancer activities. However, the antitumor activities and the underlying molecular mechanisms of 1 remain to be elucidated. Compound 1 inhibited the proliferation and tumor growth of HCT116 human colorectal cancer cells based on both in vitro cell cultures and an in vivo animal model. A cDNA microarray analysis revealed that 1 downregulated genes involved in cell cycle checkpoint control. Compound 1 also induced G0/G1 cell cycle arrest that was mediated by the regulation of S-phase kinase-associated protein 2 (Skp2)-p27 axis and minichromosome maintenance protein 4 (MCM4). Furthermore, a longer exposure of 1 exhibited an accumulation of a sub-G1 phase cell population, which is characteristic of apoptotic cells. The induction of apoptosis by 1 was also associated with the modulation of caspase family proteins. Compound 1 effectively suppressed tumor growth in a xenograft mouse model subcutaneously implanted with HCT116 cells. In addition, analysis of tumors revealed that 1 upregulated the expression of the CDK inhibitor p27 but downregulated the expression of Skp2 and MCM4. These findings demonstrate the involvement of 1 in cell cycle regulation and the induction of apoptosis in human colorectal cancer cells.

Although gemcitabine-based chemotherapy is common and effective for pancreatic cancer (PC), acquired drug resistance is one of the major reasons for treatment failure. Therefore, a novel therapeutic approach for gemcitabine-resistant PC is required. Nuclear factor erythroid 2-related factor 2 (Nrf2) is an oxidative stress-responsive transcription factor regulating antioxidant responses and plays a crucial role in chemoresistance. In the present study, the antitumor activity of periplocin, a natural cardiac glycoside, was evaluated in an established gemcitabine-resistant PC cell line (PANC-GR). Nrf2 was overexpressed in gemcitabine-resistant cells, and Nrf2 knockdown recovered gemcitabine sensitivity in PANC-GR cells. The antiproliferative activity of periplocin was highly associated with Nrf2 downregulation and Nrf2-mediated signaling pathways in PANC-GR cells. Periplocin also increased reactive oxygen species production inducing G

Stromal androgen-receptor (AR) action is essential for prostate development, morphogenesis and regeneration. However, mechanisms underlying how stromal AR maintains the cell niche in support of pubertal prostatic epithelial growth are unknown. Here, using advanced mouse genetic tools, we demonstrate that selective deletion of stromal AR expression in prepubescent Shh-responsive Gli1-expressing cells significantly impedes pubertal prostate epithelial growth and development. Single-cell transcriptomic analyses showed that AR loss in these prepubescent Gli1-expressing cells dysregulates androgen signaling-initiated stromal-epithelial paracrine interactions, leading to growth retardation of pubertal prostate epithelia and significant development defects. Specifically, AR loss elevates Shh-signaling activation in both prostatic stromal and adjacent epithelial cells, directly inhibiting prostatic epithelial growth. Single-cell trajectory analyses further identified aberrant differentiation fates of prostatic epithelial cells directly altered by stromal AR deletion. In vivo recombination of AR-deficient stromal Gli1-lineage cells with wild-type prostatic epithelial cells failed to develop normal prostatic epithelia. These data demonstrate previously unidentified mechanisms underlying how stromal AR-signaling facilitates Shh-mediated cell niches in pubertal prostatic epithelial growth and development.

Androgen/androgen receptor (AR) signaling pathways are essential for prostate tumorigenesis. However, the fundamental mechanisms underlying the AR functioning as a tumor promoter in inducing prostatic oncogenesis still remain elusive. Here, we demonstrate that a subpopulation of prostatic Osr1 (odd skipped-related 1)-lineage cells functions as tumor progenitors in prostate tumorigenesis. Single cell transcriptomic analyses reveal that aberrant AR activation in these cells elevates insulin-like growth factor 1 (IGF1) signaling pathways and initiates oncogenic transformation. Elevating IGF1 signaling further cumulates Wnt/β-catenin pathways in transformed cells to promote prostate tumor development. Correlations between altered androgen, IGF1, and Wnt/β-catenin signaling are also identified in human prostate cancer samples, uncovering a dynamic regulatory loop initiated by the AR through prostate cancer development. Co-inhibition of androgen and Wnt-signaling pathways significantly represses the growth of AR-positive tumor cells in both ex-vivo and in-vivo, implicating co-targeting therapeutic strategies for these pathways to treat advanced prostate cancer.

Omeprazole (OM), typically used for patients with gastric problems, is a proton pump inhibitor, which reduces the amount of stomach acid production effectively by regulating the proton pump. However, the main side effect of OM is the calcium indigestion, which may cause the calcium deficiency in the long term. Seaweeds are abundant in calcium (approximately 560–1200 mg per 100 g) and nutrients such as vitamins (approximately 150–400 mg per 1 kg) and minerals. In particular, high level of magnesium in the seaweed can assist the absorption of calcium. Accordingly, seaweeds can be a good source of nutrients to support OM therapy. Herein, we produced a solid dispersion (SD) which is composed of OM and seaweed calcium (SC) and can be suitable for the oral treatment, via the ultra-multiple crystal method. We also optimized the film formulation so that the SD could be absorbed quickly and efficiently by the oral cavity. Due to many advantages of using the oral thin film (OTF), we chose to fabricate OTF with our optimized dispersion method. OTF allows for both the rapid and efficient dissolution within the oral cavity and the protection of the active ingredient (i.e., OM) from rapid degradation under acidic conditions. The prepared SD samples were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), and X-ray diffraction (XRD). Thermal analysis of SD was conducted by differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA). Hydrophobicity of SD was evaluated by the contact angle. Furthermore, the dissolution test and the HPLC analysis of SD were performed.

Triple-negative breast cancer (TNBC) is the most intractable cancer in women with a high risk of metastasis. While hyper-methylation of histone H3 catalyzed by disruptor of telomeric silencing 1-like (DOT1L), a specific methyltransferase for histone H3 at lysine residue 79 (H3K79), is reported as a potential target for TNBCs, early developed nucleoside-type DOT1L inhibitors are not sufficient for effective inhibition of growth and metastasis of TNBC cells. We found that TNBC cells had a high expression level of DOT1L and a low expression level of E-cadherin compared to normal breast epithelial cells and non-TNBC cells. Here, a novel psammaplin A analog (PsA-3091) exhibited a potent inhibitory effect of DOT1L-mediated H3K79 methylation. Consistently, PsA-3091 also significantly inhibited the proliferation, migration, and invasion of TNBC cells along with the augmented expression of E-cadherin and the suppression of N-cadherin, ZEB1, and vimentin expression. In an orthotopic mouse model, PsA-3091 effectively inhibited lung metastasis and tumor growth by the regulation of DOT1L activity and EMT biomarkers. Together, we report here a new template of DOT1L inhibitor and suggest that targeting DOT1L-mediated H3K79 methylation by a novel PsA analog may be a promising strategy for the treatment of metastatic breast cancer patients. Keywords: histone methyltransferase DOT1L, H3K79 methylation, psammaplin A analog (PsA-3091), triple-negative breast cancer, metastasis, epithelial-mesenchymal transition (EMT)

Recent genome analysis of human prostate cancers demonstrated that both AR gene amplification and TP53 mutation are among the most frequently observed alterations in advanced prostate cancer. However, the biological role of these dual genetic alterations in prostate tumorigenesis is largely unknown. In addition, there are no biologically relevant models that can be used to assess the molecular mechanisms for these genetic abnormalities. Here, we report a novel mouse model, in which elevated transgenic AR expression and Trp53 deletion occur simultaneously in mouse prostatic epithelium to mimic human prostate cancer cells. These compound mice developed an earlier onset of high-grade prostatic intraepithelial neoplasia and accelerated prostate tumors in comparison with mice harboring only the AR transgene. Histological analysis showed prostatic sarcomatoid and basaloid carcinomas with massive squamous differentiation in the above compound mice. RNA-sequencing analyses identified a robust enrichment of the signature genes for human prostatic basal cell carcinomas in the above prostate tumors. Master regulator analysis revealed SOX2 as a transcriptional regulator in prostatic basal cell tumors. Elevated expression of SOX2 and its downstream target genes were detected in prostatic tumors of the compound mice. Chromatin immunoprecipitation analyses implicate a coregulatory role of AR and SOX2 in the expression of prostatic basal cell signature genes. Our data demonstrate a critical role of SOX2 in prostate tumorigenesis and provide mechanistic insight into prostate tumor aggressiveness and progression mediated by aberrant AR and p53 signaling pathways.

Androgen signaling is essential for prostate development, morphogenesis, and regeneration. Emerging evidence also indicates a regulatory role of Notch signaling in prostate development, differentiation, and growth. However, the collaborative regulatory mechanisms of androgen and Notch signaling during prostate development, growth, and regeneration are largely unknown. Hairy and Enhancer of Split 1 (Hes1) is a transcriptional regulator of Notch signaling pathways, and its expression is responsive to Notch signaling. Hes1-expressing cells have been shown to possess the regenerative capability to repopulate a variety of adult tissues. In this study, we developed new mouse models to directly assess the role of the androgen receptor in prostatic Hes1-expressing cells. Selective deletion of AR expression in embryonic Hes1-expressing cells impeded early prostate development both in vivo and in tissue xenograft experiments. Prepubescent deletion of AR expression in Hes1-expressing cells resulted in prostate glands containing abnormalities in cell morphology and gland architecture. A population of castration-resistant Hes1-expressing cells was revealed in the adult prostate, with the ability to repopulate prostate epithelium following androgen supplementation. Deletion of AR in Hes1-expressing cells diminishes their regenerative ability. These lines of evidence demonstrate a critical role for the AR in Notch-responsive cells during the course of prostate development, morphogenesis, and regeneration, and implicate a mechanism underlying interaction between the androgen and Notch signaling pathways in the mouse prostate.

Tissue engineered scaffolds, made of natural derived materials, have the potential to be used in bone regeneration fields due to the biocompatible and biodegradable features. In this study, we propose duck's feet-derived collagen (DC) sponges blended with hydroxyapatite (HAp), incorporated with different concentrations of silymarin (Smn), for improved bone regeneration. The morphological and structural properties of DC/HAp and DC/HAp loaded with 25, 50 and 100 μM of Smn sponges were analyzed using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). In vitro evaluations were carried out on rabbit bone marrow stem cells (rBMSCs) using MTT assay for cell proliferation, ALP assay for osteogenic differentiation and reverse transcription-polymerase chain reaction (RT-PCR) for expression of mRNAs. For the evaluation of new bone formation in vivo, histological analysis and micro computed tomography (μCT) were used. Preliminary results, on Smn/DC/HAp morphology and mechanical properties, showed an interconnected porosity suitable for cells ingrowth and a higher compressive strength with the presence of Smn. Similarly, the cells proliferation and ALP activity modulation were positively influenced by the Smn content. Especially, the 100 μM Smn/DC/HAp sponge efficiently enhances the rBMSCs adhesion, growth and gene expression of osteogenic markers. The enhanced osteoinductive effects of sponges blended with Smn were confirmed using μ-CT and histological evaluations. In conclusion, results suggest that collagen sponges represent an excellent environment for cells growth and proliferation, while Smn plays an important role to improve materials osteogenic properties.

BACKGROUND AND AIMS Chromophobe hepatocellular carcinoma (HCC) is a newly included subtype of HCC in the 5th edition of the WHO classification with distinctive histological features (chromophobic cytoplasm with anaplastic nuclei and pseudocyst formation) and is strongly associated with the alternative lengthening of telomeres (ALT) phenotype. However, the clinicopathologic characterization and molecular features of chromophobe HCC are unknown. METHODS To comprehensively characterize chromophobe HCC, whole exome sequencing, copy number variation, and transcriptomic analyses were performed in 224 surgically resected HCC cases. Additionally, telomere-specific fluorescence in situ hybridization was used to assess ALT. These genomic profiles and ALT status were compared with clinicopathological features among subtypes of HCC, particularly chromophobe HCC and conventional HCC. RESULTS Chromophobe HCC was observed in 10.3% (23/224) cases and, compared to conventional HCC, was more frequent in females (P = .023). The overall and recurrence-free survival outcomes were similar between patients with chromophobe HCC and conventional HCC. However, chromophobe HCC displayed significantly more upregulated genes involving cell cycle progression and DNA repair. Additionally, ALT was significantly enriched in chromophobe HCC (87%; 20/23) compared to conventional HCC (2.2%, 4/178; P

In this study, highly concentrated hydrogen nanobubble water was utilized as the blending water for cement mortar to improve its compressive and flexural strengths. Highly concentrated nanobubbles can be obtained through osmosis. This concentration was maintained by sustaining the osmotic time. The mortar specimens were cured for 28 days, in which the nanobubble concentration was increased. This improved their flexural strength by 2.25–13.48% and compressive strength by 6.41–11.22%, as compared to those afforded by plain water. The nanobubbles were densified at high concentrations, which caused a decrease in their diameter. This increased the probability of collisions with the cement particles and accelerated the hydration and pozzolanic reactions, which facilitated an increase in the strength of cement. Thermogravimetric analysis and scanning electron microscopy were used to confirm the development of calcium silicate hydrate (C-S-H) and hydration products with an increase in the nanobubble concentration. Quantitative analysis of the hydration products and the degree of hydration were calculated by mineralogical analysis.

Description, Dark matter interpretations of the DAMA signals are excluded by COSINE-100 with the same NaI(Tl) target for various models., We present new constraints on dark matter interactions using 1.7 years of COSINE-100 data. The COSINE-100 experiment, consisting of 106 kg of tallium-doped sodium iodide [NaI(Tl)] target material, is aimed to test DAMA’s claim of dark matter observation using the same NaI(Tl) detectors. Improved event selection requirements, a more precise understanding of the detector background, and the use of a larger dataset considerably enhance the COSINE-100 sensitivity for dark matter detection. No signal consistent with the dark matter interaction is identified and rules out model-dependent dark matter interpretations of the DAMA signals in the specific context of standard halo model with the same NaI(Tl) target for various interaction hypotheses.

Purpose Periodontal treatment aims at complete regeneration of the periodontium, and developing strategies for periodontal regeneration requires a deep understanding of the tissues composing the periodontium. In the present study, the stemness characteristics and gene expression profiles of cementum-derived cells (CDCs) were investigated and compared with previously established human stem cells. Candidate marker proteins for CDCs were also explored. Methods Periodontal ligament stem cells (PDLSCs), pulp stem cells (PULPSCs), and CDCs were isolated and cultured from extracted human mandibular third molars. Human bone marrow stem cells (BMSCs) were used as a positive control. To identify the stemness of CDCs, cell differentiation (osteogenic, adipogenic, and chondrogenic) and surface antigens were evaluated through flow cytometry. The expression of cementum protein 1 (CEMP1) and cementum attachment protein (CAP) was investigated to explore marker proteins for CDCs through reverse-transcription polymerase chain reaction. To compare the gene expression profiles of the 4 cell types, mRNA and miRNA microarray analysis of 10 samples of BMSCs (n=1), PDLSCs (n=3), PULPSCs (n=3), and CDCs (n=3) were performed. Results The expression of mesenchymal stem cell markers with a concomitant absence of hematopoietic markers was observed in PDLSCs, PULPSCs, CDCs and BMSCs. All 4 cell populations also showed differentiation into osteogenic, adipogenic, and chondrogenic lineages. CEMP1 was strongly expressed in CDCs, while it was weakly detected in the other 3 cell populations. Meanwhile, CAP was not found in any of the 4 cell populations. The mRNA and miRNA microarray analysis showed that 14 mRNA genes and 4 miRNA genes were differentially expressed in CDCs vs. PDLSCs and PULPSCs. Conclusions Within the limitations of the study, CDCs seem to have stemness and preferentially express CEMP1. Moreover, there were several up- or down-regulated genes in CDCs vs. PDLSCs, PULPSCs, and BMSCs and these genes could be candidate marker proteins of CDCs., Graphical Abstract

This study analyzed the effects of applying highly concentrated hydrogen nanobubble water (HNBW) on the workability, durability, watertightness, and microstructure of cement mixtures. The number of hydrogen nanobubbles was concentrated twofold to a more stable state using osmosis. The compressive strength of the cement mortar for each curing day was improved by about 3.7–15.79%, compared to the specimen that used general water, when two concentrations of HNBW were used as the mixing water. The results of mercury intrusion porosimetry and a scanning electron microscope analysis of the cement paste showed that the pore volume of the specimen decreased by about 4.38–10.26%, thereby improving the watertightness when high-concentration HNBW was used. The improvement in strength and watertightness is a result of the reduction of the microbubbles’ particle size, and the increase in the zeta potential and surface tension, which activated the hydration reaction of the cement and accelerated the pozzolanic reaction.

Androgens/androgen receptor (AR)-mediated signaling pathways are essential for prostate development, morphogenesis and regeneration. Specifically, stromal AR signaling has been shown to be essential for prostatic initiation. However, the molecular mechanisms underlying AR-initiated mesenchymal-epithelial interactions in prostate development remain unclear. Here, using a newly generated mouse model, we have directly addressed the fate and role of genetically marked AR-expressing cells during embryonic prostate development. Androgen signaling-initiated signaling pathways were identified in mesenchymal niche populations at single-cell transcriptomic resolution. The dynamic cell-signaling networks regulated by stromal AR were additionally characterized in relation to prostatic epithelial bud formation. Pseudotime analyses further revealed the differentiation trajectory and fate of AR-expressing cells in both prostatic mesenchymal and epithelial cell populations. Specifically, the cellular properties of Zeb1-expressing progenitors were assessed. Selective deletion of AR signaling in a subpopulation of mesenchymal rather than epithelial cells dysregulated the expression of the master regulators and significantly impaired prostatic bud formation. These data provide novel, high-resolution evidence demonstrating the important role of mesenchymal androgen signaling in the cellular niche controlling prostate early development by initiating dynamic mesenchyme-epithelia cell interactions.

Prostate embryonic development, pubertal and adult growth, maintenance, and regeneration are regulated through androgen signaling-mediated mesenchymal-epithelial interactions. Specifically, the essential role of mesenchymal androgen signaling in the development of prostate epithelium has been observed for over 30 years. However, the identity of the mesenchymal cells responsible for this paracrine regulation and related mechanisms are still unknown. Here, we provide the first demonstration of an indispensable role of the androgen receptor (AR) in sonic hedgehog (SHH) responsive Gli1-expressing cells, in regulating prostate development, growth, and regeneration. Selective deletion of AR expression in Gli1-expressing cells during embryogenesis disrupts prostatic budding and impairs prostate development and formation. Tissue recombination assays showed that urogenital mesenchyme (UGM) containing AR-deficient mesenchymal Gli1-expressing cells combined with wildtype urogenital epithelium (UGE) failed to develop normal prostate tissue in the presence of androgens, revealing the decisive role of AR in mesenchymal SHH responsive cells in prostate development. Prepubescent deletion of AR expression in Gli1-expressing cells resulted in severe impairment of androgen-induced prostate growth and regeneration. RNA-sequencing analysis showed significant alterations in signaling pathways related to prostate development, stem cells, and organ morphogenesis in AR-deficient Gli1-expressing cells. Among these altered pathways, the transforming growth factor β1 (TGFβ1) pathway was up-regulated in AR-deficient Gli1-expressing cells. We further demonstrated the activation of TGFβ1 signaling in AR-deleted prostatic Gli1-expressing cells, which inhibits prostate epithelium growth through paracrine regulation. These data demonstrate a novel role of the AR in the Gli1-expressing cellular niche for regulating prostatic cell fate, morphogenesis, and renewal, and elucidate the mechanism by which mesenchymal androgen-signaling through SHH-responsive cells elicits the growth and regeneration of prostate epithelium., Author summary Prostate formation, growth, and regeneration, as well as tumorigenesis, depend on androgens and androgen receptor (AR)-mediated signaling pathways. Tissue recombination assays done more than 30 years ago demonstrated a decisive role for stromal androgen signaling in prostatic epithelium development. However, in the intervening time, the identity of the mesenchymal cells in the urogenital sinus mesenchyme that convey androgen signaling and control prostate epithelium development, morphogenesis, and regeneration has not been determined. In this study, using mouse genetic tools, we demonstrate for the first time that selective deletion of AR in mesenchymal Gli1-expressing cells abolishes early development of prostate tissue and normal prostate formation, and diminishes prostate pubertal growth and regeneration. In addition, using tissue recombination assays, we directly determined an essential requirement for AR expression in mesenchymal Gli1-expressing cells during prostate epithelium development. Our results not only resolve a 30-year-old scientific puzzle by identifying the mesenchymal cell properties of androgen-responsive cells that elicit development of the embryonic prostate epithelium, but also explore a new regulatory mechanism for androgen and Shh signaling-mediated cellular niches in regulating prostatic cell fate, growth, and renewal through paracrine regulation. Given the importance of sex hormone and hedgehog signaling pathways in human development and tumorigenesis, this study extends beyond the field of prostate biology, raising new questions underlying sex hormone and SHH signaling in development and tumorigenesis.

Prostate cancer (PC) is the most common disease in men over age 50, and its prevalence rate has been gradually increasing since 1980. Taxane-derived anticancer agents are the primary agents used to treat metastatic prostate cancer patients; however, the side effects and acquired drug resistance limit the success of these therapies. Because there is no specific treatment for paclitaxel-resistant prostate cancer, it is necessary to develop new targets and therapeutic strategies to overcome the acquired resistance. In this study, the antitumor activity of a novel selenonucleoside (4′-selenofuranosyl-2,6-dichloropurine, LJ-2618), a third-generation nucleoside, and its plausible mechanisms of action in paclitaxel-resistant prostate cancer (PC-3-Pa) cells were investigated. The established PC-3-Pa cells exhibited over 100-fold resistance against paclitaxel compared to the paclitaxel-sensitive PC-3 cells. LJ-2618, however, effectively inhibited the proliferation of both cell lines with similar IC50 values in vitro. In PC-3-Pa cells, the activated PI3K/Akt signaling pathway was suppressed by LJ-2618 treatment. In addition, Skp2 was found to be over-expressed in paclitaxel-resistant cells, and the transfection of Skp2 siRNA recovered the sensitivity of paclitaxel in PC-3-Pa cells. Furthermore, LJ-2618 significantly down-regulated Skp2 expression in PC-3-Pa cells by promoting degradation and inducing destabilization of Skp2, which triggers G2/M cell cycle arrest. In a xenograft mouse model implanted with PC-3-Pa cells, LJ-2618 (3 or 10 mg/kg) effectively inhibited tumor growth with the enhancement of Skp2 degradation and induction of p27 expression in tumor tissues. These findings suggest that LJ-2618 may have potential for overcoming paclitaxel resistance via promoting Skp2 degradation and stabilizing p27 expression in PC-3-Pa cells. Therefore, the novel selenonucleoside LJ-2618 may lead to the development of a new treatment strategy for patients with paclitaxel-resistant, castration-resistant prostate cancer.